STUDIES ON THE EFFECT OF ORGANIC MATTERS AND BIOINOCULANTS ON MELOIDOGYNE INCOGNITA (KOFOID & WHITE) CHITWOOD INFECTING SOLANUM LYCOPERSICUM L.
THESIS SUMITTED FOR THE AWARD OF THE DEGREE OF DOCTOR OF PHILOSOPHY IN BOTANY
MOHD ASIF
DEPARTMENT OF BOTANY ALIGARH MUSLIM UNIVERSITY ALIGARH (INDIA) 2017
Certificate
This is to certify that the thesis entitled “Studies on the effect of organic matters and bioinoculants on Meloidogyne incognita (Kofoid & White) Chitwood infecting Solanum lycopersicum L.”, embodies a record of bonafide research work carried out by Mr. Mohd Asif at the Department of Botany, Aligarh Muslim University, Aligarh under my guidance and supervision and that no part of it has been submitted for the award of any other degree or diploma. He is allowed to submit his thesis to the Aligarh Muslim University, Aligarh for the consideration of the award of the degree of Doctor of Philosophy in Botany.
Prof. Mansoor A. Siddiqui (Research Supervisor)
Acknowledgements
In the praise of Almighty “GOD”, the most merciful and beneficent who bestowed on me the divine guidance to embark upon this task of keeping in realms of facts and events.
First and foremost I must gratefully record my indebtness to all the eminent authorities, scholars and researchers whose investigations I have consulted and whose ideas and insights have richly contributed to the shaping up of this study.
Words are insufficient to bear the depth of sincere gratitude that I owe to my esteemed supervisor, Prof. Mansoor A. Siddiqui for suggesting the problem, valuable guidance, incessant encouragement, constructive criticism, expedient advice and care during the course of investigation and preparation of this manuscript. His valuable suggestions added to the output of the research and inspired me to have a better grip over my research field. His adeptness creative thoughts, perspicuous mind and words shall forever remain in my mind. I am greatly beholden to Prof. Mohd Yunus Khalil Ansari, Chairman, Department of Botany, Aligarh Muslim University, Aligarh for his kindness and providing necessary facilities to undertake this study. I extend my thanks and deep sense of gratitude to my learned teachers Prof. Irshad Mahmood, Prof. Hisamuddin, Prof. Zaki Anwar Siddiqui, Prof. Abrar Ahmad Khan, Prof. Tabreiz Ahmad Khan, Prof. Samiullah Khan Dr.(Mrs.) Rose Rizvi and Dr. (Mrs.)Sana Choudhary for their constant help and guidance.
I feel immense pleasure to express my sincere thanks to Dr. Sartaj A.Tiyagi and Mr. Prem Raj Tyagi for their valuable guidance and helpful suggestions. I am immensely thankful for the genuine and kind help offered to me by the seniors Dr. Faheem Ahmad, Dr.Tauheed Khan, Dr. Bushra Rehman, Dr. Kavita Parihar, Dr.Safiuddin Dr. Ashraf Ghanai, Dr.Saima, Dr Irfana and Dr. Usman A. They need admiration, deepest thanks for their co-operation, healthy suggestions, extensive help and availability in the time of crisis.
Special thanks are due to my lab mates Mr. Mohd Tariq, Mr. Amir Khan, Miss Taruba Ansari and Mr. Faryad Khan for their co-operation, moral support and extensive help whenever needed to sort out the intricacies and complexity of work.
Fortunately I am blessed with a galaxy of sincere and adoring friends whose smile made my painstaking work easy. Diction is not enough to express my thanks to my friends Mr.Rafiul amin, Mr. Zishan, , Mr.Waseem, Mr. Faisal, Mr. Naushad, Mr. Anees, Mr.Gulwaiz, Mr. Danish,Mr. Shuaib, Miss Aisha, Miss Nusrat, Miss Rakhshanda, Miss Gulnaz, Mrs. Darakhshan, Miss Afsheen Miss Yawar, Mr. Malik, Dr.Khairu, Mr. Amaj, Mr. Siraj, Mr.
Ibraheem Mr. Wajahat, Mr.Zubair and Mr. Sadab and all my nears and dears who were always there to help and encourage me during the course of this work.
My eternal gratitude goes to uncle Mr. Mujahid Hussain for their consistent inspiration, encouragement and guidance regarding the agriculture practices.
I would like to acknowledge the people who mean world to me, my parents, my sisters Mrs. Aqsha, Mrs. Asma, Mrs.Trannum, Miss. Uzma, Miss. Aisha and brother Mr.Rehman, Mr Adil and Mr.Bilal. Their countless blessings, deep love and affection always work as a hidden power behind every task I perform. Without their constant inspiration, this milestone would have eluded me.
With all regards, I acknowledge the co-operation and help rendered by the non-teaching staff particularly Mr. Sharib and Mr. Imran of this department. I pay my sincere thanks to Shailesh K.Tiwari from IIVR for providing seeds of various cultivars of Tomato. Lastly the financial assistance in the form of PURSE Fellowship rendered by DST, Ministry of Science and Technology, Govt. of India, New Delhi and University Grants Commission, New Delhi is also greatly acknowledged.
“Commission and Omissions are mine in the shower of God ’s Mercy”
Mohd Asif
Contents
Page No.
Chapter I Introduction 1-19
Chapter II Review of Literature 20-60
Chapter III Materials and Methods 61-80
Chapter IV Experimental Results 81-135
Chapter V Discussion 136-165
Chapter VI Summary 166-170
Bibliography 171-239
Abstract
ABSTRACT
The southern root knot nematode (Meloidogyne incognita) is one the most destructive and virulent pest to the tomato crop affecting plant growth and yield. The present study was designed to test the response of tomato cultivars against M.incognita and to appraise the effect of various organic soil amendment viz., chopped leaves, biochar, oilcakes, sawdust, agriculture wastes and biocontrol agents viz., Pseudomonas fluorescens, Pochonia chlamydosporia Purpureocillium lilacinum, and Trichoderma viride under glasshouse conditions against root-knot nematode, Meloidogyne incognita attacking Tomato, Solanum lycopersicum L cv. ‘K-21’. The aqueous extract of some plant species, biochar, various oilcakes and bioagents were also investigated for their nematostatic and nematicidal properties under in vitro condition. Besides this all these agents were also examined for the penetration test against the M.incognita in the root of tomato cv.K-21.
Fourteen cultivars of tomato viz., EC-538380, K-21, CO-3, FEB-02, EC- 570018, NDT-3, S-22, GT-1, GT-2, GT-3, H-88-78-1, PB Barkha bahar-2, VRT- 101A, Kalyanpuri-T1 were trialed for resistant and susceptibility behaviour with the objective to see the potential sources of genes for resistance to M.incognita. All the cultivars differently varied significantly in causing reductions in growth parameters over their controls. In addition to this, all cultivars responded differently regarding formation of eggmasses, eggs, nematode population and reproductive factor. It was observed that none of the screened cultivars showed immune or highly resistant behaviour. Out of the fourteen cultivars tested two cultivar H-88-78-1 and VRT-101A were found resistant against M. incognita three were found moderately resistant (EC- 570018, GT-2 and PB Barkha bahar-2); three were found moderately susceptible (FEB-02, CO-3, and Kalyanpuri-T1); five cultivars showed susceptible behaviour (NDT-3, EC-538380, GT-3, GT-1 and S-22) and K-21 alone showed highly susceptible behaviour.
The aqueous leaves extracts of some selected plants viz., Mexican poppy (Argemone mexicana L., Family Papaveraceae), Trailing eclipta (Eclipta alba L., Family Asteraceae), Wild eggplant (Solanum xanthocarpum Schrad. & Wendl, Family Solanaceae), Black pigweed (Trianthema portulacastrum L., Family Aizoaceae), Indian mallow (Abutilon indicum L. (Sweet), Family Malvaceae), Ivy
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Abstract gourd (Coccinia grandis (L.) Voigt Family Cucurbitaceae) were assayed against the hatching of M.incognita eggs and mortality of second stage juveniles (J2) of M.incognita in vitro. The aqueous extract showed toxic effect and results in significant mortality of the juveniles of M.incognita. Similarly the aqueous dilutions of the oil cakes viz., castor (Ricinus communis L., Family-Euphorbiaceae), cotton (Gossypium arboreum L. Family- Malvaceae), mahua (Madhuca longifolia J. Konig, Sapotaceae), mustard (Brassica juncea L., Family-Brassicaceae), and soybean (Glycine max (L.) Merr. Family-Fabacaeae) were also investigated for their egg hatching inhibitory and juvenile mortality behaviour against M.incognita under in vitro conditions. The deleterious effect of the different extract increased with increase in the concentration and exposure period. The extract also inhibited the egg hatching and efficiency was found directly proportional to the concentration and inversely proportional to the exposure time. Similar results were also depicted by the biochar exudates which results in inhibition in egg hatching and mortality of second juvenile of M.incognita.
The aqueous culture filtrates of bioagents viz., Pseudomonas fluorescens, Pochonia chlamydosporia, Purpureocillium lilacinum, and Trichoderma viride were also analysed under in vitro conditions for their nematostatic and nematicidal properties. All the bioagents were found promising in the inhibition of egg hatching and juvenile mortality of M.incognita. Juvenile mortality was found directly proportional to the extract concentration and period of exposure whereas juvenile hatching was observed inversely proportional to the filtrate concentration and directly proportional to the exposure period.
Aqueous dilutions of leaf extract of selected plants showed noxious effect on the inhibition of juvenile penetration of M.incognita into the root of tomato cv.K-21. Similar results were also obtained with the aqueous dilutions of oilcakes and biochar. Maximum inhibition in the juvenile penetration in the root of tomato cv.K-21 was observed in the aqueous extract of Mexican poppy, Wild eggplant and Trailing eclipta. Among the oil cakes maximum inhibition was observed in castor cake, mustard cake and soybean cake. Similarly the exudates released by the biochar at 5% of the concentration caused most inhibitory and protective effects against juvenile penetration. All the treatments responded differently in terms of inhibition in penetration as compared to the undipped inoculated control and it was significantly 2
Abstract decreased with increase in the dip duration. In the related study all the bioagents showed significant inhibition in the juvenile penetration but among all the bioagents Pochonia chlamydosporia and Pseudomonas fluorescens was found most promising. The percent inhibition in juvenile penetration was found directly proportional to the concentration of the extract.
Aqueous leaf extract of Mexican poppy, Wild eggplant and Trailing eclipta used as bare-root dip treatment of tomato cv.K-21 seedling showed protection against root-knot nematode, M. incognita. Maximum potential for the suppression of eggmasses and root knot indices was obtained by Mexican poppy followed by Trailing eclipta and Wild eggplant. The toxicity of the juveniles increased with the increase in the concentration of the extracts and dip duration. Likewise, the aqueous culture filtrate of Pseudomonas fluorescens and Pochonia chlamydosporia showed most significant results. In a similar study application of bioagents P. fluorescens, P. lilacinum and P. chlamydosporia in combination with water hyacinth extract were also tested as bare root dip treatments on tomato cv.K-21. Bioagents along with plant extract significantly suppressed the nematode infestation thereby results in the enhancement of plant growth. But combined treatment of water hyacinth extract+ Pseudomonas fluorescens and Pochonia chlamydosporia were found most effective in reducing the root knot incidence and improvement of the plant growth characters.
In another glass house experiment, investigation was conducted to test the potential of chopped leaves of some plants viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pigweed@50g/pot integrated with the Black nightshade Seed powder@10g/pot against the impact of M.incognita and their efficacy in enhancing the plant growth characters on tomato cv.K-21. All the treatments were found significantly effective in suppressing the nematode population and preventing the infection at significantly low level. However, the potential of Mexican poppy integrated with Black nightshade Seed powder was most pronounced in reducing the root knot nematode infestation in terms of number of eggmasses/plant, eggs/eggmass and root-knot index and nematode population/250g soil and thereby exaggerating the plant growth characters in terms of length, fresh and dry weight, yield/plant followed by Trailing eclipta+ Black nightshade powder and Wild eggplant+ Black nightshade powder. A direct correlation was observed in between the reduction in nematode infestation and plant growth and yield of tomato cv.K-21. 3
Abstract
In the similar experiment where biochar alone and in combination with sawdust viz., Eucalyptus, Lebbeck, Jambul, Mango, Poplar and Babool was the most potent in restricting the root knot nematode development caused by M.incognita and exaggerating the plant growth characters of tomato cv.K-21. Biochar alone@65g/pot was not found much effective but in combination with Eucalyptus and lebbeck sawdust@20g/pot revealed highly nematicidal potential in suppressing the root knot nematode development hence results in promoting the plant growth and yield. The biochar+ babool sawdust combination was observed least effective In another experiment biochar alone@65g/pot and in combination with agriculture waste@20g/pot showed promising results in the enhancement of growth characters and reduction in the nematode infestation. But the most curative and protective effect was observed in biochar +tobacco waste which ultimately results in promoting the plant growth characters and yield of tomato cv.K-21. It was followed by biochar+ mentha waste and biochar +garlic waste while biochar alone showed least toxicity in reduction of root knot nematode infestation in terms of eggmasses, egg/eggmasses, nematode population and root knot indices and improvement of plant growth characters in terms of length, weight, pollen fertility, yield and biochemical parameters of tomato.
Another experiment was conducted under glass house conditions to test the potential of bioagents viz., Pseudomonas fluorescens, Pochonia chlamydosporia, Purpureocillium lilacinum, and Trichoderma viride as seed dressing agents against the virulent effect of M.incognita and their efficacy in improving the plant growth characters of tomato cv.K-21. Application of this strategy provides safety and defense against the root knot nematode infestation of M.incognita. Decreasing in the nematode population and root knot indices, the plant growth characters were augmented significantly. P. chlamydosporia and P. fluorescens @4g/kg seeds using 2%cellulose as a sticky agent were the most effective in reducing the nematode infestation and enhancing the plant growth. Similar results were attained in another experiment when the seeds treatments were done with the aqueous dilution of various oil seed cakes viz., castor cake, mustard cake, soybean cake, Mahua cake and cotton cake. All the treatments worked as effective promoter in the enhancement of plant growth characters and reduction of nematode development and root knot indices. However, among the treatments castor cake@21%w/w was most impressive in
4
Abstract improving the plant growth characters and arresting in nematode infestation. It was followed by mustard cake and soybean cake @21% concentration. Similar types of findings were achieved in another experiment where seeds were treated with aqueous exudates of biochar at various concentrations. Among the various concentration 2.5 and 5% concentration showed most pronounced effect in exaggerating the plant growth and yield and reduction in nematode infestation. Direct correlation was detected between the reduction in nematode infestation and improvement in plant growth character of tomato cv. K-21.
In another investigation conducted under glass house conditions the effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and biocontrol agents viz., P. fluorescens, P. chlamydosporia, P. lilacinum, and T. viride on the growth of tomato cv. K-21 in relation to root knot development and nematode multiplication. Experimental results revealed that the application of bioagents in all the treatments either individually, concomitantly and sequentially significantly improved plant growth parameters or reduced the root knot nematode infestation. The concomitant inoculation of bioagents with M. incognita significantly suppressed the root knot infestation and improved the plant growth characters as compared to untreated inoculated control. In the sequential inoculation, when bioagents were applied 15 days prior to the M.incognita results in pronounced improvement of plant growth parameters and high reduction in nematode population as compared to inoculation of M. incognita 15 days prior to the biocontrol agents. Thus the biocontrol agents have potential to improve plant growth parameters and suppressive impact on root-knot nematode reproduction and root knot indices.
In another pot experiment the integrated effect of fungal bioagents@ 2.0g/pot and bacterial bioagents @ 1.5g /pot and organic soil amendment viz., press mud, castor cake mustard cake, mahua cake, soybean cake and cotton cake@20g significantly suppressed the nematode infestation in terms of eggmasses, egg/eggmasses, nematode population and root knot indices and improved the plant growth characters in terms of length, weight, pollen fertility and biochemical parameters and yield of tomato cv. K-21. All the treatments showed nematicidal properties and results in enhancement of growth characters. However, among the various treatments P.flueroscens+ castor cake was observed most promising in the reduction of nematode infestation and improvement of plant growth characters. It was 5
Abstract followed by P. chlamydosporia+ castor cake and P.flueroscens+ pressmud. In the similar experiment fungal bioagents@ 2.0g/pot and bacterial [email protected]/pot integrated with plant straws and potato waste@15g/pot were evaluated against M.incognita under glass house conditions in tomato cv.k-21. The findings proved that the combined action of biocontrol agents and agriculture waste straw and potato waste@15g/pot significantly reduced the eggmasses/root, eggs/eggmass, and population of M.incognita and increased the plant growth parameters. The application of bioagents viz., P. fluorescens, P.chlamydosporia, P. lilacinum in combination with agriculture waste straw significantly suppressed the nematode infestation. P.flueroscens+ mustard straw was recorded highest in reducing nematode population and improvement of plant growth characters and P .lilacinum + pearl millet were least effective.
6
Introduction
CHAPTER I
INTRODUCTION
Since the inception of human civilization the vegetables have played a pivotal role in the human nutrition. Vegetables are the source of natural stock of nutrients and are one of the paramount constituents of the daily life by enabling us energy rich food. They are not only the component of our dining table but also the ample source of vitamins, minerals and fibers for the growth and development. In addition to this, short duration, economic viability, high yield, growing in every climate and ability to create on-farm and off-farm employment have developed an advantage for the growers. Vegetables are source of phytochemicals having antioxidant, antifungal, anti-viral, antibacterial and anti-carcinogenic properties (Steinmetz and Potter, 1996; Gruda, 2005). Existing per capita consumption of vegetable is still very low about to 130 g compare to the recommended dose of 250 g given by National Institute of Nutrition (NIN), Hyderabad (2011). All of this is due to the inaccessibility and production of sufficient amount of vegetables as compare to their consumption. In addition to this climate change, urbanization, shrinkage of natural resources, fragmentation of land and uneven growth are some of the key factors that have restricted the vegetable production of the country. Assuming a 3.5 to 5.5 percent Gross Domestic Product growth rate in India, the projected demand for vegetables in 2030 will be 151-193 million metric tons (Sammugasundaram, 2005). So to earn this goal, every feasible and accessible effort has been made to assure the required food production in the future. In this era of fast growing population explosion, it’s very necessary to increase the vegetable production in the form of food security, safety and nutritional security for the sustainable future. Up to now, extensive utilization of chemical pesticides, urbanization and industrialization have created havoc worldwide by the contamination of the food chain and disturbance in the ecological equilibrium of the soil and water by persistent pesticide residues. It leads to reduced nutrient and flavour contents through low-cost intensive food production. Organic farming is an approach that has created attention and interest among the growers for raising healthy and safe food in sustainable manner. The main aim of the organic farming is to provide nutritionally safe, potentially viable and high quality food, and to restore and maintain the long term fertility. Worldwide, emphasis is increasingly being put on the relationship between food, nutrition and health (WHO, 2004; WCRF, 2007). 1
Introduction
Therefore, the organic development of vegetables in sustainable manner not only increases the productivity and nutritional quality but also accomplish the challenges of population explosion and healthy and safe food. In global paradigm India is the second largest producer of vegetables, producing 162.2 million tons supplying 14% of total world vegetable production (FAO, 2014 and Indian Horticulture Database, 2013). In 1991-92 the area occupied by vegetables in India was 5593 hectares that has attained and registered a quantum jump of 9396 hectares in 2013-2014 with the total production of 162897metric tons. The trends are shown in Fig. A.
Area (HA) Production (MT) Productivity (MT/HA)
146555 162187 162897
58532 10,5 17,3 17,6 17,3
5593 8495 9205 9396 1991-92 2010-2011 2012-13 2013-14
Fig. A. Area, Production and Productivity of Vegetables (Source: Indian Horticultural Database 2014).
In agriculture based country, like India; large cultivar of vegetables are grown in tropical, subtropical and temperate regions. The most commonly cultivated vegetables are potato, cabbage, cucurbits, eggplant, okra, tomato, onion, carrot, chili and some other legumes (FAO, 2014).Vegetables are highly prone to the pathogens and continue assault make them susceptible to the disease and finally reduce the productivity of the crop. This all may be due the favourable climate that favors the reproduction. Among the major obstacles, pest and parasites occupy predominant position for restricting the productivity of the vegetables. Vegetable crops are usually the most susceptible and worst affected by the nematodes (Sharma et al., 2006; Anwar and Mckenry, 2007; Dhaliwal and Koul, 2007). Phytonematodes are ruinous
2
Introduction and economically most important pests of many cultivated crops around the world (Trifonova et al., 2009) damaging vegetables, particularly in tropical and subtropical countries (Sikora and Fernandez, 2005). The importance of nematode as a constraint on economic and ornamental crop production was realized long ago in our country. Since then, nematodes problem of national importance have appeared. In some production areas the reduction in vegetable yield due to phytonematodes has reached as high as 30% (Anwar et al., 2009).
1.1. BRIEF INTRODUCTION OF THE TEST CROP:
1.1.1. Tomato:
Classification
Division- Plantae
Order- Polemoniales
Family- Solanaceae
Genus- Solanum
Species- lycopersicum
The edible fruit of Solanum lycopersicum (Encyclopedia of Life) commonly known as a tomato plant belongs to the family Solanaceae. The word tomato was originated from the Spanish word tomate which has come from the Nahuatl word tomatl. The species originated in Central and South America. Tomato is one of the most popular and widely consumed vegetable crops grown in outdoor fields, greenhouses and net houses of the country for food and cash. It has very high potential for expansion under wide range of agro-ecologies of the country, provided that certain production constraints are alleviated (Fekadu Mariame et al., 2003). Tomato can be produced by direct sowing of the seed in the field or transplanted from seed beds. Transplanting has the advantage of economic use of seeds, selecting superior and vigorous seedlings, easiness for field establishment and early harvest (Lemma Dessalegn, 2002).
World production of tomatoes was 170.8 million tons in 2014, of all this total 31% was accounted by China followed by India, the United States and Turkey as the
3
Introduction major producers (FAO, 2014). In 2014, tomatoes accounted for 23% of the total fresh vegetable output of the European Union, with more than half of this total coming from Spain, Italy and Poland (Antonella, 2016).Tomato is a popular vegetable crop worldwide and it is grown on more than 5 × 106 ha with a production of approximately 161 × 106 metric tons. Africa and Asia account for more than 80% of the global tomato area with about 70% of world output (FAO, 2012).
The major tomato producing states are Bihar, Karnataka, Uttar Pradesh, Andhra Pradesh, Orissa, Himachal Pradesh, West Bengal, and Maharashtra. Among them, Andhra Pradesh is leading producer sharing 17.90% of the total in year 2013- 2014 (IHB-2014).
Madhya Pradesh; 10,34% Himachal Pradesh; Others; 12,82% Karnataka; Chattisgarh;3,35% 11,04% 4,35% Bihar; 5,67%
West Bengal; 6,09% Andhra Pradesh; 17,90%
Maharashtra; 6,40% Gujrat; 6,72% Odisha; 7,40% Telangana; 7,92%
Fig. B. Leading tomato producing states 2013-2014 (Source: Indian Horticultural Database 2014)
Tomato plants are dicots, requiring warm season and prefer well drained sandy loamy soil. Plants are erect or trailing pubescent herb (generally 1-3m tall) with the small yellowish flowers borne in clusters (3-12 flowers). The fruit is typically fleshy and juicy berry type. Medicinally tomato is a very important as it contains carotenoid pigment, lycopene, one of the most powerful natural antioxidants, The pigment that makes tomatoes red and has been linked to the prevention of many types of cancer, heart disease and premature aging (Cerkauskas, 2005; Wamache, 2005). Lycopene has also been shown to improve skin ability to protect harmful UV rays (BBC News 28 April 2008). In some studies lycopene, especially in cooked tomatoes, has been
4
Introduction found to help prevent prostate cancer (Nkondjock et al., 2005; Canene-Adams et al., 2007). Tomatoes are a treasure of their antioxidant benefits in terms of impressive amount of vitamin C and beta-carotene; a very good amount of the mineral manganese; and vitamin E. It helps in reduction of lipid peroxidation (oxygen damage to fats in cell membranes or in the bloodstream). Various studies have shown that high consumption of tomato is consistently correlated with a reduced risk of some type of cancer (Franceschi et al., 1994) and may account for a low incidence of ischemic heart disease (Gerster, 1997). Tomato is used in preparation of various products locally and on commercial scale; paste, soup, pickle, juice, ketchup, puree, powder and eaten as fresh salad, cooked food. A gluco-alkaloid known as tomatine found in tomato that is used as a precipitating agent for cholesterol. The fruits are rich source of some essential amino acids like citric acid and malic acid. It helps in maintaining the bone health and promote the prevention of the excessive clumping an important aspect maintaining heart health. The pulp and juice are digestible, a promoter of gastric secretion, good in chronic dyspepsia and worked as blood purifier. Ripened fruits are very helpful in healing wounds because of antibiotic properties (Conn and Stumpy, 1970), good appetizers and suitable food for diabetic patients (Myers and Croll, 1921). Its extract has a better effect on urinary acidity as compared to orange juice (Saywell and Lane, 1933). Dietary intake of tomatoes, extracts, and supplementation phytonutrients (like lycopene) has been found to enhance the profile of fats in our bloodstream. Most importantly intake of tomato caused to decreased total cholesterol, LDL cholesterol and triglyceride levels and also has the potential in suppression of accumulation of cholesterol molecules inside of macrophage cells. A branded product of tomato, Lycomato is excessively used to decrease the blood pressure. It is nutritionally very important because of its richness in vitamins and minerals (Brown and Hutchinson, 1949; Baloch, 1994).The chemical composition of tomato varies with the stage of maturity and variety. The pulp represents 85.4% of the whole fruits. The nutritive value as yielded upon the analysis of 100 g of edible portion is:
Energy-74 kJ; Carbohydrates-3.9 g; Sugars-2.6 g; Dietary fiber-1.2 g; Fat-0.2 g; Protein-0.9 g; Vitamin A equiv. -42µg; Beta-carotene 449µg; Lutein zeaxanthin 123µg; Thiamine (B1)-0.037 mg; Niacin (B3) 0.594 mg; Vitamin B6-0.08 mg; Vitamin C-14 mg; Vitamin E-0.54 mg; Vitamin K 7.9g; Magnesium 11 mg;
5
Introduction
Manganese-0.114 mg; Phosphorus-24 mg; Potassium -237 mg; Water -94.5 g; Lycopene -2573μg (USDA)
Area (HA) Production (MT) Productivity (MT/HA)
16826 18226,6 18735,9
4243,4 14,7 19,5 20,7 21,2
289,1 865 879,6 882 1991-92 2010-2011 2012-13 2013-14
Fig. C. Area, Production and Productivity of Tomato (Source: Indian Horticultural Database 2014).
1.2. CROP LOSSES BY NEMATODES
Despite of the huge efforts and investment in pest management the reduction in yield of our food, fibers and ornamental crops continues to be of utmost economic and social consequences. Beyond the shadow of doubt, plant parasitic nematodes are one of the polyphagus and virulent pest, usually one of the major threats to agriculture industry by the reduction of economic value of ornamental and other vegetable crops. For example, losses of over $100 billion per year world-wide can be attributed to infections caused by parasitic nematodes (Urwin et al., 1997). These tiny parasites obtain nutrition from the cytoplasm of living plant cells and comprise many species including ectoparasites and endoparsites (Williamson, 1999). These nematodes infect thousands of different herbaceous and woody plants and are major constraints in most agricultural cropping systems, causing both yield reductions and quality loss (Karssen and Moens, 2006). The majority of species within 50 genera of plant parasitic nematodes feed on root tissue with diverse modes of parasitism (Lilley et al., 1999).
Nematodes can cause damage to almost all kinds of crops (Sasser and Freckman, 1987; Khan et al., 2004a, 2004b; Borah et al., 2009; Serfoji et al., 2010) however, due
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Introduction to their subterranean habit, microscopic size (from 0.3 to 10 mm length), they are invisible to the naked eye (Torr et al., 2007). The degree of damage of a nematode is host and age dependent. Beside this, nature of soil, climatic and environmental conditions also influence the threshold population density, above which detectable damage occurs. More generally, Sikora and Fernández (2005) suggest that vegetable production in tropical and sub-tropical environments cannot be considered without some form of nematode management. It has widely been recognized that plant parasitic nematodes constitute one of the most devastating pest groups and are responsible for severe disease symptoms in different crops causing huge losses (Abad et al., 2008). In economic terms nematodes cause an estimated loss of about $ 157 billion annually to world agriculture (Abad et al., 2008) and yield loss of the major crops are 12.3% (Sasser and Freckman, 1987). In United states alone, the damages exceeded $10 billion / year by plant parasitic nematodes (Koenning et al., 1999).
The root-knot nematode, Meloidogyne species comprises the main nematode pathogen in developing nations, restricting the productivity of the crop. Almost 100 species of the genus Meloidogyne which are highly pathogenic and polyphagous in nature, have been identified so far (Karssen, 2002; Wesemael et al., 2011; Handoo et al., 2013; Khan et al., 2014) with four species M. incognita, M. arenaria, M. hapla and M. javanica being most commonly wide spread (Netscher, 1970; Sawadogo et al., 2000; Hunt and Handoo, 2009). Among different Meloidogyne spp., Meloidogyne incognita is the economically most important species (Hussey and Janssen, 2002), infecting to several botanical families like Solanaceae, Cucurbitaceae, Leguminosae, Liliaceae, Chenopodiaceae, Asteraceae, Apiaceae, Brassicaceae and Malvaceae (Jain et al., 2007; Grace et al., 2009). More than 3000 plant species including monocots, dicots and several weeds parasitized by root knot nematode (Hussey and Janssen, 2002; Abad et al.,2003; Grace et al., 2009) and distributed worldwide (Karssen and Van Hoenselaar, 1998).These nematode species pose particular control problems due to their wide host ranges, short generation periods and high reproduction rates (Trudgill and Block, 2001), and are thus considered as the greatest threat to the global agricultural production ( Haseeb and Pandey, 1995; Jayasinghe et al., 2003; Eyal et al., 2006).
The crop loss is defined as the difference between the attainable yield and the actual yield (Chiarappa, 1971). In the tropical and sub-tropical climates, crop 7
Introduction production losses attributable to nematodes were estimated at 14.6% compared with 8.8% in developed countries. Perhaps more importantly, only ~0.2% of the crop value lost to nematodes is used to fund nematological research to address these losses (Sasser and Freckman, 1987). Handoo (1998) estimated global crop losses due to nematode attack in the range of $80 billion. The production of tomato (Solanum lycopersicum), a major nutrient to man and income generating to its growers is impaired due to the infestation by nematodes and other factors. Reduction in yield ranging from 28% to 75% has been reported by various researchers (Ibrahim et al., 2000; Rajendran et al., 2003). The loss to Indian agriculture is estimated about Rs. 210 crore annually (Jain et al., 2007). It has been estimated that alone root-knot nematode, Meloidogyne incognita amounts to $78 billion global yield losses (Chen et al., 2004). M. incognita and M. javanica caused 24-38% loss for tomato, (Kathy, 2000). In India the loss is predicted almost 14.6% and in some could reach as high as 50-80% (Bhatti, 1992). Seshadri (1970) reported reduction in yield of tomato from 26.5 to 73.3% due to root knot nematode Meloidogyne spp. While Bhatti and Jain (1977) observed a loss of 46% yield in tomato due to root-knot nematode in India. Jain et al. (1994) reported 47.3- 71.9% avoidable yield loss due to M. javanica and M. incognita respectively.
The root-knot nematodes are obligate parasites invades plant roots and feed on root cells, causing the roots to grow into large galls to complete their life cycle. Penetration of nematode disrupts the vascular system of the plants and thereby inhibited the proper absorption of minerals and nutrients resulting in stunted growth and development of the plants and consequently crop yield is reduced. The primary infection is initiated by eggmass that persists in the soil and from which second stage juveniles (J2) hatch (Bleve-Zacheo and Melillo, 1997). The root-knot nematode juveniles infected plant roots drain photosynthates and nutrients of plants (Ashworth, 1991). The above ground symptoms of nematodes are characterized by poor-growth, severe stunting, crop patchiness, dieback, wilting and chlorosis. The underground symptoms are in the form of root galls. The infection of plants by these nematodes can also be visualized as stunted growth, wilting and susceptibility to various other pathogens (Williamson and Hussey, 1996; Black et al., 2002). Root-knot nematode infection in young plants may be fatal, while in mature plants cause reduction in yield and loss of vigour. Plant nutrients and water uptake are substantially reduced by the
8
Introduction resulting damage to root system and infested plants are therefore, weak and give low yields (Abad et al., 2003). Most root-knot nematodes reproduce by parthenogenesis (Castagnone-Sereno, 2006). Males migrate out of the plant and play no role in reproduction. After establishment of nematode selected cells become multinucleate and enlarge considerably through additional synchronous nuclear divisions in the absence of cell division (Jones and Payne, 1978). After the first molting inside the egg newly hatched juveniles emerged and remain in the short free-living stage in the rhizosphere of the host plant. It may reinvade the host plant or migrate through the soil to find a new host root. Second stage juveniles (J2) do not feed during the free living stage and then invade in the zone of elongation and become sedentary after attaining the position in cortical zone of the root. Oesophageal secretion of the second stage juveniles promote parenchyma cells near the head of the juvenile to become multinucleate and form feeding site, generally known as “giant cells”, from which the second stage juveniles (J2) and later adults feed (Sijmons et al., 1994). The first sign of giant cell induction is cell cycle activation leading to the formation of vascular binucleate cells (De Almeida Engler et al., 1999). As the giant cell formation occurs the surrounding root tissue give rise to an outgrowth in the form of gall having juveniles. Second stage juvenile (J2) undergo morphological changes and have three consecutive moults (J3, J4 and adult female). Thereafter, at last moult adult females appear pear like shape and resume feeding. Females of the species lay eggs into a gelatinous matrix (GM) which is produced by six rectal glands and secreted before and during egg laying (Maggenti and Allen, 1960).
1.3. NEMATODE MANAGEMENT STRATEGIES
Nematodes are soil born pests; interact with a wide range of microorganism inhibited in the soil rhizosphere of the plant. In contrast to the fungi and insect, nematodes are slow moving and encompasses zigzag pattern in the field during a growing season and management tactic may be in the form of individual field or even infested patch based. Control measures aim to reduce nematode feeding and invasion of roots to reduce crop damage and/or to reduce the fecundity of adult females and decrease post-crop populations left in the soil (Kerry and Hominick, 2002). Nematodes cause extensive damage to the cropping and non cropping system. Therefore, effective and environmental friendly strategy is need of the present hour for the nematode management in a sustainable manner without disturbing the 9
Introduction equilibrium of the soil and environment. Appropriate farming practices and a minimal use of nematicides/ pesticides is an important part of agriculture and horticulture worldwide and provide a basis for pest management in future (Hillocks, 2002; Feder et al., 2004). The aim of nematode management can be fulfilled by the integration of various approaches having environmentally protective and soil conditioning activity that finally enhance the soil rhizospheric nature and represent growth promoting activity.
Integrated nematode management involves the combination of various management strategies for all economically important nematodes of the agro system with the purpose of environmental sustainability, safety, quality, optimizing productivity, net returns and stability. It allows the combination of physical, cultural, chemical and biological management measures to give an environmentally safe, stable and long term nematode management. The main thematic principles involved in the integrated nematode management are (1) identifying the key nematode pests, (2) to define the management unit-the agro-ecosystem, (3) developing the management strategy, (4) developing assessment technique, (5) to evolve descriptive or predictive models (Ravichandra, 2008). The main purpose of sustainable approach is to integrate various approaches such as antagonistic crops, crop rotations, organic manure, soil solarization, organic amendments, bioagents and resistant cultivar of plants against the nematodes for the maintenance of the soil food web. Promotion and enhancement of healthy soil environment and increment in the beneficial microbes activity in the rhizosphere depend on the added amount of organic matter. Application of organic matter to the soil increase microbial abundance and the populations of free living nematodes, which increase the activity of the nematode trapping fungi to kill any plant parasitic species (Linford et al., 1938). However, it was found that the relationship between the activity of nematode trapping fungi, the nature and type of the soil organic matter was more complex and there was no simple relationship with nematode population density (Cooke, 1962).`
1.3.1 CULTURAL METHODS
In this era of modern technology, cultural practices often have limited use in suppressing nematode population densities and minimizing yield losses. Crop rotation, growing non-host, resistant or antagonistic cover crops, incorporation of
10
Introduction plant materials or animal manures, and destruction or removal of cotton stalks and roots to minimize nematode survival and reproduction have been investigated (Barker and Koenning, 1998; Davis et al., 2000, 2003; Koenning et al., 2003a, 2003b).
1.3.1.1. Crop rotation
Crop rotation, conceivably one of the most commonly applied cultural means of restricting nematode populations, requires adequate land for growing alternate crops that are non hosts to the nematode species. Seasonal rotations of susceptible crops with non-host or poor-host crops in the same area of land remain one of the most important techniques used for nematode management worldwide (Viaene et al., 2006). A number of crops and other plants have been found resistant to phytonematodes (Stefanova and Fernandez, 1995; Gomez and Rodriguez, 2005; Rehman et al., 2006). The decrease in nematode populations by intercropping mustard could be attributed to the presence of 2-propenyl isothiocynate in mustard having nematicidal activity as reported by Kowalska and Sonalinska (2001). Timper et al. (2001) demonstrated that the rotations of peanuts with 2 years of bahia grass, cotton or corn, in a field naturally infested with M. arenaria and P. penetrans the abundance of the bacterium was related to the population densities of the nematode and greatest were observed under continuous peanut cropping and next most abundant under the bahia grass peanut rotation. The crop rotation may provide a short-term suppression of nematode population densities (Starr et al., 2002). Various cover or trap crops and antagonistic plants are useful for reducing nematode populations as well as conserving soil and often improving soil texture (Alam and Jairajpuri, 1990; Abawi and Thurston, 1994). Rotation of crops with the crop such as beans, bahia grass, maize and cabbage support extensive growth of the nematophagous fungus, Pochonia chlamydosporia in their rhizosphere but support only limited reproduction of root-knot nematodes. They are used to maintain the abundance of the fungus in the soil while suppressing populations of the nematodes (Timper et al., 2001; Puertas and Hidalgo-Diaz, 2007) but the obligate parasites, like the bacterium Pasteuria penetrans, need to be introduced into the soil with a nematode susceptible crop in order to build up nematode population on which the bacterium multiply (Oostendorp et al., 1991). It has been reported by several workers that different cropping sequences reduce the populations of some harmful phytonematodes to the levels that do not cause economic losses (Alam et al., 1981; Idowu and Fawole, 1989; Haider et al., 11
Introduction
2001; Haider and Pathak, 2001). Crop rotation is difficult to use with most perennial crops. Crotalaria spp. are used as a cover crop in crop rotations and occasionally incorporated into the soil, due to their nematode suppressive effect and nitrogen- fixing ability (Wang et al., 2002). Witch-grass peanut rotation not only has beneficial effects on soil but also reduces the population of phytonematodes and causes shift in rhizosphere microbial soil ecology (Kokalis-Burelle et al., 2002). Rotations of susceptible vegetables together with poor hosts or tolerant crops have been used for the control of phytonematodes (Stefanova et al., 2005).
1.3.1.2. Organic amendments
Organic amendments have been commonly used on large scale for the management of nematodes. Expeditious decrease in nematode population levels may occur by the releasing of toxic chemical compounds from the decomposed organic matter and most importantly long-term effects might enhance nematode antagonists. Large number of studies have demonstrated the role of organic soil amendments for the management of noxious nematodes to a varying extent depending upon the type of organic matter, nematode, host plant species and prevailing ecological conditions (Akhtar and Alam, 1993; D’ Addabbo, 1995; Akhtar and Malik, 2000, Litterick et al., 2004; Oka, 2010; Ansari et al., 2016; Asif et al., 2016). Adding decomposable organic matter to the soil recognized as very efficient method for changing the environment of soil rhizosphere, thereby adversely affecting the life cycle of phytonematodes (Bhosle et al., 2006; Haseeb et al., 2006). The ability of plant parts / or products to reduce crop damage caused by root-knot nematodes, Meloidogyne spp., and population in amended soil is well documented (Begum, 2003; Youssef and Ali, 1998; Jesse et al., 2006). The activity of egg parasitic fungi has also been increased in the field by the incorporation of radish as a green manure (Schlang et al., 1988). On the other hand, application of organic substrates leads to build up of beneficial micro flora around the rhizosphere, which will help to reduce the plant parasitic nematodes in the soil (Oka, et al., 2007). Amendment of soil with non edible deoiled seed cakes of neem, mahua, sesame, karanj, castor, mustard and groundnut improve soil fertility and organic matter status. Beside this reduce the nematode population density through their decomposition products which are toxic in nature (Akhtar, 1998; Abbasi et al., 2005). The majority of the amendments used are wastes or by products of agricultural crops and industries, animal manure, compost and crop 12
Introduction residues. Alam (1990) stated that with ample supply of water, the organic matter undergoes decomposition and release many compounds like phenols, aldehydes and many gases including ammonia.
Organic amendments are assumed to affect the nature of facultative parasites of their saprotrophic phase more than obligate parasites that have bounded growth in soil. So, the reaction of organic amendments on the incorporation of organic matter to soil inoculated with root-knot nematodes with spores of P. penetrans, improved plant growth and multiplication of both nematode pest and bacterium (Gomes et al., 2002).Research on nematode trapping fungi has demonstrated that the enhancement of trapping activity resulting from the application of organic matter in soil is dependent on the fungal species, type and amount of organic material added (Jaffee, 2004).
Natural synthetic analogs based on natural compounds in plants have been isolated and identified from plants (Chitwood, 2002) such as thinyl, alkaloids, phenols, sesquiterpenes, diterpenes, pentacyclic triterpenoids and polyacetylenes have been found in healthy plant tissue (Gommers and Barker, 1988; Matsuda et al., 1989; Qamar et al., 2005). Some of this type of studies has been reported interesting metabolites against Meloidogyne incognita, such as lantanosides and lantanone from Lantana camara (Begum et al., 2000; Qamar et al., 2005).
Organic amendments directly can improve soil texture, structure and biological activity and may also reduce the populations of nematodes, indirectly it may involve in promotion of antagonistic soil organisms, stimulation of the competitive status of the non-pathogenic organisms and release of toxic substances during the decomposition process which may kill or suppress pathogens including pest-nematodes (Rodriguez-Kabana 1986; Akhtar and Malik, 2000; Oka, 2010; Thoden et al., 2011). Incorporation of biochar; the solid carbon-rich products of biomass pyrolisation into soil or potting mixes induces systemic plant defense against fungal pathogens by increasing populations of certain bacteria, actinomycetes, yeasts and fungi (Elad et al. 2010). Biochar have been shown to decrease Fusarium root rot of asparagus (Elmer and Pignatello, 2011) and Rhizoctonia damping-off in cucumber (Jaiswal et al., 2014). Although various investigations have revealed the effect of biochar on fungal and bacterial soil populations, there had been very little information
13
Introduction on the interaction of biochar with nematodes. Recent microcosm study showing that wheat straw-biochar increased the abundance of soil fungal-feeders and decreased the plant parasitic nematode populations (Zhang et al., 2013). Many of the pores within a biochar particle can accommodate soil bacteria and some fungi but exclude their larger predators including free-living non-plant parasitic nematodes (Warnock et al., 2007).
1.3.1.3. Resistant cultivar
The exploration of resistant cultivar against the root-knot nematodes has been recognized as a possible means of replacing the health hazardous chemical pesticides frequent in use. However, resistance breakdown is a limiting factor (Netcher and Sikora, 1990). Utilization of resistant cultivar is one of the environmental friendly, safe and economically viable way of managing root-knot nematode and has better yield than the susceptible crop cultivar. Plants highly resistant to nematodes are often tolerant to nematodes as well, because they endure significantly less parasitism than susceptible plants when exposed to the same soil population densities of nematodes (Evans and Haydock, 1990). Intolerant plants may appear to be resistant, if nematode feeding reduces the amount of root tissue thereby reducing potential nematode feeding sites (Young, 1998). Zhou and Starr (2003) also reported that resistance to nematodes imparted tolerance to nematodes. After entering the roots of resistant cultivars, these nematodes get trapped due to the failure of feeding site development (Dufour et al., 2003). Nematode resistance in host plant is manifested by reduced rates of nematode reproduction and, consequently, lower nematode population densities in the crop rhizosphere than that of a susceptible one (Medina-Filho and Tanksley, 1983). According to Khan (1994) genetic resistance in tomato against root knot nematodes is efficient in reducing their population densities and thereby, reducing the need for pesticide application. Approach of resistance in tomato against the Meloidogyne species is extensively exploited in India. The utilization of bioagents like fungi and bacteria may provide an environmental friendly strategy that could be more effective in combination with resistant or partially resistant cultivars by reducing the nematode reproduction enough to affect the residual nematode population density in a field (Ciancio and Mukerji, 2008). Host plant resistance remains a very important potential component of a solution to many nematode problems of tropical agriculture especially, for the low input, small-scale farmers 14
Introduction when used in combination with cultural techniques and traditionally grown crops (Luc et al., 2005). But the additional important benefits of resistant crops are their environmental compatibility that do not require specialized applications and apart from preference based on agronomic or horticulture desirability, they usually do not require an additional cost input or deficit (Starr et al., 2001). Because of these all values has been given preferity over regulatory, cultural, chemical and biological control measures of nematode management. It furnishes reliable, effective and economically sustainable approach for the management of nematodes both in low and high cropping systems.
1.3.2. CHEMICAL METHODS
Since chemical nematicides were first developed, they played a dominant role in nematode control in major crops (Minton and Baujard, 1990). Nematicides are fast acting chemicals, rapidly reduce nematode population within few days after the application. Incorporation of chemical nematicides to the soil may lead to the dismissing of microbes which even may lower down the beneficial microbes’ population that further enhance the dominant nature of pathogen and finally may cause infection in the crop and promote for the negative shift in biological equilibrium. The modification of existing agricultural practices in order to manage nematode populations is one of the most acceptable alternatives to chemical control for both the small and large scale farmers in the tropics (Starr et al., 2001). Nematicides are categorized into fumigants or non-fumigants on behalf of their volatility in soil. Majority of the fumigants are broad-spectrum pesticides and potentially efficient against eggs, juveniles and adult nematodes in comparison to other pests and diseases. Moreover, they also attack non-target organisms comprises of nematodes natural enemy. Non-fumigant nematicides involve carbamte and organophosphate showing either contact or nematostatic effects. They are applied during the planting time in soil (Dawar et al., 2008; Gitanjali devi, 2010). One of the greater environmental problems sometimes associated with nematicide usage is groundwater contamination thereby posing serious hazards to man and animals (Alam and Jairajpuri, 1990). In most of the countries, the use of some of the fumigant nematicides such as methyl bromide used on large scale, is under way while, ethylene dibromide (EDB), 1, 2- Dibromochloropropane (DBCP) and 1, 3-Dichloropropene has been suspended. The most effective and widely used fumigant is methyl bromide 15
Introduction
(Oka et al., 2000a). However, methyl bromide is implicated in the depletion of the ozone layer (Bell et al., 1996; Ristaino and Thomas, 1998). The elimination of mycorrhizae by methyl bromide can result in poorer plant growth (Klein, 1996). Moreover, many of the currently available nematicides offer no long-term suppression, often costly; having differential effects on species of nematodes and their activity is affected by many environmental factors (Schmitt, 1986; Starr et al., 2002). Aldicarb/Temik-10G (2-methyl-2-(methylthio) propionaldehyde-o-(methylcarbamoyl) oxime) a cholinesterase inhibitor, is a commonly used commercial nematicide and is no longer authorized for use in EU, and in August 2010 Bayer Crop science announced its plan to discontinue Aldicarb by 2014. Several chemical nematicides are expensive, hazardous and phytotoxic in nature and often lead to the eradication of most microbial residents (Jesse and Jada, 2004). The unavoidable consequences are the contamination of food and environment with toxic chemical residues resulting in the detrimental effects on humans, useful flora and fauna, livestock and outbreak of secondary pest infestation (Horrigan et al., 2002). Nematicidal chemicals besides being expensive and non-available when it needed (Jesse and Jada, 2004), also found to be unsafe i.e., highly toxic aldicarb and methyl bromide used to control nematodes. This create microbial vacuum, which often leads to a rebounding of pathogens (Gamliel et al., 2000).
1.3.3. BIOLOGICAL METHODS
The soil rhizosphere is the most diverse zone of passionate and intensified microbial activity where organisms of multiformity and varied diversity, along with nematodes, live together. Introduction of antagonists in the soil micro-environment has resulted in an efficient method for biological control of nematodes (Akman et al., 2002; Singh and Mathur, 2010). Biological control of nematodes can be acquired either by exploration of antagonistic organisms, promotion and conservation of indigenous antagonists, or by the combined usage of both of the approaches. The application of biological control has been changeable in suppressing nematode populations because the efficacy of antagonists is influenced by other soil organisms and the host-plant (Bhagawati et al., 2009). Welker (1988) reported that the microorganisms present in the rhizosphere provide a front line defense for root pathogen attack that can be used as an alternative to chemical control of plant pathogen and has generated much interest in recent years (Cook and Baker, 1983; 16
Introduction
Blackman and Fokkema, 1982; Vivader, 1982). Application of biological agents is more successful when harmonized with other management strategies, especially cultural control methods (Stirling, 1991; Kerry and Hominick, 2002). Trichoderma species for example, are capable of mycoparasitism; aggressive competitors for nutrients, space and producers of chemical agents such as antibiotics. For this reason these fungi have been the most thoroughly researched fungal biocontrol agent (Whipps, 2001). Antibiosis is the dominant mode of action of bacterial antagonists as well (Chet et al., 1990).
Biological control of plant-parasitic nematodes with antagonistic fungi is promising techniques which may be incorporated in integrated nematode management (Stirling, 1991; Persson and Jansson, 1999; Duponnois et al., 2001). In this method one or more organisms are used to check the population of another pest (Kroschel, 2001; Nekouam, 2004). Several soil inhabiting fungi like Paecilomyces lilacinus, Pochonia chlamydosporia and Trichoderma spp. have been reported to be aggressive parasites of sedentary stages of nematodes (Siddiqui and Mahmood, 1996; Sharon et al., 2001, 2007). In several countries Purpureocillium lilacinum strain 251 is registered for biological control of nematodes (Siddiqui and Mahmood, 1996; Atkins et al., 2005). Pochonia chlamydosporia (Goddard) Gams and Zare is a ubiquitous, facultative, nematophagous fungus with parasitic activity against eggs and sedentary females of economically important plant-parasitic nematodes and associated with soils that suppress the multiplication of cyst nematode populations (Kerry and Crump, 1977; Kerry et al., 1982). In addition to this P. chlamydosporia may promote plant growth in the initial stages of root colonization, and elicit plant defense against endophytic colonization (Bordallo et al., 2002; Lopez-Llorca et al., 2002; Mácia- Vicente et al., 2009a, b). Fluorescent Pseudomonas spp. are among the most effective rhizosphere bacteria in ameliorating diseases caused by soil-borne pathogens (Siddiqui and Shaukat, 2004). The bacteria might produce compounds antagonistic to other soil microbes, such as phenazine-type antibiotics or hydrogen cyanide (Vanloon and Bakker, 2005). Pseudomonas fluorescens can act as strong elicitors of plant defense reaction (M’Piga et al., 1997). These bacteria antagonise pathogens either by producing bacterial siderophores, antibiotics, chitinases and glucanases or by inducing systemic resistance in plants against soil-borne fungi and plant-parasitic nematodes (Mazzola et al., 1992; Cronin et al., 1997; Hasky-Gunther et al., 1998). Several
17
Introduction strains of Pseudomonas produce secondary metabolites, such as 2, 4- diacetylphloroglucinol (DAPG), phenazines and hydrogen cyanide, thereby inhibiting soil-borne pathogens (Haas and Defago, 2005). DAPG inhibits mitochondrial activity (Gleeson et al., 2010) and also helps in the control of plant-parasitic nematodes by Pseudomonas (Cronin et al., 1997; Siddiqui and Shaukat 2003a). Trichoderma spp. has been widely studied as a biological control agent against microbial diseases of crops (Cherif and Benhamou, 1990; Chet, 1987; Chet et al., 1981; Elad et al., 1980, 1983). Sharon et al. (2001) have suggested that nematicidal activity of T. viride may be due to the eggs and larvae being infected through the increase in chitinase and protease activity. According to Shebani et al. (2008), direct parasitism of eggs through increase in extra cellular chitinase activity as indicated by egg infection capability and inducing plant defense mechanism leading to systemic resistance are the two possible mechanisms for the suppression of nematodes.
Sustainable functioning approaches have been recommended, along with the integrated pest management (IPM) which allow the combination of biological, physical and cultural methods so that they act synergistically through the direct suppression of nematodes, promotion of plant growth, and facilitation of rhizosphere colonization and activity of the microbial antagonists (Akhtar, 1997; Barker and Koenning, 1998; Barker, 2003).To fulfill this aim it is necessary that the first fungi should establish, survive and proliferate finally. The establishment and proliferation of the nematophagus fungi is mainly organic matter dependent having proper nutrient availability. Organic matter serves as primary source of nutrients for nematophagous fungi and nematode as secondary source (Nicolay and Sikora, 1991). Utilization of organic matter in combination with microbes seems to be an efficient and interesting tactic for the conservation and sustainment of soil productivity. It is also beneficial for the improvement of plant growth and management of root-knot disease.
During the course of the study, screening of different cultivar of tomato against root knot nematode M.incognita to test the resistance and susceptibility behavior. Most of the cultivars were observed susceptible and moderately susceptible and only two cultivar were found resistant. Whereas variety K-21 was found most susceptible. Keeping this view, it’s desirable to select this crop with particular cultivar, K-21 which is grown locally at the large scale for the commercial purpose and to manage the nematode problem through the involvement of organic matter and 18
Introduction bioagents that are environmental friendly and safe. So following objectives were investigated and pot trials were done in Department of Botany, AMU, Aligarh (Glasshouse)
1. Screening of different cultivars of tomato against root-knot nematode Meloidogyne incognita, under glass house conditions.
2. Effect of aqueous dilutions of different plant species, oilcakes, bioagents and biochar on the juvenile hatching and mortality of Meloidogyne incognita in vitro
3. Effect of bare-root dip in various plant extracts bioagents, oilcakes and biochar on the penetration of second stage juveniles (J2) of Meloidogyne incognita into the roots of tomato cv. K-21.
4. Effect of bare-root dip in various aqueous dilutions of plant extracts, bioagents alone and in combination on root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots.
5. Effect of chopped leaves of different plant species in combination with seed powder of Black nightshade on the root-knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots.
6. Effect of biochar alone and in combination with various agriculture wastes and saw dust used as soil amendment on the root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21in pots.
7. Effect of various aqueous dilutions of oil cakes, bioagents and biochar as seed dressing agents on the root-knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots.
8. Studies on the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and bioinoculants on the root-knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots.
9. Effect of bioinoculants in combination with different organic soil amendments on the root-knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots.
10. Effect of bioinoculants in combination with composted plant straws and Potato waste on the root-knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots.
19
Review Of Literature
CHAPTER II
REVIEW OF LITERATURE
2.1. CULTIVAR RESISTANCE/SUSCEPTIBILITY
Plant parasitic nematodes are disastrous, extremely polypahgus and antagonistic to crops and agriculture system causing reduction in yield and productivity. Whitehead (1998) reported that 10% of world crop production is lost as a result of plant nematode damage. Although among the plant parasitic nematodes, root knot nematode is sedentary endparasite that considerably damage the economically important annual, biennial and perennial crops (Khan et al., 2000; Gogoi and Neog, 2003). Exploitation of resistance in crops is one of the most effective and ecofriendly components of integrated pest management and inclusion of this property ensures increased crop yield in the presence of nematode (Khan and Mukhopadhyay, 2004). Resistant varieties can produce the most dramatic increase in yields of many crops and appear to hold the solution to most nematode problems (Luc et al., 2005). Resistance in the host plant can be superiorized all the management strategies viz., amendment, cultural, chemical and biological. Since it provides environmental friendly, safe and sustainable nematode management method at low level and commercial scale cropping system. Netscher and Mauboussin (1973) suggested that application of resistant cultivar is an economical and environmentally safe method for the nematode management. Preformed defense and response are two type of resistance occurring in the plants. However, in plant nematology in relation to disease susceptibility it is defined as the potential of plants to suppress the reproduction of nematode species relative to reproduction on a plant lacking such resistant (Friedman and Baker, 2007).
Screening of crop varietiess is time-consuming, but if a stable cultivar is discovered, it covers many years’ expenses (McDonald and Linde, 2002). Mi gene 27 originally found in wild tomato species, Lycopersicon peruvianum (Mill). is one of the best characterized nematode resistance genes and has been genetically engineered into many commercial tomato varieties (Nono-womdim et al., 2002; Abad et al., 2003). Mai and Abawi (1987) reported that intensive galling significantly reduced root efficiency resulting into yield reduction.
20
Review Of Literature
Nematode feeding on the root lead to poor plant growth (Hussey and Williamson, 1997). Reproduction of species which have close relationship with their hosts can often be assessed by severity of plant symptoms, such as galling caused by root-knot nematodes (Cook and Evans,1987). However, Hirunsalee et al. (1995) observed that reproduction and galling of nematodes on plant root were favoured on tolerant and susceptible varietiess but inhibited on resistant ones. Mattos et al. (2011) who found that S. asperolanatum, S. stramonifolium and Solanum sp. were resistant to M. incognita race with reproduction factors of 0.17, 0.06 and 0.49, respectively, these values were less than 1. Root and shoot growth of five tomato verities like, Roma Holland, Roma v.f., Sunehra, Roma Holland and Tomato Anmol was significantly reduced when inoculated with second stage infective larvae of M. incognita as compared to un-inoculated tomato varieties. Also, number of larvae was significantly increased in tomato variety Roma v.f., Gola France followed by Roma Holland and that was decreased in Anmol (Khanzada et al., 2012). To identify resistant genotypes, in 2013–2014 a total of 32 genotypes were screened against M. incognita at 2000 J2/Kg inoculum. Genotypes, Motelle and H-88-78-1 have shown immune reaction. Mogor and Hisar Lalit also showed resistance reaction (Yerasu, et al., 2016).
Khan (1994) reported that nematode resistance in host plants was manifested by reduced rates of nematode reproduction, eggmasses and consequently, low nematode population densities than that of a susceptible one. However, for some crops, root galling is not a completely satisfactory indicator of root knot nematode resistance and usually a preliminary test should be conducted to determine if a strong correlation exists between galling and nematode reproduction (Hussey and Boema, 1981).
Kaur et al. (2014) screened two hundred tomato germplasm lines to identify resistant sources. PNR-7 and Punjab Upma were used as resistant and susceptible checks, respectively. Seven lines viz. Hisar Lal, EC631955, EC631956, EC631957, EC119197, 8-2-1-2-5 and 1-6-1-4 were found to possess high degree of resistance and two lines viz. EC 531804 and PAU Acc.-1 were found resistant through conventional screening. Several varietiess of tomatoes such as Montelle, Sun6082, Pik Red, Celebrity, Baja, Roma VFN, Lemon Boy, enchant men, Better boy and Beefmaster have been developed in an attempt to produce root-knot nematodes resistant varietiess (Milligan et al., 1998). Appleman (2003) reported that, galling increased dramatically 21
Review Of Literature after 42 days, with 70-80% galling being reached on the 45th day. These results confirm that lettuce can function as a rapid indicator of nematode galling and will shorten the screening time from 90 days to 45 days. Susceptibility of a plant to root- knot nematode, Meloidogyne spp. depends on the ability of juveniles (J2) to penetrate the roots of the plant and cause the formation of giant cells which appears as galls on the roots (Chen et al., 2004).
Nadary et al. (2006) and Chindo et al. (2006) observed that as initial inoculum level of M. incognita populations increased, populations that had high infection incidence and reproduction rates induced greater root galling than populations. Also, six tomato varietiess Moneymaker, Beefsteak, Roma, Summer taste, Mini Roma and Small fry were tested for their susceptibility to root-knot nematodes at inoculum levels of 200, 400, 600 juveniles (J2) per pot. All varieties were found to be susceptible to varying extent as eggmasses were present in all varities.Moneymaker and Roma were found most susceptible and Mini Roma, the least susceptible Singh and Khurma, (2007).
Fassuliotis (1979) reported that because galling occurs in most susceptible plants infected with root-knot nematodes, this can often be the sole measurement of resistance during screening.Cousins and Walker (1998), root-knot nematode eggs developed poorly on resistant accession compared to susceptible accession. Niblack et al. (1986) demonstrated that at moderate to high initial population densities, of root- knot nematodes reached their maximum levels on susceptible varieties. El-Sheriff et al. (2007a) observed the effect of fifteen population densities of M. incognita ranging from zero to 5000 eggs on yield of tomato.They found the maximum number of nematode juveniles in moderate population density (1000 to1500 eggs); and at very high population densities the reproduction potentials of root-knot nematodes on the plant declined because the population in the root system reached its peak and could not support further reproduction. Karssen and Moens (2006) reported that highly susceptible host plants allowed the juveniles to enter the roots, reached maturity and produced many eggs, while the resistant plants suppressed their development and thus, do not allow reproduction.
Maqbool and Ghazala (1986) reported that the growth of tomato seedlings that was significantly reduced when artificially inoculated with Meloidogyne spp. Darban
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(1994) observed that growth of tomato variety Roma was decreased when inoculated with different inoculum levels of M. incognita. Rao et al. (1998) found post- penetration of second stage larvae of M. incognita in tomato hybrid FM-2 and Pusa Ruby before and after transplanting. Lawerence and Clark (1986) observed that number of females, root-galls and eggmasses were increased on susceptible varieties inoculated with M. incognita. Roberts and May (1986) studied the interaction of M. incongita and M. javanica and tomato varieties on average root-knot index and number of eggs on infected plants. Nematode reproduction and host damage are both affected by the initial inoculum levels and revealed an increase in number of juveniles and egg masses of the plant as the inoculum level increased from 250 to 1000 eggs/ tomato plant (Salem et al., 2007).
Several varieties of tomatoes such as Montelle, Sun6082, Pik Red, Celebrity, Baja, Roma VFN, Lemon Boy, enchant men, Better boy and Beef master have been developed in an attempt to produce root-knot nematodes resistant varietiess (Milligan et al. 1998).The occurrence of variation in susceptibility among seven tomato genotypes to M. incognita might be due to genetic differences (Ehlers et al., 2002; Jacquet et al., 2005).The higher the number of adult feeding females the greater the stress on the plant leading to alteration in physiological functions like up-take of nutrients, photosynthesis (Williamson and Hussey, 1996) and consequent plant growth. The infected plants showed disruption of xylem elements (Jatala and Jensen, 1976; Ismail et al., 2004), phloem (Sundararaju and Mehta, 1992), collapse of vascular vessel by hyperplasia and hypertrophy of giant cell (Acosta and Negron, 1982; Vovlas et al, 2005)
Heavily root-knot nematodes-infested plants, according to Sidiqqui and Alam (1985), exhibited stunted growth and poor yield. Resistant varieties gave maximum increase in plant height and minimum increase in fresh root weight due to the less root-knot formation. The gradual reduction in plant growth coupled with increased nematode multiplication with increasing Pi on susceptible cv. Round-41 is comparable with that of bitter gourd and round melon (Pankaj and Siyanand, 1990), cucurbits (Khan et al., 2004), cowpea (Haseeb et al., 2005) and beans (Nadaryet al., 2006). Hunt et al. (2005) reported that root-knot nematodes establish specialized feeding cells in roots, redirecting photosynthates produced in the leaves to supply the energy demands of the nematode in the roots. Root weight of susceptible varieties as a 23
Review Of Literature result of nematode parasitism increases whereas shoot weight declines, shifting the root-shoot balance (Roberts, 1995).Wonang and Akueshi (1990) found that there was greater increase in the fresh weight of tomato roots with increase in the inoculum density.
Rehman et al. (2006) tested six tomato varietiess Moneymaker, Beefsteak, Roma, Summertaste, Mini Roma and Small fry for their susceptibility to root-knot nematodes at inoculum levels of 200, 400, 600 juveniles (J2) per pot. All were found to be susceptible to varying degrees as eggmasses were present in all with Moneymaker and Roma being the most susceptible and Mini Roma, the least susceptible. According to Khan (2000) the influence of nematode inoculum density on number of galls developed on tomato seedlings revealed significant increase in the number of galls with increase in the inoculum density.
2.2. ORGANIC AMENDMENTS
Nematodes are small, slender, colourless, unsegmented, multicellular and eukaryotic roundworms of the phylum, Nematoda. By their body structure they are quite simple in body organization having bilateral symmetry, triploblastic, pseudocoelomic, cell wall made up of cuticle and lacking circulatory and respiratory system. Members of this phylum are distributed worldwide in all climatic zones, occurring often in great numbers wherever suitable nutrients and environmental conditions are favourable (Caveness and Ogunfowora, 1985). Most plant-parasitic nematodes feed on root tissue and damage their host mainly by:1) reducing root nutrient and water uptake; 2) promoting microbial or fungal infections through wound sites; 3) serving as vectors for pathogenic viruses. In some cases, nematode infection intensifies the disease caused by a primary pathogen (Hollis, 1964; Ohl et al., 1997). The majority of the plant species, which account for the major world’s food supply, is susceptible to attack from phytonematodes which are capable of causing sustainable economic losses in the quantity and quality of the crops (Jain et al., 2007; Berry et al., 2008). However, over 4,100 species of plant-parasitic nematodes have been identified (Decraemer and Hunt 2006), new species are continually being described while others, previously viewed as benign or non-damaging, are becoming pests as cropping patterns change (Nicol, 2002). The crop losses caused by phytonematodes in economic terms estimated about $ 157 billion annually to the world agriculture (Abad
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Review Of Literature et al., 2008). Several management strategies are there for the nematode management in agriculture system, however, among all these management through chemical control is one of the best method but pest control strategies that solely rely on chemical control have been the subject of widespread criticism (Black and Sweetmore, 1994). Moreover, due to various short comings of nematode control by purely chemical means and environmental reasons has lead to search better and safe alternative through the involvement of resistant varieties, plants parts/products and bioagents. Which are environmental friendly, reliable and ecologically safe so that they may maintain the equilibrium of the soil and environment. The prime objective of the research work present in the thesis deals with the test of the efficacy of organic amendment, biochar, agriculture waste and bioagents for the management of root-knot nematode, Meloidogyne incognita. So therefore attempt has been drawn to summarize the review of different agriculture tactic has been employed for the management of nematode Meloidogyne spp.
2.2.1. Plant parts/Products
Plants are reservoirs of glorious and fascinating natural biopesticides. Addition of plant parts to the soil is one of the traditional agricultre practices for the nematode management that improve the soil structure, texture, nutrient content and soil flora and fauna. Organic amendment has been suggested as a promising alternative practice to improve soil quality and plant health such as suppressing plant-parasitic nematodes (Collange et al., 2011; Oka, 2010; Thoden et al., 2011) and develop sustainable crop production in small and large scale experiments (Franco-Otero et al., 2012; Bowles et al., 2014; Ninh et al., 2015).
Amending soil with different parts of neem (A. indica) has been found to be highly effective in reducing the population of various phytonematodes (Rather and Siddiqui, 2007b; Nattli et al., 2009). The addition of organic matter in the form of composed manure decreased nematode pest population and associated with the improvement of damaged crops (Oka and Yermiyahu, 2002; Walker, 2004;) who evaluate that root-gall nematode damage on lima bean decreased with increase ammonium supplied to the plant. Jiskani et al. (2005) observed that the efficacy of different doses of neem seed decoction and neem leaf extracts on the reproductive activity of M. incognita infesting tomato showed that plant growth parameter
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Review Of Literature significantly increased and root galls decreased. The degradation of neem (Azadirachta indica) leaves over a period of six weeks before transplanting the tomato seedlings significantly reduced the root-knot incidence and improved the shoot weight and length (Jain and Bhatti, 1988). Javed et al. (2008) recorded that soil treatment with neem crude formulation significantly reduced the intensity of root galling and number of egg masses caused by Meloidogyne javanica on the roots of tomato.
Baloch et al. (2013) stated that application of Ferula asafoetida, Spatoglossum variabile, Stokeyia indica and Melanothamnus afaqhusainii showed significant suppressive effect on root- knot nematode, Meloidogyne incognita attacking watermelon and eggplant. Length of vine of watermelon, shoot length of eggplant and fresh shoot weight were higher in seaweed and asafetida treated plants as compared to control or Topspin-M, a fungicide, treated plants. Soil amendment with Argemone mexicana, a tropical annua1 weed and Amaranthus spp. substantially lowered the populations of M. javanica in the roots and rhizosphere of tomato plants under greenhouse conditions (Shaukat et al., 2002).
The nematicidal properties of some plants like marigold, basil, nitta and rattle weed recorded completely (100%) inhibition of hatching of root-knot nematode eggs and also destroyed (100%) the root-knot nematode juveniles at 10% concentrations (Olabiyi and Oyedunmade, 2003). The presence of nematicidal compounds (flavonoids and saponins) in the root extracts might be responsible for significant reduction in soil population of M. incognita and gall index of tomato root. Shoot extracts of Euphorbia helioscopia, Descurainia Sophia, Gypsophila pilosa, Eruca sativa and P. lanceolata were found to suppress the root galls in tomato plants produced by M.incognita and increased the fruit yield of the plant (Hoseinpoor and Kargar, 2012).
Arshad et al. (2011) observed that leaf amendment of Azadirachta indica, Calotropis procera, Datura stramonium and Tagetes erecta at different dosages significantly improved the plant growth characteristics of okra and reduced root-knot infections compared with the untreated control. Azadirachta indica and C. procera caused the maximum reductions in number of galls, egg masses and reproduction factor (Rf) of the nematode. Sharma and Tiagi (1989) studied the efficacy of certain leaf powders against root-knot nematodes on pea and reported enhanced plant growth
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Review Of Literature and reduced galling by all leaf powders. Akhtar and Alam (1990) presented Calotropis procera as the most effective against freshly hatched juveniles of Meloidogyne incognita. It was better than Azadirachta indica, Ricinus communis and a host of other plant leaves evaluated against root knot nematodes on tomato and chilli.
Badra et al. (1979) reported that plants growing in amended soil contained greater concentrations of phenols than those growing in unamended soil and this may induce disease resistance in roots. Aqueous extract of some plants viz., Lantana (Lantana camara), Mexican marigold (Tagetes minuta), as soil drenching @ 5% concentration results in pronounced reduction in final nematode population density, root-knot index and a significant increase yield per plant and total yields of tomato (Taye et al., 2012). Application of Garlic bulb extracts reduced root-knot infection indices on tomato. Garlic extract was found to have greater potential than neem leaf extract in the control of root knot infection in tomato (Agbenin et al., 2005). Gitanjali (2010) demonstrated that incorporation of Tithonia leaf and stem significantly increased the plant growth and decreased the host infestation by M.incognita.
Chimbekujwo et al. (2013) observed that leaf powder of neem (Azadirachta indica), kassod tree (Cassia siamea), eucalyptus (Eucalyptus gigantea) and locust bean tree (Parkia biglobosa) shows significant (p<0.001) reduction in root galling and nematode population, as well as improved plant growth and yield of cowpea. Although all treatments were effective in reducing root galls and nematode population, application of Azadirachta indica leaf powder gave the highest reduction in root galls (0.293) and nematode population (24), followed by Cassia siamea, Eucalyptus gigantea and Parkia biglobosa.Parihar et al. (2012) claimed that the soil treated with Datura stramonium (100g) leaves were found most effective in reducing the reproductive potential of populations of root-knot nematode and increased chlorophyll content and plant growth parameters viz., length, fresh and dry weight of shoot and root as compared to other plant species viz., Argemone mexicana, Lantana camara, Parthenium hysterophorus and Withania somnifera. Pakeerathan et al. (2009) revealed that all green leaf manures viz., Gliricidia maculata, Thespesia populnea, Calotropis gigantia, Azadiracta indica and Glycosmis pentaphylla cause significant improvement in the plant growth and reduced the nematode infestation in
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Review Of Literature tomato fields. Among all Gliricidia maculate shows best results compared to other treatments.
Marigold and sea ambrosia plants have been reported to suppress infection and damage caused by Meloidogyne incognita, when incorporated as a green manure (El-Hamawi et al., 2004). Dried powder of turmeric rhizome, neem cake, neem leaf, neem seed kernel and root powder of some plants viz., Tagetes erecta, Ocimum sanctum, Calendula officinalis, Carica papaya and Azadirachta indica were used to suppress the nematode effectively and consequently improved the plant growth (Chakraborti, 2001; Rajendran and Saritha, 2005).
Tariq et al. (2008) stated that Powder from all plant parts, like leaves, stem and pneumatophore, from Avicennia marina and Rhizophora mucronata were effective in suppression of root knot nematode. He also observed that all the tested parts of Avicennia marina and Rhizophora mucronata as the organic amendments @ 0.1, 1 and 5% (v/v) caused significant increase in germination of seeds, shoot length, shoot weight, root length, root weight. Ismail et al. (2009) reported that waste residues from black cumin seed (Nigella sativa L.) and jojoba (Simmondsia chinensis) applied significantly reduces soil and root population of M.incognita. Different concentrations of the spent mushroom compost were applied to the M. incognita eggs. Oyster mushroom spent compost was found more effective in reducing the egg hatching and killing juveniles of root-knot nematodes than button compost. Oyster spent compost was found more effective in inhibiting the galling and production of eggmasses in the roots followed by button compost thereby stimulating the plant growth as compared to button compost (Sabina and Saifullah, 2013).
El-Nagdi et al. (2013) claimed that aqueous extracts of garlic (Allium sativum) cloves and castor bean (Ricinus communis) seeds, was found to be highly effective and significantly (p ≤ 0.05) reduced nematode criteria including number of galls and eggmasses on roots of tomato and number of juveniles in roots and soil, compared to nematicides and non-treated plants. The use of marine algae and garlic as control agents against plant-parasitic nematodes has been studied by many workers (Paracer et al., 1987; Anter et al., 1994). Previous studies have also been showen that the addition of asparagus, Egyptian clover, marigold, marguerite, neem, wormwood and olive dried plant materials caused significant reduction in galls and eggmasses of
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Meloidogyne spp. on sunflower plants (EL-Nagar et al., 1993; Abadir et al., 1994).Combined application of wild spinach powder along with the fresh chopped leaves of all the plants suppressed pathogenic effect of nematode and thereby resulted in significant reduction in Meloidogyne incognita infestation and population density of Meloidogyne incognita in soil. The highest reduction in Meloidogyne incognita infestation was reported in plants employed with 10 g of wild spinach powder combined with 50 g of Mexican poppy leaves. It was followed by 50 g of Trailing eclipta, 50 g of Wild eggplant, 50 g of Black pigweed, 50 g of Indian mallow, 50 g of Ivy gourd in the descending order. However the lowest reduction was recorded with the application 10 g of wild spinach powder plus 50 g of fresh chopped Ivy gourd. (Asif et al., 2016)
2.2.2. Nemato-toxic Potential of Plants
Plants are rich source of biochemicals which may serve as safer alternatives for synthetic nematicides and research in this area has increased manifolds (Ujvary, 2002). These compounds are known as alkaloids, diterpenes, fatty acids, glucosinolates, isothiocyanates, phenols, polyacetlenes, sesquiterpenes and thinly (Ma et al., 1998; Bruneton, 1999; Chitwood, 2002).
The inhibitory effect of crude extracts of T. violacea on nematode population densities in the roots was high while in the soil was low, suggesting that the material had the ability to penetrate through the roots. The inhibitory effect might be due to the chemical properties present in the extract that possess nematicidal properties (Agbenin et al., 2005). Most reports on the use of garlic and neem leaf extracts in nematode control used high application rates and concentrations (Sukul et al., 1974; Agbenin et al., 2005). Aqueous and ethanolic extracts of T. violacea tubers have previously been shown to have anthelmintic activity (McGraw et al., 2000). Natural compounds accumulated in plant extracts serve as growth regulators or have ecological role such as protection against fungal, viral and bacterial diseases (Pretorious et al., 2003). The increased plant growth under crude extracts of T. violacea was different to that in plants infected with Meloidogyne species (Siddiqui and Alam, 1987). All the manures were effective in suppressing nematode activities as manifested in enhanced growth observed in both organic and inorganic manure treatments compared with the control. Egg plants in control treatment were heavily
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Review Of Literature galled by Meloidogyne incognita resulting in poor growth while the organic and inorganic manure treated eggplant recorded better growth and were significantly different from the control (Abolusoro et al., 2013). Increased plant growth observed in this study suggested that the concentrations of crude extract levels of T. violacea were still in the stimulation range for growth of tomato plants. Similar responses were observed when tomato plants were exposed to fermented crude extracts of fruits from wild Cucumis species (Pelinganga, 2013).
In vitro and pot studies, extracts of Calendula officinalis, Enhydra fluctans and Solanum khasianum reduced galling and inhibit hatching (Goswami and Vijayalakshmi 1986). Khan et al. (2011) pointed out that the curative effect of plant extracts viz., neem, tobacco, Aloe vera, chilli, clove, garlic and onion against M. incognita. Garlic (Allium sativum) leaf extract has been successfully used to increase Tylenchulus semipenetrans mortality at high concentrations in laboratory conditions (Ayazpour et al., 2010). Garlic has indirect effects on nematode populations because it disrupts their mobility, food absorption and reproduction (Fadzirayi et al., 2010).
Costa et al. (2003) reported that Artemisia vulgaris rhizome extracts inhibited egg hatch, caused second stage juvenile mortality and reduced root gall of Meloidogyne megadora on Phaseolus vulgaris. Oka et al. (2001) observed that mixed leaf powder of Inula viscosa at a concentration of 0.1% (w/w) greatly reduced the number of second-stage juveniles of M. javanica and organic solvent extracts were found to be more toxic than aqueous extracts in in vitro experiment. Similar observation was reported by Javed et al. (2008b) who suggested that the 10 % extracts of neem leaf and cake caused 83 % and 85 % immobility and 35 % and 28 % mortality, respectively. Aza caused neither immobility nor mortality of juveniles. Goswami (1993) obtained a significant reduction in root gall index where soil was treated with P. lilacinus with castor leaves and fertilizer.Results revealed that considerable inhibition in hatching of eggs occurs in the aqueous extracts of vermicompost produced from wastes of menthol mint (Mentha arvensis), chamomile (Matricaria recutita), geranium (Pelargonium graveolens), qinghao (Artemisia annua) followed by pyrethrum(Chrysanthemum cinerariaefolium), isabgol (Plantago ovata), African marigold (Tagetes minuta), Boerhavia (Boerhavia diffusa), mustard (Brassica compestris), lemongrass (Cymbopogon flexuosus) and garden mint (Mentha viridis) (Pandey and Kalra 2010). 30
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Botanicals used by various workers to manage root-knot nematode (eggs and juveniles in vitro and in field conditions)
Botanicals Parts used Target Reference nematode Cassia tora and Morus alba Leaf M. incognita Tanweer et al.(2009) (juveniles) Nicotiana tabacum; Leaf M. incognita Wiranto et al.(2009) Syzygium aromaticum; Piper ( juveniles) betle; Acorus calamus Andrographis panniulata Leaf M.incognita Devi (2009) (juveniles and eggs) Cassia tora; Morus alba; Leaf M. incognita Tanweer and Musa paradisica; Psidum (juveniles) Hisamuddin (2010) gujava Melia azadirachta and Leaf and M. incognita Katooli et al. (2010) Brassica spp. seed oil (juveniles) Lantana camara leaf M.incognita Faheem et al. (2010) (eggs and juveniles) Anagallis arvensis; Curcuma Leaf, fruits M.incognita Haidar and Askary longa; Mentha viridis; and seeds (juveniles) (2011) Moringa oliefera and Ocimum sanctum Castor bean, Chinaberry, Leaf M.incognita Katooli et al. (2011) sweet wormwood and (juveniles) rapeseed Red chilli Fruit M. incognita Khan et al. (2011) (eggs and females) Jatropha curcas; Parkia Leaf M.incognita Ugwuoke et al. biglobosas; (juveniles) (2011) Newbouldia laevis; Ficusex asperate and Cassia alata P. hysterophorus, N. Leaf M incognita Usman and plumbaginifolia, A. fatua, C. (Eggs and Siddiqui(2011a) album, A. retroflexus, C. juveniles) murale,A. spinosus and O. corniculata Garlic, castor beans and Bulbs, leaf Meloidogyne Tibugari et al. (2012) marigold and flowers javanica (juveniles)
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Tithonia diversifolia; Leaf M.incognita Akpheokhai et Azadirachta indica; ( juveniles) al.(2012) Zanthoxylum zanthoxyloides and Datura metel Glycosmis pentaphylla and Leaf and M. incognita Bhattacharya et al. Holarrhenaantidys enterica bark ( juveniles) (2012) Couroupita quianensis; Leaf Meloidogyne Pavaraj et al. (2012) Nepeta cataria and incognita Pentanema indicum (eggs) Datura stramonium Leaf Meloidogyne Parihar et al.(2012) javanica (juveniles) Chrysanthemum coronarium, Leaf and M.javanica Moosavi (2012) Azadirachta indica, Nerium seed (eggs and oleander juveniles) Vernonia amygodalina, leaf M.incognita Taye et al. (2012) Lantana camara,Tagetes (eggs) minuta, Millettia ferruginea Bitter leaf and Cashew Leaf, seed Meloidogyne Umar and Aji (2013) kernel incognita Eichhornia crassipes Leaf M. incognita Umar and (juveniles) Mohammed (2013) Calotropis procera Fresh leaf M.incognita Chedekal (2013) (eggs and juveniles) Rauvolfia tetraphylla Root, leaf M.incognita Mandal and Nandi and fruit (juveniles) (2013) extract Mimusops elengi Leaf Meloidogyne Azhagumurugan and incognita Rajan (2013) (eggs) Calotropis procera Leaf M.incognita Eunice et al. (2013) (eggs) Allium sativum and Ricinus Clove and M.incognita Youssef (2013) communis seed (eggs and juveniles) Castor bean Seed Meloidogyne Adomako and spp.(eggs and Kwoseh (2013) juveniles) Datura stramonium Leaf M. incognita Chaudhary et al. (juveniles) (2013) Hunteriaum bellata and Leaf M.incognita Okeniyi et al. (2014) Mallotus oppositifolius (eggs and
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juveniles) Gmelina asiatica Leaf M.incognita Azhagumurugan and (eggs and Rajan (2014) juveniles) Jatropha Leaf M.incognita Asif et al. (2013) pandurifolia; Polyalthia (eggs and longifolia;Wedelia juveniles) chinensis; Nerium indicum;Duranta repens and Cassia fistula Mentha piperata leaf M.incognita El-Deen et al. (2014) (eggs and juveniles) Melia azedarach; Humulus Fruit, M.incognita Akyazi (2014) lupulus; Sambucus nigra; leaf,stem, (eggs and Conium maculatum inflorescence juveniles) Euphorbia heterophylla, Leaf M. incognita Umar and Adamu Richardia bransiliensis and (juveniles) (2014) Scoparia dulcis Bridelia micrantha; Mallotus Leaf M. incognita Okeniyi et al. (2014) oppositifolius; Hunteriaum (eggs and bellata and Citrus medica juveniles) Nigella sativa, Ferula assa- Seeds and M. incognita Sholevarfard and foetida, Peganum harmala, oleo- gum (eggs and Moosavi (2015) and Portulaca oleracea resin juveniles) Adhatoda vasica; Leaf M.incognita Singh et al. (2015) Andrographis affinis (eggs and juveniles) Wedelia chinensis, Colocasia Leaf M.incognita Ansari et al.(2016) esculenta, Euphorbia hirta, (eggs and Coccinia grandis, Abutilon juveniles) indicum and decaffeinated tea Amaranthus spinosus L., Leaf M.incognita Asif et al.(2016) Ranunculus pensylvanicus (eggs and L.f., Cassia tora L., Oxalis juveniles) stricta Adapted and modified from Mnaju and Sankari (2015)
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2.2.3 Agricultural waste
The addition of residue and waste from plants and animals to soil has been explored as one of the best substitute for the nematode management. These products comprise agricultural wastes in the form of dried residues of crops, green manures, remaining of crops and vegetables in general and industrial by-products such as press mud, slurry, oilcakes, sawdust, sugar-cane bagasse and cellulosic waste in particular. Besides these, other biological wastes such as chitin, wastes-bone meal, meat meal, blood meal, horn and hoof meal, fish wastes, sewage-sludge, municipal refuse and livestock wastes have been proved successful in controlling nematodes for several years (Akhtar and Alam, 1993).
Hassan et al. (2010) conducted field experiments to test the efficacy of three organic wastes, namely refuse dump (RD), rice husk (RH) and sawdust (SD), @ of 15, 30 and 45 metric tons per hectare. Furadan (3G) was applied @ of 16, 32 and 64 kg/ha for the management of root knot nematodes, Meloidogyne spp. on tomato. The results showed that refuse dump treatment gave significantly (P = 0.05) highest reduction in the nematode population compared to non-amended treatment. It resulted in a significant (P = 0.05) increase in the yield of tomato by 17–100% for RD, 13– 84% for SD and 21–63% for RH. Sikora et al. (1973) reported 22 % reduction in the number of root galls induced by Meloidogyne spp. through soil amendment with sugarcane bagasse, at the rate of 4 t/ha, just before tomato transplantation; the reduction reached 90 % when tomato was transplanted 100-150 days after amendment. Ogwulumba et al. (2010) showed that soil amended with grass ash and rice husk ash was good for optimum growth performance and control of M. javanica infecting tomato. The use of composted dry cork, dry-grape (fruit residue after pressing) and a 1:1 mixture of dry-olive marc +dry-rice husk as an amendment to potting mixtures was assessed for the management of Meloidogyne species. Amending the potting mixture with composted dry cork at rates of 0%, 25%, 50%, 75% and 100% v/v, reduced the root galling and final populations of M. incognita (Akhtar, 1998).
Akhtar (1993) pointed out that press mud waste of plant origin such as vegetable and fruit processing and tobacco wastes were the most effective in reducing the incidence of root knot and the development of plant-parasitic nematodes on
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Review Of Literature tomato compared with other sources. Amendments to soil with tea, wheat straw, paddy husk, paddy straw, sugarcane trash, domestic garbage, dead vegetation and pigeon pea stubble also gave some level of control. Effect of bidi tobacco dust on root-knot nematode attacking okra cv. Parbhani Kranti showed that application of tobacco dust, 10 days prior to sowing (DPS); either at 1, 5 or 10 per cent significantly reduced the plant growth of okra indicating its adverse/toxic effect. However, it significantly reduced the root-knot disease and nematode multiplication compared to control. Increase in doses of tobacco dust further gradually reduced the growth of okra, root-knot disease and nematode multiplication (Tapre and Patel, 2015).
According to Berkerlaar (2001) nematode problems are worse in soils with low amounts of organic matter content than in soils with high amounts. Organic soil amendments should ideally be by-products and wastes from industrial and other activities and include oil cakes, sawdust, plant composites, green manures and agro- industrial wastes ( Ibrahim and Ibrahim, 2000; Umar and Jada, 2000).
Poultry litter results significant improvement appeared especially when combined with sorghum cover crop (Everts et al., 2006).Cassava peelings, cocoa pod husk and rice husk significantly reduced Meloidogyne spp. population infecting cowpea (Egunjobi, 1985; Egunjobi and Olaitan, 1986). Cassava leaf and tuber rind applied as soil amendment @100g or 50 g/pot, significantly reduced the population of M. incognita and improved plant growth parameters of okra (Ramakrishnan et al., 1999). Amending soil with 1:1 mixture of composted dry-olive marc and dry-rice husk did not reduce the root galling and final populations of M. incognita and M. javanica (Nico et al., 2004).
Directly amending mushroom compost into soil containing low organic matter resulted in poor performance in nematode suppression (Marrush, 2007). In fact the numbers of nematodes/g root were higher (P< 0.05) in plants with mushroom compost amendment than the unamended treatment. It is possible that poor establishment of the mushroom mycelium in the sand: soil mix with limited organic matter resulted in no suppression of the nematodes. This result would resemble a scenario where oyster mushroom compost was to be amended directly into a field soil with low organic matter. Agriculture soils generally contain less than 1-6% organic matter (McClellan et al., 2007).
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Application of Tithonia diversifolia, pyrethrum marc, vegetable waxy resins (VWR) and tea residue significantly reduced the nematode population and disease severity thereby help in improvement in plant growth characters (Nchore et al., 2012).The olive mill waste was more effective but slightly inhibited egg hatch (30%). These results are contradictory to reports that sterile water extracts of two phase olive mill waste strongly inhibited egg hatch and J2 motility of M. incognita (Cayuela et al., 2008). Root galling and egg masses on tomato roots induced by M. incognita and M. javanica were significantly reduced by addition of waste water from the olive oil industries to the soil (Vouyoukalou and Stefanoudaki, 1998).
The treatments rice straw and/or biofertile significantly increased nitrogen, phosphorus, potassium and organic matter contents of the soil, increased soil biological activity and reducing the appearance of infected root-knot nematode population and actualized prodigious depletion in egg production. Whereas rice straw compost application in faba bean rhizosphere resulted reducing root-knot nematode population by 74.9%. The rice straw extract reduced root-knot nematode (Abdelazzez Heba and Tewfike, 2014). Rice straw is agricultural waste produced in large amounts from rice cultivation and consists of different biopolymers such as cellulose (32- 37%), hemicellulose (29-37%) and lignin (5-15%) (Conrad, 2007). Incorporation of four by products namely poonac, tea waste, cattle manure and sugar molasses significantly reduced the nematode population on tomato thereby increase the growth (plant height, fresh and dry weights) of tomato and yield. The highest yield and lowest nematode population (galling index) was observed from plots treated with poonac (Nishantha and Ekanayake, 2000).
Plants treated with poultry and farmyard manures gave significantly higher yields than those of other organic manures. This was because rare root-galls occurred at poultry and farmyard manure applications. Plants with fewer root-galls would translocate more nutrients to vegetative organs than heavily galled roots (Otiefa and Elgindi, 1962).The addition of various rice straw composts on the rhizosphere soil microorganisms showed a high fertilizer value when applied @ of 5% (w/w). Various rice straw composts reduced root-knot nematode population and egg production (Rashid et al., 2011).
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An in vitro experiment involving five organic waste extracts, citrus waste, cocoa bean testa, compost and oil palm bunch waste were evaluated for their hatching inhibitory potential to the southern root-knot nematode, M. incognita eggs and best result was obtained with citrus waste extract (Osei et al., 2011). The reduction of soil pH by crude extracts of fever tea (Lippia javanica) used as a phytonematicide (Mashela et al., 2010).The reduced stem diameter in tomato plants was not simillar to the plants which were infected by Meloidogyne species, but not treated with crude extracts of garlic. Generally, nematode infection (Eisenback et al., 1991; Mashela, 2002; Mashela and Nthangeni, 2002a).
Hassan et al. (2001) tested powder and extract of ginger against root-knot nematodes and observed a better growth of plant with lower root galling index on brinjal. Zasada et al. (2010) reported that garlic suppressed nematode numbers, while phytotoxicity on tomatoes was not observed. However, the aqueous extract and volatile compounds of the wild garlic bulbs were found to be strong inhibitors of seed germination and seedling growth compared to those in the leaves (Djurdjevic et al., 2004). Nico et al. (2004) used decomposed agro-industrial waste products, namely composed dry cork, dry grape marc, dry olive marc and rice husk, as soil amendments for the management of Meloidogyne spp. Amending the potting mixture with dry- grape marc reduced the root galling and final populations of M. incognita race 1 and M. javanica in tomato, though there reductions in root galling (24.4% and 25.6%, respectively) and final nematode populations (34.2% and 34.7%, respectively). Gong et al. (2013) revealed that the application of garlic straw results in reductions of up to 50% nematode population compared to control.
Various rice straw composts @ 5, 7.5 resulted in reducing root-knot nematode population of 79, 84% respectively and actualized prodigious depletion in egg production (Rashad et al., 2011). Results indicated that all bio preparations, ground neem seed treatment suppressed the root-knot nematode to the best level expressing least number of root-knots and parasitic nematodes. Tobacco waste treatment was found to be the second best followed by the treatment with P. fluorescens (Motha et al., 2010). Vegetables and fruit processing waste and tobacco waste were most effective in reducing root-knot nematode.
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2.2.4. Oil cakes
It is well documented that incorporation of organic additives into the soil increased the organic matter contents and availability of other plant nutrients (Brar et al., 2004) and subsequently increased the predaceous nature of rhizospheric population of microbes thereby maintain the beneficial soil rhizosphereic population microbes.The soil application of organic amendments viz., neem cakes, castor cakes, groundnut cakes, sunflower cakes and Farm yard manures have significantly reduced the nematode population and increased the plant growth (Jagadeeswaran and Singh, 2011). Rehman et al.( 2012) claimed that non edible seed oil cakes can be used in the bio management of root-knot nematode, M. incognita affecting chickpea for pollution free and sustainable environment. Seed dressing with karanj, neem, mustard, castor, mahua cakes and neem cake was found the most effective followed by karanj @ 20% (w/w) in improving plant growth characters of chickpea and suppress M. incognita (Yadav et al., 2006).
Singh and Singh (1996) observed the effect of application of neem cakes, mahua cakes, saw dust and gobar gas products on root-knot nematodes. Neem cakes were found to be most effective followed by Mahua cakes. Saw dust only reduced root-knot formation whereas gobar gas product enhanced the growth weight of shoot and root. Organic amendment with neem cake, pongamia cake, farmyard manure, parthenium, castor cake etc. cause complete inhibition of egg hatching at 100% concentration (Naidu and Sudheer, 2007).
El-Sherif et al. (2008c) stated that when certain oil cakes viz., fennel and sesame significantly improved the growth of eggplant and suppressing the number of galls, females and eggmasses of M. incognita. The efficacy of oil seed cakes viz., Azadirachta indica (Neem), Brassica campestris (Mustard) and Gossypium hirsutum (Cotton) @50 g and 100 g as soil amendment gives satisfactory results in significantly reducing the root galls and enhance all the plant growth characters of C.arietinum. Higher dose of neem oil cake was found to be most effective as compared to other treatments (Frauk et al., 2011).
Neem cake extract was found to be most effective in killing M. incognita larvae (Gowda and Setty, 1978; Gowda and Gowda, 1999) whereas the larval hatching of M. incognita was suppressed significantly by boiled extracts of 38
Review Of Literature mustard and cotton oil cake up to 99.92 and 99.38% in water (Lanjeswar and Shukla, 1986) .Water soluble fractions of oil-cakes extracted from neem, mahua, groundnut and castor inhibited the larval hatching of M. incognita (Khan et al., 1974a).
Yang et al. (2015) showed that Camellia seed cake extracts under low concentration (2 g/L) showed a strong nematicidal effect. After treatment for 72 h, the eggs of M. javanica were gradually dissolved, and the intestine of the juveniles gradually became indistinct. Nematicidal compounds, including saponins identified by HPLC-ESI-MS and 8 types of volatile compounds identified by GC-MS, exhibited effective nematicidal activities, especially 4-methylphenol. The pot experiments demonstrated that the application of Camellia seed cake suppressed M. javanica and promoted the banana plant growth. Akhtar and Mahmood (1996) reported that the efficacy of oil seed cakes of neem(Azadirachta indica) and castor (Ricinus communis), composted manure and urea as a soil amendment gives satisfactory results in significant decrease of plant-parasitic nematodes, whereas predatory and free-living nematodes increased.
Singh et al.(1980) treated tomato seeds (cv. Marglobe) with either oilcakes of castor, mustard, neem, mahua and groundnut (2 g oilcake/10 g seeds) (0.1 or 0.2 g nematicides/10 g seeds) and planted in soil infested with Tylenchorhynchus brassicae, Hoplolaimus indicus, Helicotylenchus indicus and Meloidogyne incognita. Growth was promoted by oilcake treatment (mustard > neem > groundnut > castor > mahua). Radwan et al. (2009) in a pot experiment, oil cakes of cotton, flax, olive, sesame and soybean mixed with soil at the rate of 5, 10, 15, 20 or 50 g/kg soil. The results showed that M. incognita populations in the soil and root galling were significantly suppressed. The highest reduction in galls was noted in plants treated with sesame cake, whereas the lowest reduction was observed in plants treated with olive cake.Poornima and Vedivalu (1993) reported that the oil cakes of neem, castor and mahua alone and in combination with different plant extracts and nematicides were effective in reducing the populations of M. incognita on brinjal. Zahir (2004) reported that sesame oil cake achieved the highest increase in plant fresh weight (78.92%) with reduction percentage of galls (90.29%) and second stage juveniles /250g soil (80.39%), whereas oil cakes gave lowest values for root galling (84.29%) and J2/250g soil (60.13%) compared to nematode alone.
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2.2.5. Sawdust
Sawdust is generated from the timber industry and made of very fine particles of natural origin of the wood because of various massive chemical constituents presnt in sawdust has been suggested for the pest management and control of plant-parasitic nematodes. A significant reduction in the intensity of root galling was reported (Srivastava et al., 1971) when sawdust applied in a field planted with okra, eggplant and tomato. However, various other workers have pointed out the reduction in nematode intensity by amending the soil with sawdust (Bora and Phukan, 1983; Singh et al., 1986; Osunlaja, 1990; Akhtar, 1998).
The combined effect of sawdust and ammonium sulphate was greater than for either of the separate treatments both with respect to nematode control and to the improvement in plant growth. Sawdust of neem was more efficacious than that of mango. The phytotoxicity of sawdust was effectively eliminated by supplementing the sawdust with ammonium sulphate (Siddiqui and Alam, 1990).Amendments with low C: N ratio such as animal manure, oil cake and green manure, result in better nematode control than those with wide ratio, especially grassy hay, stubbles and cellulosic materials such as paper and sawdust (Akhtar and Malik, 2000).
Soil amended with spent tea, sugarcane bagasse, paddy husk, wheat straw, domestic garbage, saw dust, fly ash, cellulosic waste, waste water and sewage sludge etc also decreased the root-knot population (Eyal et al., 2006). Results showed that severe root galls occurred on plants treated with sawdust and rare galls on those treated with poultry and farmyard manures. Growth and yield characteristics of the plant were also affected by root-galls damage at the various organic manure treatments (Agu, 2008)
All the sawdust at all the concentrations and the bioinoculants either singly or in combination, improved plant growth parameters in terms of plant length, fresh weight, dry weight and number of nodules per plant and suppressed root-knot nematode infection in terms of number of galls/plant and nematode population. Among the four sawdusts, neem was found to be most effective followed by sheesham, teak and chir. The effectiveness of pigeon manure and all the four sawdusts was proportional to their doses (Aisha et al., 2015; Hassan et al., 2009)
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The soil amendments with organic additives except gram and rice husks significantly reduced the multiplication of M. incognita and the root galling caused by root-knot nematode which consequently increased the plant growth. The greatest improvement in plant growth and reduced reproduction factor and root galling was recorded in soil amendment with leaves of Calotropis procera whereas the least was in kail saw dust (Khan and Husain, 1988). Neem products, including leaf, seed kernel, seed powders, seed extracts, oil, saw dust and particularly oil cakes, have been reported as effective for the control of several nematode species (Egunjobi and Afolami, 1976; Akhtar, 1998).
Organic amendment viz., rice husk, saw dust and neem cakes have been significantly effective against M. incognita (Singh and Khurma, 2007; Nagaraju et al., 2010). The production of egg masses by the adult female was 60.6% less on the roots of plants in neem amended soil. Neem amendment was more effective than sugarcane bagasse, kaner leaves, saw dust and castor cake. The plants in the soil amended with neem and saw dust produced 58.4% and 33.7% less root galls. The lower number of galling and egg masses in neem amended soil (Hayat et al., 2012).
2.2.6 Biochar
Biochar is the pyrolyzed carbon rich charred biomass produced through the crop residue, wood, manures and waste of natural origin. It has gain immense attenation and interest as soil amendment for the improvement of soil function and for the nematode disease management. Few studies, however, investigated the effects of biochar on plant parasitic nematodes. Zhang et al. (2013) reported a decrease in the population of several nematodes, e.g., Hirschmanniella and Pratylenchus in wheat fields amended with biochar. Perry and Beane (1988) who reported no suppressive effect of activated charcoal on the final population density of G. rostochiensis in soil. Most importantly, Viger et al. (2015) used higher concentrations (5 %), and it is known that an excessive activation of plant growth causes a negative effect on plant defense because of resource-limited trade-off effects.
Perry and Beane (1988) who demonstrated a delay in hatching of J2 of G. rostochiensis when potatoes were planted in soil amended with activated charcoal. Nematode genera belonging to fungivores preferred biochar-enriched environment to get more microbial resource as food (Atkinson et al., 2010), while those belonging to
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Review Of Literature plant parasitic group favoured environment with no or low biochar to avoid the competition of microbes (Hominick, 1999). Biochar produced from citrus wood using traditional charcoal-making techniques was found to induce systemic resistance on foliar fungal pathogen, Botrytis cinerea (gray mold) on pepper and tomato and to the broad mite pest (Polyphagotarsonemus latus) on pepper (Elad et al., 2010). Fusarium infection of Asparagus was found to decrease after addition of coconut biochar and was similar to the benefits derived from manure made from coffee residue (Matsubara et al., 2002). A decrease in Fusarium infection of Asparagus was also reported after addition of biochar made by fast pyrolysis of wood powder (Elmer and Pignatello, 2011). Infection of tomato with another soil borne disease, bacterial wilt (Ralstonia solanacearum), was significantly reduced by adding wood biochar in some experiments and consistently by adding biochar made from municipal biowaste (Nerome et al., 2005).
A 1.2 % concentration of biochar added to the potting medium of rice was found to be the most effective at reducing nematode development in rice roots, whereas direct toxic effects of biochar exudates on nematode viability, infectivity or development. The increased plant resistance was associated with biochar-primed H2O2 accumulation as well as with the transcriptional enhancement of genes involved in the ethylene (ET) signaling pathway. Significant differences in nematode mortality were not observed between biochar exudates (6.6 ± 0.7 %) and water (7.2 ± 0.6 %) 24 h after initiation of the bioassay at doses ranging from 0.3 to 5 % biochar. Similar results were also observed when the nematodes were incubated in biochar exudates for 72 h. These data suggest that biochar exudates do not have a direct nematicidal effect on M. graminicola at the doses tested (Huang et al., 2015).
Makoto et al. (2010) showed not only a significant increase in root biomass (47%) but also root tip number (64%) increased within a layer of char from a forest fire with larch twigs. The number of storage roots of asparagus also increased with coconut biochar additions to a tropical soil (Matsubara et al., 2002). Root length of rice was shown to increase with biochar additions (Noguera et al., 2010). Germination and rooting of fir embryos (Abies numidica) significantly increased from 10 to 20% without additions to 32-80% of embryos when activated carbon was added to various growth media (Vookova and Kormutak, 2001). A single application of 20 t/ha biochar to a Colombian savanna soil resulted in an increase in maize yield by 28 to 140% as 42
Review Of Literature compared with the unamended control in the 2nd to 4th years after application (Major et al., 2010). Soil biochar incorporation has a positive impact on beneficial fungi (Atkinson et al., 2010) which would support more fungivorous nematodes. When comparing the root knot nematode infected plants with no infected plants, slight but significant reductions in the root length and total plant height were observed in the root knot nematode infected plants. The number of adult females in biochar-amended roots was slightly lower than that of non-amended plants (Huang et al., 2015). The amendment of poultry-litter biochar to the soil generally decreased the number of plant-parasitic nematodes while increasing the amount of free-living nematodes in the soil (Rahman et al., 2014). Certain biochar additions to soil have been shown to significantly improve the soil tilth, nutrient retention and availability to plants, and crop productivity (Steiner, 2007; Elad et al., 2010; Lehman et al., 2003).
Ebrahimi et al. (2016) demonstrated that although biochar significantly delayed hatching of both potato cyst nematode species but it did not reduce the final number of juveniles that hatched and penetrated the roots during the growing period. On the contrary, when plants were inoculated with hatched J2, fewer juveniles were detected in the potato roots in soil amended with pig slurry,cattle slurry, mineral nitrogen, wood chip compost and crab shell compost, but not with biochar. Adding biochar at 0.3 and 1% did not reduce the survival or the reproduction of any of potato cyst nematode species; moreover, it inhibited the suppressing effect of wood chip compost and pig slurry on potato cyst nematode reproduction when added together with these amendments.
Bio-assays were developed to assess the effect of biochar soil amendments on the soil-borne plant-parasitic nematodes Meloidogyne chitwoodi on bean and carrot, Globodera sp. on potato. Incorporation of 3% (DW) biochar produced from holm oak at 650◦C in peat significantly reduced the incidence of Botrytis cinerea on strawberry leaves; whereas 1% (DW) amendment had no effect. Compost amendment reduced the plant-parasitic nematode population on carrot and potato, but this effect was absent in case of additional biochar application (Debode et al., 2014). The experiments have shown that the addition of biochar to a grape vineyard reduced the incidence of plant parasitic nematodes by a factor of eight compared to the control (Rahman, 2014). The biochar treatment had 82.5% less nematodes compared to the highest population achieved and it may contain bio-toxic compounds. There was a 43
Review Of Literature sharp drop in total nematode population compared to the control on tomato (Abdulrahman MEA Al-Fraih, 2015).
Vaccari et al. (2015) found that the use of biochar on tomatoes increased plant growth compared to non-amended control for processing tomatoes. An increase in cherry tomatoes was also seen with the addition of biochar, which increased production by 64% compared to control (Hossain et al., 2010). Biochar soil application resulted in higher upland rice (Oryza sativa) grain yields at sites in northern Laos with low P availability, and improved the response to N and NP chemical fertilizer treatments (Asai et al., 2009).
Five different soil amendments (Nostoc calicola amended compost, bokashi, biochar, garlic straw, and compost) were applied at 500g /pot. Plant performance, and nematode incidence and diversity were analyzed in Lycopersicon esculentum cv. Capppricia. The results indicated that there were no significant differences (p>0.05) in plant height, stem diameter, leaf sap analysis (N, K, P, Ca, NO3, NH4, Mg, Zn, Mo, Cu, Fe, Mo, Se, Cl, Al, and Mo), tomato fruit production, and root knot index among treatments. Root knot index (RKI) values were unexpectedly low which seems to be indicative of relatively low nematode pressure. Significant differences were observed in leaf count; stem final fresh weight, leaf final fresh weight, with the garlic straw treatment having the highest average means. In terms of total nematode populations, bokashi + garlic straw showed the highest values compared to the other treatments (Abdurrahman MEA Al-Fraih, 2015). Breazeale (1906) have explained the pronounced increase in root growth after additions of carbon black (soot) to soil with sorption of allelopathic compounds that were phytotoxic. In several cases, not only root and shoot biomass increased after biochar additions, but the shoot-to-root ratio increased, as well (Lehman et al., 2011) Such an increase in the shoot-to-root ratio may indicate improved resource supply that requires fewer roots to sustain the same above-ground biomass production (Wilson, 1988).
2.3. BARE ROOT DIP/PLANT EXTRACT
Seedlings are more prone to the infection with the nematode and it premirily attack on the root and disturb the function of the plant. That is why application of plant based biopesticides and bioagents may be safe, economical to lower the nematode infection. Significant reduction was observed in the population of plant 44
Review Of Literature parasitic nematodes, Meloidogyne incognita in eggplant, when the seedlings were given the root-dip treatment in leaf extracts of Argemone maxicana and Solanum xanthocarpum (Ajaz and Tiyagi, 2003). The applications of methanolic extract of botanicals are very effective in the control of phytonematodes (Usman and Siddiqui, 2011a,b). Hussain et al. (1984) showed that root dip treatments of eggplant seedling with Margosa and Marigold leaf extracts considerably reduced root knot nematode development.
Tomato seedlings (root dip treatment) were dipped in alcoholic leaf extracts of neem, karanj, castor and Jatropha with two concentration (10 and 20% each) with carbosulfan 25EC (2%) for 4hours. Leaf extracts of neem @ 20% was found best treatment in improving plant growth characters and reduced nematode reproduction followed by other plant extracts (Meena et al., 2013). Akhtar and Alam (1993a) also recorded maximum total plant length in nimin in bare-root-dip treated tomato plants and increase in the length was pronounced as compared to neem oil, castor oil, rocket- salad oil and mustard oil. Usman and Siddiqui (2011a) evaluated the leaf extract of Murraya koenigii L. and Vitex negundo L. as bare-root dip treatment for the management of M. incognita infecting tomato (Lycopersicon esculentum) and chilli (Capsicum annum) plants.
Tiyagi et al. (2009) studied the effect of leaf extracts of two latex-bearing plants such as Calotropis procera and Thevetia peruviana as a bare-root dip treatment for the management of phytonematodes, M. incognita and R. reniformis infecting tomato and plants. Khan et al. (2011) showed that the curative effect of plant extracts viz., neem, tobacco, Aloe vera, chilli, clove, garlic and onion against M. incognita.
Khan et al. (1997) have reported that the efficacy of culture filtrates of M. vaginatus against egghatching and mortality of M. incognita was dependent on its concentration and period of exposure. Seed soaking treatment of okra (Abelmoschus esculentum) with graded concentrations of M. vaginatus showed significant increase in plant growth and inhibit root galling and population of M. incongnita. Saravanapriya and Sivakumar (2005) demonstrated that bare-root dip treatment with oak leaf extract caused lowest gall index of 1.7 followed by leaf extract of neem (2.4), marigold (2.9), seed extract of betel nut (3.4) and watermelon (3.5). Similar results by Tariq and Siddiqui (2005) demonstrated that bare-root-dip treatment of tomato with
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Review Of Literature neem cake + carbofuran caused highest value of total plant length (50.8 cm) as compared to neem leaves, neem cake, carbofuradan and neem leaves + carbofuran. Carbofuradan 3-G was applied as bare-root-dip treatment caused 249.4% increase (19.5g) in total plant weight as compared to the inoculated control (5.6g).
Akhtar and Alam (1993b) reported that bare-root-dip treatment with standard concentration of nimin caused significantly maximum total plant weight of tomato i.e. 69.7g followed by S/2 and S/10 concentration i.e. 65 g and 62.7 g respectively over the inoculated control. Similar observations were also given by Javed et al. (2007) that bare-root-dip treatment with neem cake and aza (refined neem product) was more effective and gave minimum number of galls in tomato.
Khan et al. (2011) reported that bare-root-dip treatment of tomato plant with fruit extract of chilli caused maximum reduction in mature females i.e. 10.6% as compared to the control. In the present study, neem leaf extract used as bare-root-dip treatment caused 31.3% reduction in number of females/gall as compared to the inoculated control. Javed et al. (2007) also reported that least number of females/root system i.e. 38.9% in aqueous neem leaf extract used as bare-root-dip treatment as compared to the inoculated control.Akhtar and Mahmood (1993) reported that the neem based product Nimin, applied as a bare-root dip treatment resulted in a decrease in nematode population and an improvement in plant growth responses. Bare root dip in castor and mustard oil cake and neem leaf extracts effectively controlled M. javanica on tomato and eggplant (Abid and Maqbool, 1991).
Akhtar et al. (1992) documented the reduction in the development of root-knot on tomato caused by M. incognita, with bare root dip in leaf extracts of Melia azedarach and Calotropis procera. Root-dip treatment of eggplant seedling with margosa and marigold leaf extract, mustard cake considerably reduced root knot development as compared to treatment with cine, piperazine citrate, chenopodium oil and ground nut cake. Treatments with “S/2” concentrations of test materials were better for plant growth improvement but for disease control “S” concentrations were more effective (Hussain et al., 1984).
Olaniyi (2012, 2014) investigated the effects of rhizome dip of pared plantain in crude water extracts of Acalypha wilkesiana leaves and at different duration of exposure in comparison with the conventional hot water dip under field conditions. 46
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The most effective treatment in terms of reduction in nematode population density and related damage was obtained with 15 mins exposure to the extract on the field. Hussain et al. (1984) showed that root dip treatments of eggplant seedling with Margosa and Marigold leaf extracts considerably reduced root-knot nematode development as compared to treatment with Cina, Piprazine citrate, Chenopodium oil and Ground nut cake. Neem oil based formulation, when used as seed treatment and bare root dip controlled the population of root knot nematode, Meloidogyne incognita in tomato and chickpea (Majumdar and Basu, 1999; Javed et al., 2008).
Abid and Maqbool (1991) showed bare root dip treatment in the leaf extracts and neem leaves significantly reduced root-knot infection caused by M. javanica on tomato and eggplant. The damaging effects of the nematode were masked by bare root dip treatments as shown by improved plant growth in both the test plants. Usman and Siddiqui (2011a) evaluated the leaf extracts of Murraya koenigii L. and Vitex negundo L. as bare-root dip treatment for the management of M. incognita infecting tomato (L. esculentum) and chilli (C. annuum) plants. Significant reduction was observed when leaf extracts of Murraya caused relatively higher inhibition in root knot development and nematode multiplication than Vitex. Tiyagi et al. (2009) studied the effect of leaf extracts of two latex-bearing plants such as Calotropis procera and Thevetia peruviana as a bare-root dip treatment for the management of phytonematodes, M. incognita and R. reniformis infecting tomato plants.
Khan et al. (2011) showed that the curative effect of plant extracts viz., neem, tobacco, Aloe vera, chilli, clove, garlic and onion against M. incognita. Similarly significant reduction was observed in the population of plant parasitic nematodes, M. incognita infesting eggplant and cauliflower, when the seedlings were given the root- dip treatment in leaf extracts of Argemone mexicana and Solanum xanthocarpum (Ajaz and Tiyagi, 2003). Leaf extracts of neem @ 20% was found best treatment in improving plant growth characters and reduced nematode reproduction followed by other plant extracts (Meena et al., 2013).
2.4. BIOLOGICAL CONTROL
The International Organization of Biological Control (IOBC, 2014) founded in 1956, is set to promote environmentally, socially and economically efficient methods to eliminate the diseases and pests from agriculture and forests. It defines biological
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Review Of Literature control as “the use of living organisms or their products to eliminate or reduce the damages or losses due to pests ". Biological control involves the use of beneficial organisms, of their genes and/or of products such as metabolites that reduce the negative effects of plant pathogens and promote positive responses from the plant (Junaid et al., 2013). Biological control was discovered by trial and error and then practiced in agriculture long before the term itself came into use (Baker and Cook, 1974). Biological control has become a promising alternative to the use of chemicals (Stirling, (1991), but not act as a real substitute, because their potential is governed by environment, multiplication rate, time of usage and it is slow acting as well. However, according to Stirling (1991) biological control refers to "A reduction of nematode populations which is accomplished through the action of living organisms other than the nematode-resistant host plant, which occurs naturally or through the manipulation of the environment or the introduction of antagonists".
The fungus, P. lilacinus, is an egg parasitic fungus which infects by direct hyphal penetration. The hyphae branches grows across the egg shell (Khan et al., 2006).The egg pathogenic fungus P. lilacinus is one of the most widely tested soil Hyphomycetes for the biological control of plant parasitic nematodes (Atkins et al., 2004). According to Atkins et al. (2004) the egg- pathogenic fungus Purpureocillium lilacinum is the most widely tested biological measure for plant parasitic nematodes, and has shown promising potential as an alternative to chemical control at both pre- planting and planting applications. P. lilacinus significantly reduced M. incognita soil and root population and increased tomato production (Kalele et al., 2010). Lara Martez et al. (1996) demonstrated that P. lilacinus significantly reduced M. incognita soil and root populations and increased yield of tomato.
Hashem and Abo-elyousr (2011) suggested that application of different biocontrol agents (P. fluorescens and P. lilacinus) in combination and alone not only has a lethal effect on nematode and enhances the plant growth, supplying many nutritional elements and induction of the systemic resistance in plants. Treatments with P. fluorescens and P. lilacinus caused mortality of M. incognita as 45% and 30% of juveniles after 48 h of exposures respectively.
The maximum reduction in root galling and the soil population, occurred in soil treated with both fungi in combination with mustard cake. T. viride used alone
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Review Of Literature responded least as compared to P. lilacinus which was also observed by Khan and Goswami (2000). The fungal bioagents viz., P. lilacinus and Trichoderma viride alone or in combination with mustard cake and furadan promoted plant growth, reduced number of galls/plant, eggmasses/root system, eggs/egg mass and nematodes reproduction factor as compared to untreated infested soil (Goswami et al., 2006).
The secondary metabolites of Trichoderma include chitinase enzyme which is considered to be the most effective component against pathogenic fungi. Chitin comprises the outer shell of nematode’s eggs so that nematode eggs are affected greatly by Trichoderma species treatment (Haggag and Amin, 2001; Jin et al., 2005). Stephan et al. (2002) reported that T. harzianum and animal organic matters reduced the numbers of root-knot nematodes. Siddiqui et al. (2000) has reported the reduction of M. javanica infection on tomato by P. lilacinus.
Several studies (Susan et al., 2000; Haggag and Amin, 2001; Siddiqui and Shaukat, 2004; Santhosh et al., 2005) showed the use of Trichoderma for inhibiting the growth of plant parasitic nematodes. Faruk et al. (2002) indicated the effectiveness of T. harzianum on the biocontrol of root knot nematodes on tomato. Saifullah (1996a, b) showed the death of 100% of Globodera rostochiensis and G. pallida by using poisoning compounds from T. harzianum on the medium after 24 h of exposure. Cannayane and Sivakumar (2001) reviewed the biocontrol efficacy of P. lilacinus and listed several reports where root-knot nematodes were successfully controlled by this egg-pathogenic fungus. Among numerous organisms that have shown antagonism against root knot nematodes, P. chlamydosporia (Kerry, 2000; Siddiqui, et al., 2009), P. lilacinus (Jatala, 1985; Kiewnick and Sikora, 2006), and T. harzianum (Siddiqui and Shoukath, 2004; Bokhari, 2009) have been found to be highly suppressive to plant nematodes, especially under greenhouse conditions.
Usman and Siddiqui (2012a) showed that Trichoderma culture filtrate was more significant on root-knot nematode, M. incognita. Trichoderma controls nematode genera by a direct effect on toxic metabolites and inhibits nematode penetration and development (Bokhari, 2009). Root colonization by Trichoderma spp. frequently enhances root growth and development, crop productivity, resistance to abiotic stresses and uptake and use of nutrients (Harman et al., 2004).Siddiqui and Shaukat (2003) reported production of metabolites, including 2,4-
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Review Of Literature diacetylphloroglucinol (DAPG) and hydrogen cyanide (HCN) by P. fluorescens strains CHAO that inhibited egghatch and induced mortality in J2 of root-knot nematodes. El-Sherif et al. (1999d) reported that culture filtrates of certain bacterial species, such as Pseudomonas and Bacillus, caused inhibition of egg hatching and juvenile survival of plant-parasitic nematodes in soil. Son et al. (2007) reported that Paenibacillus strains inhibited egg hatching and caused mortality of M. incognita juveniles and other plant parasitic nematodes.
El-Sherif et al. (1994b) proved that culture of bacteria inhibited hatching of Meloidogyne incognita and was highly toxic to juveniles. Different fungal strains isolated from nematodes, soil and plants were shown to produce substances that inhibit nematode egg hatch or kill nematodes (Khan and Saxena, 1997). P. fluorescens produced the greatest mortality of nematodes. These results are in agreement with the findings of many scientists, who showed that P. fluorescens and P. lilacinus were lethal to juveniles of M. incognita (Khan et al., 2005; Kiewnick and Sikora, 2006; Siddiqui et al., 2009). Antibiotics and other toxic compounds produced by the bacterial species as well as direct interaction might be responsible for the J2 mortality.
Khan and Park (1999) have observed that culture filtrate of Microcoleus vaginatus inhibited hatching of second-stage juveniles of M. incognita and killed hatched juveniles. Production of metabolites by rhizosphere bacteria causes lysis of nematode eggs (Wetscott and Kluepfel, 1993) affects vitality of second-stage juveniles (Becker et al., 1988) and degrades specific root exudates resulting in reduced attraction and penetration (Oostendrop and Sikora, 1989).
2.5. SEEDLING BARE ROOT DIP/BIOAGENTS
Nematode is a root parasites entering through root tip from the zone of elonagtaion and causing major damage to the root that may disrupt the physiology of the plant. Culture filterate of various biocontrol agents through involvement of spores and colonies offer sustainable alternative for the nematode management. Application of bare root dip through P. lilacinus fungus in chilli as seedling bare root-dip treatment showed significant difference in shoot weight and root weight when compared to the untreated control. Seedling root-dip treatment of chilli in fungal suspension containing 8 x 106 spores/ml reduced gall formation of M. incognita by 50
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47.9% and in vitro conditions eggs of Meloidogyne incognita undergo necrosis. The infected eggs became vacuolated and filled with fungal hyphae (Cannayane, 2006)
Siddiqui et al. (1999) revealed that bare root dip treatment with Pseudomonas aeruginosa along with and without Trichoderma harzianum, Trichoderma koingiia and T.hamatum significantly controlled the infection of M. javanica on Chilli. Combined use of Trichoderma harzianum with Pseudomonas aeruginosa caused the greatest reduction in root galling of M.javanica. Bare root dip treatment of tomato seedlings in a suspension of P. fluorescens was reported effective against M. incognita (Shanti and Sivakumar, 1995).
Sahebani and Hadavi (2008) stated that bare-root-dip treatment of tomato plants treated with filamentous fungi, Trichoderma harzianum caused minimum number of galls/plant at concentration 107, 108 and 109 spore /ml over the control. Seven spp. of Aspergillus have been investigated as bare root dip treatment and soil drenching against root knot nematode, M.javanica in tomato. Among all Aspergillus niger was found to be most successful in minimizing the root and soil densities of nematode and improvement of plant growth parameters (Zareen et al., 2001).
The effects of two strains of Pseudomonas aeruginosa (IE-6 and Pa-7) and an isolate of Bacillus subtilis on egg hatching, mortality and infection of Meloidogyne javanica in tomato roots was evaluated under laboratory, greenhouse and field conditions. Cell-free culture filtrates of the bacterial isolates significantly reduced egghatching and caused mortality of second stage juveniles. Introduction of rhizobacteria as bare root gave better results than stem injection or foliar application. Under field conditions, the isolates also significantly reduced nematode population densities and subsequent gall formation there by increase in plant growth character and yield (Siddiqui, 2002). Siddiqui and Shaukat (2002) reported that two rhizobacteria Pseudomonas aeruginisa strain IE-6S+ and Pseudomonas fluorescens strain CHA0, used as bare root dip treatment and soil drenching, substantially reduced M. javanica juveniles penetration into the tomato roots under glass house condition.
Seed treatment combined with soil drenching through Pseudomonas putida strain Mt-19 caused significant reduction in nematode potential and improvement in plant growth parameters of tomato (Munif et al., 2001). Kempester et al. (2001) found that application of pacteolytic Pseudomonas fluorescens strain P29 and P80 as a soil
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Review Of Literature drench reduce the fecundity of Heterodera trifolii infecting white clover and increased the proportion of distorted females and female with few eggs compared with control. S/2, S/10 and S/20 dilutions of Pseudomonas fluorescens when applied as soil drench/dipping against root-knot nematode in brinjal was found to be highly effective in reducing nematode reproduction and improvement of enzyme activity such as peroxidase, polyphenol oxidase and chitinase (Osman et al., 2011)
Kaur et al. (2015) reported the nematicidal potential of neem and Bacillus thuringiensis in controlling M. incognita as bare root dip treatment on tomato. Reduction in root galling and final nematode population of Meloidogyne incognita were observed in brinjal seedlings which were given bare root dip treatment in 10% neem leaf suspension mixed with spores of Paecilomyces lilacinus (at 4x105 spores/ml) for 30 minutes. Significant increase were also observed in root colonization, parasitization of eggs and spore density of Paecilomyces lilacinus in soil (Rao et al., 1997b). Fresh leaf extracts of Azadirachta indica, Allium sativum (Garlic) and Tagetes erecta (African marigold) and bacterial suspensions of Pseudomonas fluorescens significantly reduced the root galling, nematode population, and enhanced the plant growth and yield (Abo-Elyousr et al., 2010).
2.6. SEED TREATMENT
Investigation of seed coating of okra cv. UV with Pseudomonas fluorescens alone and in combination with soil application of carbofuran @ 1 and 2 kg per ha and neem seed powder @ 1 % (w/w) have been done against Meloidogyne incognita. This treatment showed reduced penetration, least number of eggmasses per plant, minimum soil population after 60 days and significantly higher shoot-root ratio (Sharma et al., 2008). Davide and Batino (1985) on undelinted and delinted cotton seeds treated with P.lilacinus the number of galls, nematode and egg masses were significantly reduced. Paecilomyces lilacinus enhanced the growth of tomato and lettuce infected by M. incognita and also increased parasitization of egg masses when the bio-nematicide was used for seed, nursery bed treatment and also FYM enrichment (Goswami et al., 2006; Prakob et al., 2009).
Kumar et al. (2012) evaluated the seeds of okra cv. A-4 treated with Trichoderma harzianum, Trichoderma viride, Pochonia chlamydosporia, Paecilomyces lilacinus and Pseudomonas fluorescens at 20g/kg seed. All treatments 52
Review Of Literature significantly reduced the soil population density of the nematode and improved the plant growth, although the reduction with T. harzianum was not significant. Seed coatings with T. viride and P. lilacinus were equally effective in reducing the nematode soil population. Toxic Aspergillus spp., and egg parasitic Paecilomyces spp., was found more effective than a single bioagents against M. incognita, resulting in better plant growth (Verma et al., 2009).
Dhawan and Singh (2009b) evaluated the effect of seed treatment and soil application of P. chlamydosporia at various doses under in vivo and micro plots. A 96% egg parasitism of M.incognita and highest recovery upto 81% in plant vigor was recorded in the treatment that received soil application of the P.chlamydosporia @ 3% w/w under pot trial. Seed treatment/Soil drench with Pseudomonas aeruginosa (strain 78) and Pseudomonas spp. (Strain 82 and strain 51) significantly reduced population of M. javanica and subsequently decreased root knot disease severity with enhanced protein content and yield of mungbean (Ali et al., 2002).
Vinod and Jain (2010) evaluated some fungal and bacterial antagonists as seed dressing treatment against root-knot nematode, M. incognita infecting okra. The seeds of okra variety A-4 were treated with T. harzianum, T. viride and Pseudomonas fluorescens each @ 10g/kg seed. The observations recorded 45 days after sowing indicated that the growth parameters of okra plants were better and root–knot nematode populations were reduced in all the treatments compared to inoculated control. Dawar et al. (2008) observed the potentialities of seed treatment with fungal and bacterial antagonists viz., B. thuringiensis, R. meliloti, A. niger, T. harzianum and nematicides in the suppression of root knot nematode and shoot length, shoot weight, root length and root weight were significantly increased in sunflower and okra.
2.7. BIOAGENTS COMBINED WITH ORGANIC AMENDMENT
Integration of two or more stragies for the management is one the parmount method for nematode management. Single approach for the nematode management is not too much effective. The combined application of certain botanicals and P. lilacinus is a better tactic in the reduction of nematode population and in enhancing the plant growth and yield when used alone (Zaki and Bhatti, 1990; Zaki, 1998; Walia and Gupta, 1997). P. lilacinus and green manuring of Zea maize and Sesbania
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Review Of Literature aculeata were very effective for the management of Rotylenchulus reniformis (Mahmood and Siddiqui, 1993).
Paecilomyces lilacinus has an almost worldwide distribution occurring most frequently in warmer climates (Domsch et al., 1980; Dunn et al., 1982). Paecilomyces lilacinus has been reported to be very effective in controlling root-knot nematodes M.incognita by several workers (Shahzad and Ghaffar, 1987, Sharma and Trivedi, 1989, Zaki and Maqbool, 1992, Sosamma and Koshy, 1997)
Azam et al. (2009) reported that the combination of leaf powder C. tora and P. lilacinus was most effective in managing the root knot disease. Fruit wastes of apple, banana, papaya, pomegranate and sweet orange @ 20g/plant and the fungal biocontrol agent P. lilacinus @ 2g (mycelium+spores) /plant, alone and in combination were very effective for the management of R. reniformis (Ashraf and Khan, 2008).
Perveen et al. (2007) reported that application of T. viride and P. fluorescens with farmyard manure effectively control M. incognita and plant growth. These findings are in confirmation by many workers on other crops (Pant and Pandey, 2002; Kumar et al., 2009; Abuzar and Haseeb, 2010). Application of P.fluorescens (108 cfu/g) and T.harzianum (106cfu/g) each at 5g/kg soil in papaya seeds reduced significantly the eggs/eggmasses of M.incognita and 4ml of fungal suspension (8×106 spore/3kg) was found to be optimum dose for effective reduction of M.incognita population (Rao, 2007; Priya and Kumar, 2006).
Sundraraju and Kiruthika (2009) demonstrated that the integration of P. lilacinus with neem cake or anyone of the botanicals viz., Tagetes spp., S. torvum, can be effectively used in the management of root knot nematode. The combined application of biocontrol agents and various oil cakes have been suggested to minimize the crop losses caused by various plant-parasitic nematodes (Tiyagi et al., 2002; Borah and Phukan, 2004; Zareena and Kumar, 2005). The combined application of neem sawdust with the biocontrol fungus P. lilacinus was more promising in increasing the plant growth and decreasing the reproduction factor and root-galling. The inhibitory effect of kail sawdust on the parasitism of P.lilacinus could in fact be due to the toxic principles contained in kail sawdust and released in soil, which might have inhibited the activity of P. lilacinus (Ahmad and Khan, 2004). Research on nematode trapping fungi has revealed that the improvement of trapping is 54
Review Of Literature controlled by the fungal species and the type and amount of organic material added (Jaffee et al., 1994; Jaffee, 2004).
Combined application of biocontrol agents, organic amendment and nematicides treatment was found to be best among all treatments which significantly increased the growth parameters including yield and reduced the population of M. incognita (Das and Sinha, 2005).
P. fluorescens, Asprgillus niger, T. harzianum and P.lilacinus @ 50kg/ha (2×108 cfu /g) each and organic matters viz., fresh chopped and murraya leaves, farmyard manure, mint manure and neem seed powder @ 1q/ha, carbofuran 3G and Topspin M [email protected] kg a.i./ha. T.harzianum was found superior among all the treatments followed by P.fluorescens, carbofuran (Saikia and Borah 2008; Tripathi and Singh, 2006).
Udo et al. (2014) reported that the application of the bionematicide Paecilomyces lilacinus and L. camara leaf extract alone significantly (P≤ 0.05) inhibited root galling and egg production. However, the severity of root galling and eggmass production was more significantly (P< 0.05) suppressed with the application of P.lilacinus than L. camara leaf extract. Combined application of both results in maximum reduction in root galling and improvement plant growth characters. Trivedi (1990) evaluated the fungus P. lilacinus for the biological control of root-knot nematode, M. incognita on Solanum melongena. Better reductions in gallings, final soil nematode population and number of eggs per egg mass were noted in fungus inoculated plants. Nagesh et al. (1997) conducted a field experiment for the management of root-knot nematode, M. incognita infecting tuberose (Polianthes tuberosa) by integrating the use of the antagonistic fungus, P. lilacinus with leaf extracts of castor and neem as bulb treatment and soil drenches. Combination of P. lilacinus with neem leaf extracts resulted in significantly higher plant fresh weight and flower yield. Root gall index was least under P. lilacinus plus neem leaf extract combination followed by P. lilacinus plus castor leaf extract treatment.
The fresh leaf extracts of Azadirachta indica, Allium sativum (Garlic) and Tagetes erecta and bacterial suspensions of Pseudomonas fluorescens and P. aeruginosa were examined against M. incognita on tomato in vitro and in vivo conditions (Abo-Elyouser et al., 2010). All treatments immobilized juveniles (J2), the
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Review Of Literature highest effect caused by neem leaves extract after 24 and 48 h of exposure. Bhat et al. (1998) studied the combined application of P. lilacinus and oil cakes for protection of chickpea against M. incognita. The P. lilacinus at 5 or 10 ml (3.5 × 107 spores/ml) differently limited the damage caused by M. incognita and increased the plant growth and nodulation of chickpea. The best protection of chickpea against M. incognita was obtained with the combined application of P. lilacinus (10 ml) and neem cake (1.0 g/Kg).The incorporation of neem cake along with P. lilacinus reduced the root-knot nematode, the number of galls and egg masses. Plant growth parameters were better in the neem cake treatment. Combining P. lilacinus, neem cake and chemicals did not further reduce the root-knot-nematode population significantly than the neem cake alone (Sharma et al., 2007).
Kumar et al. (2008) studied the effect of culture filtrates of P. lilacinus isolates on the mortality and hatching of root-knot nematode, M. incognita. Culture filtrates of all isolates of P.lilacinus showed toxic effect against M. incognita at varying degree. Isolate PIT-3 was found to be more effective among all isolates in both mortality and hatching inhibition.
Devi and Sharma (2002) studied the effect of T. harzianum and T. viride @ 1g/kg soil against M. incognita on tomato. They reported that both the treatments improved the plant growth and reduced the nematode population. However, variation among the treatments was not significant. Nama et al. (2015) stated the effect of Trichoderma harzianum, T. viride and Pseudomonas fluorescens @ 1.5 g/kg soil and @ 5 g/kg seed against root-knot nematode, Meloidogyne incognita infecting cowpea. The result revealed that T. harzianum @ 1.5 g/kg soil + T. harzianum @ 5 g/kg seed were found most effective in improving plant growth and reduction of nematode reproduction over control.
Islam et al. (2005) tested the efficacy of three antagonistic fungi, T. harzianum isolates (GT1, TV1 and W120) and two organic matters (poultry refuse and mustard oil cake) against root-knot nematode disease (Meloidogyne spp.) of tomato. Results showed that combined application of poultry refuse and T. harzianum (PR+GT1, PR+TV1 and PR+W120) gave higher disease reduction and increased plant growth. Kienwnicks and Sikora (2004) observed that soil treated with P. lilacinus before planting, increased the tomato fruit yield and reduced the number of galls and
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Review Of Literature nematodes in the soil. Applications of P. lilacinus significantly reduced M. incognita soil and root population and increased tomato production (Kalele et al., 2010).
Atkins et al. (2005) emphasised that the egg- pathogenic fungus Paecilomyces lilacinus is the most promising bioagent as an alternative to chemical control at both pre-planting and planting. Parihar et al. (2015) observed the effect of Pochonia chlamydosporia, oil cakes of neem, mustard and cotton in the management of root knot nematode, Meloidogyne javanica infecting brinjal under glasshouse conditions. All the treatments were effectively suppressed the nematode population and kept the infection at significantly low levels. Combined treatment, neem cake + P. chlamydosporia were more effective in the management of M. javanica followed by mustard cake + P. chlamydosporia and cotton cake + P. chlamydosporia. However, the efficacy of P. chlamydosporia increased in the presence of oilcakes. Similarly Cannayane and Rajendran (2001) recorded that single applications of P. lilacinus, P. chlamydosporia and oil cakes suppressed M. incognita.
Dhawan and Singh (2009b) reported that the combination of P. chlamydosporia, carbofuran and neem cake gave highest yield and suppress number of eggs and root galling in okra. Individual treatment of neem cake was found to be more effective in the management of M. javanica followed by linseed cake, castor cake, groundnut cake and mahua cake. However, the efficacy of biocontrol agents increased in the presence of oil cakes. The highest improvement in plant growth and best protection against M. javanica was obtained by the integration of P. lilacinus with groundnut cake followed by neem cake, linseed cake, castor cake and mahua cake Ashraf and Khan (2010). Combination of P. penetrans with the organic amendments, particularly with different oil cakes, showed cumulative effect on the efficacy of the bacterium and exhibited the greatest nematode suppression in chilli (Chaudhary and Kaul, 2013).The integration of oil cakes (except mahua-cake), bone and horn meals with P. lilacinus, resulted in increased plant growth of tomato and reduced population buildup of nematodes and root galling. The groundnut-cake with P. lilacinus was most effective (Khan, 1997).
Karthikeyan et al. (2001) used antagonistic organisms Trichoderma viride and T. harzianum (talc-based formulation as seed treatment at 4 g/kg seed) against the pathogen and Paecilomyces lilacinus (as seed treatment at 1g of culture/kg seed)
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Review Of Literature against the nematode in aubergine. The dry shoot and root weights were higher in T. viride+ farmyard manure treatment, followed by T. viride + neem cake treatment and the nematode gall index was very much reduced in Paecilomyces lilacinus + farmyard manure and in Paecilomyces lilacinus + neem cake treatment. Sundraraju and Kiruthika (2009) demonstrated that the integration of P. lilacinus with neem cake or anyone of the botanicals viz., Tagetes spp., S. torvum, can be effectively used in the management of root knot nematode. Field experiment was to test the efficacy of Acacia compost individually and in integration with bioagents viz., Pochonia chlamydosporia and Paecilomyces lilacinus. All the treatments were significantly superior over untreated check. Acacia compost enhanced the growth parameters with drastic reduction in root-knot index. Among the integrated treatments, P. lilacinum with acacia compost recorded maximum growth parameters and yield with least root- knot index. In contrast the two bioagents, P. lilacinus + P. chlamydosporia with acacia compost registered high root-knot index and decreased growth parameters and yield among the other integrated treatment (Ravindra et al., 2014).
Rizvi et al. (2012) claimed that the efficiency of different oil-seed cakes of neem (Azadirachta indica), castor (Ricinus communis), groundnut (Arachis hypogaea), linseed (Linum usitatissimum), sunflower (Helianthus annuus) and soybean (Glycine max) with association of Pseudomonas fluorescens in relation to growth parameters of chickpea. Significant improvement was observed in plant- growth parameters such as plant weight, percent pollen fertility, pod numbers, root- nodulation and chlorophyll content of chickpea.It can be due to reduction in disease incidence and might be due to growth promoting substances secreted by P. fluorescens.Pandey and Kalra (2003) evaluated the efficacy of various organic materials (neem compound, Mentha distillate, Murraya koengii distillate, Artemisia annua marc and vermicompost) and biological control agents (Glomus aggregatum and T. harzianum) alone or in combinations for the management of M. incognita on Withania somnifera. Highest root-knot suppression was noticed in vermicompost and T. harzianum combination over Mentha distillate and G. aggregatum. Goswami et al. (2006) observed that the maximum reduction in root galling caused by Meloidogyne incognita on tomato plants, as well as the nematode population occurred in soil, treated with both fungi (T. viride and P. lilacinus) in combination with mustard cake. However, mustard cake alone also showed adverse effects on the root-nodulation.
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Kumar et al. (2006) reported the effect of bioagents viz., Paecilomyces lilacinus and Trichoderma viride alone or in combination with mustard cake and furadan promoted plant growth, reduced number of galls/plant, egg masses/root system and eggs/eggmass. The fungal bioagents along with mustard cake and nematicide showed least nematodes reproduction factor as compared to untreated infested soil. Similar results have been given by Ashraf and Khan (2010) on the efficacy of biocontrol agents (P. lilacinus) and oil-cakes such as castor, linseed, groundnut, mahua and neem for the management of root-knot nematode, M. javanica infecting eggplant under glasshouse conditions. All the treatments effectively suppressed the nematode population and kept the infection at significantly low level. The highest improvement in plant growth and best protection against M. javanica was obtained by the integration of P. lilacinus with groundnut cake followed by neem cake, linseed cake, castor cake and mahua cake.
Khan and Goswami (2000) reported that maximum reduction in root galling occurred in soil treated with fungi, P lilacinus and T.viride in combination with mustard cake and furadan. The combined application of various oil cakes and biocontrol agents have been reported to be an effective approach to minimize the losses caused by various plant parasitic nematodes (Tiyagi et al., 2002; Borah and Phukan, 2004; Zareena and Kumar, 2005). Kumar and Khanna (2006) tested T.harzianum, with or without neem cake against M.incognita on tomato. The fungus was highly effective against M.incognita when eggmasses were inoculated 15 days prior to transplanting of tomato seedlings and fungus at transplanting time in absence or presence of neem cake.
2.8. HYPOTHESIS
The hypothesis which was put to trial in this study was that the application of Bioinoculants/ Biocontrol agents and organic matters will ameliorate the hazardous effects of root-knot nematode on the growth and yield characteristics of the test plants, Tomato (Solanum lycopersicum L. cv. K-21) that are widely cultivated and accepted by the local farmers as cash crops. The addition of organic soil amendments in the form of chopped leaves, agriculture waste, saw dust, biochar and oil cakes should positively influence crop performance with an addition of organic matters. As the decomposition of organic matters release some nitrogen compounds, organic
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Review Of Literature acids, or other compounds such as ammonia, tannins and phenols which are toxic and may be helpful in reducing the root knot nematode infection. In addition to this they may release nutrients that may be beneficial for the enhancement of the crop productivity. These chemical compounds may also act as inhibitor of egg hatching and reproduction. Organic soil amendment may be expected to change the activities of facultative parasites during their saprophytic phase more than the obligate parasites that have limited growth in the soil. So, the effect of organic matters in combination with nematode trapping fungi and bacterium may suppress the root knot nematode infection and multiplication and improved the growth and yield of the plants.
The exploration of bioagents influence in the suppression of nematode population because the efficacy of the antagonist may be varied by other soil organisms and the host plants. When bioagents were applied in the soil might cause direct parasitism, produce compounds such as enzymes, antibiotics or inducing plant defense mechanism leading to systemic resistance to antagonistic are the possible mechanisms for the suppression of nematodes.
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Materials And Methods
CHAPTER- III
MATERIALS AND METHODS
3.1. SITE FOR THE EXPERIMENT
All the glasshouse experiments described in the thesis were conducted in the Department of Botany, Aligarh Muslim University, Aligarh located in between the Ganga and Yamuna rivers. Aligarh, is a district of western Uttar Pradesh (North India) and is located 130 km East of Delhi at 27.88° N latitude and 78.08° E longitude at an average elevation of 178 m (587 feet) above sea level. It has average temperature ranges from 28–38 °C (82–100 °F) with the scanty rainfall of 650 to 750 mm recorded throughout the year with the average relative humidity ranges from 32 to 82%. The city experiences tropical monsoon influenced humid subtropical climate. Different types of soils, such as sandy, loamy, sandy-loam and clay loam are found in the district.
3.2. Test plant and pathogen
Tomato (Solanum lycopersicum L. cv. K-21, Family Solanaceae) was selected as a test plant and Meloidogyne incognita (Kofoid and White, 1919; Chitwood 1949) was chosen as test pathogen to evaluate the effect of various organic soil amendments on the management of M. incognita.
3.3. Preparation and sterilization of soil mixture
A mixture of soil and organic manure was prepared in the ratio of 3:1. The clay pots (15 cm in diameter) were filled with this mixture at the rate of 1 kg/pot. The pots with soil were sterilized at 15 lbs pressure for 20 minutes in autoclave. However, for raising seedlings of test plants, pots (30 cm diameter) were filled with autoclaved soil-manure mixture (compost @1g N/kg soil).
3.4. Collection of infected root
Meloidogyne incognita infected roots were collected from vegetable fields near Sasni, Agra Road, Hathras, UP, India. Infected roots of the plants were gently removed from the soil and kept in polythene bags and then labelled. Further all these
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Materials And Methods roots were brought to the laboratory for the examination. The infected root samples were washed with tap water and examined for the presence of galls and eggmasses.
3.5. Source of Meloidogyne incognita juveniles
Juveniles of Meloidogyne incognita (Kofoid and White, 1919; Chitwood, 1949) were prepared from a pure culture that was previously cultured by eggmasses and propagated on eggplant (Solanum melongena L.) in the glasshouse of Section of Plant Pathology and Plant Nematology, Department of Botany, Aligarh Muslim University, Aligarh, India. Surfacely attached eggmasses were detached by using sterilized forceps from the infected roots. These eggmasses were placed in 15 mesh sieves (8 cm in diameter) having crossed layer of tissue paper. These were kept in petridishes full of water so that eggmasses may remain in contact with water. These petridishes were then incubated at 28 ±2˚C for hatching and for freshly hatched second stage juveniles (J2) of Meloidogyne incognita.
3.6. Meloidogyne species identification
Meloidogyne incognita was identified on the basis of perineal pattern from infected root samples and was examined under stereoscopic microscope. .After species identification, single eggmass culture of Meloidogyne incognita was raised on eggplant (Solanum melongena L.) and maintained for experimental purpose.
3.7. Growing and maintenance of seedlings
The seeds of tomato cv. K-21 were surface sterilized in 0.01% mercuric chloride for three minutes and then rinsed with Distilled Water (DW) three times. For nursery preparation seeds were sown in autoclaved clay pots (30cm diameter) along with the soil. Three weeks after germination of proper seedlings of tomato cv. K-21 were transplanted to each15 cm diameter clay pots filled with 1 kg autoclaved soil. These pots were treated with fresh chopped leaves of different plants viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed applied 50g of fresh chopped leaves of different plants combined with 10g seed powder of Black nightshade per pot. The pots were watered regularly for proper decomposition of the organic additives for two weeks. As the roots of seedlings get established in pots each of them was inoculated with 1500 hatched second stage juveniles (J2) of Meloidogyne incognita. There were four replicate of each treatment.
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3.8. Screening of different cultivar of tomato to test the resistance and susceptibility against root knot nematode, M.incognita.
The main objective of the study was to test the resistance and susceptibility behaviour, if any, of different cultivar of tomato against root knot nematode M.incognita.
The Seeds of fourteen cultivar of tomato were procured from Indian Institute of Vegetable Research (IIVR), Varanasi. Seeds of all these cultivar of tomato were surface sterilized in 0.01% mercuric chloride for three minutes and then rinsed with Distilled Water (DW) three times and sown separately in clay pots (30 cm diameter) filled with steam sterilized soil and river sand in the ratio of 3:1. Three weeks after germination of proper seedlings of tomato were transplanted to each 15 cm diameter clay pots filled with 1 kg autoclaved soil. Four replications of each tomato cultivar were used for root-knot nematode-resistance screening in completely randomised design(CRD). After the acclimatization of the plants and roots get stabilized, they were inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M. incognita by making 3-4 holes in the pots without disturbing the root system. After three months of inoculation, the plants were uprooted crop and different plant growth parameters like plant length (cm), fresh and dry weight (g), pollen fertility (%) and yield (g) were estimated. The effect on different biochemical parameters such as chlorophyll content (mg/g), carotenoid (mg/g) were determined besides this, the degree of root knot infestation was examined in terms of eggmasses/plant and eggs/eggmass, juvenile population/250g soil, root knot indices. Reaction in terms of resistance and susceptibility were also calculated at the termination of the experiment. Rating scale for the assessment of level of resistance or susceptibility of tomato cultivars based on number of galls given by Taylor and Sasser (1978).
Number of galls Resistance rating 0 Immune (I) 1-2 Resistant (R) 3-10 Moderately resistant (MR) 11-30 Moderately susceptible (MS) 31-100 Susceptible (S) >100 Highly susceptible (HS)
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3.9. Preparations of Talc Based Formulation of Biocontrol Agents
3.9.1. Talc based formulation of fungal biocontrol agents
The cultures of the fungal biocontrol agents viz., Pochonia chlamydosporia (ITCC No. 6898) and Purpureocillium lilacinum (ITCC No. 6064) were obtained from the Indian Agriculture Research Institute (IARI), New Delhi while Trichoderma viride (MTCC No. 3833) was obtained from Institute of Microbial Technology (IMTECH)-Chandigarh, India. The method of Vidhyasekaran and Muthamilan (1995) with some modification was followed for the preparation of talk based formulations of fungal bioagents. The fungal bioagents were prepared and maintained on Potato Dextrose Agar (PDA) medium and Potato Dextrose Broth (PDB) was used for mass culturing. The constituents of PDA (Riker and Riker, 1936) are given below:
Agar 17.00 g
Potato peeled and sliced 200.00 g
Dextrose 20.00 g
Doubled Distilled water 1000.00 ml
PD Broth (PDA minus agar) with amount of five hundred (500) ml was kept in 1000 ml capacity conical flasks and autoclaved at 15 lbs for 20 minutes. It was cooled at room temperature and each flask was transferred under aseptic conditions with the tested fungal isolate for mass multiplication. To prevent the bacterial contamination antibiotic streptomycin at 1ml/L was added. The flasks were then kept in incubator for incubation at 25±2°C for three weeks. After that, the fungal mat and broth were mixed properly with an electric mixer and grinder. Thereafter, 1 kg of sterilized talc as carrier material was mixed with fungal biomass thus bulk up the material for field application. The 1 % (w/w) carboxymethyl cellulose a carrier material was also added as a sticky agent. All the prepared contents were thoroughly mixed manually and dried at room temperature. The material (formulation) was then packed in polythene bags and used for field trials. The formulations packed in polythene bags were applied as broadcast with farm yard manure (FYM). At the time of application the colony forming unit counts (CFU) of P. lilacinum, P. chlamydosporia and T. viride were 2.5 × 106, 2.2 × 106 and 2.7 ×106/g talc formulation, respectively.
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3.9.2. Talc based formulation of bacterial biocontrol agent
Pure culture of Pseudomonas fluorescens (ITCC No. BE0004) was obtained from IARI, New Delhi. The method of Vidhyasekaran and Muthamilan (1995) was used for the preparation of formulation by using a mixture of 10g of carboxymethyl cellulose and 1 kg of talc. The 15g of calcium carbonate was added to maintain the pH at 7.0 and the mixture was autoclaved for 30 minutes for two consecutive days. The culture of P. fluorescens was grown on liquid King’s B medium (KBM) for 48 hours at room temperature (25±2°C) as a shake culture in an Infors AG shaker at 150 rpm. Thereafter, 1 kg of talc was mixed with 400 ml of bacterial suspension, containing 2×106 CFU/ml, under sterile conditions and shed dried to lower down the moisture content less than 20%. Then the formulation (mixture) was packed in polythene bags, sealed and kept under room temperature and for field application it was mixed with Farm Yard Manure.
3.10. Antinemic action of aqueous extracts of plants, oil cakes, bioagents and biochar (In vitro studies)
3.10.1. The objective of the study was to evaluate the nematicidal potential of aqueous leaf extracts of different plant species
Different plant species tested for their nematicidal potential were Mexican poppy (Argemone mexicana L., Family Papaveraceae), Trailing eclipta (Eclipta alba L., Family Asteraceae), Wild eggplant (Solanum xanthocarpum Schrad. & Wendl, Family Solanaceae), Black pigweed (Trianthema portulacastrum L., Family Aizoaceae), Indian mallow (Abutilon indicum L. (Sweet), Family Malvaceae), Ivy gourd (Coccinia grandis (L.) Voigt Family Cucurbitaceae). They were taken from the campus Aligarh Muslim University, Aligarh. Fresh leaves 25g of each sample were dissolved in 100 ml distilled water and kept for 12 hours and then samples were macerated separately in a warring blender and centrifuged. The obtained supernatant was then filtered through Whatman’s filter paper No. 1 and the filtrate was termed as Standard (S) represented as 100%. Standard (S), concentration was further diluted to S/2, S/10 and S/100 by adding requisite amount of distilled water.
For mortality experiment, 100 freshly hatched second stage juveniles were transferred separately to 40 mm diameter petridishes containing 10ml of the various
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Materials And Methods dilutions of water extracts, according to the method of Alam (1985). The eggmasses placed in sterilized distilled water served as control and each treatment was replicated in quadruples. The observations were made on mortality of juveniles after 12, 24 and 48 hours. The mean percentage of mortality was calculated. Death of nematodes was ascertained after the juveniles were transferred to plain water for 1 hour.
For hatching experiments, five healthy eggmasses of average-sized of root knot nematode, Meloidogyne incognita were taken with the help of sterilized forceps and placed in 40 mm petridishes containing 10 ml of above test aqueous extracts of various dilutions. There were four replicates of each treatment including distilled water as control. Petridishes were left for 5 days at 28 ± 2˚C temperatures. Hatched juveniles were counted after five days with the help of counting dish under stereoscopic microscope and the percent inhibition over distilled water control was calculated.
3.10.2. Effect of aqueous dilutions of different oil cakes on egghatching and juvenile mortality of Meloidogyne incognita.
The various oil seed cakes of castor, cotton, mahua, mustard and soybean were grounded into the powder form with the help of grinder. Twenty gram of finely grounded powder was mixed with 100 ml of distilled water to prepare the aqueous extract of oil cakes and kept for overnight and then centrifuged at 5000 g for 10 minutes. The pallet was discarded and supernatant was filtered through Whatman filter paper no. 1. The filtrate was passed through a bacterial filter (2µ) and stored in a refrigerator at 5 °C. The filtrate obtained was 100 % and was denoted as standard ‘S’. To avoid mould growth few drops of freshly prepared copper sulphate solution were also added. The other dilutions and their antinemic properties were assessed as described in 3.10.1.
3.10.3. Effect of culture filtrates of biocontrol agents on hatching and mortality of M. incognita juveniles
Studies were carried out to elucidate the potential of culture filtrates of P. lilacinum, P. chlamydosporia, T. viride and P. fluorescens on juvenile hatching and mortality of M. incognita in vitro. The fungal biocontrol agents cultured and maintained on Potato Dextrose Agar (PDA) at 28 ± 2° C as expressed in 3.9 worked
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Materials And Methods as a source of inoculum. Fungal inoculation was performed under sterilized conditions by adding one disc (1 cm diameter) for each 500 ml Erlenmeyer flask containing 100 ml of Potato Dextrose (PD) broth media for 15 days in darkness at 28 ± 2° C. Thereafter, the culture was centrifuged (6000 rpm for 10 min.) and then filtered through Whatman’s filter paper no. 1. The obtained filtrate was designated as Standard concentration ‘S’, and further dilutions viz., S/2, S/10 and S/100 were prepared by the addition of requisite amount of distilled water. P. fluorescens was cultured in liquid King’s B Medium (KBM) as classified in 3.9. To separate bacterial cells from the culture medium 48 hour old culture was centrifuged at 10000 g for 10 minutes. The obtained pellets (bacterial cells) were washed by centrifugation thrice with distilled water (DW) and finally suspended in distilled water. The obtained culture filtrate was designated as Standard ‘S’, and remains procedure was same as followed for other fungal bioagents. Mortality test of biocontrol agents viz., P. lilacinum, P. chlamydosporia, T. viride and P. fluorescens were conducted in the concentration grades i.e., S, S/2, S/10, S/100 kept in cavity blocks @ 10ml having 100 juveniles (J2) of M. incognita in each block as obtained in 3.9 were added to each block. Numbers of mobile and immobile nematodes were counted after12, 24and 48 hours of exposure period. To avoid bacterial contamination two drops of 0.1% streptomycin sulphate were added to each block containing fungal filtrates. There were four replicate of each treatment including the water control where no treatment has been given.
For examining the effect of biocontrol agents on juvenile hatching, 10 ml of each dilution of fungal and bacterial filtrate were pipette to each cavity block. To avoid bacterial contamination two drops of 0.1% streptomycin sulphate was mixed to each cavity block. Five healthy and same size eggmasses of M. incognita were transferred to each block. Distilled water containing cavity block served as control. There were four replicates of each treatment. The cavity blocks were kept in incubator at 25 °C. The number of hatched juveniles in each block was counted after 5 days under stereo-binocular microscope and percent inhibition was calculated.
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3.10.4. Effect of aqueous exudates of biochar on egghatching and juvenile mortality of Meloidogyne incognita.
Biochar prepared from Prosopis juliflora wood through pyrolysis was procured from Greenfield Eco solutions Pvt. Ltd Jodhpur. Finely grounded powder of biochar with the amount of 25g was mixed with 100 ml of distilled water to prepare the aqueous extract of biochar. The aqueous extract was kept for overnight and then centrifuged at 5000 rpm for 10 minutes. The pallet was removed and supernatant was filtered through Whatman filter paper no. 1. Thereafter, the obtained filtrate was passed through a bacterial filter (2µ) and stored in a refrigerator at 5 °C. The filtrate was 100 % and was termed as standard ‘S’. This S concentration was further diluted to S/2, S/10 and S/100 concentration by adding necessary amount of water. To avoid mould growth few drops of freshly prepared copper sulphate solution were also added. Antinemic properties were determined as described in 3.10.1
3.11. Bare-root-dip and juveniles penetration (root dipping study)
The chopped leaves viz., Mexican poppy, Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd, various oil cakes viz., cakes of castor, cotton, mahua, mustard and soybean, bioagents viz., P. lilacinus, P. chlamydosporia, T. viride and P. fluorescens and biochar were tested to evaluate the potential of their extracts against the penetration of second stage juveniles of root knot nematode, Meloidogyne incognita into the roots of tomato cv.K-21.The aqueous dilutions of chopped leaves and oil cakes were obtained by the same procedure 3.10.1.and 3.10.2. The aqueous extract of bioagents and biochar was prepared by the same procedure 3.10.3 and 3.10.4 respectively.
Three week old seedlings root of tomato were raised in sterilized pots having manure supplemented autoclaved soil, were dipped in the different aqueous dilutions of various plants extract, oil cakes, bioagents and biochar for 50 and 100 minutes respectively. Each treatment was replicated four times. After dipping the roots were thoroughly washed and the seedlings were transplanted to 5 cm pots filled with thoroughly washed river sand. The seedlings were inoculated with 1500 second stage juveniles of Meloidogyne incognita. After 72 hours of the dipped treatment, inoculated seedlings were removed safely and the sand was isolated by using Cobb’s sieving and decanting technique followed by modified Baermann’s funnel technique, 68
Materials And Methods for determining the number of juveniles which could not penetrated the roots of the test plants. The extent of penetration and percent inhibition was assessed by deduction of this value from the initial number of inoculated juveniles.
3.12. Management of nematodes through plant extracts and biocontrol agents alone and in combination as bare root dip treatment
3.12.1. The objective of the study was to evaluate the effect of bare-root dip treatment by aqueous extracts of leaves of different plants against root knot nematode, Meloidogyne incognita.
The standard aqueous extracts of leaves of Mexican poppy, Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd were prepared in the same manner described by the method in the 3.10. 1. The prepared standard was diluted to S/2 and S/10 by adding required amount of water. Three week old seedlings roots of tomato cv. K-21, grown in clay pots 30 cm containing sterilized soil were dipped for 45 and 90 minutes in S, S/2 and S/10, concentration of leaf extracts of Mexican poppy, Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd respectively. After the completion of root dipping treatment, the seedlings roots were washed with distilled water and transplanted in earthen pots (15 cm) filled with 1 kg of autoclaved soil. Each treatment was replicated four times. Undipped uninoculated and undipped inoculated plants served as control. The roots of the acclimatized plants were inoculated with 1500 freshly hatched second stage juveniles (J2) of root-knot nematode, M. incognita by making 4-5 holes nearby to the rhizosphere of the root system without disturbing the plant. After three months of inoculation different plant growth parameters like shoot and root length (cm), fresh and dry weight (g) pollen fertility (%), physiological parameters chlorophyll content (mg/g), carotenoid (mg/g) and nematode infestation in terms of eggmasses/root and root knot indices was observed.
3.12.2 Management of nematode by Bioagents
The method of culture of bioagents viz., P. fluorescens, P. lilacinum, P. chlamydosporia and T. viride was done as described in 3.9. The 48 h-old-culture in King’s medium B (KBM) was centrifuged at 10,000 g for 10 min to separate bacterial cells from culture media. After centrifugation, supernatants were discarded and pellets
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Materials And Methods
(bacterial cells) were washed by centrifugation three times with distilled water (DW) and finally suspended in distilled water. The number of Pseudomonas fluorescens culture solution was adjusted to 2x106CFU/ml and was considered as slandered S concentration. Which was diluted to S/2 and S/10 by adding requisite amount of the water. For the fungal bioagents preparation, fifteen days old culture of bioagents viz., P. chlamydosporia, T. viride, and P. lilacinum grown on standard dextrose potato agar medium was scraped and comminuted in distilled water with 0.01% Triton X-100 in a warring blender and the spores concentration was adjusted to 2x106 spores/ml and it was considered as S concentration and was further diluted to S/2, S/10 by adding necessary amount of water. Roots of three week old seedlings of tomato cv. K-21, raised in sterilized soil, were washed and dipped for 45 and 90 minutes in S, S/2 and S/10 concentration of P. fluorescens, P. lilacinus, P. chlamydosporia and T. viride respectively. After washing the roots in water the seedlings were transplanted in earthen pots (15 cm) containing 1 kg of sterilized soil. There were four replicates of each treatment. Undipped uninoculated and undipped inoculated plants served as control. Five days after transplanting, plants were inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M. incognita by making 4-5 holes nearby to the root system without disturbing the plant. After three months of inoculation, the plants were carefully uprooted and the roots were gently washed in running tap water to remove the adhering soil particles and different plant growth parameters, physiological and pathological parameters were calculated.
3.12.3. Management of nematode by combined treatment of plant extract with Bioagents
The bioagents viz., P. fluorescens, P. lilacinum, P. chlamydosporia and T. viride were cultured as classified in 3.9. The P. fluorescens culture grown on liquid King’s B medium (KBM) for 48 hours was centrifuged at 10,000 g for 10 min to separate bacterial cells from culture media. The supernatants were discarded and pellets (bacterial cells) were washed by sterilized distilled water through centrifugation three times and finally suspended in sterile distilled water. The number of pseudomonas fluorescens in bacterial suspension was adjusted to 1x105CFU/ml and was considered as standard S concentration. This was further diluted to S/2 and S/10 by adding requisite amount of the water. Fifteen days old culture of fungal bioagents viz., P. chlamydosporia, T. viride, and P. lilacinus were grown on standard 70
Materials And Methods dextrose potato agar medium was scraped and comminuted in distilled water with 0.01% Triton X-100 in a warring blender and the spores concentration was adjusted to 1x105 spores/ml and it was considered as ‘S’ concentration. It was further diluted to S/2, S/10 by adding necessary amount of water. Roots of three week old seedlings of tomato cv. K-21, raised in sterilized soil were washed and dipped for 45 and 90 minutes in S, S/2 and S/10 concentration of P. fluorescens, P. lilacinus, P. chlamydosporia and T. viride respectively. The standard aqueous extract of the leaves of water hyacinth was prepared as described by the method in the 3.10. 1. Equal amount of S concentration of the plant extract was mixed with S/2 dilution (1x105CFU/ml/2) and S/10 dilution (1x105CFU/ml/10). Thereafter, the roots were washed in water and seedlings were transplanted in earthen pots (15 cm) containing 1 kg of autoclaved soil. Each treatment was replicated four times. Undipped uninoculated and undipped inoculated plants served as control. After acclimatization, the plant were inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M. incognita by making 4-5 holes nearby to the rhizosphere of the plant without disturbing the root system. Three months of post inoculation, the plants were carefully uprooted and the roots were washed in running tap water to remove the adhering soil particles and different plant growth parameters, physiological and pathological parameters were calculated.
3.13. Nematode management with chopped leaves of plant in combination with seed powder of black nightshade
3.13.1. The objective of the study was to evaluate the potential of chopped leaves of different plants when used and in combination with seed powder of black nightshade For assessing the efficacy and nematostatic potential of organic soil amendments, chopped leaves of different plants viz., Mexican poppy, Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd were applied @ 50g combined with seed powder of Black nightshade @ 10 g per pot and thoroughly mixed with soil. Untreated uninoculated and untreated inoculated served as control. Each treatment was replicated four times. The clay pots (15 cm in diameter) were watered at regular interval for ensuring proper decomposition of the organic additives for two weeks. After two week of waiting period, three week old seedlings of tomato cv. K-21 were transplanted singly into each pot. As the plants get acclimatized they were inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M. 71
Materials And Methods incognita by making 3-4 holes in the pots soil nearby to the roots at the same distance in the manner so that root don’t get damage. Then requisite amount of suspension having necessary number of second stage juveniles was procured into the holes and then covered them with the soil. After three months of the inoculation, roots of tomato cv. K- 21 were uprooted gently from the pots and were washed in running tap water to wash off the soil particles adhered with the root. The plant growth parameters, physiological and pathological parameters were assessed.
3.13.2 Nematode management with biochar alone and in combination with sawdust
Biochar were used alone and in combination with sawdust of different plants viz., Eucalyptus, Lebbeck, Jambul, Mango, Poplar and Babool against root-knot nematode M.incognita. The biochar was separately mixed with soil @ 65g/kg soil and then these pots were amended with sawdust of different plants @ 20g/pot and thoroughly mixed with soil. These pots were kept in an open field exposed to natural sunlight. The plants were irrigated with tap water on alternate days to keep the soil moist and facilitate the complete decomposition of sawdust. Untreated uninoculated and untreated inoculated served as control. Each treatment was replicated four times. The treated pots of the above dose with soil in clay pots (15 cm in diameter) were allowed two weeks for decomposition of organic amendments. Thereafter, three week old seedlings of cv. K-21 were transplanted singly into each pot. As the plant get acclimatized they were inoculated with 1500 freshly hatched second stage juveniles (J2) of root-knot nematode, M. incognita by making 3-4 holes nearby to the root system without disturbing the plant. Necessary watering, weeding and care was done as per requirement of the experiment. After three months the experiment was terminated. The different plant growth parameters like length (shoot and root in cm), fresh and dry weight of shoot and root in grams, yield/plant in grams and Percent pollen fertility in percent were estimated. The degree of root knot infection in terms of eggmasses/plant, eggs/eggmass and nematode population (J2)/250g soil and root knot indices was determined at the experiment termination.
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3.13.3. Nematode management with biochar alone and in combination with agriculture waste
For evaluating the efficacy and nematostatic potential of organic soil amendments in the form of biochar were applied @ 65g/pot. Then these pots were amended with agriculture waste viz., tobacco, garlic waste, mentha waste, decomposed potato, black gram waste and bagasse applied @ 20g/pots and thoroughly mixed with soil. There were four replicate of each treatments. Untreated uninoculated and untreated inoculated were served as control. Immediate watering was done for the proper decomposition of the tested dose of the amendment. The pots treated with different dose with soil in clay pots (15 cm in diameter) were allowed for two weeks for decomposition of organic matter. After proper decomposition, three week old seedlings of tomato cv. K-21 were transplanted singly into each pot. After acclimatization, the plants were inoculated with 1500 freshly hatched second stage juveniles (J2) of root-knot nematode, M. incognita by making 3-4 holes in the rhizosphere of the plant without damaging the root system. Three months after inoculation, the experiment was terminated and different plant growth parameters, physiological and pathological parameters were estimated.
3.14. Management of nematode through bioagents, oil cakes and biochar as seed dressing
3.14.1. Seed dressing with bioagents
The seeds of tomato cv.K-21 were surface sterilized with 1% sodium hypochlorite (NaOCl) for three minutes, rinsed thoroughly with running tap water and dried aseptically. The bioagents viz., P. chlamydosporia, P. lilacinum, P. fluorescens and T. viride were cultured as described in 3.9. and applied @ 4 g/kg seed using 1% and 2% cellulose and molasses as sticking agents. The seed were thoroughly mixed manually to get uniform and smooth coating. Coated seeds were spread in enamel tray and allowed to shade dried before sowing. These treated seeds were sown in autoclaved soil in clay pots (30cm diameter) for nursery preparation. Seedlings of tomato cv. K-21, three week after germination were transplanted to each clay pot singly filled with 1 kg of autoclaved soil for further experiment. Untreated uninoculated and untreated inoculated served as control. Each treatment was replicated four times. After acclimatization, the plants were inoculated with 1500
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Materials And Methods second stage juveniles (J2) of root-knot nematode, M. incognita with the help of sterilized pipette by making 3-4 holes around the plant without disturbing the root. These holes covered carefully with the sterilized soil. The plants were carefully uprooted after three months of inoculation from the pots and the roots were washed gently in running tap water so that eggmasses may not detached and root adhered soil particle can be removed. The plant growth parameters like plant length (shoot and root) in centimeters, weight (fresh and dry) in gram, pollen fertility (%), yield (g), physiological parameters (chlorophyll and carotenoid content) in mg/g and beside this nematode infestation in terms of eggmasses/plant, eggs/eggmass and juvenile population/250g soil and root knot indices were evaluated at the end of the experiment.
3.14. 2. Seed dressing with oil seed cakes
The experiment was conducted to test the efficacy of deoiled powdered cakes of castor, cotton, mahua, mustard and soybean as seed dressing agents @ 7, 15 and 21 % w/w on seed using gum as adhesive and chalk as drier against root knot nematode infestation. Seeds were mixed properly to form a smooth and uniform coating and then dried in shed in enamel tray before sowing. Seeds which were untreated served as control. These treated and untreated seeds were sown in autoclaved soil in clay pots (30cm diameter) for nursery preparation. As the seedlings of tomato cv.K-21 germinated properly, were transplanted to clay pot containing 1 kg autoclaved soil singly for the experiment. Each treatment was replicated quadruples. After three months, the plants were terminated and different plant growth parameters viz., shoot and root length in centimeters, fresh and dry weight in gram, pollen fertility (%), yield (g), physiological parameters (chlorophyll and carotenoid content) in mg/g and beside this, nematode infestation in the form of eggmasses/plant, eggs/eggmass and juvenile population/250g soil and root knot indices were observed at the end of the experiment.
3.14.3. Seed dressing with biochar
Biochar prepared from Prosopis juliflora wood through pyrolysis was purchased from Greenfield Eco solutions Pvt. Ltd Jodhpur. Biochar was dissolved in distilled water at the concentrations of @ 0.2, 0.4, 0.8, 1.6, 3.2, 6.4% (v/v) for 1 week. The suspension was centrifuged at 12,000 rpm for 5 min, and the obtained supernatant
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Materials And Methods was used to test the direct toxic effect of biochar exudates on the root knot nematode as seed dressing agents. A 5% solution of gum arabic was added to the supernatant and stirred mechanically until the gum dissolved completely and it was used as a sticker. The treated seeds were then spread in an enamel tray and allowed drying in the shade before sowing. Untreated seeds were served as control. Both treated and untreated seeds were sown in autoclaved soil in clay pots (30cm diameter) for nursery preparation. Three week after germination the single seedlings of tomato cv.K-21 were transplanted to clay pot containing 1 kg autoclaved soil for the experiment. Each treatment was replicated four times. Three months after inoculation, the plants were harvested and different plant growth parameters, physiological parameters and pathological parameters were observed at the end of the experiment.
3.15. Effect of individual, sequential and concomitant application of biocontrol agents on Meloidogyne incognita
The experiments were carried out in the Department of Botany, Aligarh Muslim University, Aligarh, under glasshouse conditions. Individual, Sequential and Concomitant inoculation of biocontrol agents viz., P. chlamydosporia, T. viride, P. lilacinum, and P.fluorescens were tested against M. incognita in pots. At two leafy stage of tomato cv. K- 21were transplanted singly into each clay pots (15 cm in diameter) filled with autoclaved soil. After acclimatized plants were inoculated with M.incognita and bioagents viz., P. chlamydosporia, P. lilacinum, P. fluorescens, and T. viride were done according to the following experimental design:
Untreated Uninoculated Control (UUC)
T. viride - Inoculated with T. viride @ 3.0g/pot alone
T. viride + Mi - Inoculated with T. viride @ 3.0g + M. incognita (1500 J2) simultaneously
Mi →T. viride - Inoculated with M. incognita (1500 J2) followed by T. viride 3.0g after 15 days
T. viride→ Mi - Inoculated with T. viride @ 3.0g followed by M. incognita (1500 J2) after 15 days.
Untreated Inoculated control (UIC) - inoculated with M. incognita alone (UIC)
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Other bioagents were also applied on similar lines viz., P. chlamydosporia (3.0g/pot), P. lilacinum (3.0g/pot) and P. fluorescens (2.0g/pot). Each clay pot having 1 kg autoclaved soil was inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M. incognita by making 3-4 holes nearby the rhizosphere of the plant without disturbing the root system. The plants were harvested after three months of inoculation and different plant growth parameters, viz., shoot and root length (cm), weight fresh and dry (g) of shoot and root, pollen fertility (%) and yield (g) were taken into consideration. The impact on different biochemical parameters like chlorophyll content in mg/g , carotenoid (mg/g), and nitrate reductase (NRA) (nmol/h/g) besides this root knot infestation in terms of eggmasses/plant and eggs/eggmass, nematode population/250g soil and root knot indices were also determined at the experiment termination.
3.16. Management of nematodes through biocontrol agents in combination with various organic amendments
3.16.1. The objective of the study was to elucidate the potential of various biocontrol agents in combination with various organic soil amendments against root knot nematode under pot conditions
Press mud and various oil cakes viz., Castor (Ricinus communis L. Family Euphorbiaceae), Mustard (Brassica juncea L. Family-Brassicaceae), Mahua (Madhuca longifolia L. Sapotaceae), Soybean (Glycine max L. Fabaceae), cotton (Gossypium arboreum L. Family- Malvaceae), were used in combination with biocontrol agents against root-knot nematode, M. incognita. The pots were treated with press mud and different oil cakes viz., castor, mustard, cotton, mahua and soybean applied @ 20g/pot separately. The pots were left for 2 weeks and watered at regular interval for the proper decomposition of oil cakes. Seedlings were raised in 30 cm pots filled with autoclaved soil. Three week old seedlings of tomato cv. K-21 were transplanted singly into each pot containing decomposed oil cakes. As the plants get acclimatized in pots then treated with the biocontrol agents viz., P. chlamydosporia, T. viride, and P. lilacinum, @ 2.0g/pot and P.fluorescens @ 1.5g /pot separately. Untreated uninoculated and untreated inoculated represented as control. Each treatment was replicated four times. After 5 days each pot inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M. incognita by making 3-4 holes in the rhizosphere of the plants in a manner so that the root don’t get 76
Materials And Methods damage. Three months after inoculation, the experiment was terminated and various growth parameters like length of shoot and root (cm), fresh and dry weight of shoot and root (g), yield/plant (g) and pollen fertility (%) were estimated. The root knot infection level in terms of eggmasses/plant, eggs/eggmass and nematode population (J2)/250g soil and root knot indices was determined at the termination of the experiment.
3.16.2. Management of nematodes through biocontrol agents alone and in combination with potato waste and composted plant straws
Potato waste and plants straw of various plants viz., Pigeon pea, Pearl millet, Maize, Mustard, and Sorghum were allowed to decompose in the container. Water was added at 7 days of interval for the proper decomposition of the plants straw. The pots filled with autoclaved soil were treated with potato waste and different composted straws viz., Pigeon pea, Pearl millet Maize, Mustard, and Sorghum applied @ 15g/pot separately. Seedlings were raised in 30 cm pots filled with autoclaved soil. Seedlings of tomato cv. K-21 at two leafy stage were transplanted singly into each pot containing composted straw and potato waste. After the acclimatization of plants in pots then treated with the biocontrol agents viz., P. chlamydosporia, T. viride, and P. lilacinum @ 2.0g/pot and P. fluorescens @ 1.5g /pot separately. Each treatment was replicated four times. Untreated inoculated and untreated uninoculated plants served as control. After 5 days as the roots get stabilized each pot was inoculated with 1500 second stage juveniles (J2) of root-knot nematode, M.incognita by making 3-4 holes in the rhizosphere of the plants nearby to the roots. Three months after inoculation, the experiment was terminated and various growth parameters, physiological and pathological parameters were calculated.
3.17. Chlorophyll Estimation
The chlorophyll and carotenoid content in the fresh leaves were determined by the method of Mackinney (1941) and MacLachlan and Zalik (1963) respectively.
One gram of finely cut fresh leaves of test plants tomato cv.k-21were ground to a fine pulp with the help of mortar and pestle after adding 20 cm3 of 80 % acetone. The obtained mixture was then centrifuged at 5000 rpm for 5 minutes and supernatant was collected in 100 cm3 volumetric flask. The residue was washed three times by
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Materials And Methods using 80 % acetone and each washing was collected in the same volumetric flask and the final volume was made up to the mark using 80 % acetone. The absorbance was observed at the wavelength of 645 nm and 663 nm against blank (80 % acetone) on spectrophotometer (UV 1700, Shimadzu, Japan). The chlorophyll content present in the extract (mg g-1 tissue) was calculated by using the following equation: mg chlorophyll ‘a’ g-1 tissue = 12.7 (A663)-2.69 (A645) × V/1000×W mg chlorophyll ‘b’ g-1 tissue = 22.9 (A645)- 4.68(A663) × V/1000×W mg total chlorophyll g-1 tissue = 20.2 (A645)- 8.02 (A663) × V/1000×W
Carotenoids (mg g-1FW) = . A − . A D× ×W × V A = Absorbance at specific wavelength ( )
V = Final volume of extract in 80 % acetone
W = Fresh weight of tissue extracted
3.18. Nitrate Reductase Activity (NRA)
The activity of nitrate reductase in fresh leaves was estimated by following Jaworski (1971) method.
The chopping of the leaves was done and 200 mg of these chopped leaves were weighed and transferred to plastic vials. To each vial, 2.5 ml of phosphate buffer pH 7.5 and 0.5 ml of potassium nitrate solution was added followed by the addition of 2.5 ml of 5 % of iso-propanol. These vials were kept in BOD incubator for incubation for 2 hours at 30 ± 2 °C in dark. To the 0.4 ml of incubated mixture taken in a test tube 0.3 ml each of sulphanilamide solution and NED-HCl were added and left for 20 minutes, for the colour development. The mixture was diluted to 5 ml by Distilled Water (DW). The absorbance was read at 540 nm using spectrophotometer (UV 1700, Shimadzu, Japan).
A blank sample was preceded simultaneously with each sample. Standard curve was plotted by compared with the calibration curve and nitrate reductase activity expressed at nm (nmolh-1 g-1).
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Materials And Methods
3.19. Pollen Fertility
Young and fresh flowers from plants were used each treatment and in the control. 2% acetocarmine was used for the staining of the pollen grain to estimate the pollen fertility. Stain attaining pollen grains with the regular outline were considered as fertile, whereas, empty, unstained and shrunken were considered as sterile. The pollen fertility was calculated from the following formula.
Pollen fertility (%) =Number of fertile pollen/Total number of pollen ×100
3.20. Assessment of Nematode Parameters (eggmass and eggs / eggmass).
The eggmasses were estimated by following the method of Daykin and Hussey (1985). The roots of the infected plants were immersed in Phloxine B solution (0.015%) for 20 min and then washed with tap water so that rest of the Phloxine B may remove. Its gives pink colour to the egg-masses and the roots remain colourless or stain very lightly. The number of eggs/eggmass was estimated by choosing 10 healthy uniform size egg-masses from each root system and shaking was done in 1% NaOCI solution for 3 min. Sieving of the egg suspension was done with 200 and 500 mesh (75 and 26 m) with tap water to remove the debris on the first sieve and eggs were collected on the second one (Hussey and Barker, 1973). Released eggs were collected in 50 ml water suspension and number of eggs was counted in 1 ml by the help of a light microscope under low power (10X). Average number of eggs/egg-mass was calculated. The extent of root-knot infestation was assessed according to the rating scale of Taylor and Sasser (1978).
Root-knot index Number of galls/root system 0 0 1 1-2 2 3-10 3 11-30 4 31-100 5 >100
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Materials And Methods
3.21. Experimental Design and Statistical Analysis
The glasshouse pots study was set up in completely randomised design (CRD) with four replicates in Department of Botany, Aligarh Muslim University, Aligarh. Each treatment was represented by four pots where each pot was considered as a replicate. The data of the experiments was statistically analyzed and standard errors were calculated. Analysis of variance was performed on the data by using SPSS software (version12.00 Inc., Chicago, IL, USA) to determine the significance at P = 0.05. Least Significant Difference (L.S.D.) was calculated for the significant data to identify difference in the mean of treatment. Duncan's Multiple Range Test (DMRT) was employed to test for significant difference between the treatments. Where means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05
pathological parameters (eggmasses, eggs/eggmass and nematode population/250g soil) are in round figures.
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Experimental Results
CHAPTER IV
EXPERIMENTAL RESULTS
4.1. Screening of different cultivars of tomato against root-knot nematode, Meloidogyne incognita in pots
Fourteen cultivars of tomato (Solanum lycopersicum L.) viz., EC-538380, K- 21, CO-3, FEB-02, EC-570018, NDT-3, S-22, GT-1, GT-2, GT-3, H-88-78-1, PB Barkha bahar-2, VRT-101A, Kalyanpuri-T1 were appraised for their resistant and susceptibility behaviour against the root knot, Meloidogyne incognita in pots under glasshouse conditions. The parameters were studied in terms of plant growth characters viz., plant length (cm), fresh and dry weight (g), chlorophyll content (mg/g), carotenoid content (mg/g), yield (g) and pollen fertility (%). The disease intensity was studied in terms of number of eggmasses/root, eggs/eggmass, nematode population and root knot index. Among the fourteen cultivars screened considerable variation was observed in resistance level against M.incognita. The overall reduction in plant growth characters was observed in all the tested cultivars of tomato which may be due the infestation of nematode. The variety H-88-78-1 showed highest resistance against the root knot nematode by the presence of number of eggmasses, eggs and root-knot indices and variety K-21 was found highly susceptible. On the basis of resistance, tomato cultivars can be arranged in the following descending order: VRT-101A> EC-570018> GT-2 > PB Barkha bahar-2> FEB-02> CO-3> Kalyanpuri-T1> NDT-3> EC-538380> GT-3> GT-1> S-22> K-21 (Table-1a).
All the cultivars of tomato inoculated with M.incognita responded differently in terms of plant length and exhibited significant difference in comparison to the cultivars which were untreated inoculated. Maximum plant length (82.0 cm) was observed in cultivar H-88-78-1 in uninoculated control pots and 78.0 cm in inoculated control pots. It was followed by 80.0 and 75.0 cm; 78.5 and 70.5 cm; 77.3 and 65.2 cm; 80.7 and 66.3 cm in VRT-101A, EC-570018, GT-2 and PB Barkha bahar-2 both uninoculated and inoculated control respectively. In K-21 cultivar, the plant length was (77.6 cm) in uninoculated control and 39.0 cm in inoculated control pots.
It was observed that root knot nematode; M.incognita significantly reduced the fresh weight of various cultivars of tomato as against inoculated control. Different
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Experimental Results cultivars of tomato represent varying degree of reaction on fresh weight of tomato against M.incognita. Minimum reduction in fresh weight (73.0 g) was observed in H- 88-78-1 in uninoculated control and (69.5 g) in inoculated control. It was followed by 71.6 and 67.2 g; 69.0 and 62.1 g; 67.7 and 56.5; 72.0 and 58.7 g in VRT-101A, EC- 570018, GT-2, PB Barkha bahar-2 both uninoculated and inoculated control respectively. The maximum reduction was observed in weight K-21 (71.0 g) in uninoculated control and (32.4 g) in inoculated control.
Root knot nematode, M.incognita significantly reduced the dry weight of tested cultivars of tomato as against uninoculated control. Minimum reduction in dry weight (24.3 g) was observed in H-88-78-1 in uninoculated control and (24.0g) in inoculated control. It was followed by 24.4 and 22.8g; 22.9 and 20g; 22.0 and 18.7; 24.0 and 19.5g in VRT-101A, EC-570018, GT-2 and PB Barkha bahar-2 both uninoculated and inoculated control respectively. The maximum reduction was observed in weight K-21 (24.0g) in uninoculated control pots and (11.3g) in inoculated control pots. Reduction in the dry weight may be one of the features representing the susceptibility of cultivars of tomato. On the basis of dry weight the other cultivars can be arranged in the in the following descending order; VRT-101A> EC-570018> GT-2> PB Barkha bahar-2> FEB-02> CO-3> Kalyanpuri-T1> NDT-3> EC-538380> GT-3> GT-1> S-22 (Table-1a).
The chlorophyll content of different tomato cultivars was also reduced due to the infestation of M.incognita, however, to varying degree. Lowest reduction in chlorophyll content in the tested cultivar was observed in H-88-78-1 (2.82 mg/g) in uninoculated control and (2.70 mg/g) in inoculated control but the reduction was insignificant. It was followed by 2.74 and 2.53 mg/g, 2.69 and 2.47 mg/g, 2.64 and 2.26 mg/g, 2.78 and 2.30 mg/g in VRT-101A, EC-570018, GT-2, and PB Barkha bahar-2 both in uninoculated and inoculated control respectively. Highest decrease among all the screened cultivars was detected in K-21 and it was 2.78 mg/g in uninoculated and1.45 mg/g in inoculated respectively (Table-1a).
The carotenoid content of different tomato cultivars was also reduced due to the infestation of M.incognita however, to varying extent. Minimum reduction in carotenoid content was observed in H-88-78-1 was 0.938 mg/g in uninoculated and0.889 mg/g in inoculated control. It was followed by 0.920 and 0.862 mg/g, 0.912
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Experimental Results and 0.800 mg/g, 0.902 and 0.770 mg/g in VRT-101A, EC-570018, GT-2, and PB Barkha bahar-2 both uninoculated and inoculated control respectively. Most pronounced decrease in all the tested cultivar was detected in K-21 and it was 0.869 mg/g in uninoculated control pots and 0.285 mg/g inoculated control pots respectively (Table-1a).
Significant reduction in yield was noticed among all the screened cultivars of tomato against M.incognita although up to a varying degree. Minimum reduction in yield in all the tested cultivar was analyzed in H-88-78-1 was 394 g in uninoculated control pots and 374 g in inoculated control pots. It was followed by 382 and 351 g, 372 and 327 g, 365 and 309 g, 389 and 320 g in VRT-101A, EC-570018, GT-2, PB Barkha bahar-2 both uninoculated and inoculated control pots respectively. Most remarkable decrease in yield among all the tested variety was observed in K-21and it was 386 g in uninoculated and 182 g in inoculated control respectively. It reflects a sense towards the susceptibility of the cultivar (Table-1a).
Percent pollen fertility of different tomato cultivars was also reduced due to the infestation of M.incognita, however, to varying degree. Lowest reduction in percent pollen fertility in the tested cultivar was observed in H-88-78-1 and it was 90.2 % in uninoculated control and 85.4 % in inoculated control. It was followed by 89.2 and 83 %, 87.8 and 67.3 %, 86.0 and 75.5 %,89.6 and 74.6 in VRT-101A, EC- 570018, GT-2, PB Barkha bahar-2 both in uninoculated and inoculated control pots respectively. The reduction in percent pollen fertility was found significant, but most significant reduction was observed in K-21 and it was 89.0 % in uninoculated and 48.2 % in inoculated control pots respectively.
Significant difference in eggmass production was observed on the root of each tested cultivars against M.incognita infection. Minimum number of eggmasses/root (4.0) was observed in cultivar H-88-78-1 as against to all other tested cultivar. It was followed by 6.0, 13.0, 20.0 and 24.0 eggmasses/root in cultivar VRT-101A, EC- 570018, GT-2 (20) and PB Barkha bahar-2 respectively. Maximum number of eggmasses/root (176) in all the tested cultivar was produced in K-21 in inoculated control pots (Table-1b).
Significant difference in eggs production was observed on the root of each tested cultivars against M.incognita infection. Minimum number of eggs/eggmasses in
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Experimental Results the cultivar H-88-78-1 was observed 6.0. It was followed by 9.0, 19.0, 29.0, and 36.0 in VRT-101A, EC-570018, GT-2 and PB Barkha bahar-2 respectively. Maximum number of eggs/eggmasses in all the screened cultivars was detected in K-21 in inoculated control pots (Table-1b).
The root knot nematode M.incognita population varied in the soil rhizosphere of all the screened cultivars of tomato. Population of nematode was directly concerned with the susceptibility of the variety and it showed negative correlation with growth parameters of the plant. Reduction in nematode population in the soil rhizosphere of all the screened cultivars was recorded. The minimum nematode population in H-88-78-1 was 223.0.It was followed by 328.0, 695, 744 and 814 in VRT-101A, EC-570018, GT-2 and PB Barkha bahar-2 respectively. Maximum number of nematode population after harvesting observed in K-21was 1632. All other cultivar showed less number of nematodes in their soil rhizosphere (Table-1b).
The root knot indices were significantly varied in all the tested cultivars of tomato against M.incognita infection. Root knot indices show indirect relationship with the reduction in plant growth characters and demarcated susceptibility of the tomato. High root knot indices are one of the indications of increase of reproduction rate favoured by the susceptible cultivars and suppressed in resistant one. Lowest magnitude of the root knot indices was detected in H-88-78-1was 1.2. It was followed by 1.4, 2.2, 2.5 and 2.7 in VRT-101A, EC-570018, GT-2 and PB Barkha bahar-2 respectively. Highest root knot indices in all the tested cultivar were observed in K-21 was 5.0. (Table-1b).
All the cultivars of tomato screened against root knot nematode illustrate differential response in terms of susceptibility and resistance behaviour. Out of the fourteen cultivars screened two (H-88-78-1, VRT-101A) showed resistant behaviour, three were found moderately resistant (EC-570018, GT-2 and PB Barkha bahar-2); three were found moderately susceptible (FEB-02, CO-3, Kalyanpuri-T1). Rest of the six cultivars showed susceptible behaviour (NDT-3, EC-538380, GT-3, GT-1, S-22, K-21) (Table-1b).
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Experimental Results
4.2. IN VITRO EXPERIMENTS
4.2.1. Effect of aqueous extract of fresh chopped leaf of some plant species on the larval hatching of Meloidogyne incognita in vitro
The aim of the experiment was to test the antinemic potential of aqueous extract of leaf of some selected plant species viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed under in vitro conditions against the second stage juvenile (J2) of root-knot nematode, Meloidogyne incognita. From the data it was clear that aqueous leaf extract of all the treated plants results in toxic effect to the egg, however, up to varying extent. Inhibitory effect on juvenile emergence was found to be directly proportional to the concentration of extract and exposure period (Table- 2, Figure-1).
From the results it was revealed that average number of juveniles hatched in S, S/2, S/10 and S/100 concentrations of Indian mallow leaf extract were 5, 36, 295 and 486 respectively. The corresponding values for various concentrations of Mexican poppy leaf extract were 0, 0, 185, 402; for leaf extract of Ivy gourd were 12, 46, 386, 514; for leaf extract of Trailing eclipta were 0, 2, 210, 430; for leaf extract of Wild eggplant were 0, 0, 238, 445 and for leaf extract of Black pig weed were 0, 17, 270, 462 respectively as compared to distilled water control showed maximum juvenile hatching were 560 (Table-2, Figure-1).
4.2.2. Effect of Water soluble fractions of various deoiled cakes on hatching of Meloidogyne incognita in vitro
This experiment was performed in vitro conditions to examine the effect of aqueous extracts of various oil cakes viz., castor cake, cotton cake, mahua cake, mustard cake, soybean cake on second stage juvenile hatching of root knot nematode, Meloidogyne incognita. All the dilutions of tested oil cakes cause inhibitory effect on juvenile development. However, inhibitory effect varied with concentration of the extract. The concentration was directly proportional to the inhibitory effect. (Table- 3, Figure-2).
It was clear from the results that minimum numbers of juveniles hatched in S, S/2, S/10 and S/100 concentrations of castor cake were 0, 30, 190 and 440 respectively. It was followed by mustard cake 0, 50, 216 and 457; soybean cake 2, 76,
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Experimental Results
242 and 478. While maximum number of juvenile hatched in cotton cake were 20, 97, 300 and 510. The distilled water (DW) control showed maximum juveniles hatching, where number of juveniles hatched was 535.
Percent inhibition of juvenile hatching in S, S/2, S/10 and S/100 concentrations of castor cake were 100 %, 94.4 %, 64.5 % and 17.7 %. The corresponding figures in the dilutions of mustard cake were 100%, 90.65%, 59.63% and 14.58%. The cotton cake showed least potential where the inhibition of juvenile hatching were 96.26 %, 82.06 %, 43.93 %, 4.670 % respectively (Table-3, Figure-2).
4.2.3. Effect of cultural filtrate of biocontrol agents on the larval hatching of juvenile of root-knot nematode Meloidogyne incognita in vitro
The experiment was carried out to evaluate the antinemic potential of culture filtrate of Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum and Trichoderma viride on larval hatching of second stage juvenile of Meloidogyne incognita. All the cultural filtrate was found nematotoxic to the larval emergence. However nematotoxic effect of various cultural filtrates varied with the concentration (Table- 4, Figure-3).
Complete inhibition in egghatching was observed in S concentration of the extract of Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum, Trichoderma viride. S/2 concentration of Pochonia chlamydosporia, Purpureocillium lilacinum causing 100% inhibition in juvenile hatching while Pseudomonas fluorescens and Trichoderma viride causing 95.36% and 99.64% inhibition at the same. At S/10 concentration the corresponding values of larval emergence were P. chlamydosporia (82.14 %), P. fluorescens (71.79 %), P. lilacinum (79.46 %), and T. viride (66.07 %) respectively.
The inhibition range was least in S/100 concentration of cultural filtrate viz., Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum, Trichoderma viride were 35.71, 28.75, 31.79, 24.82 % respectively (Table-4, Figure- 3).
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Experimental Results
4.2.4. Effect of aqueous dilutions of biochar and water hyacinth extract on hatching of Meloidogyne incognita in vitro
The experiment was conducted to evaluate the antinemic potential of biochar and water hyacinth extract on larval hatching of second stage juvenile of Meloidogyne incognita. Both the tested agents results in inhibitory response to the larval emergence. However inhibitory effect of both biochar and water hyacinth extract varied with the concentration (Table-5, Figure-4).
S concentration of water hyacinth extract causes complete inhibition in egghatching and biochar showed 96.64% inhibition in juvenile hatching. However the observed percent inhibition in S/2, S/10 concentration was 95.14 % and 57 %, 82.06 % and 45.05 % respectively. The inhibition range was least in S/100 concentration of both water hyacinth extract and biochar were 22.05% and 11.21% respectively (Table-5, Figure-4).
4.3.1. Effect of aqueous extracts of fresh chopped leaves of some plant species on the mortality of root knot nematode Meloidogyne incognita in vitro
The experiment was performed to test the nematicidal behaviour of chopped leaf extract of some plants viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed against second stage juvenile of Meloidogyne incognita in vitro. The results present in the table-6 clearly revealed that leaf extract, of all the tested plants cause toxicity to the nematode up to a varying extent. However, the leaf extract of Mexican poppy and Trailing eclipta was found to be highly toxic to the juveniles, pursued by the aqueous extract of Wild eggplant and Black pig weed respectively. Mortality was found to be proportional to the concentration of the extract and exposure period (Table-6, Figure-5).
The results present in the table-6 clearly depicted that the aqueous leaf extracts of Mexican poppy results in juveniles percent mortality of 92, 87, 63, 18 in S, S/2, S/10, and S/100 concentrations as compared to 0 in Distilled Water control (DW) after 12hours of exposure period. In the leaf extract of Trailing eclipta the percent mortality were 89, 82, 58, 11, and 0; in the leaf extract of Wild eggplant were 84, 78, 56, 13, and 0; in Black pig weed were 80, 75, 51, 9, 0; in the leaf extract of Indian mellow were 72, 60, 45, 7,0. While the least mortality was observed in the aqueous
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Experimental Results extract of leaf of Ivy gourd were 65, 54, 37, 5, and 0 as compared to the 0 in Distilled Water control.
The highest percent mortality at 24 hours of the exposure in the aqueous extract of Mexican poppy was 100, 94, 70 15 and 0 followed by Trailing eclipta it was observed 100, 91, 64, 13, and 0; followed by the leaf extract of Wild eggplant were 97, 87, 63, 14 and 0 followed by Black pig weed were 92, 84, 58, 11, 0 followed by in the leaf extract of Indian mellow were 88, 72, 51, 9, 0. While the least mortality was observed in the aqueous extract of leaf of Ivy gourd were 71, 62, 45, 8 and 0 as compared to the 0 in Distilled Water control. (Table-6,Figure-5).
The percent mortality at 48 hour of exposure was found maximum in Mexican poppy were 100, 100, 74 13, 0 followed by Trailing eclipta were 100, 98, 69, 15, and 0 followed by the leaf extract of Wild eggplant were 100, 93, 67, 14, 0 followed by Black pig weed were 100, 91, 63, 13, 0 followed by the leaf extract of Indian mellow were 94, 85, 57, 12, 0. While the least mortality was observed in the aqueous extract of leaf of Ivy gourd were 80, 70, 52, 10, 0 as compared to the 0 in Distilled Water control (Table-6, Figure-5).
4.3.2. Effect of aqueous extracts of various oil cakes on the mortality of root knot nematode, Meloidogyne incognita in vitro
The experiment was carried out with the aim to examine the nematicidal potential of aqueous extracts of oil cakes on juveniles mortality of root knot nematode Meloidogyne incognita in vitro. Data presented in the table-7 indicate that extract of oil cakes viz., castor cake, cotton cake, mahua cake, mustard cake, soybean cake were proved to be effective against root knot nematode up to varying degree. Among all the tested oil cakes castor cake was found to be most potent in increasing mortality while cotton cake was least toxic to the juveniles of Meloidogyne incognita (Table-7, Figure-6).
Toxicity to the juvenile increase with increase in the concentration of aqueous extract of oil cake and dip duration. From the data presented in the table-7 it was revealed that maximum percent toxicity at 12 hours of the exposure in S, S/2, S/10, S/100 concentration in castor cake were 85,79,58,16; extract of mustard cakes were 82, 77, 53, 14; the aqueous extract of soybean cake were 76,72,48,10; Mahua cake
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Experimental Results
72, 67, 51, 8. While the minimum percent mortality was observed in cotton cake were 70, 65, 46, 6 as compared to distilled water control (0)
The average percent mortality at 24 hours of the duration in S, S/2, S/10, S/100 concentrations of castor cake were 96, 90, 66, 18; aqueous extract of mustard cakes were 90, 88, 61, 15; aqueous extract of soybean cake were 87, 79, 59, 13; Mahua cake were 82, 70, 61, 12. Lowest mortality observed in cotton cake was 76, 72, 54, 8 as compared to (0) in distilled water control.
The highest percent mortality at 48 hour of the exposure were 100, 95, 71, 20 in S, S/2, S/10, S/100 concentration of castor cake extract. It was followed by 100, 93, 68, 18 in mustard cake extract followed by 94, 88, 64, 16 in soybean cake extract followed by 91, 83, 66, 14, in mahua cake. Lowest was observed in cotton cake were 89, 80, 60, and 11 as compared to 0 in distilled water control (Table- 7, Figure-6).
4.3.3. Evaluation of nematotoxic potential of culture filtrates of biocontrol agents on the mortality of root knot nematode, Meloidogyne incognita in vitro
The experiment was conducted with the purpose to evaluate the nematicidal efficacy of cultural filtrate of Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum, Trichoderma viride on juveniles mortality of root knot nematode, Meloidogyne incognita in vitro. Results present in the table-8 indicate that cultural filtrate of all the screened bioagents were proved to be toxic against root knot nematode up to variable degree. However among all applied culture filtrate P. chlamydosporia, results in paramount toxicity to juveniles while Trichoderma viride recorded least toxicity. Mortality of the juveniles was found to be directly concerned with concentration of cultural filtrate and dip duration of the juveniles (Table-8, Figure-7).
S concentration of the culture filtrates of all the tested bioagents results in 100% mortality at 24 and 48 hours of the exposure except Trichoderma viride (96%) at 24 hours. At 12 hours of the duration the average percent mortality in S/2, S/10, S/100 concentrations of Pochonia chlamydosporia were 88, 65 and 18. It was followed by P. lilacinum 85, 60, 12; P. fluorescens 82, 55, 14; T. viride 80, 52, 11 as compared to (0) Distilled water control.
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Experimental Results
The percent mortality for 24 hour of duration of P. chlamydosporia in S/2, S/10, S/100 were 96, 75, 23; P. lilacinum 92, 69, 14 followed by P. fluorescens 91, 64, 18 while the least was observed in T. viride 88, 65, 15 as compared to (0) Distilled water control.
At 48 hour of the duration in S/2, S/10, S/100 concentration of P. chlamydosporia the percent mortality were 100, 83, and 26. It was followed by P. lilacinum 100, 78, 18; P. fluorescens 99, 75, and 24. However T. viride caused least mortality were 96, 71, 17 as compared to (0) distilled water control (Table-8, Figure- 7).
4.3.4. Evaluation of antinemic efficacy of aqueous extracts of biochar and water hyacinth extract on the mortality of root knot nematode, Meloidogyne incognita in vitro
This experiment deals to test the nematicidal efficacy of biochar and water hyacinth extracts against second stage juveniles on mortality of root knot nematode, Meloidogyne incognita in vitro. Both the tested agents biochar and water hyacinth extract cause mortality at all the dilutions of the extract. Mortality of the juveniles varied with the dilutions and time exposure (Table-9, Figure-8).
The S concentration of the extract of water hyacinth and biochar results in 100% mortality at 48 hour of exposure. Water hyacinth cause 92, 83, 50, 7 percent mortality in S, S/2, S/10, S/100 concentrations at 12 hours of the duration. While at the same duration the mortality of juveniles in presence of biochar at S/2, S/10, and S/100 concentration were 91, 66, 41, and 4 respectively.
At 24 hours of the duration in S/2, S/10, S/100 concentrations of water hyacinth extract the percent mortality of juveniles were 100, 91, 64, 11.While at the same the mortality found in biochar extract were 96, 78, 52, 7. However highest mortality was observed at 48 hours of the exposure of juveniles in S/2, S/10, S/100 concentrations of water hyacinth extract were 96, 71, 14 but at the same duration and concentrations the percent mortality in biochar were 90,60,10 as compared to 0 in distilled water control (Table-9, Figure-8).
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Experimental Results
4.4.1. Effect of bare-root dip in various plant extracts on the penetration of second stage juveniles (J2) of Meloidogyne incognita into the roots of tomato cv. K-21
The present experiment of root dipping was carried out with the aim to test the efficacy of aqueous extracts of various plants viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed against the second stage juvenile of Meloidogyne incognita penetration into the root of tomato cv.K-21 at two different time intervals (Table-10).
All the plant extracts inhibited the juvenile emergence into the roots of the tested plants. Penetration of juveniles significantly decreased with increase in the dip duration and differentially varied from treatment to the treatments as compared to the undipped inoculated control (Table-10).
Results presented in the table-10 clearly revealed that dip duration at (50 and 100 min) significantly reduced the juvenile penetration in the roots of tomato cv.K-21. When the tomato seedling inoculated with 1500 juveniles at 100 minutes of dip duration the number of juveniles penetrated in aqueous extract of Ivy gourd was 661; in Indian mallow was (616); in Black pig weed was (580); in Trailing eclipta was (538); in Wild eggplant was (508) and Mexican poppy was (472) respectively as compared to undipped inoculated control was 1356 and after the 50 minutes of the dip duration the corresponding values was 770, 740, 715, 689, 660, 626 in Ivy gourd, in Indian mallow, Black pig weed, Trailing eclipta, Wild eggplant, and Mexican poppy respectively.
The percent inhibition of juvenile penetration at 100 minutes of duration in aqueous extract of Ivy gourd was 51.5%; in Indian mallow was 54.5; in Black pig weed was 57.0%; in Trailing eclipta was 60.32%; in Wild eggplant was 62.53% and Mexican poppy was 65.19% respectively and after 50 minutes of the dip duration the percent inhibition in juveniles penetration was 43.21, 45.42, 47.2, 49.18, 51.32, 53.38% in Ivy gourd, in Indian mallow, Black pig weed, Trailing eclipta, Wild eggplant, and Mexican poppy respectively (Table-10).
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Table10: Effect of bare root dip in the leaf extract of different plant species on the penetration of root knot nematode, juveniles in the roots of tomato cv. K-21 Dip treatment Dip duration(min) Number of juveniles penetrated/plant %inhibition in penetration over control Indian mallow 50 740c 45.42 100 616g 54.57 Mexican poppy 50 626g 53.38 100 472k 65.19 Ivy gourd 50 770b 43.21 100 661f 51.25 Trailing eclipta 50 689e 49.18 100 538i 60.32 Wild eggplant 50 660f 51.32 100 508j 62.53 Black pig weed 50 715d 47.21 100 580h 57.22 Undipped Inoculated Control - 1356a - Each value is the mean of four replicates
Initial inoculum = 1500 (J2) of Meloidogyne incognita
Experimental Results
4.4.2. Effect of bare-root dip in various extract of oil cakes on the penetration of second stage juveniles (J2) of Meloidogyne incognita into the roots of tomato cv. K-21
This experiment was conducted on the same lines as in 4.4.1. This experiment was conducted to test the efficacy of the extracts of oil cake viz., castor, cotton, mahua, mustard and soybean against the penetration of Meloidogyne incognita juveniles in the root of tomato cv.K-21 inoculated with 1500 juveniles for the dip duration of 50 minutes and 100minutes. All the treatment results in significant suppression in the penetration of juvenile in the root of tomato at both of the durations (Table-11).
Cotton cake was found to be least effective with reference to the maximum penetration of second stage juveniles in the root of tomato at 100 minutes of exposure 650. It was followed by 616, 560, 520 and 500 in mahua, soybean, mustard and castor respectively as compared to the undipped inoculated control. However at 50 minutes of dip duration the penetrated juveniles were 775, 742, 712, 680 and 656.
Percent inhibition of second stage juvenile of M.incognita directly depends upon dip duration and varied with concentration of the oil cakes extract. All the treatment were found to be significantly effective in inhibiting the juvenile penetration in the root of tomato. Maximum percent inhibition was observed in castor oil cake (63.12%) at 100 minutes of dip duration. It was followed by 61.65, 58.70, 54.57 and 52.06% mustard, soybean, mahua and cotton cake extracts respectively. While the corresponding values of percent inhibition of juveniles in tomato root at 50 minutes of duration were (51.7, 49.85, 47.49, 45.28, and 42.84 %) (Table-11).
4.4.3. Effect of bare-root dip in various extracts of biocontrol agents on the penetration of second stage juveniles (J2) of Meloidogyne incognita into the roots of tomato cv. K-21
The experiment was conducted on the same procedure as given in 4.4.1. Form the tabulated data it was observed that root dipped seedling of tomato in various bioagents viz., Trichoderma viride, Purpureocillium lilacinum, Pseudomonas fluorescens, and Pochonia chlamydosporia for 100 minutes and inoculated with 1500 juvenile, recorded 528, 482, 452, 400 juveniles penetration in tomato root.
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Table 11: Effect of bare root dip in the extract of oil cake on the penetration of root knot nematode juveniles in the roots of tomato cv. K-21 Dip treatment Dip duration(min) Number of juveniles penetrated/plant %inhibition in penetration over control Castor cake 50 656f 51.7 100 500i 63.12 Cotton cake 50 775b 42.84 100 650f 52.06 Mahua cake 50 742c 45.28 100 616g 54.57 Mustard cake 50 680e 49.85 100 520i 61.65 Soybean cake 50 712d 47.49 100 560h 58.70 Undipped Inoculated Control - 1356a - Each value is the mean of four replicates
Initial inoculum = 1500 (J2) of Meloidogyne incognita
Table 12: Effect of bare root dip in the extract of bioagents on the penetration of root knot nematode juveniles in the roots of tomato cv. K-21 Dip treatment Dip duration Number of juveniles/plant %inhibition in penetration over control Purpureocillium lilacinum 50 681c 49.77 100 482g 64.45 Pochonia chlamydosporia 50 580e 57.22 100 400i 70.50 Pseudomonas fluorescens 50 638d 52.94 100 452h 66.66 Trichoderma viride 50 720b 46.90 100 528f 61.06 Undipped Inoculated Control - 1356a - Each value is the mean of four replicates
Initial inoculum = 1500 (J2) of Meloidogyne incognita
Experimental Results
However at 50 minutes of duration caused 720, 681, 638, 580 juveniles penetration in tomato root (Table-12).
Fungal spores of Pochonia chlamydosporia cause maximum percent inhibition of second stage juvenile of M. incognita in tomato at 100 minutes duration was 70.5% it was followed by 66.66, 64.45, 61.06% in Pseudomonas fluorescens, Purpureocillium lilacinum and Trichoderma viride. While at 50 minute of dip duration the percent inhibition was 57.22, 52.94, 49.77 and 46.90 respectively (Table- 12).
4.4.4. Effect of bare-root dip in various aqueous dilutions of biochar on the penetration of second stage juveniles (J2) of Meloidogyne incognita into the roots of tomato cv. K-21
This experiment was carried out on the same procedure as given in 4.4.1. This experiment was conducted to test the potential of biochar at various dilutions of 0.4, 0.8, 1.6, 3.2, and 5.0% against M incognita juveniles penetration on tomato cv.K-21 inoculated with 1500 juveniles. All the treatment significantly reduced the juvenile penetration in the root of tomato. Minimum number of juveniles penetration at 100 minutes of dip duration was observed in 5.0% of biochar (692). It was followed by 713, 773, 850, 998 in 3.2, 1.6, 0.8, 0.4 %biochar respectively (Table-13).
M. incognita juveniles penetration was found to be directly proportional to the concentration and exposure period. Biochar at various concentrations significantly reduced the penetration of juveniles in the root of tomato. Biochar at 5.0 % shows highest 49.11 % penetration inhibition at 100 minute of exposure. It was followed by 47.57, 43.61, 37.5, 26.6 % in 3.2, 1.6, 0.8, 0.4 % biochar respectively. The corresponding figure for the same concentration at 50 minute of duration was 36.57, 32.35, 26.47, 23.01, and 19.11 respectively (Table-13).
4.5.1. Effect of bare-root dip with various aqueous dilutions of Wild eggplant leaf extracts on root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots
Organic amendment in the form of plant products and plant parts offers safer and reliable approach for the management of nematode and improvement in plant growth parameters. Organic soil amendment with fresh chopped leaves was found
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Table13: Effect of bare root dip in biochar extract on the penetration of root knot nematode juveniles in the roots of tomato cv. K-21
Dip treatment Dip duration(min) Number of juvenile penetrated/plant %inhibition in penetration over control .4%biochar 50 1100b 19.11 100 998d 26.6 .8%biochar 50 1047c 23.01 100 850f 37.5 1.6%biochar 50 1000d 26.47 100 773g 43.61 3.2%biochar 50 920e 32.35 100 713h 47.57 5.0%biochar 50 860f 36.67 100 692h 49.11 Undipped Inoculated Control - 1360a - Each value is the mean of four replicates
Initial inoculum = 1500 (J2) of Meloidogyne incognita
Experimental Results beneficial for nematode suppression. Thus the aim of the present study was to test whether the bare root in the extract of Wild eggplant will be advantageous in managing the root-knot nematode infestation caused by Meloidogyne incognita (Table-14).
The bare-root dip treatment of tomato cv. K-21 seedlings in the leaf extract S, S/2, S/10 concentrations of Wild eggplant significantly reduced the nematode damage in tomato plants by reducing the root knot development. Root-knot indices were gradually decreased with increase in dip duration and concentration of the leaf extracts of Wild eggplant (Table-14).
The inhibition of root knot nematode development was more pronounced in S concentration of the leaf extract at 90 minute of exposure. The growth of the plants was significantly improved with increase in the concentration of the extract. The length of tomato K-21 was 46.7, 44.2 and 41.4 cm respectively when the roots of the plant were dipped in Wild eggplant leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita/pot. The similar trending figures for root dipping were 45.4, 42.8 and 40.0 cm at 60 minutes of the duration and as against 36.5 cm in undipped inoculated and 78.2 cm in undipped uninoculated control (Table-14).
The fresh weight of tomato K-21 was observed 35.5, 34.2 and 31.3 g respectively when the roots of the plant were dipped in Wild eggplant leaf extract of S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 34.4, 32.5 and 30.2g at 60 minutes of the duration and as against 28.0g in undipped inoculated and 72.0g in undipped uninoculated control (Table-14).
The total chlorophyll content of tomato cv. K-21 was observed 1.65, 1.57 and 1.51 mg/g respectively when the roots of the plant were dipped in Wild eggplant leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 1.60, 1.54 and 1.49 mg/g for 60 minutes of the duration and as against 1.12 mg/g in undipped inoculated and 2.78 mg/g in undipped uninoculated controls (Table-14).
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Table 14: Effect of bare-root dip in leaf extracts of Wild eggplant on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Pollen Root Duration Concentration Shoot Root Total Shoot Root Total content Content Eggmasses/root fertility knot (minutes) (mg/g) (mg/g) (%) Index S 31.1b 14.3bc 45.4b 24.4bc 10.0 b 34.4b 1.60 c 0.400c 129de 61.3b 2.8e 45 S/2 29.2c 13.6bcd 42.8cd 23.3cd 9.2bcd 32.5bc 1.54 de 0.380 d 134bc 57.7cd 3.4cd S/10 26.9d 12.5de 40.0e 21.8e 8.4 e 30.2d 1.49 f 0.350 f 137 b 54.5 e 3.9b S 32.0 b 14.7b 46.7b 25.2b 10.3 b 35.5b 1.65 b 0.415 b 127 e 62.6 b 2.3f 90 S/2 30.2bc 14.0bc 44.2bc 24.5bc 9.7cde 34.2b 1.57cd 0.392 c 132cd 59.4 c 3.2de S/10 28.4cd 13.0cde 41.4de 22.6de 8.7 de 31.3cd 1.51 f 0.368 e 135bc 56.0 de 3.7bc Undipped Uninoculated Control 53.6 a 25.4a 78.2a 52.0 a 20.0 a 72.0 a 2.78 a 0.880 a 0 f 89.5 a 0 g Undipped Inoculated Control 24.7 e 11.8e 36.5f 20.0f 8.0 f 28.0e 1.12 0.252g 175 a 47.0 f 5.0 a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
The total carotenoid content of tomato cv. K-21 was observed 0.415, 0.392 and 0.368 mg/g respectively when the roots of the plant were dipped in Wild eggplant leaf extract of S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 0.400, 0.380 and 0.350 mg/g for 45 minutes of the duration and as against 0.252 mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated controls (Table-14)
Number of eggmasses/root was significantly reduced in all the dilution of the extracts. The total number of eggmasses/root of tomato cv. K-21 were 127, 132 and 135 respectively when the roots of the plant were dipped Wild eggplant leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 129, 134 and 137 for 45 minutes of the duration and as against 175 in undipped inoculated and 0 in undipped uninoculated controls (Table-14)
The percent pollen fertility of tomato cv.K-21 was observed 62.6, 59.4 and 56.0 % respectively when the roots of the plant were dipped in Wild eggplant leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 61.3, 57.7 and 54.5 % for 60 minutes of the duration and as against 47.0 % in undipped inoculated and 89.5% in undipped uninoculated controls (Table-14)
The root-knot indices was significantly reduced to 2.3, 3.2 and 3.7 respectively when the roots of the plant were dipped in Wild eggplant leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 2.8, 3.4 and 3.9 for 60 minutes of the duration and as against 5.0 in undipped inoculated control (Table-14).
4.5.2. Effect of bare-root dip with various aqueous dilutions of Trailing eclipta leaf extracts on root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots
This experiment was conducted on the same procedure as in 4.1.9. to evaluate the effect of leaf extract of Trailing eclipta as bare-root dip treatment on tomato cv. K- 21 against the root-knot development caused by M. incognita. The inhibition of root
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Experimental Results knot nematode development was more outstanding in S concentration of the leaf extract at 90 minutes of exposure. All the dilutions of the extracts significantly improved the plant growth parameters with increase in the concentration of the extract (Table-15).
The length of tomato cv.K-21 was 48.0, 44.5 and 41.7 cm respectively when the roots of the plant were dipped for 90 minutes in S, S/2 and S/10 concentrations of leaf extracts of Trailing eclipta and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 45.9, 43.5 and 40.4 cm at 60 minutes of the duration and as against 36.5cm in undipped inoculated and 78.2 cm in undipped uninoculated control.
The fresh weight of tomato cv. K-21 was observed 37.5, 35.8 and 32.2 g respectively when the roots of the plant were dipped in Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 36.0, 34.2 and 31.0 g at 45 minutes of the duration and as against 28.0g in undipped inoculated and 72.0g in undipped uninoculated control (Table-15)
The total chlorophyll content of tomato cv. K-21 was observed 1.69, 1.61 and 1.55 mg/g respectively when the roots of the plant were dipped Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 1.65, 1.58 and 1.52 mg/g for 60 minutes of the duration and as against 1.12 mg/g in undipped inoculated and 2.78 mg/g in undipped uninoculated controls (Table-15).
The total carotenoid content of tomato cv. K-21 was observed 0.435, 0.408 and 0.388 mg/g respectively when the roots of the plant were dipped Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 0.422, 0.397 and 0.378 mg/g for 45 minutes of the duration and as against 0.252 mg/g in undipped inoculated and .880 mg/g in undipped uninoculated controls (Table- 15).
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Table 15: Effect of bare-root dip in leaf extracts of Trailing eclipta on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggma Pollen Root Duration Concentration Shoot Root Total Shoot Root Total content content sses/root fertility Knot (minutes) (mg/g) (mg/g) (%) index 45 S 30.7 c 15.2bc 45.9bc 26.0bc 10.2b 36.2bc 1.65bc 0.422c 126de 62.7bc 2.7d S/2 29.4cd 14.1cd 43.5cd 24.6cd 9.6bcd 34.2cd 1.58 de 0.397e 130cd 60.2cd 3.3bc S/10 27.6 e 12.8d 40.4e 22.7 f 8.7d 31.4e 1.52f 0.378g 135b 57.0e 3.7b 90 S 32.4 b 15.6b 48.0b 26.9b 10.6b 37.5b 1.69b 0.435b 123e 64.2 b 2.2e S/2 30.0cd 14.7bc 44.7c 25.8bc 10.0bc 35.8bc 1.61cd 0.408d 128cd 61.0cd 3.1cd S/10 28.5de 13.0d 41.5de 23.2de 9.0cd 32.2de 1.55ef 0.388f 132bc 58.4de 3.5bc Undipped 53.6a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a 0.880a 0f 89.5a 0f Uninoculated Control Undipped Inoculated Control 24.7f 11.8e 36.5f 20.0g 8.0e 28.0 f 1.12g 0.252h 175a 47.0f 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
Number of eggmasses/root was significantly reduced in all the dilutions of the extracts with increase in the concentration. The total number of eggmasses /root of tomato cv. K-21 were 123, 128 and 132 respectively when the roots of the plant were dipped Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 126, 130 and 135 for 60 minutes of the duration and as against 175 in undipped inoculated controls (Table-15)
The percent pollen fertility of tomato cv. K-21 was observed 64.2, 61.0 and 58.4 % respectively when the roots of the plant were dipped in Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 62.7, 60.2 and 57.0 % for 45 minutes of the duration and as against 47.0 % in undipped inoculated and 89.5% in undipped uninoculated controls (Table-15)
The root-knot indices was significantly reduced to 2.2, 3.1 and 3.5 respectively when the roots of the plant were dipped in Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 2.7, 3.3 and 3.7 for 45 min of the duration and as against 5.0 in undipped inoculated control (Table-15)
The root-knot indices was significantly reduced to 2.1, 2.5 and 3.0 respectively when the roots of the plant were dipped Trailing eclipta leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 2.3, 2.8 and 3.5 for 60 min of the duration and as against 5.0 in undipped inoculated control (Table-15).
4.5.3. Effect of bare-root dip with various aqueous dilutions of Mexican poppy leaf extracts on root knot development caused by Meloidogyne incognita and plant growth character of tomato cv. K-21 in pots
This experiment followed the same procedure as in 4.1.9, to elucidate the effect of leaf extract of Mexican poppy as bare-root dip treatment on tomato cv. K-21 against the root-knot development caused by M. incognita. The inhibition of root knot
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Table 16: Effect of bare-root dip in leaf extracts of Mexican poppy on growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmasses/root Pollen Root Duration Concentration Shoot Root Total Shoot Root Total content content fertility Knot (minute) (mg/g) (mg/g) (%) Index 45 S 33.2c 16.2bc 49.4bc 27.3b 10.7bc 37.8b 1.67bc 0.452c 121de 64.7bc 2.3e S/2 31.2de 15.4 cd 46.6de 25.2cd 9.8bcd 35.0bc 1.61de 0.438d 126bc 62.1de 2.8cd S/10 29.6 e 14.6d 44.2e 24.1d 9.2d 33.3d 1.55f 0.422e 130b 59.0f 3.5b 90 S 35.0 b 16.6b 51.6b 27.9b 11.0b 38.9b 1.71b 0.460b 117 e 66.2b 2.1e S/2 32.3cd 15.7bcd 48.0cd 26.3bc 10.3bcd 36.6bc 1.64cd 0.446c 123cd 63.4cd 2.5de S/10 30.0 e 15.0d 45.0e 24.7cd 9.5cd 34.2d 1.58ef 0.426e 128b 60.6ef 3.0c Undipped Uninoculated Control 53.6a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a 0.880a 0f 89.5a 0f Undipped Inoculated Control 24.7 f 11.8e 36.5f 20.0e 8.0e 28.0e 1.12g 0.252f 175a 47.0g 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results nematode development was more outstanding in S concentration of the leaf extract at 90 minutes of exposure. All the dilutions of the extracts significantly improved the plant growth parameters with increase in the concentration of the extract (Table-16)
The length of tomato K-21 was 51.6, 48.0 and 45.0 cm respectively when the roots of the plant were dipped for 90 minutes in S, S/2 and S/10 concentrations of leaf extracts of Mexican poppy and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 49.0, 46.6 and 44.2 cm at 60 minutes of the duration and as against 36.5 cm in undipped inoculated and 78.2 cm in undipped uninoculated control (Table-16).
The fresh weight of tomato K-21 was observed 38.9, 36.6 and 34.2 g respectively when the roots of the plant were dipped in Mexican poppy leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 37.8, 35.0 and 33.3 g at 60 minutes of the duration and as against 28.0 g in undipped inoculated and 72.0 g in undipped uninoculated control (Table-16).
The total chlorophyll content of tomato cv. K-21 was observed 1.71, 1.64 and 1.58 mg/g respectively when the roots of the plant were dipped Mexican poppy leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 1.67, 1.61 and 1.55 mg/g for 60 minutes of the duration and as against 1.12 mg/g in undipped inoculated and 2.78 mg/g in undipped uninoculated controls (Table-16).
The total carotenoid content of tomato cv. K-21 was observed 0.460, 0.446 and 0.426 mg/g respectively when the roots of the plant were dipped Mexican poppy leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 0.452, 0.438 and 0.422 mg/g for 60 minutes of the duration and as against 0.252 mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated controls (Table-16)
Number of eggmasses/root was significantly reduced in all the dilutions of the extracts with increase in the concentration. The total number of eggmasses /root of tomato cv. K-21 were 117, 123 and 128 respectively when the roots of the plant were
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Experimental Results dipped in A. mexicana leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 121, 126 and 130 for 60 minutes of the duration and as against 175 in undipped inoculated and 0 in undipped uninoculated controls (Table- 16).
The percent pollen fertility of tomato cv. K-21 was observed 66.2, 63.4 and 60.6% respectively when the roots of the plant were dipped in Mexican poppy leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 64.7, 62.1 and 59.0 % for 60 minutes of the duration and as against 47.0 % in undipped inoculated and 89.5% in undipped uninoculated controls (Table-16).
All the treatment significantly inhibited the root-knot indices. The root-knot indices was significantly reduced to 2.1, 2.5 and 3.0 respectively when the roots of the plant were dipped in Mexican poppy leaf extract in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for dipping were 2.3, 2.8 and 3.5 for 60 min of the duration and as against 5.0 in undipped inoculated control (Table-16).
4.5.4 Effect of bare-root dip with different aqueous dilutions of fungal spores of Purpureocillium lilacinum on root knot development caused by Meloidogyne incognita and plant growth character of tomato cv. K-21 in pots
Nematode management through the involvement of bioagents is considered as one of the safer and effective methods. Thus with this aim the present experiment was assessed to investigate the nematicidal potential of fungal spores of Purpureocillium lilacinum as bare root dip treatment against M incognita in tomato cv. K-21 (Table- 17).
Aqueous dilution of fungal spore of P. lilacinum in S, S/2, S/10 concentrations as bare root dip treatment of tomato seedling significantly reduced the nematode infestation in tomato by suppressing the root knot development. With increase in the concentration of aqueous dilutions of P. lilacinum and dip duration significantly reduced the root-knot indices (Table-17).
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Experimental Results
The S concentration of the fungal spores of P. lilacinum shows most remarkable inhibition in root knot nematode development at 90 min of the exposure period. Aqueous dilution of all the fungal spores significantly ameliorates the plant growth characters with increase in the concentration. However the highest plant length (52.7cm) of tomato cv.K-21 was recorded in S concentration of the fungal spores of P. lilacinum at 90 min of the exposure, with the inoculation of 1500 second stage juveniles of M. incognita. It was followed by 49.7, 47 .2 in S/2, S/10 of the fungal spores. Similar trends of figures for dipping at 60 minutes duration were 49.0, 46.6 and 44.2 cm as compared to 36.5 cm in undipped inoculated and 78.2 cm in undipped uninoculated control. The increase in plant growth was most prominent in S concentration of the fungal spore (Table-17).
Total fresh weight of the plant was significantly increased with increase in the concentration of fungal spores and dip duration. S concentration of the fungal spores of P. lilacinum showed most promising increment (40.2 g) in the fresh weight of tomato cv. K-21 at 90 min of dip duration and 1500 juveniles inoculation. It was followed by 38.2, 35.9 g at S/2, S/10 concentration of the fungal spores. The similar trend for improvement in growth parameter at 60 min of dip duration were 39.0, 37.2 and 34.8 g respectively as compare to 28 g in undipped inoculated and 72 g in undipped uninoculated controls (Table-17).
All the dilutions of the fungal spores significantly enhanced the chlorophyll content. Although highest chlorophyll content (1.73 mg/g) in tomato cv.K-21 was detected in S concentration of aqueous dilution of fungal spores P. lilacinum at 90 min of dip duration after inoculating 1500 juveniles. It was followed in descending order of 1.67, 1.61 mg/g at S/2 and S/10 concentration of aqueous dilution of fungal spores. The corresponding figures for the chlorophyll at 60 min of root dip duration were 1.69, 1.64 and 1.58 mg/g respectively as compare to 1.12 mg/g in undipped inoculated and 2.78 mg/g in undipped uninoculated controls (Table-17).
The carotenoid content of tomato cv. K-21 was significantly observed were .486, .472 and .458 mg/g respectively when the fungal spore of P. lilacinum in S, S/2 and S/10 concentrations for 90 min were used as root dip treatment and inoculated with 1500 second stage juveniles of M. incognita. The similar trends for root dipping were
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Table 17: Effect of bare-root dip in fungal spores of Purpureocillium lilacinum on the growth of tomato cv. K-21 in relation to root- knot development caused by Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmasses/root Pollen Root content content fertility Knot Duration Concentration Shoot Root Total Shoot Root Total (mg/g) (mg/g) (%) Index (minutes) 45 S 34.6bc 16.9bc 51.5b 27.7bc 11.3b 39.0bc 1.69bc .479bc 119de 64.7bc 2.2de
S/2 33.0de 16.0cd 49.0c 26.7cd 10.5cd 37.2cde 1.64cd .466de 125bc 62.6c 2.8c
S/10 30.4f 15.0e 45.4e 25.0e 9.8d 34.8e 1.58e .450f 128b 60.5d 3.5b
90 S 35.4b 17.3b 52.7b 28.5b 11.7b 40.2b 1.73b .486b 115f 66.0b 2.0e
S/2 33.4cd 16.3cd 49.7c 27.2bc 11.0bc 38.2bcd 1.67c .472cd 121cd 63.8bc 2.5cd
S/10 31.8ef 15.4de 47.2d 25.7de 10.2d 35.9de 1.61de .458ef 126bc 61.6cd 3.2b
Undipped Uninoculated Control 53.6 a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a .880a 0g 89.5a 0f
Undipped Inoculated Control 24.7g 11.8f 36.5f 20.0f 8.0e 28.0f 1.12f .252g 175a 47.0e 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
0.479, 0.466 and 0.450 mg/g for 60 minutes of the duration and as compared to 0.252 mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated controls (Table-17).
Aqueous dilution of the fungal spores significantly reduced number of eggmasses/root. Number of eggmasses /root of tomato cv. K-21 dipped with P. lilacinum fungal spores having various dilutions S, S/2 and S/10 at 90 min were 115, 121 and 126 respectively when the root was inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for 60 min dipping were 119, 125 and 128 and as compared with 175 in undipped inoculated and 0 in undipped uninoculated controls (Table-17).
When the roots of the tomato plant cv.K-21 were dipped in fungal spores of P. lilacinum at S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita the observed percent pollen fertility were 66.0, 63.8 and 61.6 % respectively. The corresponding figures for 45 minutes dip duration were 64.7, 62.6 and 60.5 % and as against 47.0% in undipped inoculated and 89.5 % in undipped uninoculated controls (Table-17)
Aqueous dilutions and dip duration significantly reduced the root-knot indices in tomato cv.K-21.The root knot indices were reduced to 2.0 2.5 and 3.2 respectively when the P. lilacinum fungal spores were used as bare root dip in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures of 45 minutes of dipping were 2.0, 2.8 and 3.5 respectively and as against 5.0 in undipped inoculated control (Table-17).
4.5.5. Effect of bare-root dip with different aqueous dilutions of fungal spores of Pochonia chlamydosporia on root knot development caused by Meloidogyne incognita and plant growth character of tomato cv. K-21 in pots
This experiment was conducted on the same trends as in 4.5.4. to investigate the effect of fungal spores of Purpureocillium lilacinum as bare root dip treatment against M. incognita in tomato cv. K-21. Bare root dip treatment of tomato seedling cv.K-21 with aqueous dilution of fungal spore of Pochonia chlamydosporia (S, S/2 and S/10) significantly reduced the nematode infestation in tomato cv. K-21 by
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Experimental Results deflating the root-knot development. Dip duration and concentration of aqueous dilutions of P. chlamydosporia significantly reduced the root knot indices (Table-18).
S concentration of the fungal spores of P. chlamydosporia shows most notable inhibition in root knot nematode development at 90 min of the exposure period. With increase in the concentration of aqueous dilution of all the fungal spores significantly enhanced the plant growth parameters. However the highest plant length (54.0 cm) of tomato cv.K-21 was recorded in S concentration of the fungal spores of P.chlamydosporia at 90 min of the exposure, with the inoculation of 1500 second stage juveniles of M. incognita. It was followed by 51.8, 48.6 cm in S/2, S/10 of the fungal spores. Similar trends of figures for dipping at 45 minutes duration were 53.0, 51.0 and 46.0 cm as compared to 36.5 cm in undipped inoculated and 78.2 cm in undipped uninoculated control. The increase in plant growth was most prominent in S concentration of the fungal spore (Table-18).
Total fresh weight of the plant was significantly increased with increase in the concentration of fungal spore and dip duration. S concentration of the fungal spores of P.chlamydosporia shows most promising increment (41.8 g) in the fresh weight of tomato cv. K-21 at 90 min of dip duration and 1500 juveniles’ inoculation. It was followed by 38.8, 36.3 g at S/2, S/10 concentration of the fungal spores. The similar trend for improvement in growth parameter at 45 min of dip duration were 39.9, 36.6 and 35.0 g respectively as compare to 28g in undipped inoculated and 72g in undipped uninoculated controls (Table-18).
P. chlamydosporia fungal spores significantly improved the chlorophyll content. Although highest chlorophyll content (1.75 mg/g) in tomato cv.K-21 was detected in S concentration of aqueous dilution of fungal spores of P. chlamydosporia at 90 minutes of dip duration after inoculating 1500 juveniles. It was followed in descending order of 1.69, 1.65 mg/g at S/2 and S/10 concentration of aqueous dilution of fungal spores. The similar trending figures for the chlorophyll at 45 minutes of root dip duration were 1.71, 1.66 and 1.62 mg/g respectively as compare to 1.12 mg/g in undipped inoculated and 2.78 mg/g in undipped uninoculated controls (Table-18).
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Table 18: Effect of bare-root dip in fungal spores of Pochonia chlamydosporia on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmasse Pollen Root content content s/root fertility Knot Duration Concentration Shoot Root Total Shoot Root Total (mg/g) (mg/g) (%) index (minutes) 45 S 35.7bc 17.3b 53.0bc 28.4bc 11.5bc 39.9c 1.71bc .484c 116de 68.0bc 2.0e
S/2 34.3c 16.7bc 51.0d 26.9cd 10.7cd 37.6de 1.66def .469d 122bc 65.4cd 2.5cd
S/10 31.2e 15.2d 46.4f 25.0e 10.0d 35.0f 1.62f .456e 126b 62.0e 3.3b
90 S 36.4b 17.6b 54.0b 29.8b 12.0a 41.8b 1.75b .494b 112e 69.2b 1.9e
S/2 34.8c 17.0b 51.8cd 27.6c 11.2bcd 38.8cd 1.69cde .478c 119cd 66.5c 2.2de
S/10 32.8d 15.8cd 48.6e 26.0de 10.3cd 36.3ef 1.64ef .464d 124b 63.2de 2.8c
Undipped Uninoculated Control 53.6a 24.6a 78.2a 52.0a 20.0e 72.0a 2.78a .880a 0f 89.5a 0f
Undipped Inoculated Control 24.7f 11.8e 36.5g 20.0f 8.0e 28.0g 1.12g .252f 175a 47.0e 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
The total carotenoid content of tomato cv. K-21 was significantly observed were .494, .478 and .464 mg/g respectively when the fungal spore of P. chlamydosporia in S, S/2 and S/10 concentrations for 90 minutes were used as roots dip treatment and inoculated with 1500 second stage juveniles of M. incognita. The corresponding figures for root dipping were 0.484, 0.469 and 0.456 mg/g for 45 minutes of the duration and as compared to 0.252 mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated controls (Table-18).
Aqueous dilution of the fungal spores significantly reduced number of eggmasses/root. Number of eggmasses /root of tomato cv. K-21 dipped with P.chlamydosporia fungal spores having various dilutions S, S/2 and S/10 at 90 minutes were 112, 119 and 124 respectively when the root was inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for 45 minutes dipping were 116, 122 and 126 and as compared with 175 in undipped inoculated and 0 in undipped uninoculated controls (Table-18).
When the roots of the tomato plant cv.K-21 were dipped in fungal spores of P.chlamydosporia at S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita the observed percent pollen fertility were 69.2, 66.5 and 63.2 % respectively. The corresponding figures for 45 minutes dip duration were 68.0, 65.0 and 62.0 % and as against 47.0 % in undipped inoculated and 89.5 % in undipped uninoculated controls (Table-18).
Aqueous dilutions and dip duration significantly reduced the root knot indices in tomato cv.K-21.The root knot indices were reduced to 1.9 2.2 and 2.8 respectively when the P.chlamydosporia fungal spores were used as root bare root dip in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures of 45 minutes of dipping were 2.0, 2.5 and 3.3 respectively and as against 5.0 in undipped inoculated control (Table- 18).
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Experimental Results
4.5.6. Effect of bare-root dip with different aqueous dilutions of bacterial suspension of Pseudomonas fluorescens on root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots
This experiment was conducted on the same lines as given in 4.5.4. to examine the effect of bacterial suspension of Pseudomonas fluorescens as bare root dip treatment against M incognita in tomato cv. K-21. Bare root dip treatment of tomato seedling cv.K-21 with aqueous dilution of bacterial suspension of Pseudomonas fluorescens (S, S/2, and S/10) significantly reduced the nematode infestation in tomato cv. K-21 by reducing the root knot development. Dip duration and concentration of aqueous dilutions of P. fluorescens significantly reduced the root knot indices (Table-19).
The S concentration of the bacterial suspension of P. fluorescens showed most remarkable inhibition in root knot nematode development at 90 minutes of the exposure period. With increase in the concentration of aqueous dilution and dip duration of all bacterial suspension significantly enhanced the plant growth parameters. The highest plant length (55.6 cm) of tomato cv. K-21 was recorded in S concentration of the bacterial suspension of P. fluorescens at 90 minutes of the exposure, with the inoculation of 1500 second stage juveniles of M. incognita. It was followed by 52.7, 50.2 cm in S/2, S/10 of the bacterial suspension. The corresponding figures for dipping at 45 minutes duration were 53.8, 51.5 and 48.5 cm as compared to 36.5 cm in undipped inoculated and 78.2 cm in undipped uninoculated control. S concentration of the bacterial suspension showed pronounced improvement in plant growth (Table-19).
The S concentration of the bacterial suspension of P. fluorescens showed most promising improvement (43.6 g) in the fresh weight of tomato cv. K-21 at 90 minutes of dip duration and 1500 juveniles inoculation. It was followed by 41.0, 38.5 g at S/2, S/10 concentration of bacterial suspension. The similar trend for improvement in growth parameter at 45minutes of dip duration were 42.0, 39.6 and 36.8 g respectively as compare to 28 g in undipped inoculated and 72 g in undipped uninoculated controls (Table-19).
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Experimental Results
P. fluorescens bacterial suspension significantly improved the chlorophyll content. Although highest chlorophyll content (1.79 mg/g) in tomato cv.K-21 was detected in S concentration of aqueous dilutions of bacterial suspension of P.chlamydosporia at 90 min of dip duration after inoculating 1500 juveniles. It was followed in descending order of 1.71, 1.67 mg/g at S/2 and S/10 concentration of aqueous dilution of bacterial suspension. The corresponding figures for the chlorophyll were 1.74, 1.69 and 1.65 mg/g respectively at 45 minutes root dip duration as compare to 1.12 mg/g in undipped inoculated and 2.78 mg/g in undipped uninoculated controls.
The total carotenoid content of tomato cv. K-21 was significantly observed were; 0.500, 0.484 and 0.473 mg/g respectively when the bacterial suspension of P. fluorescens in S, S/2 and S/10 concentrations for 90 minutes were used as roots dip treatment and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for root dipping were 0.490, 0.478 and 0.468 mg/g for 45 minutes of the duration and as compared to 0.252mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated controls (Table-19).
Bacterial suspension significantly reduced number of eggmasses/root. Number of eggmasses /root of tomato cv. K-21 dipped with P. fluorescens suspension having various dilutions viz., S, S/2 and S/10 at 90 minutes were 111, 118 and 123 respectively when the root was inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures for 45 minutes dipping were 115, 121 and 125 and as compared with 175 in undipped inoculated and 0 in undipped uninoculated controls (Table-19).
When the roots of the tomato plant cv. K-21 were dipped in bacterial suspension of P. fluorescens at S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita the observed percent pollen fertility were 71.0, 68.0 and 65.0 % respectively. The corresponding figures for 45 minutes dip duration were 69.5, 66.6 and 64.0 % and as against 47.0 % in undipped inoculated and 89.5% in undipped uninoculated controls (Table-19).
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Table 19: Effect of bare-root dip in bacterial suspension of Pseudomonas fluorescens on the growth of tomato cv. K-21 in relation to root- knot development caused by Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmasss Pollen Root content content es/root fertility Knot Duration Concentration Shoot Root Total Shoot Root Total (mg/g) (mg/g) (%) Index (minutes) 45 S 35.2bcd 18.6b 53.8c 30.0bc 12.0bc 42.0c 1.74bc .490c 115de 69.5bc 1.9e
S/2 34.8cd 16.7d 51.5de 28.3de 11.0de 39.6d 1.69cd .478de 121bc 66.6cd 2.3d
S/10 32.8e 15.7d 48.5f 26.6f 10.2f 36.8e 1.65d .468f 125b 64.0d 3.2b
90 S 36.4b 19.2b 55.6b 31.2b 12.4b 43.6b 1.79b .500b 111e 71.0b 1.8e
S/2 35.5bc 17.2c 52.7cd 29.4cd 11.6cd 41.0c 1.71cd .484c 118cd 68.0cd 2.1de
S/10 34.0de 16.2d 50.2e 27.9e 10.6ef 38.5d 1.67d .473ef 123bc 65.0d 2.7c
Undipped Uninoculated Control 53.6a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a .880a 0g 89.5a 0f
Undipped Inoculated Control 24.7f 11.8e 36.5g 20.0g 8.0g 28.0f 1.12e .252g 175a 47.0e 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
Dip duration and concentration of bacterial suspension significantly reduced the root knot indices in tomato cv. K-21.The root knot indices were reduced to 1.8, 2.1 and 2.7 respectively when the P. fluorescens suspension were used as bare root dip in S, S/2 and S/10 concentrations for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita. The similar trending figures of 45 minutes of dipping were 1.9, 2.3 and 3.2 respectively and as against 5.0 in undipped inoculated control (Table-19).
4.5.7. Effect of bare root-dip treatment with various aqueous dilutions of Water hyacinth extract mixed with Purpureocillium lilacinum spores on root knot development caused by Meloidogyne incognita and plant growth character of tomato cv. K-21 in pots
Any of the single approach either through the involvement of bioagents or through the organic amendment are not self sufficient for the management of the nematode. So the management of nematode with the involvement of more than one technique can be done. Thus an experiment was conducted to elucidate the potential of bioagents in combination with plant extract as a bare root dip treatment against root knot nematode Meloidogyne incognita on tomato cv. K-21. The present experiment was conducted under glasshouse condition in autoclaved soil filled in the pots (Table- 20).
Combined application of both bioagents and plant extract significantly abate the nematode severity by reducing the root knot index. Number of eggmasses/root and root knot index was found to be directly concerned with the concentration of both and dip duration.
In combined treatment, Purpureocillium lilacinum along with extract of water hyacinth plant length of tomato were 58.8, 55.7, 52.5 cm respectively for 90 minutes of dip duration in S concentration of P. lilacinum + water hyacinth extract, S/2 concentration of P. lilacinum + water hyacinth extract, S/10 concentration of P. lilacinum + water hyacinth extract which were inoculated with 1500 second stage juveniles /pot. The trending figures for 45 minutes of dip duration were 57.0, 54.4 and 51 cm as compare to 36.5 cm in undipped inoculated and 78.2 in undipped uninoculated controls (Table-20).
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Experimental Results
The total weight of tomato cv.K-21 in S concentration of P. lilacinum + water hyacinth extract, S/2 concentration of P. lilacinum + water hyacinth extract S/10 concentration of P. lilacinum + water hyacinth extract were 45, 0, 42.5, 40.0 g when roots were dipped for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita/pot. The corresponding figures for 45 minutes of root dipping were 43.7, 41.3, 39.1 g respectively as compared to 28 g in undipped inoculated and 78.2 g in undipped uninoculated controls (Table-20).
In combined treatment, application of bioagents and plant extract significantly improved the chlorophyll content compare to inoculated control. The chlorophyll content of tomato cv.K-21 for 90 minutes of root dipping in S concentration of P. lilacinum + water hyacinth extract, S/2 concentration of P. lilacinum + water hyacinth extract, S/10 concentration of P. lilacinum + water hyacinth extract were 1.85, 1.76, 1,70 mg/g respectively with the inoculation of 1500 second stage juveniles of M. incognita /pot. The trending figures for dip duration of 45 minutes was 1.80, 1.73 and 1.67 mg/g as compared to 1.12 mg/g undipped inoculated and 2.78 mg/g in undipped uninoculated controls.
Combined treatment of both plant extract with the biocontrol agents significantly enhanced the carotenoid content. Although highest carotenoid content (0.561mg/g) in tomato cv.K-21 was detected in S concentration of P. lilacinum extract mixed with P. lilacinum for 90 minutes of dip duration after inoculating 1500 second juveniles/pot. It was followed in descending order of 0.546, 0.534 mg/g at S/2 concentarion of P. lilacinum extract+ P. lilacinum and S/10 concentration of P. lilacinum extract + P. lilacinum. The corresponding figures for the carotenoid were 0.553, 0.540 and 0.528 mg/g respectively for 45 minutes of root dipping as compared to 0.252mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated control (Table-20).
Combined application of both Water hyacinth extract mixed with Purpureocillium lilacinum spores was found to be potent in reducing the number of eggmasses/plant. Highest reduction in number of eggmasses/root (105) was observed when S concentration of water hyacinth extract mixed with P. lilacinum spores for 90 minutes of dip duration inoculating with 1500 second stage juveniles/pot. It was followed by 110, 114 in S/2 concentration of P. lilacinum + water hyacinth extract,
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Table 20: Effect of bare root- dip in aquatic weed Water hyacinth leaf extract mixed with Purpureocillium lilacinum spores on plant growth of tomato cv. K-21 infested with Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmas Pollen Root content content ses/root fertility Knot Duration Concentration Shoot Root Total Shoot Root Total (mg/g) (mg/g) (%) Index (minutes) 45 S 37.6bc 19.4b 57.0bc 31.2bc 12.5bc 43.7c 1.80bc 0.553c 108de 71.0b 1.8de
S/2 36.6c 17.8c 54.4de 29.6d 11.7cd 41.3d 1.73cde 0.540d 112bcd 68.2c 2.3d
S/10 34.5d 16.5d 51.0f 28.0e 11.1e 39.1e 1.67e 0.528e 116b 65.6d 3.1b
90 S 38.8b 20.0b 58.8b 32.0b 13.0b 45.0b 1.85b 0.561b 105e 72.2b 1.6e
S/2 37.5bc 18.2c 55.7cd 30.4cd 12.1bcd 42.5cd 1.76cd 0.548c 110cd 69.5bc 2.0e
S/10 35.3d 17.2cd 52.5ef 29.2d 11.8cd 40.0e 1.70de 0.534de 114bc 66.8cd 2.6c
Undipped Uninoculated Control 53.6a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a 0.880a 0f 89.5a 0f Undipped Inoculated Control 24.7e 11.8e 36.5g 20.0f 8.0f 28.0f 1.12f 0.252f 175a 47.0e 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
S/10 concentration of P. lilacinum + water hyacinth extract respectively. The numbers of trending figures for 45 minutes dip duration were 108, 112, 116 respective in combined concentration of both and as compared with 175 in undipped inoculated and 0 in undipped uninoculated controls (Table-20).
The percent pollen fertility of tomato cv.K-21 was significantly enhanced in all treatments. Combined application of S concentration of the P. lilacinum + water hyacinth extract was found highest in increasing percent pollen fertility (72.2 %) of tomato followed by S/2 concentration of P. lilacinum + water hyacinth extract (69.5 %), S/10 concentration of P. lilacinum + water hyacinth extract (66.8 %), respectively for the dip duration of 90 minutes when the root were inoculated with 1500 second stage juvenile of M. incognita /pot. The trending figures were 71.0, 68.2 and 62.6 % respective at 45 minutes dip duration as compared to 47 % in undipped inoculated and 89.5 % in undipped uninoculated controls (Table-20).
Application of aqueous extract of water hyacinth in with combination Purpureocillium lilacinum spores was found to be effective in reducing nematode severity and root knot indices. The root knot indices for the dip duration of 90 minutes were 1.6, 2.0 and 2.6 respectively in combined concentration of P. lilacinum (S) with water hyacinth extract, P. lilacinum (S/2) + water hyacinth extract, P. lilacinum (S/10) + water hyacinth extract inoculated 1500 second stage juvenile /pot. The corresponding values were 1.8, 2.3 and 3.1 respectively as compared to 5.0 in undipped inoculated control (Table-20).
4.5.8. Effect of bare root-dip treatment with various aqueous dilutions of Water hyacinth extract mixed with Pochonia chlamydosporia spores on root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots
This experiment was conducted on the same procedure as in 4.1.9. Combined application of both bioagents and plant extract significantly reduce the nematode damage by reducing the root knot index. With increase in the dip duration and mixed concentration of both significantly reduced number of eggmasses/root and root knot index
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Experimental Results
In combined treatment, extract of water hyacinth along with Pochonia chlamydosporia length of tomato cv. K-21 were 62.0, 59.0, 55.4cm respectively for 90 minutes of dip duration in S concentration of P. chlamydosporia + water hyacinth extract, S/2 concentration of P.chlamydosporia + water hyacinth extract, S/10 concentration of P.chlamydosporia + water hyacinth extract which were inoculated with 1500 second stage juveniles/pot. The corresponding figures for 45 minutes of dip duration were 60.4, 57.5 and 53.9 cm as against 36.5 cm in undipped inoculated and 78.2 cm in undipped uninoculated controls (Table-21).
The total weight of tomato cv.K-21 in S concentration of P.chlamydosporia + water hyacinth extract, S/2 concentration of P.chlamydosporia + water hyacinth extract, S/10 concentration of P.chlamydosporia + water hyacinth extract observed were 46.6, 44.7, 42.5 g when root were dipped for 90 minutes and inoculated with 1500 second stage juveniles of M.incognita /pot. The corresponding figures were 45.9, 44.0, 42.0 g respectively when the root were dipped for 45 minutes of duration as compared to 28 g in undipped inoculated and 78.2 g in undipped uninoculated controls (Table-21).
In combined treatment, application of water hyacinth extract significantly improved the chlorophyll content compare to inoculated control. The chlorophyll content of tomato cv.K-21 for 90 minutes of root dipping in S concentration of P.chlamydosporia + water hyacinth extract, S/2 concentration of P.chlamydosporia + water hyacinth extract, S/10 concentration of P.chlamydosporia + water hyacinth extract were 1.88, 1.81, 1.73 mg/g respectively with the inoculation1500 second stage juveniles of M.incognita/pot. The trending figures for dip duration of 45 minutes was 1.84, 1.77 and 1.71 mg/g as compared to 1.12 mg/g undipped inoculated and 2.78 mg/g in undipped uninoculated controls.
Application of both water hyacinth extract with P.chlamydosporia spores significantly improved the carotenoid content. Although highest carotenoid content (0.568 mg/g) in tomato cv.K-21 was detected in S concentration of P.chlamydosporia mixed with water hyacinth extract for 90 min of dip duration after inoculating 1500 second juveniles/pot. It was followed in descending order of 0.554, 0.541 mg/g at S/2 concentration of P.chlamydosporia + water hyacinth extract and S/10 concentration of P.chlamydosporia + water hyacinth extract. The corresponding
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Experimental Results figures for the carotenoid were 0.559, 0.546 and 0.532 mg/g respectively for 45 minutes of root dipping as compared to 0.252 mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated control (Table-21).
Combined application of both Water hyacinth extract mixed with P. chlamydosporia spores was found to be effective in reducing the number of eggmasses/plant. Highest reduction in number of eggmasses/root (100) was observed when S concentration of P.chlamydosporia mixed with water hyacinth extract spores for 90 minutes dip duration inoculating with 1500 second stage juveniles/pot. It was followed by 106, 112 in S/2 concentration of P.chlamydosporia + water hyacinth extract, S/10 concentration of P.chlamydosporia + water hyacinth extract respectively. The number of trending figures for 45 minutes of dip duration was 103, 110, 114 respectively in combined concentration of both as compared with 175 in undipped inoculated control (Table-21).
The percent pollen fertility of tomato cv.K-21 was significantly enhanced in all treatments. Root dipping in 90 minutes of duration in S concentration of the P.chlamydosporia + water hyacinth extract was found highest in increasing percent pollen fertility (74.0 %) of tomato followed by S/2 concentration of the P.chlamydosporia + water hyacinth extract (71.6 %), S/10 concentration of P.chlamydosporia+ water hyacinth extract (69.0 %) respectively when the root were inoculated with 1500 second stage juvenile of M.incognita/pot. The trending figures were 73.0, 70.5 and 67.0 % respective at 45 minutes dip duration as compared to 47 % in undipped inoculated and 89.5 % in undipped uninoculated controls (Table-21).
Root dip treatment in the extract water hyacinth in combination with P. chlamydosporia spores was found to be effective in reducing nematode severity and root knot indices. The root knot indices for the dip duration of 90 minutes were 1.5, 1.8 and 2.3 respectively in combined concentration of P.chlamydosporia (S) with water hyacinth extract spores, P.chlamydosporia (S/2) + water hyacinth extract, P.chlamydosporia (S/10) + water hyacinth extract inoculated 1500 second stage juvenile/pot. The corresponding values were 1.7, 2.0 and 2.9 respectively as compared to 5.0 in undipped inoculated control (Table-21).
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Table 21: Effect of bare root- dip in aquatic weed Water hyacinth leaf extract mixed with Pochonia chlamydosporia spores on the plant growth of tomato cv. K-21 infested with Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmass Pollen Root content content es/root fertility knot Duration Concentration Shoot Root Total Shoot Root Total (mg/g) (mg/g) (%) Index (minutes) 45 S 40.4b 20.2b 60.4c 32.3bc 13.6b 45.9bc 1.84bc .559c 103de 73.0bc 1.7cd
S/2 38.2c 18.8c 57.5e 31.0de 13.0bc 44.0de 1.77de .546d 110bc 70.5c 2.0cd
S/10 36.5d 17.4e 53.9g 29.6f 12.4c 42.0f 1.71e .532e 114b 67.0d 2.9b
90 S 41.2b 20.8b 62.0b 32.8b 13.8b 46.6b 1.88b .568b 100e 74.4b 1.5e
S/2 38.4c 19.0c 59.0d 31.5cd 13.2bc 44.7cd 1.81cd .554c 106cd 71.6c 1.8de
S/10 36.6d 18.2cd 55.4f 30.0ef 12.5c 42.5ef 1.73e .541d 112b 69.0cd 2.3c
Undipped Uninoculated Control 53.6a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a .880a 0f 89.5a 0f
Undipped Inoculated Control 24.7f 11.8f 36.5h 20.0g 8.0d 28.0g 1.12f .252f 175a 47.0g 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
4.5.9. Effect of bare root-dip treatment with various aqueous dilutions of Water hyacinth extracts mixed with Pseudomonas fluorescens suspension on root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21 in pots
This experiment was carried out on the same lines as given in 4.1.9. Application of both bioagents and plant extracts significantly reduce the nematode damage and improvement of plant growth parameters. Dip duration and mixed concentration of both are the factors which directly influence the number of eggmasses/root and root knot indices (Table-22).
When extract of water hyacinth along with Pochonia chlamydosporia spores were used the length of tomato cv. K-21 were 64.6, 61.6, 58.8 cm respectively for 90 minutes of dip duration in S concentration of P. fluorescens + water hyacinth extract, S/2 concentration of P. fluorescens + water hyacinth extract, S/10 concentration of P. fluorescens + water hyacinth extract which were inoculated with 1500 second stage juveniles /pot. The corresponding figures for 45 minutes of dip duration were 63.2, 61.0 and 57.0 cm as against 36.5 cm in undipped inoculated and 78.2 in undipped uninoculated controls (Table-22).
The total fresh weight of tomato cv.K-21 in S concentration of P. fluorescens + P. fluorescens, S/2 concentration of P. fluorescens + water hyacinth extract, S/10 concentration of P. fluorescens + water hyacinth extract observed were 48.0, 45.7, 43.5 g when root were dipped for 90 minutes and inoculated with 1500 second stage juveniles of M. incognita /pot. The corresponding figures were 47.2, 45.0, 42.6 g respectively when the root were dipped for 45 minutes of duration as compared to 28 g in undipped inoculated and 78.2 g in undipped uninoculated controls (Table-22).
Application of water hyacinth extract in combination with bioagents significantly improved the chlorophyll content compare to inoculated control. The chlorophyll content for 90 minutes of root dipping of tomato cv. K-21 in S concentration of P. fluorescens + water hyacinth extract, S/2 concentration of P. fluorescens + water hyacinth extract, S/10 concentration of P. fluorescens + water hyacinth extract were 1.92, 1.85, 1,76 mg/g respectively with the inoculation of 1500 second stage juveniles of M. incognita /pot. The trending figures for dip duration of
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Experimental Results
45 minutes was 1.88, 1.79 and 1.74 mg/g as compared to 1.12 mg/g undipped inoculated and 2.78 mg/g in undipped uninoculated controls.
Combined use of P. fluorescens with water hyacinth extract significantly enhanced the carotenoid content. Highest carotenoid content (0.568 mg/g) at 90 minutes dip duration in tomato cv.K-21 was detected in S concentration of P. fluorescens mixed with + water hyacinth extract after inoculating 1500 second juveniles /pot. It was followed in descending order of 0.564, 0.548 mg/g at S/2 concentration of P. fluorescens + water hyacinth extract and S/10 concentration of P. fluorescens + water hyacinth extract. The corresponding figures at 45 minutes of root dipping were 0.571, 0.556 and 0.542 mg/g respectively as compared to 0.252mg/g in undipped inoculated and 0.880 mg/g in undipped uninoculated control (Table-22).
All the treatments showed significant results in reducing the number of eggmasses/plant. Highest reduction in number of eggmasses /root (96) was observed when S concentration of P. fluorescens + water hyacinth extract for 90 minutes dip duration inoculating with 1500 second stage juveniles/pot. It was followed by 103,107 in S/2 concentration of P. fluorescens + water hyacinth extract, S/10 concentration of P. fluorescens + water hyacinth extract respectively. The corresponding figures for 45 minutes of dip duration were 99, 105, 109 respectively in combined concentration of both and as compared with 175 in undipped inoculated control (Table-22).
The percent pollen fertility of tomato cv.K-21 was significantly enhanced in all treatments. Root dipping in 90 minutes of duration in S concentration of the P. fluorescens + water hyacinth extract was found highest in increasing percent pollen fertility (76.0 %) of tomato followed by S/2 concentration of the P. fluorescens + water hyacinth extract (72.5 %), S/10 concentration of the P. fluorescens + water hyacinth extract (70.0 %) respectively when the root were inoculated with 1500 second stage juvenile of M. incognita /pot. The trending figures were 74.2, 71.2 and 68.6 % respectively at 45 minutes of dip duration as compared to 47 % in undipped inoculated and 89.5% in undipped uninoculated controls (Table-22).
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Table22: Effect of bare root- dip in aquatic weed Water hyacinth leaf extract mixed with Pseudomonas fluorescens colony on the plant growth of tomato cv. K-21 infested with Meloidogyne incognita in pots Dip Treatment Length(cm) Fresh weight(g) Chlorophyll Carotenoid Eggmass Pollen Root content content es/root fertility Knot Duration Concentration Shoot Root Total Shoot Root Total (mg/g) (mg/g) (%) Index (minutes) 45 S 42.5bc 20.7bc 63.2bc 33.0bc 14.2b 47.2bc 1.88bc .571c 99e 74.2bc 1.6de
S/2 41.0a 20.0bc 61.0d 31.5de 13.5bcd 45.0de 1.79de .556d 105cd 71.2c 1.9d
S/10 38.3e 18.7d 57.0f 30.0f 12.6d 42.6f 1.74e .542e 109b 68.6d 2.7b
90 S 43.4b 21.2b 64.6b 33.6b 14.4b 48.0b 1.92b .580b 96e 76.0b 1.4e
S/2 41.4cd 20.2bc 61.6cd 32.0cd 13.7bc 45.7cd 1.85cd .564c 103d 72.5c 1.8d
S/10 39.5e 19.3cd 58.8e 30.5ef 13.0cd 43.5ef 1.76e .548e 107bc 70.0cd 2.2c
Undipped Uninoculated Control 53.6a 24.6a 78.2a 52.0a 20.0a 72.0a 2.78a .880a 0f 89.5a 0f
Undipped Inoculated Control 24.7f 11.8e 36.5g 20.0g 8.0e 28.0g 1.12f .252f 175a 47.0e 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; S- Standard extract; S/2, S/10, S/100 are dilutions of S.
Experimental Results
Water hyacinth extract in combination with P. chlamydosporia spores as a bare root dip treatment was found to be effective in curtailing nematode severity and root knot indices. The root knot indices for the dip duration of 90 minutes were 1.4, 1.8 and 2.2 respectively in combined concentration of P. fluorescens (S) + water hyacinth extract, P. fluorescens (S/2) + water hyacinth extract, P. fluorescens (S/10) + water hyacinth extract inoculated 1500 second stage juvenile /pot. The corresponding values were 1.6, 1.9 and 2.7 respectively as compared to 5.0 in undipped inoculated control (Table-22).
4.6.1. Effect of chopped leaves of different plant species in combination with seed powder of Black nightshade on the root-knot development caused by Meloidogyne incognita and plant growth character of tomato cv. K-21 in pots
The present experiment was carried out under glasshouse conditions to test the effectiveness and nematostatic potential of organic soil amendments with chopped leaves of Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed applied @ 50g combined with seed powder of Black nightshade @ 10 g per pot on the root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv. K-21. It was noted that there was significant improvement in plant growth parameters in all the treatments but chopped leaves of Argemone mexicana combined with Black nightshade seed powder elucidate highest improvement in plant growth parameters and maximum reduction in nematode population and root knot indices (Table-23a).
The total length of tomato cv. K -21 was found highest (53.5 cm) when tomato plants treated with leaves of Mexican poppy. It was followed by 51.7, 50.0, 48.2, 46.4 and 44.7 cm in Wild eggplant, Black pig weed, Indian mallow and Ivy gourd respectively as compared to 36.5 cm in untreated inoculated and 78.2 cm in untreated uninoculated control plants (Table-23a).
The total fresh weight of tomato cv. K -21 was found highest (41.0 g) when tomato plants treated with leaves of Mexican poppy. It was followed by 39.6, 38.4, 37.2, 36.0 and 35.2 g in Trailing eclipta, Wild eggplant, Black pig weed, Indian mallow and Ivy gourd respectively as against to 36.5 g in untreated inoculated and 78.2 g in untreated uninoculated control plants (Table23a)
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Experimental Results
The total dry weight of tomato cv. K -21 was found highest (14.2 g) when tomato plants treated with leaves of Mexican poppy. It was followed by 13.5, 12.9, 12.3, 11.5 and 11.0 g in Trailing eclipta, Wild eggplant, Black pig weed, Indian mallow and Ivy gourd respectively as compared to 8.5g in untreated inoculated and 24.0 g in untreated uninoculated control plants (Table-23a).
The total chlorophyll and carotenoid content was significantly increased. The chlorophyll and carotenoid content was 1.73 and 0.495 mg/g, 1.68 and 0.474 mg/g, 1.62 and 0.446 mg/g, 1.55 and 0.418mg/g, 1.51 and 0.400 mg/g and 1.45 and 0.390 in Mexican poppy, Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd respectively as compared to 1.12 and 0.252 mg/g in untreated inoculated and 2.78 and 0.880 mg/g in untreated uninoculated control (Table-23b).
The percent pollen fertility was significantly increased in all the treatments. Maximum pollen fertility (67.0 %) when the tomato was treated with Mexican poppy. It was followed by as 65.6, 64.0, 63.5, 62.0 and 61.0 % in Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd respectively as against 47.0 % in untreated inoculated and 89.5 % in untreated uninoculated control (Table-23b).
The chopped leaves along with the seed powder significantly increased the yield. Maximum yield 228 g was observed when the tomato plants were treated with Mexican poppy. It was followed by as 200, 186, 167.0, 160.0 and 144.0 g in Trailing eclipta, Wild eggplant, Black pig weed, Indian mallow and Ivy gourd respectively as against 112.0 g in untreated inoculated and 355.0 g in untreated uninoculated control (Table-23b).
Significant reduction was observed in eggmasses, eggs and nematode population of M. incognita when tomato was treated with chopped leaves combined with seed powder of different plant species. Highest reduction in eggmasses, eggs and nematode population 114.0, 188.0 & 1042 was observed in Mexican poppy leaves. It was followed by 118.0, 193.0 and 1069 in Trailing Eclipta, 123.0, 197.0 and 1100 in Wild eggplant, 128, 202 and 1142 in Black pig weed, 132, 209 and 1170 in, Indian mallow and 140,215 and 1204 in C. grandis respectively as against 148.0, 254.0 and 1558.0 in untreated inoculated control (Table- 23b).
114
Experimental Results
Fresh chopped leaves in combination with seed powder significantly reduced the root knot development. The highest reduction in the root-knot development was detected when tomato plants were treated with leaves of Mexican poppy and root knot index was recorded as 2.0.It was followed by 2.2, 2.3, 2.5, 2.8 and 3.0 in Trailing eclipta, Wild eggplant, Black pig weed, Indian mallow and Ivy gourd respectively. In untreated inoculated pots the root-knot development was highest (5.0) (Table-23b).
4.6.2. Effect of biochar alone and in combination with saw dust of different plants used as soil amendment on the root knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21
The present experiment deals with the potential of biochar alone and in combination with saw dust of different plants viz., eucalyptus, lebbeck, jambul, mango, poplar and babool under glass house condition for the management of root knot nematode and plant growth development of tomato cv. K-21(Table-24a).
The efficacy of biochar amended with agriculture waste on different plant growth characters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield significantly enhanced in tomato cv. K-21 as compared to the untreated inoculated control plants (38.5 cm, 31.5g, 9.3 g, 47.2 %, 158 g). Among all the treatments applied, application of biochar integrating with eucalyptus sawdust results in paramount improvement in plant growth characters plant length, fresh weight, dry weight, percent pollen fertility and yield were 75.4 cm, 67.2g, 21.9g, 81%, 302 g. It was followed by biochar+ lebbeck sawdust 74.2cm, 65g, 21.3g, 79.6%, 299g; biochar + jambul sawdust 72.6 cm, 63 g, 20.7 g, 79 %, 294 g. While the biochar alone recorded to have least promoting activity for the enhancement of plant growth characters were 63.5 cm, 55.2 g, 18.4 g, 71%, 240 g (Table-24a,b).
Biochemical parameters significantly improved when biochar was applied alone and in integrated manner with saw dust. Significant improvement was observed in chlorophyll and carotenoid content in all the treatments was 2.30 & 0.786 mg/g, 2.24 & 0.779 mg/g, 2.17 & 0.7700 mg/g in Biochar+ eucalyptus sawdust, Biochar + lebbeck sawdust, and biochar + jambul sawdust as compared to the 1.12&0.273mg/g in untreated inoculated and 2.79 & 0.878 mg/g in untreated uninoculated control. However singly application of biochar results in lowest amount of chlorophyll and carotenoid content 1.84 and 0.680 mg/g (Table-24b).
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Experimental Results
Significant reduction in root knot nematode development and severity has been observed by the incorporation of all the treatment alone application and combined treatments. Application of biochar in combination with eucalyptus sawdust caused most paramount reduction in eggmass/root, egg/eggmass, nematode population and root knot indices were 61, 115, 597, and 1.5. It was followed by biochar+ lebbeck sawdust 65, 121, 608, 1.7 biochar + jambul sawdust 68, 129, 620, 1.9 respectively as compared to untreated inoculated control plants (174, 266, 1608, 5.0). Moreover in treated plants maximum nematode infestation was observed in alone application of biochar were 106, 178, 955, 3.2 (Table-24b).
4.6.3.Effect of biochar alone and in combination with various agriculture waste used as soil amendment on the root knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21
This experiment was conducted with the purpose to test the potential of pyrolyzed product, biochar alone and in combination with agriculture waste against root knot nematode multiplication and plant growth of tomato cv. K-21 (Table-25a).
As a result of the application of biochar in combination with agriculture waste viz., tobacco, garlic waste, mentha waste, decomposed potato, black gram waste and bagasse reduces the root knot nematode infestation and thereby enhances the plant growth characters of tomato cv. K-21. With respect to the effect of biochar on various plants growth parameters alone and in combinations (plant length, fresh weight, dry weight, percent pollen fertility and yield) on tomato cv.K-21was recorded to be significant enhancement in growth character of tomato as compared to untreated inoculated control plants 38.5 cm, 31.5 g, 9.3 g, 47.2 % 158 g. The integrated utililiztion of biochar+ tobacco waste was found to be most promising in improving the plant growth parameters. In biochar +tobacco waste combination the plant growth parameters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield were 76.4 cm, 68 g, 22.1 g, 81.5 %, 306 g. It was followed by biochar+ garlic waste 75 cm, 66.8 g, 21.6 g, 80.6 % 300 g; biochar+ mentha waste 73.5 cm, 65.5 g, 21.2 g, 79.7 %, 296 g; biochar+ decomposed potato 72.3cm, 64g, 20.8g, 78.5% 293g and lowest in biochar alone were 63.5 cm, 55.2 g, 18.4 g, 71 %, 240 g (Table-25a,b).
Pyrolyzed product, biochar singly and in combination with agriculture waste exaggerate the biochemical parameters of tomato cv. K-21. Application of biochar in 116
Experimental Results combination with tobacco waste caused most paramount improvement in chlorophyll and carotenoid content were 2.32 mg/g, 0.790 mg/g. It was followed by biochar+ garlic waste were 2.25 mg/g, 0.779 mg/g; biochar+ mentha waste were 2.18 mg/g, 0.772 mg/g; biochar+ decomposed potato waste 2.12 mg/g, 0.763 mg/g as compared to untreated inoculated control plants 1.12 mg/g, 0.269 mg/g. However, the isolated application of biochar results in least improvement in chlorophyll and carotenoid content were 1.84 mg/g, 0.680 mg/g (Table-25b).
Application of biochar in combination with agriculture waste caused significant suppression in root knot nematode severity viz., eggmass/root, egg/eggmass, nematode population and root knot indices were 174, 266, 1608, 5.0 as compared to untreated inoculated control plants. However, among all the treatments applied singly or in combination, biochar + tobacco waste recorded minimum root knot nematode infestation viz., eggmass/root, egg/eggmass, nematode population and root knot indices were 59, 112, 593, 1.4 followed by biochar+ garlic waste were 61, 120, 610, 1.6; biochar+ menthe waste were 65, 129, 628, 1.9. More over maximum number of eggmass/root, egg/eggmass, nematode population and root knot indices were observed in alone application of biochar were106, 178, 955, 3.2 (Table-25b).
4.7.1. Effect of seed dressing with biochar on the root knot development caused by Meloidogyne incognita and plant growth characters of tomato cv.K-21
The present experiment deals with the efficacy to test the potential of biochar as a seed dressing agents at various concentrations @ 0.2, 0.4, 0.8, 1.6, 3.2, 6.4% (v/v) for the management of root knot nematode, M.incognita and plant growth parameters of tomato cv.K-21. Data presented in the table-26 clearly revealed that biochar as a seed dressing agent in the tested concentrations of biochar @ 0.2, 0.4, 0.8, 1.6, 3.2, 6.4% (v/v) significantly enhanced the plant growth parameters viz., plant length, fresh weight, dry weight, percent pollen fertility, yield/plant were 38.5 cm, 31.5 g, 9.3 g, 47%, 142 g as compared to untreated inoculated control The 3.2 and 6.4% concentration of biochar showed most pronounced improvement in plant growth characters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield/plant were 51 cm, 43.6 g,13.7 g, 60 %, 196 g and 52.6 cm, 45 g, 14.6 g 62 %, 204 g respectively. Among both of the 3.2 % and 6.4 %, 3.2 was found to be statistically more significant in terms of plants growth parameters. While 0.2 %
117
Experimental Results biochar results in least promoting behaviour in plant growth characters and is was 41.0cm in length, 33.7g in fresh weight, 10.8g in dry weight, 49 % pollen fertility and 121g yield/plant. Although seed dressing with 0.2 % was not statistically significant for percent pollen fertility (Table-26a, b).
Biochar at various concentrations @ 0.2, 0.4, 0.8, 1.6, 3.2, 6.4% (v/v) enhanced the biochemical parameters of tomato cv.K-21. Maximum enhancement in chlorophyll and carotenoid content was observed at concentration of 3.2 and 6.4 % were 1.58 mg/g, 0.460 mg/g and 1.64 mg/g, 0.477 mg/g respectively as compared to untreated uninoculated control plants (1.12 mg/g, 0.275 mg/g).While the least enhancement was observed 0.2 % concentration were 1.19 mg/g, 0.325 mg/g. Moreover, 0.2 % concentration was not statistically significant for chlorophyll content (Table-26b).
Tested biochar at various concentrations @ 0.2, 0.4, 0.8, 1.6, 3.2, 6.4 % (v/v) suppressed the root knot nematode infestation viz., eggmasses/root, eggs/eggmass, nematode population and root knot index as compared to untreated inoculated control ( 175, 266, 1608, 5.0). Concentration of biochar at 3.2 and 6.4 % showed highest potential for the suppression of eggmasses/root, eggs/eggmass, nematode population and root knot index were 133,199, 1152, 3.0 and 130, 191, 1120, 2.8 respectively. While @ 0.2 % concentration cause the minimum suppression in root knot nematode severity viz., eggmasses/root, eggs/eggmass, nematode population and root knot index were 168, 250, 1389, 4.8 but none of them except nematode population was found to be significant at the same concentration (Table-26b).
4.7.2. Effect of seed dressing with bioinoculants on the plant growth of tomato cv.K-21 in relation to root knot development Meloidogyne incognita in pots
The present experiment deals with the efficacy of different bioinoculants Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum, Trichoderma viride as a seed dressing agents each @ 4 g/kg seeds against M.incognita using cellulose and molasses in 1 and 2% concentrations as a sticky agent under glass house conditions on tomato cv. K-21. Data present in the table-27 clearly represents that the screened microbial antagonist as seed dressing of tomato using 1 and 2 % of cellulose and molasses significantly enhanced the plant growth parameters 118
Experimental Results viz., plant length (cm), fresh weight (g), dry weight (g), and percent pollen fertility (%) and yield (g). Though the biocontrol agents applied, P.chlamydosporia adopting 2% cellulose as sticky agents results in most significant improvement in plant growth characters. The P.chlamydosporia adopting 2% cellulose as sticky agents the plant growth parameters viz., plant length (cm), fresh weight (g), dry weight (g), and percent pollen fertility (%) and yield (g) observed were 64.2 cm, 48.2 g, 17.0 g, 74.5 %, 248 g; 2 % cellulose as sticky agent with P. fluorescens were 63.2 cm, 46.6 g,16.2 g, 73.8 %, 243 g; 2 % celluloses sticky agent with P. lilacinum were 61 cm,45.2 g,15.7 g,72%, 237 g. While at 2% cellulose as a sticky agent the least effectiveness was recorded when T. viride was applied (58.4 cm, 44.5g, 14.5 g, 71 %, 226 g) (Table-27a,b).
Tested microbial antagonist viz., Pochonia chlamydosporia, Pseudomonas fluorescens, and Purpureocillium lilacinum, Trichoderma viride @ 4 g/kg as a seed dressing agents with the 1 and 2% cellulose and molasses as sticker significantly enhanced biochemical parameters viz., chlorophyll and carotenoid content in tomato cv. K-21. Among all the biocontrol tested P. chlamydosporia using 2% cellulose was the most efficient in augmenting the chlorophyll and carotenoid content were 1.93 mg/g and 0.615 mg/g. Though the biocontrol agents tested P.chlamydosporia was the most efficient biocontrol agents but Trichoderma viride was least. Moreover, among the tested sticker 2% concentration cellulose was most effective as a sticker to adhere the bioinoculants (Table-27b).
All the treatments viz., P. chlamydosporia, P. fluorescens, P. lilacinum, T. viride when applied @4g/kg seeds incorporating cellulose and molasses @ 1 and 2% as a sticker significantly suppressed root knot nematode infestation viz., eggmasses/root, eggs/eggmass, nematode population and root knot index. Among all the treatments 2% cellulose as sticker with P. chlamydosporia elucidate most significant reduction in eggmasses/root, eggs/eggmass, nematode population and root knot index were 97, 180, 827, and 1.7. It was followed by P.flueroscens were 100 183, 856, 1.9; P. lilacinum were 102, 185, 889, 2.6 . While Trichoderma viride was found to be least effective as biocontrol agents in reducing nematode severity and infestation. Of both of the coating material tested@1 and @ 2%, concentration of 2% cellulose record most significant results in reduction of nematode severity and nematode infestation (Table-27b). 119
Experimental Results
4.7.3. Effect of various aqueous dilutions of oil cakes as seed dressing agents on the plant growth of tomato cv.K-21 in response to nematode multiplication and root knot nematode development
The experiment was carried out to evaluate the potential from deoiled powdered cakes of castor, cotton, mahua, mustard and soybean as seed dressing agents on tomato cv.K-21 @7, 14, 21% w/w under glass house conditions for the management of root knot nematode, M. incognita and their infestation. Results indicate that all the treatments brought about the significant improvement in plant growth characters viz., plant length, fresh weight, dry weight, yield/plant and percent pollen fertility of tomato cv.K-21 in compared to untreated inoculated control (38.5 cm, 31.5 g, 9.3 g, 47 %, 142 g). Castor cake @ 21 % recorded most significant improvement in plant growth parameters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield/plant were 56.8 cm, 44 g, 15.5 g, 74 %, 212 g followed by the mustard cake were 55 cm, 43.4 g, 15 g, 72%, 209 g. While the least at the same concentration given by cotton cake was 47 cm, 37.5 g, 13.2 g 67%, 184 g. However among all the given doses of 7, 14 and 21 %., 21 % was found to be most effective in enhancing the plant growth parameters and are statistically at a par (Table-28a,b).
Seed treatment with various oilcakes significantly results in suppression of root knot nematode severity viz., number of eggmasses/root, egg/eggmass, nematode population and root knot index as compared to the untreated inoculated control (175, 266, 1608, 5.0). Among the applied various deoiled cakes, Castor cake@ 21% (w/w) results in maximum potential for the suppression of eggmasses /root, egg/eggmass, nematode population and root knot index were 112, 185, 1125, 1.8 followed by mustard cake 115, 190, 1142, 2.4. While the lowest reduction was observed by cotton cake at the same concentration were 124, 204, 1190, 3.2. Lower dose of oilcakes 7 and 10% are not too much efficient for the reduction of nematode potential a severity while higher concentration @ 21% are at a par in reducing nematode infestation (Table-28b).
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Experimental Results
4.8.1. Studies on the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Trichoderma viride on the plant growth of tomato cv. K- 21 in pots in relation to root knot development and nematode multiplication.
The experiment was carried with the aim to elucidate the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Trichoderma viride on the plant growth characters of tomato cv. K-21 in response to root knot development and M.incognita multiplication. Finding of the experiment clearly stated that all the treatments result in significant reduction in root knot development caused by M. incognita and also enhanced the plant growth characters of tomato cv.K-21. In the experiment when one pathogen is inoculated 15days before another pathogen, inoculation of M. incognita 15 days prior to the fungal pathogen results in more reduction compare to the prior fungal application with the nematode (Table-29 a).
Significant improvement in plant growth characters viz., plant length (cm), fresh weight (g) and dry weight (g) was observed when individual application of Trichoderma viride was done (83cm, 78g and 28g) as against to the untreated uninoculated plants(81.2cm, 73.0g, 26.4g). Fifteen days (15days) prior application of T. viride to the M.incognita inoculation caused greatest improvement in plant growth characters were (66.0 cm, 57.5 g and 19.5 g) as compared to the M incognita inoculation prior to the T. viride (46.0 cm, 34.5 g and 12.4 g) in tomato cv.K-21. Simultaneous inoculation of T. viride and M. incognita enhanced the length, fresh weight and dry weight (55.0 cm, 46.7 g, 15.6 g). Most significant reduction in plant growth parameters was recorded when M.incognita at 1500 inoculum was inoculated lonely (38.5 cm, 31.5 g and 9.3 g) (Table-29a).
Application of T. viride and inoculation of M. incognita either alone or with various combinations significantly reduced all the biochemical parameters such as chlorophyll, carotenoid and nitrate reductase content in the leaf of tomato. Highest increase in biochemical parameters was found when T. viride was applied alone were 2.82 mg/g, 0.890 mg/g and 317 nM .g-1.h-1 as against to the untreated uninoculated control (2.78 mg/g, 0.880 mg/g and 307 nM .g-1.h-1) but chlorophyll value was not statistically significant. Sequential and concomitant inoculation of T. viride and M.incognita enhanced the biochemical parameters. The application of T. viride and
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Experimental Results
M.incognita concomitantly bring about a significant improvement in chlorophyll, carotenoid and nitrate reductase content 1.98 mg/g, 0.660 mg/g and 230 nM .g-1.h-1. Sequential inoculation of T. viride 15 days prior to the inoculation of M.incognita was found to be effective in enhancing chlorophyll, carotenoid and nitrate reductase content (2.15mg/g, 0.780 mg/g, 266 nM g-1.h-1) as compare to the 15 days prior inoculation of M. incognita to the T.viride (1.50 mg/g, 0.520 mg/g, 209 nM g-1.h- 1).The outmost significant reduction in biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase content (1.12mg/g, 0.272mg/g, 142nM g-1.h-1) was obtained when M.incognita was applied alone with 1500 juveniles (Table-29b).
Application of Trichoderma viride singly enhanced the percent pollen fertility and yield was (89% and 360g) as compared to the untreated uninoculated control plants (89.5% and 355g) but the value of increment was not significant. Application of T. viride and M.incognita concomitantly and sequentially escalate the percent pollen fertility and yield. Concomitant application of both T. viride and M. incognita cause improvement in percent pollen fertility and yield (69% and 230g). Significant enhancement was detected when sequential inoculation of T.viride was done 15 days prior to M.incognita were 74.5 %, 280 g followed by concomitant inoculation of M. incognita and T. viride (58.0% and 176g). Most significant reduction in yield and percent pollen fertility was examined when M.incognita was inoculated alone at 1500 inoculum (47.0% and 148g) (Table-29b).
The total number of eggmasses/root, eggs/eggmass and Meloidogyne incognita population was significantly reduced in all the interactive treatments. Sequential application of P. chlamydosporia 15days prior to M.incognita inoculation showed outstanding reduction in eggmasses, eggs and nematode population (90, 160, 760) followed by concomitant application of both T. viride and M. incognita (108, 184, 860).While M. incognita inoculation 15 days prior to the T. viride shows least decrease in eggmasses, eggs and nematode population (132, 220, 985) as compared to plants inoculated with 1500 J2 of M. incognita alone (175, 266 and 1608).
Sequential application of T. viride 15 days prior to M.incognita inoculation showed magnificent reduction in root knot indices development (1.8) followed by concomitant inoculation of M.incognita and T. viride (2.6) and sequential inoculation
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Experimental Results of M.incognita 15 days prior to the T. viride (3.0) as compared to untreated inoculated control plant alone with 1500 J2 of M. incognita (5.0) (Table-29b).
4.8.2. Studies on the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Purpureocillium lilacinum on the plant growth of tomato cv. K-21 in pots in relation to root knot development and nematode multiplication.
The purpose of the present experiment was to assess the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Purpureocillium lilacinum on the plant growth parameters of tomato cv. K-21 in relation to root knot development and M.incognita multiplication. It was observed that all the treatments caused significant improvement in plant growth parameters of tomato K-21 and reduction in nematode population and root knot indices (Table-30a).
From the tabulated data it was clear that the application of Purpureocillium lilacinum alone significantly improved all the plant growth characters. The plant growth parameters viz., plant length (cm), fresh weight(g), Dry weight (g), percent pollen fertility (%) and yield (g) were 84.5cm, 89.5g, 28.8g, 90%, 367g as against untreated uninoculated control plants (81.2cm, 73g, 26.4g, 89.5%, 355g) but the pollen fertility was found to be insignificant. Sequential and concomitant application of M.incognita and P. lilacinum significantly strengthen the plant growth parameters and reduction in nematode potential. Most pronounced increase in plant growth parameters viz., plant length (cm), fresh weight (g), dry weight (g), percent pollen fertility (%) and yield (g) were 68.5cm, 58.4g, 20,77%, 282g when sequential inoculation of P. lilacinum was done 15 days prior to M.incognita against to the M.incognita inoculation 15days prior to the P. lilacinum (46.5 cm, 34.8 g, 12.6 g, 59%, 181 g). Concomitant application of both P.lilacinum and M.incognita also significantly amplify the plant growth parameters viz., plant length (cm), fresh weight (g), Dry weight (g), percent pollen fertility (%) and yield (g) were 56.4 cm, 47.4 g, 16.0 g, 70.5 %, 236 g. While the sole application of M.incognita significantly reduced the plant growth parameters viz., plant length (cm), fresh weight (g), dry weight (g), percent pollen fertility (%) and yield (g) were 38.5cm, 31.5g, 9.3g, 47%, 148g (Table- 30a,b).
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Experimental Results
Biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase content significantly increased with the application of P.lilacinum alone were 2.86 mg/g, 910 mg/g, 322 nM .g-1.h-1 as compared to untreated uninoculated control plants (2.78 mg/g, 0.880 mg/g, 307 nM .g-1.h-1) but the chlorophyll figures were not statistically significant. While the most significant reduction in biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase was observed when M. incognita was applied alone were 1.12 mg/g, 0.272 mg/g, 242 nM .g-1.h- 1.Concomitant and sequential application of both nematode and fungus significantly enhanced the biochemical parameters. Application of M.incognita and P.lilacinum concomitantly results significantly increment in biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase were 2.00 mg/g, 0.675 mg/g, 238 nM .g- 1.h-1. However, application of P.lilacinum 15 days prior to the M.incognita inoculation recorded effective improvement in chlorophyll, carotenoid and nitrate reductase content (2.20 mg/g, 0.800 mg/g, 272 nM .g-1.h-1) as compared to the P.lilacinum applied at 15 days after M.incognita inoculation were 1.56 mg/g, 0.532 mg/g, 213 nM .g-1.h-1 (Table-30b).
In the inoculation of M.incognita alone the number of eggmasses/root, eggs/eggmasses, nematode population and root knot indices were 175, 266, 1608 5.0 respectively. Concomitant and sequential application of M.incognita and P.lilacinum significantly suppressed the eggmasses, eggs, nematode population and root knot index .Concomitant application of P.lilacinum and M.incognita reduced the eggmasses/root, eggs/eggmasses, nematode population and root knot index were 104, 175, 833, 2.3. Sequential inoculation of P.lilacinum 15 days prior to the M.incognita records most significant reduction in the eggmasses/root, eggs/eggmasses, nematode population and root knot index were 82, 152, 732, 1.6 respectively as compared to the 15 days prior inoculation of M.incognita to the P.lilacinum were (131, 216, 955, 2.8) (Table-30b).
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Experimental Results
4.8.3. Studies on the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Pochonia chlamydosporia on the plant growth characters of tomato cv. K-21 in pots in relation to root knot development and nematode multiplication
The present experiment was conducted to test the effect of Meloidogyne incognita and P.chlamydosporia as individual, sequential and concomitant treatment on plant growth of tomato K-21 in response nematode multiplication and root knot development. All the treatments represent significant improvement in plant growth parameters of tomato cv.K-21 and reduction in nematode population and root knot indices. Inoculation of M.incognita alone significantly suppressed the plant growth characters and increases the nematode reproduction. While in case of sequential application of nematode and fungus, when fungus was preceded by 15 days to the nematode results in pronounced improvement of plant growth parameters and high reduction in nematode population compare to prior nematode inoculation to the fungus records reduction in plant growth and increment in nematode population (Table-31a).
Significant improvement in plant growth characters viz., plant length (cm), fresh weight (g) and dry weight (g) was observed when P.chlamydosporia was applied alone were 83cm,78g and 28g as against to the untreated uninoculated plants (86.5 cm,80.4 g,29.4 g). Simultaneous application of P.chlamydosporia and M. incognita result in improvement of the length, fresh weight and dry weight were 58.6 cm, 48.0 g, 16.4 g. In case of sequential application 15days prior application of P.chlamydosporia to the M.incognita recorded greatest improvement in plant growth parameters (70.5 cm,59.2 g and 20.8 g) as compared to the P.chlamydosporia application after M incognita inoculation were 46.8 cm,35.5 g and 13.0 g in tomato cv.K-21.Significant reduction in plant growth parameters was observed when M.incognita was inoculated alone at 1500 inoculum level were 38.5 cm, 31.5 g and 9.3 g as compared to the untreated uninoculated control plants (Table-31a).
Application of P.chlamydosporia and inoculation of M.incognita either alone or different combination significantly reduced all the biochemical parameters such as chlorophyll, carotenoid and nitrate reductase content in tomato. Application of P.chlamydosporia alone significantly enhanced the biochemical parameters viz.,
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Experimental Results chlorophyll, carotenoid and nitrate reductase content 2.90 mg/g, 0.918 mg/g and 326 nM .g-1.h-1 as compared to the untreated uninoculated control 2.78mg/g, 0.880mg/g and 307nM .g-1.h-1. Sequential and concomitant employment of P.chlamydosporia and M.incognita improved the biochemical parameters. The application of P.chlamydosporia and M.incognita concomitantly results in the significant improvement in chlorophyll, carotenoid and nitrate reductase content 2.05mg/g 0.682mg/g and 241nM .g-1.h-1. However, sequential application of P.chlamydosporia 15 days prior to the inoculation of M.incognita found to be effective in enhancing chlorophyll, carotenoid and nitrate reductase content (2.25mg/g, 0.810mg/g, 275nM g-1.h-1) as against to the 15 days after to the of M.incognita (1.60mg/g, 0.537mg/g, 215nM g-1.h-1). The most significant reduction in biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase content observed were (1.12mg/g, 0.272mg/g, 142 nM g-1.h-1) was recorded when M.incognita was applied alone at 1500 inoculum (Table-31b).
Application of P.chlamydosporia alone increased the percent pollen fertility and yield (90.5% and 376g) as compared to the untreated uninoculated control plants (89.5% and 355g) but the percent pollen fertility was not statistically significant. Concomitantly and sequentially application of P.chlamydosporia and M.incognita enhanced the percent pollen fertility and yield. Concomitant application of both P.chlamydosporia and M.incognita results in significant improvement in pollen fertility and yield (71.4 %and 242 g). Sequential inoculation of P.chlamydosporia 15 days prior to the M.incognita recorded (78.5%, 295g) pollen fertility and yield followed by M.incognita 15 days prior to P.chlamydosporia ( 58.0 % and 176 g).Maximum reduction in yield and percent pollen fertility was detected when M.incognita was inoculated alone at 1500 inoculum ( 47.0 % and 148 g) (Table-31b).
The total number of eggmasses/root, eggs/eggmass Meloidogyne incognita population and root knot index was significantly reduced in all the interactive treatments. Sequential application of P. chlamydosporia 15days prior to the M.incognita inoculation, results in most significant reduction in eggmasses, eggs nematode population and root knot index (73, 134, 700, 1.5) followed by concomitant application of both P. chlamydosporia and M.incognita (102, 172, 822, 2.2). However, M.incognita inoculation 15 days prior to the P. chlamydosporia showed least decrease in eggmasses, eggs, nematode population and root knot index (128, 126
Experimental Results
200, 928, 2.5) as compared to plants inoculated with M. incognita solely at 1500 inoculum (175, 266 and 1608,5.0) (Table-31b).
4.8.4. Studies on the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Pseudomonas fluorescens on the plant growth of tomato cv. K-21 in pots in relation to root knot development and nematode multiplication.
The aim of the present experiment was to evaluate the effect of individual, sequential and concomitant inoculation of Meloidogyne incognita and Pseudomonas fluorescens on the plant growth characters of tomato cv. K-21 in relation to root knot nematode development and nematode multiplication. It was found that significant improvement in terms of plant growth parameters of tomato K-21 and reduction in nematode population and root knot indices was observed in all the treatments. (Table- 32a).
The application of Pseudomonas fluorescens alone significantly improved all the plant growth characters viz., plant length (cm), fresh weight(g), Dry weight(g), percent pollen fertility(%) and yield(g) were 87.5 cm, 81.2 g, 29.8 g, 91.6 %, 385 g as against to the untreated uninoculated control plants (81.2 cm, 73 g, 26.4 g, 89.5 %, 355 g). Sequential and concomitant application of M.incognita and P. fluorescens significantly enhanced the plant growth characters and reduction in nematode development. Highest increase in plant growth parameters viz., plant length (cm), fresh weight (g), Dry weight (g), percent pollen fertility (%) and yield (g) observed were 71.6 cm, 60 g, 21,79.4 %, 300 g when sequential inoculation of P. fluorescens 15 days prior to the M.incognita as compared to the P. fluorescens 15 days after M.incognita inoculation were (47.5 cm, 35.8 g, 13.4 g, 61 %, 194 g). However, concomitant application of both P. fluorescens and M.incognita also significantly increase the plant growth parameters viz., plant length (cm), fresh weight (g), dry weight (g), percent pollen fertility (%) and yield (g) were 62 cm, 49.5 g, 16.7 g, 72 %, 250 g. Whereas application of M.incognita alone significantly suppressed the plant growth parameters viz., plant length (cm), fresh weight (g), Dry weight (g), percent pollen fertility (%) and yield (g) were 38.5 cm, 31.5 g, 9.3 g, 47%, 148 g (Table- 32a,b).
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Experimental Results
Individual application of P. fluorescens significantly increased biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase content were 2.95 mg/g, 0.924 mg/g, 332 nM .g-1.h-1 as compared to untreated uninoculated control plants(2.78 mg/g, 0.880 mg/g, 307 nM .g-1.h-1).While the individual application of M. incognita results in most significant reduction in biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase were 1.12 mg/g, 0.272 mg/g, 242 nM .g- 1.h-1.Concomitant and sequential application of nematode and fungus significantly enhanced the biochemical parameters. Concomitant application of M.incognita and P. fluorescens results significantly enhancement in biochemical parameters viz., chlorophyll, carotenoid and nitrate reductase were 2.12 mg/g, 0.689 mg/g, 250 nM .g- 1.h-1. While application of P. fluorescens 15 days prior to the M.incognita inoculation estimated effective improvement in chlorophyll, carotenoid and nitrate reductase content (2.32 mg/g, 0.816 mg/g, 285 nM .g-1.h-1) as compared to the M.incognita preceded 15 days with the P. fluorescens were 1.69 mg/g, 0.540mg/g, 220nM .g-1.h- 1 (Table-32b).
Maximum number of eggmasses/root eggs/eggmasses, nematode population and root knot index were when alone inoculation of M.incognita was done (175, 266, 1608 5.0) respectively. Concomitant and sequential application of M.incognita and P. fluorescens significantly reduced the eggmasses, eggs/eggmass, nematode population and root knot index. Application of P. fluorescens and M.incognita concomitant suppressed the eggmasses/root, eggs/eggmasses; nematode population and root knot index were 94, 160, 790, 2.0. Sequential inoculation of P. fluorescens 15 days before the M.incognita results in most significant reduction in the eggmasses/root, eggs/eggmasses, nematode population and root knot index were 64, 118, 640, 1.4 respectively as compared to the application of P. fluorescens 15 day after M.incognita were 118, 189, 907, 2.4 (Table-32b).
4.9.1. Effect of bioinoculants Purpureocillium lilacinum in combination with different organic soil amendment on the root-knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21 in pots
Application of Purpureocillium lilacinum integrating with organic amendment viz., press mud, cotton cake, mahua cake, mustard cake, castor cake and soybean cake augmenting significant improvement in plant growth parameters and reduction in root
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Experimental Results knot nematode infestation on tomato cv. K-21 under glass house conditions (Table- 33a).
Plant growth characters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield recorded significant improvement when P. lilacinum+ castor cake was incorporated were 75cm, 68.2g, 21.8g, 81%, 308g; P. lilacinum + press mud were 73.7 cm, 67 g, 21.3g, 80.2 %, 294g; P. lilacinum + mustard cake were 72.5 cm, 65.8 g, 21 g, 79 %, 290 g; P. lilacinum + soybean cake were 71 cm, 64.5 g, 20.4 g, 78.2 %, 285 g; P. lilacinum +mahua cake were 69.8 cm, 63 g, 19.6 g, 77 %, 287; lowest in P. lilacinum + cotton cake were 68.5 cm, 61.7 g, 18.8 g, 76 %, 275 g as compared to the untreated inoculated control plants (38.5 cm, 31.5 g, 9.3 g, 47.2 %, 154 g) (Table-33a,b).
Bioagents in combination with organic amendment enhanced the biochemical parameters.
The total chlorophyll and carotenoid content significantly improved when P. lilacinum+ castor was applied were 2.30 mg/g, 0.788 mg/g. It was followed by P. lilacinum+ press mud (2.12mg/g, 0.779 mg/g) P. lilacinum + mustard cake (2.15 mg/g, 0.768 mg/g) as compared to untreated inoculated control (1.12mg/g, 0.270mg/g). Moreover, least in treated was observed in P. lilacinum + cotton cake were (1.89 mg/g, 0.732 mg/g) (Table-33b).
All the treatments significantly reduced the root knot nematode infestation in terms of eggmasses/root, egg/eggmass, and nematode population and root knot indices. Minimum root knot nematode infestation was observed by the combined usage of P. lilacinum+ castor was (63, 117, 597, 1.7). It was followed by P. lilacinum+ press mud (66, 126, 612,1.9) P. lilacinum + mustard cake were (70,136, 633, 2.1) as compared to as compared to the untreated inoculated control plants (176, 266, 1608, 5) (Table-33b).
4.9.2. Effect of bioinoculant Pochonia chlamydosporia in combination with different organic soil amendment on the root-knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21 in pots
The perusal of the data of the experiment demonstrates that application of Pochonia chlamydosporia in combination with various organic amendments viz. press
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Experimental Results mud, cotton cake, mahua cake, mustard cake, castor cake and soybean cake significantly exaggerate the plant growth parameters and reduced the nematode infestation on tomato.cv.K-21 (Table-34a).
The potential of P. chlamydosporia integrating with organic amendment on different plant growth parameters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield significantly improved in tomato cv.K-21 as compared to the untreated inoculated control plants (38.5 cm, 31.5 g, 9.3 g, 47.2 %, 154 g). However, application of P. chlamydosporia in combination with castor cake showed most pronounced improvement in plant growth characters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield were 76.5 cm, 68.4 g, 22.4 g, 81.8 %, 308 g. While the P. chlamydosporia + cotton recorded least promoting activity for the enhancement of plant growth characters were 70 cm, 62.6 g, 19.5 g, 77 %, 287 g (Table-34a, b).
Biochemical parameters significantly improved in all the treatments. Greatest increase in chlorophyll and carotenoid was recorded in P. chlamydosporia+ castor cake were 2.35 mg/g, 0.793 mg/g; P. chlamydosporia+ press mud 2.25 mg/g, 0.779 mg/g; P. chlamydosporia+ mustard cake (2.18 mg/g, 0.770 mg/g. However, lowest was expressed by P. chlamydosporia+ cotton cake (1.96 mg/g, 0.746 mg/g). Moreover, untreated inoculated control results highest pathogenic potential in reduction of chlorophyll and carotenoid content (1.12 mg/g, 0.270 mg/g) (Table-34b).
Root knot nematode infestation significantly reduced in all the treatments. P. chlamydosporia+ castor cake recorded most significant impact in the reduction of root knot nematode in eggmasses/root, egg/eggmass, nematode population and root knot indices were 58, 109, 590, 1.4 as compared to untreated inoculated control plants (176, 266, 1608, 5).However, application of P. chlamydosporia + cotton results in least reduction of root knot nematode severity (77, 147, 662, 2.4) (Table-34b).
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Experimental Results
4.9.3. Effect of bioinoculant Pseudomonas fluorescens in combination with different organic soil amendments on the root-knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21 in pots
This experiment was aimed to clarify the potential of P. fluorescens integrated with organic soil amendment viz., press mud, cotton cake, mahua cake, mustard cake, castor cake and soybean cake on root knot nematode infestation due to Meloidogyne incognita and plant growth characters of tomato cv.K-21.From the results it was reported that all the treatments significantly enhanced the plant growth parameters and reduced the nematode severity in tomato (Table-35a).
Different plant growth parameters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield significantly enhanced in all the treatments compared to untreated inoculated control plants (38.5cm, 31.5g, 9.3 g, 47.2 %, 154 g). Highest enhancement in plant growth characters was observed when Pseudomonas fluorescens was applied in combination with castor cake were 77.4 cm, 69.6 g, 22.8 g, 82.5 %, 312 g. It was followed by P. fluorescens+ press mud 75.6 cm, 67.4 g, 22.2 g, 81.6 %, 306 g; P. fluorescens + mustard cake 74.5 cm, 67 g, 21.4 g, 81 %, 302 g. While the least was observed in P. fluorescens + cotton cake were 72.2 cm, 65.7 g, 20.2 g, 78.4 %, 292 g statistically significant in promoting growth of tomato cv.K-21 (Table-35a,b).
The total chlorophyll and carotenoid content was significantly enhanced in all the treatments. Combined application of P. fluorescens+ castor cake showed highest enhancement in chlorophyll and carotenoid content were 2.38 mg/g, 0.800 mg/g; P. fluorescens+ press mud were 2.30 mg/g, 0.385 mg/g; P. fluorescens + mustard cake were 2.24 mg/g, 0.778 mg/g. While the least among the treated was observed in P. fluorescens + cotton cake (72.2 cm, 65.7 g, 20.2 g, 78.4 %, 292 g) as compared to untreated inoculated control plants(38.5 cm, 31.5 g, 9.3 g, 47.2 %, 154 g) (Table-35b).
The root knot nematode, represent highly noxious effect on the tomato in untreated inoculated control plants where root knot severity in eggmasses/root, egg/eggmass, nematode population and root knot indices were 176, 266, 1608, 5. As compared to untreated control plants, P. fluorescens+ castor cake showed minimum average number of eggmass/root, , egg/eggmass, nematode population and root knot indices were 55, 104, 582, 1.3; P. fluorescens+ press mud 59, 110, 594, 1.4; P.
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Experimental Results fluorescens + mustard cake 62, 118, 614, 1.6.While in treated plants the maximum was recorded in P. fluorescens + cotton treatment (73, 145, 644, 2.3) (Table-35b).
4.9.4. Effect of bioinoculant Purpureocillium lilacinum in combination with composted plant straws and Potato waste on the root-knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21 in pots
This experiment was conducted on the same lines as given in 4.9.4. to evaluate the effect of bioinoculants amended with plant straw and potato waste on the root knot indices caused by Meloidogyne incognita and plant growth parameters of tomato cv.K-21. Root knot nematode infestation significantly reduced by the amendment of Purpureocillium lilacinum with straw and waste thereby improved the plant growth of tomato cv.K-21 (Table-36a).
Various plant growth parameters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield significantly improved all the treatments compared to untreated inoculated control plants (38.5 cm, 31.5 g, 9.3g, 47.2 %, 152 g). Most pronounced improvement in plant growth characters was observed when P. lilacinum was applied in combination with mustard straw were 74.2 cm, 66.2 g, 21.3 g, 80.5 %, 300 g; P. lilacinum + potato waste 72.6cm, 65.0 g, 20.6 g, 79.6 %, 294 g; P. lilacinum + maize straw 71.4 cm, 64 g, 20 g, 78.7 %, 289 g. While the least was observed in P. lilacinum + pearl millet were 67.5 cm, 61 g, 18.2 g, 76 %, 273 g statistically significant in promoting growth of tomato (Table-36a, b).
Bioagents when applied in combination with various straws and potato waste significantly enhanced the biochemical parameters. Maximum increase in chlorophyll and carotenoid was observed in P. lilacinum + mustard straw were 2.28 mg/g, 0.785 mg/g; P. lilacinum + potato waste 2.20 mg/g, 0.744 mg/g; P. lilacinum + maize straw 2.09 mg/g, 0.760mg/g. However, minimum was represented by P. lilacinum + pearl millet (1.86 mg/g, 0.728mg/g). Moreover, untreated inoculated control caused maximum pathogenic behaviour in reduction of chlorophyll and carotenoid content (1.12 mg/g, 0.270 mg/g) (Table-36b).
All the treatments result significant reduction in the root knot nematode infestation in terms of eggmasses/root, egg/eggmass, and nematode population and
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Experimental Results root knot indices. Minimum root knot nematode infestation was found when the combined application of P. lilacinum + mustard straw was 65, 120, 600, and 1.7. It was followed by P. lilacinum+ potato waste 70, 129, 615, 1.9 followed by P. lilacinum + maize waste 73, 134, 634, 2.0 as compared to as compared to the untreated inoculated control plants (174, 266, 1608, 5.0). However, the maximum number of eggmasses/root, egg/eggmass, and nematode population and root knot indices was observed when P. lilacinum + pearl millet (84, 163, 685, 3.0) (Table- 36b).
The root knot nematode, represent highly noxious effect on the tomato in untreated inoculated control plants where root knot severity in eggmasses/root, egg/eggmass, nematode population and root knot indices were 176, 266, 1608, 5 as compared to untreated control plants, P. fluorescens+ castor cake showed minimum average number of eggmass/root, , egg/eggmass, nematode population and root knot indices were 55, 104, 582, 1.3P. fluorescens + press mud 59, 110, 594, 1.4; P. fluorescens + mustard cake 62, 118, 614, 1.6.While in treated the maximum was recorded in P. fluorescens + cotton treatment (73, 145, 644, 2.3) (Table-36b).
4.9.5. Effect of bioinoculant Pochonia chlamydosporia in combination with composted plant straws and Potato waste on the root-knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21 in pots
This experiment was carried out on the same procedure to elucidate the effect of bioinoculats amended with plant straw and potato waste supplemented with autoclaved soil on the root knot indices caused by Meloidogyne incognita and plant growth characters of tomato cv.K-21 (Table-37a).
The application of various treatments reduced the root knot nematode incidence thereby affect the plant growth parameters viz., plant length, fresh, dry weight, percent pollen fertility and yield of tomato cv.K-21. However, among all the treatments tested Pochonia chlamydosporia in combination with mustard straw was recorded to be most effective in enhancing the plant growth parameters viz., plant length, fresh, dry weight, percent pollen fertility and yield were 75.5 cm 67.6 g, 22 g, 81.2 %, 306 g respectively. It was followed by P. chlamydosporia + potato waste (74 cm, 66.4 g, 21.3 g, 80 %, 300 g), P. chlamydosporia + maize straw (72.2 cm, 65.4 g,
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Experimental Results
20.7 g, 79 %, 295 g). Moreover, P. chlamydosporia + pearl millet results in lowest improvement (69 cm, 62 g, 19 g, 76.6 %, 280 g) as compared to the untreated inoculated control plants (38.5 cm, 31.5 g, 9.3 g, 47.2 %, 152 g) and untreated uninoculated control plants (81.2 cm, 73 g, 26.4 g, 89.6 %, 356 g) (Table-37a,b).
The root knot nematode caused pathogenic effect on tomato cv.K-21 in untreated inoculated control plants in which root knot nematode severity viz., eggmasses/root, egg/eggmass, and nematode population and root knot indices were 174, 266, 1608, 5.0 . P. chlamydosporia +mustard straw in combination caused least infestation viz., eggmasses/root, egg/eggmass, and nematode population and root knot indices were 61, 114, 594, 1.7 as against to the untreated inoculated plants. It was followed by (66, 125, 608, 1.9) and (70, 132, 624, 2.3) in P. chlamydosporia + potato waste and P. chlamydosporia + maize straw in combination respectively. However, maximum infestation eggmasses/root, egg/eggmass and nematode population and root knot indices (79, 156, 670, 2.9) was shown by P. chlamydosporia + pearl millet as compared to the untreated inoculated (174, 266, 1608, 5) (Table-37b).
The chlorophyll and carotenoid significantly improved in all the treatments. Application of P. chlamydosporia + mustard straw results in paramount improvement in chlorophyll and carotenoid content were 2.33 mg/g, 0.790 mg/g; P. chlamydosporia + potato waste 2.24 mg/g, 0. 776 mg/g; P. chlamydosporia + maize straw 2.19 mg/g, 0.766 mg/g. However, the least was observed in P. chlamydosporia + pearl millet (192 mg/g, 0.738 mg/g) as compared to untreated inoculated 1.12& 0.272 mg/g and untreated uninoculated were 2.79& 0.878 mg/g (Table-37b).
4.9.6. Effect of bioinoculant Pseudomonas fluorescens in combination with composted plant straws and Potato waste on the root-knot development caused by Meloidogyne incognita and plant growth of tomato cv. K-21 in pots
This experiment was conducted under glasshouse conditions to test the nematicidal potential of P. fluorescens integrating with plant straw of Pigeon pea, Pearl millet Maize, Mustard, Sorghum and potato waste on plant growth parameters and root knot nematode reproduction and nematode severity. Application of Pseudomonas fluorescens in combination with composted plant straw and potato
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Experimental Results waste significantly enhance the plant growth parameters results in reduction in nematode infestation in tomato cv.K-21 (Table-38a).
Significant enhancement in the plant growth parameters (plant length, fresh, dry weight, percent pollen fertility, yield) was observed due to the various treatments. Among all the treatments the best plant growth characters viz., plant length, fresh weight, dry weight, percent pollen fertility and yield were 76.7 cm, 68.6 g, 22.5 g, 82 %, 309 g when the pots were treated with P. fluorescens in combination with mustard straw. It was followed by P. fluorescens+ potato waste were 75 cm, 67.3 g, 22 g, 80.7 %, 303 g and P. fluorescens + maize straw were 73.8 cm, 66 g, 21.6 g, 80.3 %, 298 g and least in P. fluorescens+ pearl millet were 70.5 cm, 63.4 g, 20.6 g, 78 % 289 g as against to the untreated inoculated control plants (38.5cm, 31.5 g, 9.3 g, 47.2 %, 152 g) (Table-36a) (Table-38a,b).
Biochemical parameters viz., chlorophyll and carotenoid content was significantly improved in all the treatment were 2.36 and 0.796, 2.28 and 0.789, 2.22& 0.780 mg/g in P. fluorescens + mustard straw, P. fluorescens+ potato waste, P. fluorescens + maize straw. However, the least was observed in P. fluorescens+ pearl millet 1.98 and 0.749mg/g as compared to the untreated inoculated 1.12& 0.272 mg/g and untreated uninoculated were 2.79 and 0.878mg/g (Table-38b).
The root knot nematode infestations viz., eggmasses/root, egg/eggmass, nematode population and root knot indices was significantly suppressed through the application of various treatments in combinations as compared to untreated inoculated control (174, 266, 1608, 5.0). The combined application of P. fluorescens + mustard straw results in pronounced suppression of eggmasses/root, egg/eggmass, and nematode population and root knot indices were 57, 106, 585, 1.4; P. fluorescens+ potato waste were 60, 112, 596, 1.7; P. fluorescens + maize straw were 65, 120, 607, 2.1.While the lowest suppression was observed in P. fluorescens+ pearl millet (75, 147, 650, 2.7 ) as compared to the as compared to the untreated inoculated (174, 266, 1608, 5) (Table-38b).
135
Table 1a: Effect of root-knot nematode, Meloidogyne incognita on different cultivars of tomato in relation to plant growth parameters in pots Cultivar Treatment Length Fresh Dry %reduction Chlorophyll Carotenoid Yield(g) (cm) weight(g) weight(g) over control content(mg/g) content(mg/g) EC-538380 UUC 67.4hij 59.7gh 18.5kl - 2.65cdef 0.886ef 337fg UIC 42.3n 37.9m 11.5qr 37.84 1.73kl 0.576n 216l K-21 UUC 77.6bcd 71.0ab 24.0bc - 2.72abcd 0.869gh 386bc UIC 39.0o 32.4n 11.3qr 52.92 1.45mn 0.285r 182n CO-3 UUC 72.0efg 64.2ef 20.7fgh - 2.55fg 0.880fg 343f UIC 54.5l 47.3jk 15.5no 25.13 1.93ij 0.673k 258jk FEB-02 UUC 74.3de 65.5de 21.2fg - 2.59defg 0.882fg 345f UIC 58.5k 50.0j 16.7mn 21.23 2.00i 0.686k 269j EC-570018 UUC 78.5abc 69.0bc 22.9cde - 2.69abcd 0.912cd 372cd UIC 70.5fgh 62.1fg 20.0ghi 12.67 2.47g 0.800i 327g NDT-3 UUC 69.2ghi 62.4fg 19.7hijk - 2.69abcd 0.896ef 341fg UIC 46.2m 42.7l 13.2p 33.0 1.82jk 0.608m 226kl S-22 UUC 67.0hij 59.5gh 18.0lm - 2.55fg 0.862h 339fg UIC 36.2o 31.2n 9.2s 48.89 1.38n 0.432q 177no GT-1 UUC 68.2hij 60.0gh 19.2jkl - 2.58efg 0.869gh 334fg UIC 37.4o 32.3n 10.3rs 46.36 1.52m 0.490p 186n GT-2 UUC 77.3bcd 67.7cd 22.0ef - 2.64cdef 0.902de 365de UIC 65.2j 56.5i 18.7jkl 15.0 2.26h 0.770j 309hi GT-3 UUC 70.0fgh 63.0ef 20.0ghi - 2.69abcd 0.882 332fg UIC 45.1mn 38.5m 12.2pq 39.0 1.66l 0.554o 204lm H-88-78-1 UUC 82.0a 73.0a 25.5a - 2.82a 0.938a 394a UIC 78.0bc 69.5bc 24.3ab 4.71 2.70abcd 0.889ef 373cd PB Barkha bahar-2 UUC 80.7ab 72.0ab 24.0bc - 2.78ab 0.930ab 389ab UIC 66.3ij 58.7hi 19.5ijk 18.75 2.30h 0.768j 320gh VRT-101A UUC 80.0ab 71.6ab 24.4ab - 2.74abc 0.920bc 382bc UIC 75.0cde 67.2cd 22.8cde 6.56 2.53fg 0.862h 351ef Kalyanpuri-T1 UUC 73.0ef 64.8def 21.0fg - 2.64cdef 0.890ef 348ef UIC 53.2l 46.5k 15.0o 28.58 1.91ij 0.652l 251jk Each value is the mean of four replicates; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC-Untreated Inoculated Control. Table 1b: Effect of root-knot nematode, Meloidogyne incognita on different cultivars of tomato in relation to nematode development and nematode multiplication in pots Cultivar Treatment Pollen fertility Eggmasses/ Egg/eggmass Nematode Root knot Reaction (%) root population(250g) Index EC-538380 UUC 87.8abc - - - - S UIC 62.3i 69e 135e 1290e 4.1cd K-21 UUC 89.0abc - - - - HS UIC 48.2l 176a 262a 1632a 5.0a CO-3 UUC 87.2abc - - - - MS UIC 69.0g 42g 72h 1005h 3.6e FEB-02 UUC 87.3abc - - - - MS UIC 69.5g 34h 56i 922i 3.5e EC-570018 UUC 88.2abc - - - - MR UIC 81.0e 13jk 19k 695l 2.2g NDT-3 UUC 86.5 - - - - S UIC 64.5hi 64e 121f 1246f 4.2bcd S-22 UUC 85.8bcd - - - - S UIC 49.7l 128b 223b 1462b 4.6b GT-1 UUC 87.0abcd - - - - S UIC 53.5k 94c 178c 1430c 4.5b GT-2 UUC 86.0abcd - - - - MR UIC 75.5f 20ij 29j 744k 2.5fg GT-3 UUC 85.3cd - - - - S UIC 57.5j 82d 150d 1350d 4.3bcd H-88-78-1 UUC 90.2a - - - - R UIC 85.4bcd 4.0l 6.0l 235n 1.2h PB Barkha bahar-2 UUC 89.6ab - - - - MR UIC 74.6f 24i 36j 814j 2.7f VRT-101 UUC 89.2abc - - - - R UIC 83.0de 6.0kl 9.0l 328m 1.4h Kalyanpuri-T1 UUC 86.5abcd - - - - MS UIC 66.2gh 53f 94g 1075g 3.8de Each value is the mean of four replicates; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table2: Effect of aqueous leaf extracts of some selected plants species against egghatching of Meloidogyne incognita in vitro
Number of juvenile emerged in different concentrations(Within 5 days) Treatment S S/2 S/10 S/100 DW Indian mallow 5(99.10) 36(93.6) 295(47.32) 486(13.21) 560 Mexican poppy 0(100) 0(100) 185(66.96) 402(28.21) 560 Ivy gourd 12(97.85) 44(92.14) 386(31.10) 514(8.21) 560 Trailing eclipta 09(100) 2(99.64) 210(62.5) 430(23.21) 560 Wild eggplant 0(100) 0(100) 238(57.5) 445(20.54) 560 Black pig weed 0(100) 17(96.96) 270(51.79) 462(17.5) 560 Each value is the mean of four replicates; S= Standard extract; S/2, S/10, S/100 are dilutions of S; DW=Distilled water Values given in parentheses represent percent inhibition in juvenile emergence over distilled water control.
100 100 100 80 80 60 60 50 40 40 20 20 0 0 0 S S/2 S/10 S/100 DW S S/2 S/10 S/100 DW Trailing eclipta Indian mallow Ivy gourd
100 100 100 80 80 80 60 60 60 Percent inhibition inhibition Percent 40 40 40
20 20 20 0 0 0 S S/2 S/10 S/100 DW S S/2 S/10 S/100 DW Wild eggplant Mexican poppy Black pig weed
Treatments
Fig.1. Effect of different concentrations of aqueous extracts of leaves of some selected plants on the hatching of Meloidogyne incognita eggs in vitro.
Table3: Effect of different aqueous dilutions of various oil cakes against egghatching of Meloidogyne incognita in vitro
Number of juvenile emerged in different concentrations(Within 5 days) Treatment S S/2 S/10 S/100 DW Castor cake 0(100) 30(94.4) 190(64.5) 440(17.75) 535 Cotton cake 20(96.26) 97(82.06) 300(43.93) 510(4.67) 535 Mahua cake 12(97.8) 85(84.11) 276(48.41) 495(7.5) 535 Mustard cake 0(100) 50(90.65) 216(59.63) 457(14.58) 535 Soybean cake 2(99.63) 76(85.8) 242((54.8) 478(10.65) 535 Each value is the mean of four replicates; S= Standard extract; S/2, S/10, S/100 are dilutions of S; DW=Distilled water Values given in parentheses represent percent inhibition in juvenile emergence over distilled water control.
100 100 100 80 80 80 60 60 60 40 40 40 20 20 20 0 0 0 S S/2 S/10 S/100 DW S S/2 S/10 S/100 DW Mahua cake Castor cake Cotton cake
100 100 80 80 60 60
Percent inhibition inhibition Percent 40 40
20 20 0 0 S S/2 S/10 S/100 DW S S/2 S/10 S/100 DW Mustard cake Soybean cake
Treatments
Fig.2.Effect of different concentrations of aqueous extracts of various oil cakes on the hatching of Meloidogyne incognita eggs in vitro
Table4: Effect of various dilutions of different bioinoculants against egghatching of Meloidogyne incognita in vitro Number of juvenile emerged in different concentrations(Within 5 days) Treatment S S/2 S/10 S/100 DW Purpureocillium lilacinum 0(100) 0(100) 115(79.46) 382(31.79) 560 Pochonia chlamydosporia 0(100) 0(100) 100(82.14) 360(35.71) 560 Pseudomonas fluorescens 0(100) 26(95.36) 158(71.79) 400(28.75) 560 Trichoderma viride 0(100) 2(99.64) 190(66.07) 421(24.82) 560 Each value is the mean of four replicates; S= Standard extract; S/2, S/10, S/100 are dilutions of S; DW=Distilled water Values given in parentheses represent percent inhibition in juvenile emergence over distilled water control.
100 100 80 80 60 60 40 40 20 20 0 0 S S/2 S/10 S/100 DW S S/2 S/10 S/100 DW Purpureocillium lilacinum Pochonia chlamydosporia
100 100 80 80
Percent inhibition inhibition Percent 60 60 40 40 20 20 0 S S/2 S/10 S/100 DW 0 S S/2 S/10 S/100 DW Trichoderma viride Pseudomonas fluorescens
Treatments
Fig.3. Effect of different concentrations of biocontrol agents on the hatching of Meloidogyne incognita eggs in vitro.
Table5: Effect of various dilutions of biochar and aquatic weed water hyacinth against egghatching of Meloidogyne incognita in vitro Number of juvenile emerged in different concentrations(Within 5 days) Treatment S S/12 S/10 S/100 DW Biochar 18(96.64) 96(82.06) 294(45.05) 475(11.21) 535 Water hyacinth 0(100) 26(95.14) 230(57.0) 417(22.05) 535 Each value is the mean of four replicates; S= Standard extract; S/2, S/10, S/100 are dilutions of S; DW=Distilled water Values given in parentheses represent percent inhibition in juvenile emergence over distilled water control
100 90 80 70 60 50 40 30 20 10 0 S S/2 S/10 S/100 DW Biochar
inhibition
100 90 80 Percent 70 60 50
40 30 20 10 0 S S/2 S/10 S/100 DW Water hyacinth
Treatments
Fig.4. Effect of different concentrations of Biochar and Water hyacinth on the hatching of Meloidogyne incognita eggs in vitro.
Table 6: Effect of aqueous extract of different plants on the mortality of root knot nematode, Meloidogyne incognita juveniles (J2) in vitro Exposure Juvenile(J2) mortality in various dilutions Regression equation Treatment period S S/2 S/10 S/100 DW (Hrs) Indian mallow 12 72(76.2) 60(56.5) 45(36.8) 7(17.10) 0(-2.5) Y=36.8+19.7(X-2) 24 88(91.8) 72(67.9) 51(44.0) 9(20.1) 0(-3.8) Y=44.0+23.9(X-2) 48 94(101.8) 85(75.7) 57(49.6) 12(23.5) 0(-2.6) Y=49.6+26.1(X-2) Mexican poppy 12 92(103.4) 87(77.7) 63(52.0) 18(26.3) 0(0.6) Y=52.0+25.7(X-2) 24 100(111.6) 94(83.7) 70(55.8) 15(27.9) 0(0) Y=55.8+29.7(X-2) 48 100(123.32 100(90.36) 74(57.4) 13(24.44) 0(-8.52) Y=57.4+32.96(X-2) Ivy gourd 12 65(68.0) 54(50.1) 37(32.2) 5(14.3) 0(-3.6) Y=32.2+17.9(X-2) 24 71(76.8) 62(57.0) 45(37.2) 8(17.4) 0(-2.4) Y=37.2+19.8(X-2) 48 80(86.4) 70(64.4) 52(42.4) 10(20.4) 0(-1.6) Y=42.4+22.0(X-2) Trailing eclipta 12 89(97.8) 82(72.9) 58(48.0) 11(23.1) 0(-1.8) Y=48.0+24.9(X-2) 24 100(109.2) 91(81.4) 64(53.6) 13(25.8) 0(-2.0) Y=53.6+27.8(X-2) 48 100(113.0) 98(84.7) 69(56.4) 15(28.1) 0(-0.2) Y=56.4+28.3(X-2) Wild eggplant 12 84(92.8) 78(69.5) 56(46.2) 13(22.9) 0(-0.4) Y=46.2+23.3(X-2) 24 97(105.6) 87(78.9) 63(52.2) 14(25.5) 0(-1.2) Y=52.2+26.7(X-2) 48 100(110.6) 93(82.9) 67(55.2) 16(27.5) 0(-0.8) Y=55.2+27.7(X-2) Black pigweed 12 80(88.2) 75(65.6) 51(43.0) 9(20.4) 0(-2.2) Y=43.0+22.6(X-2) 24 92(100.4) 84(74.7) 58(49.0) 11(23.3) 0(-2.4) Y=49.0+25.7(X-2) 48 100(109.0) 91(81.2) 63(53.4) 13(25.6) 0(-2.2) Y=53.4+27.8(X-2) Each value is the mean of four replicates; S = Standard Concentrations S/2, S/10 and S/100 are dilutions of S; DW = Distilled Water Values given in parentheses represent nematode mortality (%) calculated from regression equations.
Lower Line(12hrs) Middle Line (24hrs) Upper Line (48hrs) 120 150 100 Indian mallow Mexican poppy Ivy gourd 100 80 80 100 60
(J2) (J2) 60 40 50 40 20 20 0 0 DW S/100 S/10 S/2 S DW S/100 S/10 S/2 S 0 -20 DW S/100 S/10 S/2 S -50 -20 120 120 120 Trailing eclipta Wild eggplant Black pig weed
Meloidogyne incognita Meloidogyne 100 100 100 80 80 80 60 60 60 40 40 40 20 20 20 0 0 0 -20 DW S/100 S/10 S/2 S -20 DW S/100 S/10 S/2 S -20 DW S/100 S/10 S/2 S Percent mortality of of mortality Percent Concentrations of different leaf extracts Fig.5. Regression lines showing linear relationship between different concentrations of aqueous extracts of various plants and mortality of Meloidogyne incognita juveniles (J2)
Table7: Effect of aqueous extract of various oil cakes against root knot nematode, Meloidogyne incognita juveniles (J2) in vitro Exposure Juvenile(J2) mortality in various dilutions Regression equation Treatment period S S/2 S/10 S/100 DW (Hrs) Castor cake 12 85(94.2) 79(70.9) 58(47.6) 16(24.3) 0(1.0) Y=47.6+23.3(X-2) 24 96(106.8) 90(80.4) 66(54.0) 18(27.6) 0(1.2) Y=54.0+26.4(X-2) 48 100(112.2) 95(84.7) 71(57.2) 20(29.7) 0(2.2) Y=57.2+27.5(X-2) Cotton cake 12 70(77.2) 65(57.3) 46(37.4) 6(17.5) 0(-2.4) Y=37.4+19.9(X-2) 24 76(85.2) 72(63.6) 54(42) 8(20.4) 0(-1.2) Y=45.0+21.6(X-2) 48 89(97.4) 80(72.2) 60(48.0) 11(23.3) 0(-1.4) Y=48.0+24.7(X-2) Mahua cake 12 72(80.2) 67(59.9) 51(39.6) 8(19.3) 0(-1.0) Y=39.6+20.3(X-2) 24 82(89.4) 70(67.2) 61(45.0) 12(22.8) 0(0.6) Y=45.0+22.2(X-2) 48 91(101) 83(75.9) 66(50.8) 14(25.7) 0(0.6) Y=50.8+25.1(X-2) Mustard cake 12 82(90.6) 77(67.9) 53(45.2) 14(22.7) 0(0) Y=45.2+22.7(X-2) 24 92(102.6) 88(76.9) 61(51.2) 15(25.5) 0(-0.2) Y=51.2+25.7(X-2) 48 100(110.8) 93(83.3) 68(55.8) 18(28.3) 0(0.8) Y=55.8+27.5(X-2) Soybean cake 12 76(84.0) 72(62.6) 48(41.2) 10(19.8) 0(-1.60) Y=41.2+21.4(X-2) 24 87(95.6) 79(71.6) 59(47.6) 13(23.6) 0(-0.4) Y=47.6+24.0(X-2) 48 94(104.4) 88(78.4) 64(52.4) 16(26.4) 0(0.4) Y=52.4+26.0(X-2) Each value is the mean of four replicates; S = Standard Concentrations S/2, S/10 and S/100 are dilutions of S; DW = Distilled Water Values given in parentheses represent nematode mortality (%) calculated from regression equations.
Lower Line (12hrs) Middle Line (24hrs) Upper Line (48hrs) 120 120 120 Castor cake Cotton cake Mustard cake 100 100 100
80 80 80 2)
(J 60 60 60 40 40 40 20 20 20 0 0 0 -20 DW S/100 S/10 S/2 S DW S/100 S/10 S/2 S DW S/100 S/10 S/2 S -20
120 120 Mahua cake Soybean cake
Meloidogyne incognita Meloidogyne 100 100 80 80 60 60 40 40 20 20 0 0
Percent mortality of of mortality Percent DW S/100 S/10 S/2 S DW S/100 S/10 S/2 S -20 -20 Concentrations of various dilutions of oil cakes Fig.6. Regression lines showing linear relationship between different concentrations of aqueous extracts of oil cakes and mortality of Meloidogyne incognita juveniles (J2)
Table 8: Effect of aqueous extract of various biocontrol agents against root knot nematode, Meloidogyne incognita juveniles (J2) in vitro Treatment Exposure Juvenile(J2) mortality in various dilutions Regression Equation period S S/2 S/10 S/100 DW (Hrs) Purpureocillium lilacinum 12 98(104.8) 85(77.9) 60(51.0) 12(24.1) 0(-2.8) Y=51.0+26.9(X-2)
24 100(110.8) 92(82.8) 69(55.0) 14(27.2) 0(-0.6) Y=55.0+27.8(X-2)
48 100(115.6) 100(87.4) 78(59.2) 18(31.0) 0(-2.8) Y=59.2+28.2(X-2)
Pochonia chlamydosporia 12 100(108.2) 88(81.2) 65(54.2) 18(27.2) 0(-0.2) Y=54.2+27.0(X-2)
24 100(111.0) 96(86.0) 75(58.8) 23(31.5) 0(-4.2) Y=58.8+27.3(X-2)
48 100(116.6) 100(89.2) 83(61.8) 26((34.4) 0(-7.0) Y=61.8+27.4(X-2)
Pseudomonas fluorescens 12 95(100.8) 82(75.0) 55(49.2) 14(23.4) 0(-2.4) Y=49.2+25.8(X-2)
24 100(109.2) 91(81.9) 64(54.6) 18(27.3) 0(0) Y=54.6+27.3(X-2)
48 100(114.6) 99(87.1) 75(59.6) 24(32.1) 0(-4.6) Y=59.6+27.5(X-2)
Trichoderma viride 12 89(105.8) 80(71.1) 52(46.4) 11(21.7) 0(-3.0) Y=46.4+24.7(X-2)
24 96(105.8) 88(79.3) 65(52.8) 15(26.3) 0(-0.2) Y=52.8+26.5(X-2)
48 100(112.6) 96(84.7) 71(56.8) 17(28.9) 0(-1) Y=56.8+27.9(X-2)
Each value is the mean of four replicates; S = Standard Concentrations S/2, S/10 and S/100 are dilutions of S; DW = Distilled Water Values given in parentheses represent nematode mortality (%) calculated from regression equations.
Lower Line (12hrs) Middle Line (24hrs) Upper Line (48hrs)
150 150 Purpureocillium lilacinum Pochonia chlamydosporium
100 100 (J2) (J2) 50 50
0 0 DW S/100 S/10 S/2 S DW S/100 S/10 S/2 S -50 -50
120 Trichoderma viride 140 Pseudomonas fluorescens 100 120 Meloidogyne incognita Meloidogyne 80 100 80 60 60 40 40 20 20 0 0 -20 DW S/100 S/10 S/2 S -20 DW S/100 S/10 S/2 S
Percent mortality of of mortality Percent Concentrations of various dilutions of culture filtrate Fig.7. Regression lines showing linear relationship between different concentrations of aqueous extracts of culture filtrate and mortality of Meloidogyne incognita juveniles (J2)
Table9: Effect of various dilutions of biochar and aquatic weed Water hyacinth against root knot nematode, Meloidogyne incognita juveniles (J2) in vitro Exposure Juvenile(J2) mortality in various dilutions period Regression Equation Treatment S S/2 S/10 S/100 DW (Hrs) 12 91(89.6) 66(69.80) 41(40.4) 04(16.00) 0(-8.40) Y=40.4+24.4(X-2) Biochar 24 96(99.2) 78(72.90) 52(46.6) 07(20.30) 0(-6.00) Y=46.6+26.3(X-2) 48 100(108) 90(80.00) 60(52.0) 10(24.00) 0(-4.00) Y=52.0+28.0(X-2) 12 92(105.7) 83(76.06) 50(46.4) 07(16.74) 0(-12.92) Y=46.4+29.66(X-2) Water hyacinth 24 100(109.4) 91(84.98) 64(53.2) 11(21.42) 0(-10.36) Y=53.2+31.7(X-2) 48 100(112.6) 96(84.40) 71(56.2) 14(28.00) 0(-0.20) Y=56.2+28.2(X-2) Each value is the mean of four replicates; S = Standard Concentrations S/2, S/10 and S/100 are dilutions of S; DW = Distilled Water Values given in parentheses represent nematode mortality (%) calculated from regression equations.
Lower Line(12hrs) Middle Line (24hrs) Upper Line(48hrs)
120 Biochar 100
80
60
(J2) (J2) 40
20
0 DW S/100 S/10 S/12 S -20
Meloidogyne incognita Meloidogyne 120 Water hyacinth 100
80
60
40
Percent mortality of of mortality Percent 20
0 DW S/100 S/10 S/2 S -20
Concentrations of various dilutions of Biochar and Water hyacinth
Fig.8. Regression lines showing linear relationship between different concentrations of aqueous extracts of biochar and Water hyacinth and mortality of Meloidogyne incognita juveniles (J2).
Table 23a: Effect of chopped leaves of different plant species in combination with leaf powder of Black nightshade on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight(g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Indian mallow 31.4ef 15.0cd 46.4de 25.7e 10.3bc 36.0ef 8.3de 3.2cd 11.5ef Mexican poppy 36. b 17.3b 53.5b 29.0b 12.0b 41.0b 10.0b 4.2b 14.2b Ivy gourd 30.5f 14.2d 44.7 e 25.2e 10.0c 35.2f 8.0e 3.0d 11.0f Trailing eclipta 35.0bc 16.7bc 51.7bc 28.0bc 11.6 b 39.6bc 9.5bc 4.0b 13.5bc Wild eggplant 33.8cd 16.2bc 50.0bcd 27.2cd 11.2b 38.4cd 9.2bc 3.7bc 12.9cd Black pigweed 32.4de 15.8bc 48.2cde 26.de 10.7bc 37.2 de 8.8cd 3.5bc 12.3de UUC 53.6a 24.6a 78.2 a 52.0a 20.0a 72.0a 18.0 a 6.0a 24.0a UIC 24.7g 11.8e 36.5f 20.0f 8.0d 28.0g 7.1f 1.4g 8.5g
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 23b: Effect of chopped leaves of different plant species in combination with leaf powder of Black nightshade on tomato cv. K-21 in relation to root knot development and multiplication of Meloidogyne incognita in pots
Chlorophyll Carotenoid Pollen Yield Eggmasses/ Eggs/ Nematode Root Treatment content content fertility /plant(g) root Eggmass Population Knot (mg/g) (mg/g) (%) (250g) Index Indian mallow 1.51cd 0.400 e 62.0c 160ef 132bc 209bc 1170 c 2.8bc Mexican poppy 1.73 b 0.495 b 67.0b 228b 114e 188e 1042 g 2.0d Ivy gourd 1.45 d 0.390f 61.0c 144g 140 b 215 b 1204 b 3.0b Trailing eclipta 1.68b 0.474 c 65.6b 200 c 118de 193de 1069 f 2.2cd Wild eggplant 1.62bc 0.446 d 64.0bc 186 d 123cd 197de 1100e 2.3cd Black pigweed 1.55cd 0.418 e 63.5bc 167e 128c 202cd 1142 d 2.5bcd UIC 1.12e 0.252g 47.0f 112.0g 175 a 266 a 1608 a 5.0 a UUC 2.78 a 0.880 a 89.5 a 355 a 0 f 0 f 0 g 0 e
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table24a: Effect of Biochar alone and in combination with sawdust of different plants on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots
Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Biochar 43.0f 20.5e 63.5g 38.8g 16.4f 55.2f 14.2d 4.2e 18.4e Biochar+ Eucalyptus Sawdust 50.8b 24.6b 75.4b 47.0b 20.2b 67.2b 16.7b 5.2b 21.9b Biochar+Jambul Sawdust 49.0bc 23.6bcd 72.6cd 44.0d 19.0cd 63.0cd 15.9cd 4.8bcd 20.7bcd Biochar+ Babool Sawdust 45.0e 22.0de 67.0f 40.7f 17.9e 58.6ef 15.0cd 4.3de 19.3ef Biochar+ Mango Sawdust 47.7cd 23.1bcd 70.8de 42.8e 18.6de 61.4cde 15.6bcd 4.6cde 20.2cde Biochar+ Poplar Sawdust 46.4de 22.6cd 69.0ef 42.0e 18.2de 60.2de 15.2cd 4.5cde 19.7de Biochar+ Lebbeck Sawdust 50.0b 24.2bc 74.2bc 45.3c 19.7bc 65.0bc 16.3bc 5.0bc 21.3bc UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2g 12.3f 38.5h 23.0h 8.5g 31.5g 7.9e 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 24b: Effect of Biochar alone and in combination with sawdust of different plants on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Nematode Root Eggmasses/ Eggs/ fertility plant content content population knot Treatment root eggmass (%) (g) (mg/g) (mg/g) (250g) Index Biochar 71.0f 240e 1.84f 0.680f 106b 178b 955b 3.2b Biochar+ Eucalyptus Sawdust 81.0b 302b 2.30b 0.786b 61g 115g 597g 1.5f Biochar+Jambul Sawdust 79.0bcd 294bc 2.17c 0.770c 68ef 129f 620d 1.9de Biochar+ Babool Sawdust 75.5e 277d 1.89f 0.739e 83c 159c 675c 3.0bc Biochar +Mango Sawdust 78.0cde 290bc 2.07d 0.762c 72de 138e 635e 2.2d Biochar+ Poplar Sawdust 76.7de 285cd 1.98e 0.751d 77d 147d 653d 2.7c Biochar+ Lebbeck Sawdust 79.6b 299b 2.24bc 0.779b 65fg 121g 608f 1.7ef UUC 89.6a 356a 2.79a 0.878a 0h 0 0h 0g UIC 47.2g 158f 1.12g 0.273g 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 25a: Effect of biochar alone and in combination with agriculture waste on the growth of tomato cv. K-21 in relation to root- knot development caused by Meloidogyne incognita in pots Weight (g) Length(cm) Fresh Dry Treatment Shoot Root Total Shoot Root Total Shoot Root Total Biochar 43.0e 20.5g 63.5f 38.8f 16.4f 55.2f 14.2e 4.2e 18.4e Biochar + Bagasse 45.8d 22.4f 68.2e 41.8e 18.2e 60.0e 15.4d 4.6de 20.0d Biochar + Decomposed potato 48.8bc 23.5de 72.3cd 44.7cd 19.3bcd 64.0cd 15.9cd 4.9bde 20.8bcd Biochar + Garlic waste 50.4b 24.6bc 75.0bc 47.0b 19.8bc 66.8bc 16.4bc 5.2bc 21.6bc Biochar + Mentha waste 49.5bc 24.0cd 73.5bc 45.8bc 19.7bc 65.5bcd 16.1bcd 5.1bcd 21.2bcd Biochar + Tobacco waste 51.4b 25.0b 76.4b 47.7b 20.3b 68.0b 16.7b 5.4b 22.1b Biochar + Black gram waste 47.0cd 23.0ef 70.0de 43.5d 18.8de 62.3de 15.7cd 4.8cd 20.5cd UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2f 12.3h 38.5g 23.0g 8.5g 31.5g 7.9f 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 25b: Effect of biochar alone and in combination with agriculture waste on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Nematode Eggmasses/ Eggs/ Root knot fertility plant content content population root eggmass Index Treatment (%) (g) (mg/g) (mg/g) (250g) Biochar 71.0f 240e 1.84g 0.680f 106b 178b 955b 3.2b Biochar + Bagasse 76.7e 288cd 1.92f 0.742e 80c 152c 668c 2.9bc Biochar + Decomposed potato 78.5cde 293cd 2.12de 0.763d 72d 137e 642e 2.0de Biochar+ Garlic waste 80.6bc 300bc 2.25c 0.779c 61e 120g 610g 1.6ef Biochar + Mentha waste 79.7bcd 296cd 2.18d 0.772c 65e 129f 628f 1.9de Biochar + Tobacco waste 81.5b 306b 2.32b 0.790b 59e 112h 593h 1.4f Biochar + Black gram waste 77.5de 290cd 2.06e 0.756d 75cd 144d 655d 2.4cd UUC 89.6a 356a 2.79a 0.878a 0f 0i 0i 0g UIC 47.2g 158f 1.12h 0.269g 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 26a: Effect of biochar as a seed dressing agent on plant growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total 0.2%biochar 28.0de 13.0de 41.0e 24.5cd 9.2de 33.7e 9.0d 1.8de 10.8e 0.4%biochar 28.8de 13.2de 42.0de 25.0c 9.3de 34.3de 9.1d 1.9d 11.0de 0.8%biochar 30.2cde 13.8cd 44.0d 26.2c 9.8d 36.0d 9.4d 2.2cd 11.6de 1.6%biochar 32.1bcd 14.6c 46.7c 28.0c 11.0c 39.0c 9.9cd 2.5c 12.4cd 3.2%biochar 35.0bc 16.0b 51.0b 31.0b 12.6b 43.6b 10.7bc 3.0b 13.7bc 6.4%biochar 36.2b 16.4b 52.6b 32.0b 13.0b 45.0b 11.3b 3.3b 14.6b UUC 53.5a 24.5a 78.0a 52.0a 20.0a 72.0a 18.3a 6.2a 24.5a UIC 26.2e 12.3e 38.5f 23.0d 8.5e 31.5f 7.9e 1.4e 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 26b: Effect of biochar as a seed dressing agent on the plant growth of tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Yield/ Pollen Chlorophyll Carotenoid Eggmasses/ Eggs/ Nematode Root-knot Treatment plant fertility Content content root Eggmass population Index (g) (%) (mg/g) (mg/g) (250g) 0.2%biochar 121f 49.0e 1.19e 0.325g 168ab 250ab 1389b 4.8ab 0.4%biochar 148e 51.0de 1.30d 0.358f 152c 228c 1325c 4.6bc 0.8%biochar 162d 53.4d 1.39d 0.382e 144cd 224cd 1290d 4.3cd 1.6%biochar 180c 56.7c 1.50c 0.405d 138de 214de 1260e 3.9d 3.2%biochar 196b 60.0b 1.58bc 0.460c 133e 199ef 1152f 3.0e 6.4%biochar 204b 62.0b 1.64b 0.477b 130e 191f 1120g 2.8e UUC 355a 89.0a 2.89a 0.880a 0f 0g 0h 0f UIC 142g 47.0e 1.12e 0.275h 175a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 27a: Effect of seed dressing with various bioagents on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Fresh weight(g) Dry weight(g) Treatment Shoot Root Total Shoot Root Total Shoot Root Total 1% cellulose Pc 42.6bc 20.2bcd 62.8bcd 32.6bcd 13.9bcd 46.5bcd 12.7bcd 3.2cde 16.0bc 2% cellulose Pc 43.5b 20.7b 64.2b 33.7b 14.5b 48.2b 13.4b 3.6bc 17.0b 1% molasses Pc 41.7bcd 19.7bcde 61.4cde 32.3cde 13.7bcde 46.0bcde 12.4cde 3.1cde 15.5cd 2%mollases Pc 43.0bc 20.5bc 63.5bc 32.8bc 14.2bc 47.0bc 13.0bc 3.4bc 16.4bc 1% cellulose Pl 39.0fgh 18.6ghi 57.6gh 31.2efg 13.1bcde 44.3defg 11.3f 2.4g 13.7fg 2% cellulose Pl 41.4bcde 19.6cdef 61.0cdef 31.8bcde 13.4bcde 45.2cdef 12.7bcd 3.0def 15.7cd 1% molasses Pl 38.2ghi 18.3hi 56.5hi 30.7fgh 12.8cde 43.5fgh 11.1f 2.3g 13.4g 2% molasses Pl 40.3defg 19.2def 59.5efg 31.4efg 13.2bcde 44.6defg 11.5ef 2.5fg 14.0fg 1% cellulose Pf 41.0cdef 19.5cdef 60.5def 31.6def 13.4bcde 45.0cdef 12.3cde 3.0def 15.3cd 2% cellulose Pf 42.8bc 20.4bc 63.2bcd 32.6bcd 14.0bcd 46.6bcd 12.9bc 3.3cde 16.2bc 1% molasses Pf 40.3defg 19.1efg 59.4efg 31.4efg 13.3bcde 44.7defg 12.0def 2.8efg 14.8de 2% molasses Pf 42.0bcd 20.0bcde 62.0bcde 31.8bcde 13.6bcde 45.4cdef 12.6bcd 3.2cde 15.8cd 1% cellulose Tv 37.8hi 18.2hi 56.0hi 30.4gh 12.6de 43.0gh 11.3f 2.5fg 13.8fg 2% cellulose Tv 39.4efgh 19.0efg 58.4fgh 31.5efg 13.0bcde 44.5defg 13.0bc 4.4b 14.5def 1% molasses Tv 36.7i 17.8i 54.5i 29.7h 12.3e 42.0h 11.2f 2.4g 13.6fg 2% molasses Tv 38.6ghi 18.6ghi 57.2gh 31.0fg 12.8cde 43.8efgh 11.5ef 2.5fg 14.0ef UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.6a 7.0a 26.4a UIC 26.2j 12.3j 38.5j 23.0i 8.5f 31.5i 7.9g 1.4h 9.3h Each value is the mean of four replicates; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; Pf=Pseudomonas fluorescens; Pl=Purpureocillium lilacinum; Pc=Pochonia chlamydosporia; Tv=Trichoderma viride; UUC- Untreated Uninoculated Control; UIC-Untreated Inoculated Control.
Table 27b: Effect of seed dressing with various bioagents on the growth of tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Yield/plant Pollen Chlorophyll Carotenoid Eggmasses/root Eggs/ Nematode Root-knot Treatment (g) fertility (%) content(mg/g) content(mg/g) eggmass population Index (250g) 1% cellulose Pc 239cd 72.0cde 1.85cde 0.593cd 103ghi 188efg 846i 2.0gh 2% cellulose pc 248b 74.5b 1.93b 0.615b 97.0j 180i 827j 1.7h 1% molasses Pc 235de 71.0de 1.81cdef 0.578fg 106efg 192cdef 859h 2.3fg 2%mollases Pc 243bc 73.4bc 1.87bc 0.600c 99ij 186ghi 839i 1.9h 1% cellulose Pl 227fg 69.6fgh 1.76fgh 0.554jk 113cd 195bcd 902de 3.0cde 2% cellulose Pl 237cde 72.0cde 1.86cd 0.588de 102ghij 185ghi 889f 2.6ef 1% molasses Pl 222ghi 69.0gh 1.73ghi 0.546k 116bc 197bc 908cde 3.3bc 2% molasses Pl 230ef 71.6de 1.80def 0.575fgh 108defg 189efg 898ef 2.8e 1% cellulose Pf 236cde 70.5ef 1.79defg 0.568hi 107efg 190defg 876g 2.7e 2% cellulose Pf 243bc 73.8bc 1.85cde 0.592cd 100hij 183hi 856h 1.9h 1% molasses Pf 232def 70.0ef 1.77fgh 0.560ij 111cdef 194bcd 890f 3.0cde 2% molasses Pf 239cd 72.4cde 1.82cdef 0.580ef 105fgh 187ghi 865h 2.8e 1% cellulose Tv 215ij 67.8gh 1.72hi 0.530l 116bc 197bc 914bc 3.2bcd 2% cellulose Tv 226fgh 71.0de 1.78efgh 0.570gh 110cdef 190defg 904cde 2.9de 1% molasses Tv 210j 66.5h 1.70i 0.522l 120b 200b 920b 3.5b 2% molasses Tv 219hi 69.6fgh 1.79defg 0.550k 112cde 195bcd 910bcd 3.0cde UUC 355a 89.5a 2.78a 0.880a 0k 0j 0k 0i UIC 142k 47.0i 1.12j 0.275m 175a 266a 1608a 5.0a Each value is the mean of four replicates; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; Pf=Pseudomonas fluorescens; Pl=Purpureocillium lilacinum; Pc=Pochonia chlamydosporia; Tv=Trichoderma viride; UUC- Untreated Uninoculated Control; UIC-Untreated Inoculated Control.
Table 28a: Effect of different oil cakes as a seed dressing agents on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Dose/Pot Length(cm) Fresh weight(g) Dry weight(g) Treatment (%) Shoot Root Total Shoot Root Total Shoot Root Total 7 36.3cd 17.1bc 53.4cd 29.6de 11.9cd 41.5bcd 11.2bcd 3.2bc 14.4cd Castor cake 14 37.0bc 17.5bc 54.5bc 30.4bcd 12.3bc 42.7bc 11.6bc 3.4b 15.0bc 21 38.5b 18.3b 56.8b 31.2b 12.8b 44.0b 12.0b 3.5b 15.5b 7 29.9j 14.7h 44.6k 25.0k 10.0h 35.0h 9.0g 2.1fg 11.0i Cotton cake 14 30.8ij 15.0gh 45.8jk 26.0jk 10.4gh 36.4gh 9.8fg 2.2fg 12.0h 21 31.6hi 15.4efgh 47.0ijk 26.8hij 10.7fgh 37.5fgh 10.7de 2.5ef 13.2efg 7 31.0hij 15.2fgh 46.2jk 26.5ij 10.5fgh 37.0fgh 9.8fg 2.2fg 12.0h Mahua cake 14 32.0ghi 15.5efgh 47.5ij 27.3ghi 10.9fg 38.2efg 10.4def 2.3efg 12.7fgh 21 33.2fg 16.2def 49.4ghi 28.2fg 11.3def 39.5def 10.9cde 2.7de 13.6ef 7 35.3d 16.7cd 52.0de 29.7de 11.8cd 41.5bcd 10.4def 2.6ef 13.0fg Mustard cake 14 36.2cd 17.2bc 53.4cd 30.0cde 12.0bcd 42.0bc 11.0cd 3.0cd 14.0de 21 37.3bc 17.7bc 55.0bc 31.0bc 12.4bc 43.4b 11.7bc 3.3bc 15.0bc 7 32.3gh 15.7efgh 48.0hij 27.7gh 11.3def 39.0efg 10.1ef 2.4ef 12.5gh Soybean cake 14 34.0ef 16.0defg 50.0efg 28.9ef 11.6de 40.5cde 10.6de 2.7de 13.3efg 21 35.0de 16.5cde 51.5def 29.8de 12.0cd 41.8bcd 11.2bcd 3.0cd 14.2cd UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.6a 7.0a 26.4a UIC 26.2k 12.3i 38.5l 23.0l 8.5i 31.5i 7.9h 1.4h 9.3j
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 28b: Effect of different oil cakes as a seed dressing agents on the growth of tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Nematode Dose/pot Yield/plant Pollen fertility Eggmasses/ Root knot Eggs/eggmass population Treatment (%) (g) (%) root Index (250g) 7 202bc 71.0cd 117bcd 192ef 1156l 2.3h Castor cake 14 207bc 72.3bc 114cd 189ef 1140m 2.1h 21 212b 74.0b 112d 185f 1125n 1.8i 7 166g 64.0h 129b 211b 1230b 3.8b Cotton cake 14 172fg 66.5fgh 127bc 208bc 1208c 3.5c 21 184ef 67.0fg 124bc 204bcd 1190e 3.2cd 7 177fg 65.0gh 126bc 207bc 1198cd 3.5c Mahua cake 14 181ef 67.4efg 125bc 205bcd 1178gh 3.2cd 21 195cde 68.1def 122bc 200bcde 1165jk 3.1d 7 199bcd 69.0def 121bc 197cde 1172hi 2.8ef Mustard cake 14 204bc 70.2cde 117bcd 194def 1160kl 2.6fg 21 209bc 72.0bc 115bcd 190ef 1142m 2.4gh 7 186def 67.2fg 123bc 205bcd 1184ef 3.0de Soybean cake 14 194cde 69.4cdef 121bc 198cde 1170ij 2.7fg 21 200bcd 71.0cd 118bcd 195def 1158l 2.4gh UUC 355a 89.5a 0l 0k 0o 0j UIC 142h 47.0i 175a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table29a: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Trichoderma viride on the plant growth parameters of tomato cv. K-21 in pots Length (cm) Weight (g) Treatment Fresh Dry weight (g) Shoot Root Total Shoot Root Total Shoot Root Total Trichoderma viride (Tv) 56.0a 27.0a 83.0a 55.3a 22.7a 78.0a 20.4a 7.6a 28.0a Tv+Mi 37.2c 17.8c 55.0c 32.7d 14.0c 46.7d 12.3d 3.3d 15.6d
Mi15 Tv 31.0d 15.0d 46.0e 24.7e 9.8d 34.5e 10.1e 2.3e 12.4e
Tv15 Mi 44.3b 21.7b 66.0d 40.3c 17.2b 57.5c 15.0c 4.5c 19.5c UUC 54.7a 26.5a 81.2b 50.7b 22.3a 73.0b 19.4b 7.0b 26.4b UIC 26.2e 12.3e 38.5f 23.0f 8.5e 31.5f 7.9f 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Trichoderma viride (Tv) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control.
Table 29b: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Trichoderma viride on nematode multiplication and biochemical parameters of tomato cv. K-21 in pots Pollen Yield/ Chlorophyll Craotenoid NRA Eggmasses Eggs/ Nematode Root Treatment fertility plant content content ((nM NO2 /root eggmass population knot (%) (g) (mg/g) (mg/g) .g-1.h-1) (250g) Index Trichoderma viride (Tv) 89.0a 360a 2.82a 0.890a 317a 0e 0e 0e 0e Tv + Mi 69.0c 230d 1.98c 0.660d 230d 108c 184c 860c 2.6c
Mi15 Tv 58.0d 176e 1.50d 0.520e 209e 132b 220b 985b 3.0b
Tv15 Mi 74.5b 280c 2.15b 0.780c 266c 90d 160d 760d 1.8d UUC 89.5a 355a 2.78a 0.880b 307b 0e 0e 0e 0e UIC 47.0e 148f 1.12e 0.272f 142f 175a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Trichoderma viride (Tv) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control.
Table 30a: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Purpureocillium lilacinum on the plant growth parameters of tomato cv. K-21 in pots Length (cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Purpureocillium lilacinum (Pl) 57.2a 27.3a 84.5a 56.3a 23.2a 79.5a 21.0a 7.8a 28.8a Pl + Mi 38.2d 18.2d 56.4e 33.2d 14.2d 47.4d 12.5d 3.5d 16.0d
Mi15 Pl 31.3e 15.2e 46.5d 24.9e 9.9e 34.8e 10.3e 2.4e 12.6e
Pl15 Mi 45.9c 22.6c 68.5c 40.8c 17.6c 58.4c 15.3c 4.7c 20.0c UUC 54.7b 26.5ab 81.2b 50.7b 22.3b 73.0b 19.4b 7.0b 26.4b UIC 26.2f 12.3f 38.5f 23.0f 8.5f 31.5f 7.9f 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Purpureocillium lilacinum (Pl) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control.
Table 30b: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Purpureocillium lilacinum on nematode multiplication and biochemical parameters of tomato cv. K-21 in pots Pollen Yield/ Chlorophyll Craotenoid NRA Eggmasses/ Eggs/ Nematode Root Treatment fertility plant content content ((nM NO2 root eggmass population knot (%) (g) (mg/g) (mg/g) .g-1.h-1) (250g) Index Purpureocillium lilacinum (Pl) 90.0a 367a 2.86a 0.910a 322a 0e 0 0e 0e Pl+ Mi 70.5c 236d 2.00c 0.675d 238d 104c 175c 833c 2.3c
Mi15 Pl 59.0d 181e 1.56d 0.532e 213e 131b 216b 955b 2.8b
Pl15 Mi 77.0b 287c 2.20b 0.800c 272c 82d 152d 732d 1.6d UUC 89.5a 355b 2.78a 0.880b 307b 0e 0 0e 0e UIC 47.0e 148f 1.12e 0.272f 142f 175a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Purpureocillium lilacinum (Pl) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control.
Table 31a: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Pochonia chlamydosporia on plant growth parameters of tomato cv. K-21 in pots Length (cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Pochonia chlamydosporia (Pc) 58.5a 28.0a 86.5a 56.9a 23.5a 80.4a 21.4a 8.0a 29.4a Pc + Mi 39.8d 18.8d 58.6d 33.6d 14.4d 48.0d 12.8d 3.6d 16.4d
Mi15 Pc 31.5e 15.3e 46.8e 25.3e 10.2e 35.5e 10.5e 2.5e 13.0e
Pc15 Mi 46.3c 23.2c 70.5c 41.4c 17.8c 59.2c 15.8c 5.0c 20.8c UUC 54.7b 26.5b 81.2b 50.7b 22.3b 73.0b 19.4b 7.0b 26.4b UIC 26.2f 12.3f 38.5f 23.0f 8.5f 31.5f 7.9f 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Pochonia chlamydosporia (Pc) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control.
Table 31b: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Pochonia chlamydosporia on nematode multiplication and biochemical parameters of tomato cv. K-21 in pots Pollen Yield/ Chlorophyll Craotenoid NRA Eggmasses Eggs/ Nematode Root Treatment fertility plant content content ((nM NO2 /root eggmass population knot (%) (g) (mg/g) (mg/g) .g-1.h-1) (250g) Index Pochonia chlamydosporia(Pc) 90.5a 376a 2.90a 00.918a 326a 0e 0e 0e 0e Pc+ Mi 71.4c 242d 2.05d 0.682d 241d 102c 172c 822c 2.2c
Mi15 Pc 60.0d 189e 1.60e 0.537e 215e 128b 200b 928b 2.5b
Pc15 Mi 78.2b 295c 2.25c 0.810c 276c 73d 134d 700d 1.5d UUC 89.5a 355b 2.78b 0.880b 307b 0e 0e 0e 0e UIC 47.0e 148f 1.12f 0.272f 142f 175a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Pochonia chlamydosporia (Pc) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control.
Table 32a: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Pseudomonas fluorescens on plant growth parameters of tomato cv. K-21 in pots Length (cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Pseudomonas fluorescens (Pf) 59.2a 28.3a 87.5a 57.4a 23.8a 81.2a 21.6a 8.2a 29.8a Pf+ Mi 42.0d 20.0d 62.0d 34.6d 14.9d 49.5d 13.0d 3.7d 16.7d
Mi15 Pf 32.0e 15.5e 47.5e 25.5e 10.3e 35.8e 10.7e 2.7e 13.4e
Pf15 Mi 47.0c 23.6c 71.6c 42.0c 18.0c 60.0c 16.0c 5.0c 21.0c UUC 54.7b 26.5b 81.2b 50.7b 22.3b 73.2b 19.4b 7.0b 26.4b UIC 26.2f 12.3f 38.5f 23.0f 8.5f 31.5f 7.9f 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Pseudomonas fluorescens (Pf) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control
Table 32b: Effect of individual, concomitant and sequential inoculation of Meloidogyne incognita and Pseudomonas fluorescens on nematode multiplication and biochemical parameters of tomato cv. K-21 in pots Pollen Yield/ Chlorophyll Carotenoid NRA Eggmasses Eggs/ Nematode Root Treatment fertility% plant content content (nM NO2 . /root eggmass population knot (g) (mg/g) (mg/g) g-1.h-1) (250g) index Pseudomonas fluorescens (Pf) 91.6a 385a 2.95a 0.924a 332a 0e 0 0e 0e Pf+ Mi 72.0d 250d 2.12d 0.689d 250d 94c 160 790c 2.0c
Mi15 Pf 61.0e 194e 1.69e 0.540e 220e 118b 189 907b 2.4b Pf15 Mi 79.4c 300c 2.32c 0.816c 285c 64d 118 640d 1.4d UUC 89.5b 355b 2.78b 0.880b 307b 0e 0 0e 0e UIC 47.0f 148f 1.12f 0.272f 142f 175a 266 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05 Indicate the sequence of inoculation of Meloidogyne incognita (Mi)/ Pseudomonas fluorescens (Pf) fungus 15 days prior. + Indicate simultaneous inoculation; UUC- Untreated Uninoculated Control; Untreated Inoculated Control
Table33a: Effect of Purpureocillium lilacinum in combination with various soil organic amendments on growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Press mud 49.5b 24.2bc 73.7bc 46.6bc 20.4b 67.0bc 16.2b 5.1b 21.3bc Cotton cake 46.0e 22.5d 68.5d 43.2f 18.5e 61.7e 14.6d 4.2d 18.8e Mahua cake 46.8de 23.0cd 69.8cd 44.0ef 19.0de 63.0de 15.1cd 4.5cd 19.6de Mustard cake 48.7bc 23.8bc 72.5bc 45.8bcd 20.0bc 65.8bc 16.0bc 5.0b 21.0bc Castor cake 50.3b 24.7b 75.0b 47.5b 20.7b 68.2b 16.5b 5.3b 21.8b Soybean cake 47.7cd 23.3cd 71.0cd 45.0de 19.5cd 64.5cd 15.6bc 4.8bc 20.4cd UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2f 12.3e 38.5e 23.0g 8.5f 31.5f 7.9e 1.4e 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 33b: Effect of Purpureocillium lilacinum in combination with soil organic amendments on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Treatment Pollen Yield/ Chlorophyll Carotenoid Eggmass/ Eggs/ Nematode Root knot fertility plant content content root eggmasses population index (%) (g) (mg/g) (mg/g) (250g) Press mud 80.2bc 294bc 2.21bc 0.779c 66de 126e 612f 1.9de Cotton cake 76.0e 275f 1.89e 0.732g 83b 160b 684b 2.8b Mahua cake 77.0de 278ef 2.00de 0.744f 77bc 148c 663c 2.5cd Mustard cake 79.0bcd 290cd 2.15bcd 0.768d 70cde 136d 633d 2.1d Castor cake 81.0b 302b 2.30b 0.788b 63e 117f 597g 1.7e Soybean cakes 78.2cde 285de 2.08cd 0.755e 73cd 140d 647e 2.2de UUC 89.6a 356a 2.79a 0.878a 0f 0g 0h 0f UIC 47.2f 154g 1.12f 0.270h 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 34a: Effect of Pochonia chlamydosporia in combination with various soil organic amendments on growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Press mud 49.8bc 25.0bc 74.8bc 46.7bc 20.4bc 67.1bc 16.8bc 5.2bc 22.0b Cotton cake 47.0d 23.0e 70.0d 43.8e 18.8e 62.6e 15.0d 4.5c 19.5d Mahua cake 47.6d 23.6de 71.2d 44.7de 19.0e 63.7de 15.3d 4.7c 20.0d Mustard cake 49.1bc 24.4bcd 73.5bcd 46.0bcd 20.0cd 66.0c 16.6bc 5.2bc 21.8bc Castor cake 50.8b 25.7ab 76.5b 47.6b 20.8b 68.4b 17.1b 5.3b 22.4b Soybean cake 48.2cd 24.0cde 72.4cd 45.5cde 19.7de 65.2cd 16.2c 5.0bc 21.2c UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2e 12.3g 38.5e 23.0f 8.5f 31.5f 7.9e 1.4d 9.3e
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 34b: Effect of Pochonia chlamydosporia in combination with soil organic amendments on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Eggmasses Eggs/ Nematode Root knot Treatment fertility plant content content /root eggmass population Index (%) (g) (mg/g) (mg/g) (250g) Press mud 80.6b 302bc 2.25c 0.779c 62ef 116f 602f 1.6cd Cotton cake 77.0c 287f 1.96e 0.746f 77b 147b 662b 2.4b Mahua cake 78.0bc 291ef 2.03de 0.753ef 72bc 141c 648c 2.1bc Mustard cake 80.0b 300cd 2.18c 0.770d 65de 125e 620e 1.6cd Castor cake 81.8b 308b 2.35b 0.793b 58f 109g 590g 1.4d Soybean cakes 78.7bc 294de 2.10cd 0.762de 69cd 133d 632d 1.9bc UUC 89.6a 356a 2.79a 0.878a 0g 0h 0h 0e UIC 47.2d 154g 1.12f 0.270h 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 35a: Effect of Pseudomonas fluorescens in combination with various soil organic amendments on growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Press mud 50.9bc 24.7bc 75.6bc 47.0bc 20.4c 67.4c 16.5bc 5.7bcd 22.2bc Cotton cake 48.4d 23.8d 72.2e 46.0d 19.7d 65.7d 15.2d 5.0d 20.2e Mahua cake 48.8d 24.0cd 72.8de 46.2cd 20.0cd 66.2cd 15.5cd 5.2cd 20.7de Mustard cake 50.0c 24.5bcd 74.5cd 46.8cd 20.3cd 67.0cd 16.1bcd 5.5bcd 21.6bcd Soybean cake 49.7c 24.3cd 74.0cd 46.6cd 20.2cd 66.8cd 15.8bcd 5.4bcd 21.2cde Castor cake 52.2b 25.2b 77.4b 48.4b 21.2b 69.6b 16.8b 6.0b 22.8b UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2e 12.3e 38.5f 23.0e 8.5e 31.5e 7.9e 1.4e 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 35b: Effect of Pseudomonas fluorescens in combination with soil organic amendments on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Eggmasses/root Eggs/eggmass Nematode Root Treatment fertility plant content content population knot (%) (g) (mg/g) (mg/g) (250g) Index Press mud 81.6bc 306bc 2.30bc 0.785c 59ef 110f 594e 1.4de Cotton cake 78.4d 292e 2.00f 0.752e 73b 145b 644b 2.3b Mahua cake 79.6cd 295de 2.07ef 0.760de 69bc 136c 630c 1.9c Mustard cake 81.0bc 302cd 2.24cd 0.778c 62de 118e 614d 1.6cd Soybean cake 80.5bc 299cd 2.17de 0.769cd 66cd 127d 620d 1.7bc Castor cake 82.5b 312b 2.38b 0.800b 55f 104g 582f 1.3e UUC 89.6a 356a 2.79a 0.878a 0g 0h 0g 0f UIC 47.2e 154f 1.12g 0.270f 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control
Table 36a:Effect of Purpureocillium lilacinum in combination with composted plant straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste on growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Pigeon pea 47.4de 22.6bcd 70.0cd 44.0cde 19.2bcd 63.2cde 14.8cde 4.4cd 19.3cde Pearl millet 45.8e 21.7d 67.5d 42.5e 18.5d 61.e 14.1e 4.1d 18.2e Maize 48.4cd 23.0bcd 71.4c 44.5bcd 19.5bcd 64.0bcd 15.3bcd 4.7b 20.0bcd Mustard 50.2b 24.0b 74.2b 46.0b 20.2b 66.2b 16.1b 5.2b 21.3b Potato 49.1bc 23.5bc 72.6bc 45.2bc 19.8bc 65.0bc 15.7bc 4.9b 20.6bc Sorghum 46.5e 22.2cd 68.7d 43.3de 18.9cd 62.2de 14.5de 4.3cd 18.8de UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2f 12.3e 38.5e 23.0f 8.5e 31.5f 7.9f 1.4f 9.3f
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 36b:Effect of Purpureocillium lilacinum in combination with composted plant straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Eggmasses/ Eggs/ Nematode Root knot Treatment fertility Plant content content root Eggmass population Index (%) (g) (mg/g) (mg/g) (250g) Pigeon pea 77.6cde 283de 2.00de 0.752e 77cd 144d 650d 2.3cd Pearl millet 76.0e 273f 1.86f 0.728g 84b 163b 685b 3.0b Maize 78.7bcd 289cd 2.09cd 0.760d 73de 134e 634e 2.0de Mustard 80.5b 300b 2.28b 0.785b 65f 120f 600g 1.7e Potato 79.6bc 294bc 2.20bc 0.774c 70ef 129e 615f 1.9e Sorghum 76.8de 280ef 1.92ef 0.742f 81bc 156c 665c 2.6c UUC 89.6a 356a 2.79a 0.878a 0g 0g 0h 0f UIC 47.2f 152g 1.12g 0.272h 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 37a:Effect of Pochonia chlamydosporia in combination with composted plant straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste on growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots
Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Pigeon pea 48.0cde 23.4bcd 71.4cde 45.0cd 19.3bcd 64.3cde 15.5bcd 4.7bc 20.2de Pearl millet 46.3e 22.7d 69.0e 43.2e 18.8d 62.0e 14.7d 4.3c 19.0f Maize 49.0bcd 23.7bcd 72.7bcd 45.6bcd 19.8bcd 65.4bcd 15.8bcd 4.9bc 20.7cd Mustard 50.9b 24.6b 75.5b 47.1b 20.5b 67.6b 16.6b 5.4b 22.0b Potato 49.8bc 24.2bc 74.0bc 46.3bc 20.1b 66.4bc 16.2bc 5.1bc 21.3bc Sorghum 47.2de 23.0cd 70.2de 44.3de 19.3bcd 63.2de 15.1d 4.5bc 19.6ef UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.00a 26.4a UIC 26.2f 12.3e 38.5f 23.0f 8.5e 31.5f 7.9e 1.4f 9.3g
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 37b:Effect of Pochonia chlamydosporia in combination with composted plant straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Eggmass/ Eggs/ Nematode Root knot Treatment fertility Plant content content root eggmass population Index (%) (g) (mg/g) (mg/g) (250g) Pigeon pea 78.2cd 291de 2.10cde 0.758e 72cd 138d 640d 2.6bc Pearl millet 76.6d 280f 1.92e 0.738g 79b 156b 670b 2.9b Maize 79.0bcd 295cd 2.19bcd 0.766d 70cd 132d 624e 2.3cd Mustard 81.2b 306b 2.33b 0.790b 61e 114f 594g 1.7e Potato 80.0bc 300bc 2.24bc 0.776c 66de 125e 608f 1.9de Sorghum 77.3d 284ef 2.00de 0.749f 75bc 147c 656c 2.7bc UUC 89.6a 356a 2.79a 0.878a 0f 0g 0 0e UIC 47.2e 152g 1.12f 0.272h 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table 38a:Effect of Pseudomonas fluorescens in combination with composted plant straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste on the growth of tomato cv. K-21 in relation to root-knot development caused by Meloidogyne incognita in pots Length(cm) Weight (g) Treatment Fresh Dry Shoot Root Total Shoot Root Total Shoot Root Total Pigeon pea 48.6de 24.0cd 72.6cd 45.4de 19.6bcd 65.0de 16.1bc 5.1bc 21.2cd Pearl millet 47.3e 23.2d 70.5d 44.3e 19.1d 63.4e 15.7c 4.9c 20.6d Maize 49.5cd 24.3bc 73.8bc 46.2cd 19.8bcd 66.0cd 16.3bc 5.3bc 21.6bcd Mustard 51.7b 25.0b 76.7b 47.9b 20.7b 68.6b 16.9b 5.6b 22.5b Potato 50.6bc 24.4bc 75.0b 47.0bc 20.3bc 67.3bc 16.6b 5.4bc 22.0bc Sorghum 47.8e 23.5cd 71.3d 44.9e 19.3cd 64.2de 16.0bc 5.0c 21.0cd UUC 54.7a 26.5a 81.2a 50.7a 22.3a 73.0a 19.4a 7.0a 26.4a UIC 26.2f 12.3e 38.5e 23.0f 8.5e 31.5f 7.9d 1.4d 9.3e
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Table38b: Effect of Pseudomonas fluorescens in combination with composted plant straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste on tomato cv. K-21 in relation to root-knot development and multiplication of Meloidogyne incognita in pots Pollen Yield/ Chlorophyll Carotenoid Eggmasses/ Eggs/ Nematode Root knot Treatment fertility plant(g) content content root Eggmass population Index (%) (mg/g) (mg/g) (250g) Pigeon pea 79.5c 295cde 2.14cd 0.772cd 67c 129c 626d 2.3c Pearl millet 78.0c 289e 1.98e 0.749f 75b 147b 650b 2.7b Maize 80.3bc 298cd 2.22bc 0.780c 65cd 120d 607d 2.1c Mustard 82.0b 309b 2.36b 0.796b 57e 106e 585f 1.4d Potato 80.7bc 303bc 2.28bc 0.789bc 60de 112e 596e 1.7d Sorghum 78.7c 292de 2.06de 0.760e 71bc 135c 635c 2.4bc UUC 89.6a 356a 2.79a 0.878a 0f 0f 0g 0 UIC 47.2d 152f 1.12f 0.272g 174a 266a 1608a 5.0a
Each value is the mean of four replicates; Initial inoculum = 1500 (J2) of Meloidogyne incognita per pot; Means in each column followed by same letter are not significantly different according to Duncan’s Multiple Range Test (DMRT) at P≤0.05; UUC- Untreated Uninoculated Control; UIC- Untreated Inoculated Control.
Discussion
CHAPTER V
DISCUSSION
India is primarily one of the country where major part of the economy depends upon agriculture. As in the era of rapidly increasing population, the main obstacle is to assure the food security and to provide safe and proper nutrition to all. The 35% population increase is projected by 2050 (World Bank 2008), an increase in food demand of 75% is assumed. Due to economic development and changes in food preferences (Keating et al., 2010).This increasing demand can be maintained by sustainable agriculture through the enhancement of the yield and productivity. Anthropogenic activities through the involvement of synthetic pesticides have raised the problems of environmental pollution and climate change. These problems have altered the composition of earth ecosystems particularly the composition of biological flora and fauna, species richness and biodiversity up to the threatened level. Consecutively, pest resurgence is emerged as an additional problem for food security. As excessive use of chemicals has narrowed downed the species richness of crop plants, this provided the selective advantage to the pests to easily infest the homogenous plant communities. Protection from the threat to food crop from damaging pests has become prime concern for food security and quality aspects. As climate change, which has given a favour to destructive diseases, has also altered the soil nature to support soil inhibiting pests resurgence such as nematodes and soil fungi. In addition to this, it has raised the serious issues of food safety concerns in developing world. Among the entire pests, phytonematodes occupy paramount position are one of the disastrous and polyphagous pest for the world crop production. Nematodes are unsegmented worm-like animals and are known to be the most common animal group on earth (Decraemer and Hunt, 2013). Nearly 90% of metazoan existing in the world are nematodes. However, approximately 25,000 nematodes species have been reported so far all over the world. But about 4,100 species of plant-parasitic nematodes have been identified (Decraemer and Hunt, 2006), new species are continually being described while others, previously viewed as benign or non-damaging, are becoming pests as cropping patterns change (Nicol, 2002). Plant parasitic nematodes (PPNs) represent 15% of the total number of nematode species described and are significant pathogens in agriculture (Decraemer and Hunt, 2013). The root-knot nematodes are the most important pests in reducing 136
Discussion agriculture yield of cereals, legumes and vegetables (Maqbool and Shahina, 2001). Among the 50% of crop losses caused by pests, 12.3% is estimated to be caused by nematodes and more damage to the crops due to the nematode infestations noticed in the developing countries than in the developed countries (Sasser and Frackman, 1987). Lots of work has been done on the crop losses caused by plant parasitic nematode. The crop losses caused by phytonematodes estimated about $ 157 billion annually to world agriculture (Abad et al., 2008). In India the losses of agriculture estimated at about Rs. 210 crore annually (Jain et al., 2007). Crop losses due to plant parasitic nematodes are estimated to be about 12.3% in developed nations and 14.6% in the developing countries (Sasser and Freckman, 1987). On the world-wide basis, the average annual loss of all the crops has been estimated to be about 10% especially when the indirect effects of nematodes are considered (Kleczkowski, 1997). Sehgal and Gaur (1999) reported 12% loss in vegetables in India. The crops are greatly affected with root-knot nematodes in all parts of India and elsewhere (Siddiqui and Shaukat, 2003;Sikora and Fernandez, 2005).Yield loss ranges from 20-43% in vegetables production was reported by ( Reddy, 1985; Jonathan et al., 2001 Aalders et al., 2009; Khan, 2009; Wesemael et al., 2011). However, maximum yield losses due to root-knot nematode in tomato have been reported up to the extent of 61% (Nagnathan, 1984). Most of the plant species belong the family Solanaceae accounting major part of food supply are susceptible to Meloidogyne spp. representing quantitative and qualitative economic losses to the crop. Tomato is the preferred host facing severe damage from root knot nematode, Meloidogyne spp. (Asif et al., 2015). Vegetable crops usually are among the most susceptible and worstly affected by the nematodes (Sharma et al., 2006; Dhaliwal and Koul, 2007; Singh and Khurma, 2007). Root-knot nematodes are one of the major pathogens of tomatoes worldwide and limit fruit production (Sikora and Fernandez, 2005). Hence a feasible management strategy is need of the hour for the management of nematode to tackle this unbearable and terrible problem bearing by the farmers globally. Plant protection against nematodes is quite difficult because nematodes cannot be eradicated completely from the field (Budai et al., 2005), a nematode management strategy, therefore is needed to involve manipulation of nematode densities to non-injurious and sub-economical threshold levels (Viaene et al., 2006).The use of synthetic nematicides one of the most decisive and potent methods for nematode management but the increased use of various nematicides under cultivation practices for the management of plant parasitic 137
Discussion nematodes has not only contaminated surface and ground water but has also disturbed the soil ecosystem, plant and microbial population (Bahadur et al., 2006). However, disturbance in the soil ecosystem due to nematicides leads imbalance of the rhizospheric micro flora and fauna, increase of residues in the fruits and vegetables, toxicity to the soil microorganisms, altering the soil structure and texture, ultimately accumulation of chemicals in the plants. In addition to this the other major hurdles in the use of nematicides are very expensive, not sustainable and affect the agro- ecosystem adversely besides having inherent difficulties in their handling (Noling and Becker, 1994; Jesse and Jada, 2004).Hence, in the modern era of globalization, climate change, industrialization and pathogen out break there is urgent need to develop the novel protective approach that can manage the disease in, a sustainable manner without disturbing the ecological niche and can maintain the equilibrium between the soil and atmospheric environment. The constructive effect of the combined approach and progression of flourishing soil health as a substitute of chemical nematicides is required for the nematode management programme. The property of the highest beneficial treatment for soil health should be economically effective. It should have a potential to compete for different species of pests and should not result in negative influence on the soil biodiversity, physical, chemical and biological parameters of the soil system. In addition to this the treatments should have capability to exaggerate nutrient status of the soil and promote the functioning and habitat for the favourable flora. There are no fixed criteria of microbial number for a soil to be healthy while the number of microbes varied with the soil and dependent upon the soil quality and added soil organic matter.
Organic farming is a branch of intense concern and interest in the aspect of healthy and safe food point of view and for the nematode management in sustainable manner. Organic farming is receiving increasing international support and the global product market reached a value of $US 38.6 billion in 2006 (Kaþkavalci et al., 2009). Researchers are developing alternative management techniques such as use of cropping systems, organic soil amendments, biological control agents and judicious use of nematicides (Serfoji et al., 2010). Several other tactics such as resistant cultivar, intercropping with non host crops, fallowing, summer ploughing, crop rotation, organic amendment and bioagents have been applied successfully (Akhtar, 1997). So the demand of the customers all over the worlds for the availability of the
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Discussion safe, healthy and qualitative food has lead to the emergence of the usage of plants/parts and their products of natural origin and bioagents as an alternative of chemical nematicides. Moreover, it has created interest in the management of nematodes in a sustainable way without affecting the environment. There is increasing interest in discovering nematicidal compounds from plant parts or/products (Chitwood, 2002; Sano, 2002).However, the effectiveness of organic amendment alone is low than integration of various approaches is one of the best tactic to achieve the required crave.
Economically, root-knot nematode is one of the important pests, hence scientist all over the world are emphasizing attention and showing curiousness for their management. Utilization and exploitation of resistant cultivars is economical, ecologically safe and efficient method for the management of root-knot nematodes, Meloidogyne spp. having dual purpose of causing reduction and suppression of nematode population and protection of crop from nematode infection and damage, besides this it is uttermost important from the food safety and environmental safety and for its economic sustainability (Djian-Caporalino et al., 2011). Involving mechanisms in resistant plants could be the production of toxic from the root exudates, the lack of an attractant or the hatching factor in the exudates, a barrier for penetration or the failure of nematodes to develop within plant tissues, the production of lignin and synthesis toxin including phytoalexines (Jenkins and Taylor, 1967; Favery et al., 2001; Jaubert et al., 2002). This present study showed that all the fourteen tested cultivars of tomato (Solanum lycopersicum L.) viz., EC-538380, K-21, CO-3, FEB-02, EC-570018, NDT-3, S-22, GT-1, GT-2, GT-3, H-88-78-1, PB Barkha bahar-2, VRT-101A and Kalyanpuri-T1 screened against root-knot nematode, Meloidogyne incognita reported varying degree of resistance/susceptibility in response to reduction in plant growth characters viz., plant length, fresh and dry weight, chlorophyll content, carotenoid content, yield and pollen fertility. (Table1a and 1b). Cultivar H-88-78-1 was found most resistant while cultivar K-21 was found to be most susceptible. There are several research findings on the response of tomato cultivar to the root-knot nematode (Darban et al., 2003; Williamson and Gleason, 2003; Pathan et al., 2004; Kamran et al., 2011; Jaiteh, 2012). During the studies in all the screened cultivars none of them was found immune or highly resistant against M.incognita, although resistance (1-2 galls), moderately resistance (3-10 galls),
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Discussion moderately susceptible (11-30 galls), Susceptible (31-100 galls) and highly susceptible (> 100 galls) behaviour were recorded by some cultivar. All the fourteen cultivars cause considerable variation in response root-knot nematode infestation in the form of eggmasses, eggs, nematode population and root-knot indices. The variation of the cultivars to nematode infection might be attributed to their genetic makeup and level of resistance mechanism possessed by a particular cultivar (Barham and Winstead, 1957; Anwar and Mckenry, 2002 and Abad et al., 2003).
Resistance in the species is created due to the segregation of specific gene within the species. On the other hand nematode cannot infest and reproduce on the resistant cultivars and non host crop due to the unavailability to host character necessary for the infection and parasitism. Besides this compatible host plant interaction is necessary for reproduction and susceptibility of the crop. The Compatible and incompatible reaction may be due to the resistant gene which are activated as a result of nematode invasion and some visible morphological observation can be detected in the plant cells (Davis et al., 2000; Williamson et al., 2006). All the cultivar showed remarkable variability at the same inoculum level against root-knot nematode Meloidogyne incognita on plant growth character viz., Plant length, fresh weight and dry weight of tomato. Among all the tested cultivars of tomato H-88-78-1, VRT-101A, EC-570018, GT-2 and PB Barkha bahar-2 results in least reduction in plant growth characters while the cultivar GT-1, S-22 and K-21 causing maximum reduction in plant growth parameters viz., plant length, fresh weight and dry weight of tomato over control. Reduction in plant growth parameters is directly related with the susceptibility. Meloidogyne incognita caused significant reduction in plant length, fresh and dry weight of tomato comparison to control.
Several workers like Caveness and Ogunforowa (1985) and Hussey (1989), reported that Meloidogyne spp. infested-plants are seriously affected by their uptake and transportation of water and nutrients, which in turn affect their shoot weight. Eisenback et al. (1991), heavily diseased plants do not respond to water, this is due to the nematodes which have severely damaged the conducting tissues and disruption in the rearrangement of tracheary elements (Divito et al., 2004) of the plant roots. The infected plants showed disruption of xylem elements (Jatala and Jensen, 1976; Ismail et al., 2004), phloem (Sundararaju and Mehta, 1992) and collapse of vascular vessel by hyperplasia and hypertrophy of giants cell (Vovlas et al., 2005). These giant cells 140
Discussion cause the reduction in the height of the plant and directly suppress the growth of the plants by disturbing the translocation of the minerals and nutrient to all the parts of the plants in proper amount. In addition to this, reduced growth is noticed due to decrease in plant shoot and root weight (Haseeb et al., 1993; Koenning et al., 2004; Ma, 2012). Lowering of the root efficiency to the absorption and deformation of the root system may the results in reduction of growth and yield in comparison to the control.
The reduced root growth is due to the damage in the cortical system (Sujatha and Mehta, 1998) and damage of xylem and phloem of the cells (Sundararaju and Mehta, 1992). Reduced growth is also due to decrease in plant shoot and root weight (Pandey et al., 1992; Haseeb et al., 1993; Vaitheeswaran et al., 2003). Reductions in plant length and weight due to the nematodes have been reported by several workers (Jonathan and Rajendran, 2000; Nehra and Trivedi, 2002; Hisamuddin et al., 2005; Azam and Hisamuddin, 2008; Azam et al., 2008). Disruption of vascular tissues upon the infection of nematodes reduces the transportation of water and nutrients to the foliar systems which reduces the photosynthetic rates in the plants (Meena et al., 2016)
All the susceptible cultivars results the significant reduction in photosynthetic pigment like chlorophyll and carotenoid due to the nematode infestation. It might have direct relation with growth parameters like height, weight and yield and pollen fertility of the plants. Decrease in chlorophyll and carotenoid content due to inoculation in susceptible plants could adversely affect the photosynthetic process; disturb metabolic functions in terms of reduced height, fresh as well as dry weights, number of fruits and delayed initiation of flowering ultimately resulting into reduced yield (Melakeberhan et al., 1985). The present results are also in agreement with Devarajan et al. (2003) Singh and Khurma (2007), Ahmed et al. (2009) and Azhagumurugan et al. (2013). Furthermore, a reduction in the water supply has been found to deleteriously influence physiological and biochemical processes in plants such as photosynthesis, respiration, translocation, ion uptake, nutrient levels, pigments composition, carbohydrate levels, growth promoters and metabolism (Jaleel et al., 2008, 2009). Various forms of abiotic and biotic stresses damage plant leaf tissue and the chloroplasts (Karpinski et al., 2003). Leaf pigment composition is sensitive to plant stress and nematode infection causes either a loss of photosynthetic pigments 141
Discussion
(e.g. chlorophylls) or higher levels of photoprotective pigments, such as zeaxanthin or β-carotene (Demming-Adams and Adams, 1992). Bird (1974) suggested that the nematodes inhibit photosynthesis in plants by interfering with the synthesis and translocation of plant growth hormones such as cytokinins and gibberellins. The reduction in chlorophyll pigments ‘a’ and ‘b’ and total chlorophyll content in root knot nematode infected plant was also observed by some scientists (Haseeb et al., 1996; Mohanty et al., 1999; Abbasi et al., 2008).
The reductions in growth parameters, showed positively correlation with the degree of resistance against nematode infection. The highly susceptible cultivars K-21 and S-22 results in maximum reductions in growth parameters. It was followed by susceptible cultivar, moderately resistant cultivar and least in the resistant cultivars. Reduction in the crop yield was found directly correlated with susceptibility and resistance of the crop. The rate of photosynthesis is a crucial parameter that influenced crop yield (Loveys and Bird, 1973; Melakeberhan et al., 1985; Poornima and Vadivelu, 1998; Patel et al., 2001). It was concluded that Meloidogyne spp. can develop a significant infestation on tomato plants with root damage. Furthermore, root gall formation disturbed seriously water transportation in infested plants, which led to water stress that substantially hindered photosynthetic carbon assimilation (Strajnar et al., 2011)
Pollen fertility was found significantly correlated with the nematode population and susceptibility of the crop. Susceptible cultivars depicted maximum reduction while lowest was observed in resistant plants. This reduction in the pollen fertility might be due to the unavailability of the proper nutrients and disturbance of some physiological processes (Siddiqui and Saxena, 1985). Root knot nematode, M. incognita significantly reduced the percent pollen fertility. The reduction might be due to certain abnormal changes in the host physiology in response to nematode infection. Jones (1981) reported that M. incognita and R. reniformis interfered with water uptake and minerals from soil due to certain cytoplasmic changes. It is likely that these changes might be affecting the pollen fertility. Significant reduction in yield was observed due to nematode infestation. This reduction might be due to the shortage of nutrients and disturbance of the physiological process leads to the preparation of the food. The results of several workers also imply that reduction in yield may be due to reduced food supply because of damaged vascular system, to the 142
Discussion fertile branches (Tiyagi and Alam, 1990; Khan et al., 2012) and due to the decaying of minerals (Melakeberham et al., 1985; Tiyagi et al., 2015).
All the cultivars in the present research showed differential degree of resistance and susceptibility against root knot nematode multiplication in terms of eggmasses, eggs/eggmasses, nematode population and root- knot index. Higher multiplication rate/reproduction rate was found in susceptible cultivar while the resistant cultivar recorded lowest reproduction rate/multiplication rate. Variability in multiplication might be due to the genetic factor of the plants which represent resistance/susceptibility and variation in the nematode population at the genetic level (Jacquet et al., 2005; Castagnone-Sereno, 2006, Fuller et al., 2008). Reproduction and development of M.incognita was represented by resistance and susceptibility of the plant (Cook and evans 1987; Khana et al., 2004). The results of the variety of our study H-88-78-1, VRT-101A, EC-570018, GT-2, PB Barkha bahar-2 and FEB-02 showed lower reproduction rate as compared to other cultivar. Status of the host plant to the nematode infection can be assessed by the number of root galls produced by the root systems of the host plants, nematode reproduction and the final nematode population per gram of root or soil at the time of harvest (Belair and Benoit, 1996; Davis et al., 2003). Of all the 14 screened cultivar, two cultivar H-88-78-1 and VRT- 101A have least reproduction factor viz., number of eggmasses/root, egg/eggmass, nematode population and root knot index whereas the roots of highly susceptible tomato cultivar S-22 and K-21 showed maximum eggmasses and egg/eggmasses, nematode population and root-knot index(5.0). The root-knot index, nematode population and damage to the crop were found directly proportional to the eggmass or nematode reproduction (Pathan et al., 2004; Sharma et al., 2005; Karssen and Moens, 2006). The differences in the susceptibility to M. incognita might be due to differences in their resistance response regulated by genetic makeup which was manifested in terms of number of root-galls (Williamson and Hussey, 1996; Williamson and Kumar, 2006). The production of eggmasses, egg formation was increased with the inoculation of M. incognita in all tomato cultivar. Nematode population and root -knot index were also greater in all inoculated tomato cultivar. However highest was found in the susceptible cultivar. There was variation in multiplication of nematode in certain tomato cultivars. The results of the study stated that all the cultivars responded differentially in terms of nematode
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Discussion multiplication/reproduction and host compatibility. The 1500 inoculum level of Meloidogyne incognita significantly reduced the plant growth characters that favoured the nematode multiplication and ultimately increase in the nematode population and root -knot indices. The two cultivars H-88-78-1and VRT-101A were thus recognized as resistant to M. incognita on the basis of their RKI values.
Effect of plant parts, oil cakes, biochar and biocontrol agents in vitro
In the present study a series of in vitro experiments was conducted to test the efficacy of plants parts, oil cake, biochar and biocontrol agents were tested in vitro. The aqueous leaf extracts of different plants viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed showed high nematicidal behaviour in vitro. Aqueous extracts showed differentially but significant nematicidal potential with the S concentration of leaf extract of Mexican poppy and Trailing eclipta represented cent percent mortality at 24 hours of the exposure. Similarly the S concentration of the leaf extracts of Mexican poppy, Wild eggplant and Black pigweed caused complete inhibition in egg hatching. This was found directly related with the concentration of the extract and exposure period. Our results are in conformation with the study of other workers (Singh and Dabur, 2004; Rajendran and Saritha, 2005; Parihar et al., 2011; Ganai et al., 2013; Asif et al., 2014). Sharma and Trivedi (2002) reported complete inhibition of egg hatching in the plant extracts of C. procera and A. indica. Similar results about inhibition hatching were also detected by Pavaraj et al. (2012) who reported that Nepeta cataria and Couroupita quianensis were found to be most effective in reducing egg hatching. It has been reported that the extracts contained alkaloids, flavonoids, saponins, amides including benzamide and ketones that singly and in combination inhibited hatching (Hackney and Dickerson 1975; Goswami and Vijayalakshmi, 1986; Adegbite and Adesiyan, 2005). This inhibitory effect of chemicals may cause the suppression of the embryonic development or killed the eggs or even dissolved the eggs. It has been reported that extracts contained amino acids single and in combination inhibited hatching (Kayani et al., 2001).
Many workers like Ansari et al. (2016) and Singh et al. (2015) demonstrated the role of extracts of plants in enhancing the percent mortality and reduction in egg hatching. This inhibition in egg hatching and juvenile mortality may be due to certain
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Discussion chemicals present in the extract such as Polythienyls in Tagetes spp. (Kyo et al., 1990), isothiocyanates and glucosinolates from Brassicaceae (Brown and Morra, 1997), alkaloids (Matsuda et al., 1989), phenolics (Evans et al., 1984) and pentacyclic triterpenoids from Lantana camara (Qamar et al., 2005) have been reported to possess nematicidal activity. Begum et al. (2000) reported that chemical constituents such as lantanoside, lantanone, camaric acid and oleanolic acid isolated from aerial parts of L. camara, possessing nematicidal activity against root-knot nematode, M. incognita. Shaukat and Siddiqui (2001) found that the extracts of various weeds caused significant mortality of M. javanica (Treub) Chitw. juveniles in vitro. The increase in juvenile mortality and inhibition in egg hatching might be due to the ovicidal and larvicidal properties caused by the extracts. Similar results were found by Jyomati et al. (2003) in ten different medicinal plant methanol extracts against second stage juveniles of M.incognita. Results of the present study also confirmed that mortality was directly proportional to the concentration of the extract and duration of the exposure was reported by Hoseinpoor and Kargar (2012) and Moosavi (2012). Taye et al. (2012) reported that aqueous extract of some plants results in pronounced reduction in root knot nematode infestation in ecofriendly manner. Elbadri et al. (2008) revealed that the five plants extracts viz., Dinbera retroflexa (leaves), Cucumis melo (fruits), Eucalyptus microtheca (leaves), Acacia nilotica (pods) and Chenopodium album (leaves) exhibited highly promising mortality of second stage juvenile of Meloidogyne incognita. El-Deen et al. (2014) reported that the peppermint, tarragon and marjoram resulted in significantly higher larval mortality percentages at 1 and 2 % of concentrations that were amounted to 100 % at 24 hrs of exposure time. Many components of the essential oils that revealed nematicidal activity, such as carvacrol, thymol, carvone, limonene, Artemisia ketone and t- anathole have also been reported for their biocidal effect on insects, fungi, bacteria and weeds (Landolt et al., 1999; Isman 2000; Rodriguez-Kabana and Simmons, 2005)
In vitro aqueous, chloroform, methanol extracts of plants viz., Tagetes erecta, Parkinja vanica, Clerodendrun indicum, C. serratum, Tectona grandis, Mussenda glabra, Melia azedarach and Xylosoma longifolia inhibited the egg hatching of M. incognita and M. javanica. Fresh plant parts viz., leaves, stem and seeds of Lantana camara and Jatropa spp. have strong nematicidal properties (Joymati, 2008; Javed et al., 2007). The mechanism of plant extracts action may include denaturing and
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Discussion degrading of proteins, inhibition of enzymes and interfering with the electron flow in respiratory chain or with ADP phosphorylation. In the similar experiment on the same procedure aqueous extracts of biochar were tested in vitro for the nematotoxic potential. Biochar at all the dilutions showed deleterious effect on the hatching inhibition and juvenile mortality of M.incognita up to a varying extent. Concentration and exposure periods were the dependent factors that directly relate the mortality and hatching inhibition. This is in agreement with findings of Perry and Beane (1988) who demonstrated a delay in hatching of J2 of G. rostochiensis when potatoes were planted in soil amended with activated charcoal but the results are in contradiction with Ebrahimia et al. (2016) who demonstrated that although biochar significantly delayed hatching of both potato cyst nematode species and it did not reduce the final number of juveniles that hatched and penetrated the roots during the growing period. Huang et al. (2015) reported that significant differences in nematode mortality were not observed between biochar exudates and water 24 h after initiation of the bioassay at doses ranging from 0.3 to 5 % biochar. Similar results were also observed when the nematodes were incubated in biochar exudates for 72 h. Deleterious and inhibitory effect of aqueous extracts of the biochar might be due to release of some toxic chemicals that may cause inhibitory effect on the hatching inhibition and juvenile mortality. Graber et al. (2010) identified a number of biochar compounds that are known to adversely affect microbial growth and survival. These includes, ethylene glycol and propylene glycol, hydroxy-propionic and butyric acids, benzoic acid and o- cresol, the quinines (recorsinol and hydroquinone), and 2-phenoxyethanol. Beside this many other organic compounds were identified in organic solvent extracts of the biochar n-alkanoic acids, hydroxy and acetoxy acids, benzoic acids, diols, triols and phenols.
The extracts of different oil cakes viz., castor cake, cotton cake, mahua cake, mustard cake and soybean cake were tested for their nema-toxic and nematicidal behaviour on second stage juvenile hatching of M. incognita in vitro. The results of the study revealed that all the treatments caused deleterious effect against the egg hatching and juvenile mortality of M.incognita up to a varying degree. The inhibition of egg hatching and juvenile mortality both were concentration and time dependent. Where inhibition in egghatching was observed inversely proportional to the exposure period and directly proportional to the concentration of the extract. Percent juvenile
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Discussion mortality of M.incognita were directly proportional to both concentration of the extract and time duration. Our observations are supported by the various workers (Sitaramaiah and Singh, 1978; Hussain et al., 1992; Pandey, 2000; Radwan et al., 2007) who stated that the aqueous extracts of oil cakes caused toxicity to a variety of plant-parasitic nematodes. The inhibition of juvenile hatching by the oil cakes probably may be because of they contain varying amounts of phenols, aldehydes, fatty acids and some other chemicals of unknown composition (Mishra et al., 1989; Singh et al., 2001). Lazzari et al. (2004) reported that the hydrolysis products of glucosinolates (essentially isothiocyanates) present in brassica oil cakes resulted in highly biocidal activities. Belay et al. (2013) reported that the rapeseed cake extract preferably inhibited the egg hatching as well as juvenile motility of M. incognita. Similar results were reported by Sudheer et al. (2007) where deoiled cake extract of Jatropha was found to reduce the egg hatching of M. incognita due to toxic compounds present in the deoiled cake extract of Jatropha. Alam et al. (1982) reported that the water soluble fractions of various oil cakes became progressively more toxic to nematodes and inhibitory to larval hatching of the root-knot nematode during the course of decomposition. It may be due to the facts that the oil cakes are rich in nutrients and other nematicidal compounds, such as ammonia, which may be generated during degradation of oil cakes (Abbasi et al., 2005). It was proposed that water soluble fractions of oil cakes release some inhibitory and toxic compounds which either may disrupt the embryonic development and caused paralysis and even death of the juveniles as well.
On the same procedure some other experiment were conducted in vitro. In another experiment biocontrol agents were tested for the inhibition of egg hatching and juvenile mortality of M.incognita under in vitro conditions. All the biocontrol agents showed toxic effect against the egg hatching and juvenile mortality of M.incognita. Where Pochonia chlamydosporia showed highest nematicidal effect and least was shown by Trichoderma viride. Mortality of the juveniles represents a strong relationship with the concentration and exposure time. The finding of the study was in agreement with several workers. Various parameters have been taken into consideration for the in vitro activity of the nematode eggs parasitic fungi were egg parasitic index (EPI),severity of the egg infection, effect of cultural filtrates on the hatching, mobility and mortality of second stage juveniles (Chen and Chen, 2002;
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Discussion
Mukhtar and Pervaz,2003;Khan et al., 2004;Adekunle and Akinsanmi, 2005). Tariq and Ijaz (2003) reported that different dilutions of the culture filtrates of P. chlamydosporium significantly inhibited the eggs hatching of M. javanica. Extracellular enzymes proteases and chitinases secreted by P. chlamydosporia play an important role in the infection process of eggs as they enable the fungus to degrade the nematode eggshell (Tikhonov et al., 2002; Esteves et al., 2009a). These worked in the infection process and serve as virulence factors (Yang et al., 2007; Mi et al., 2010; Palma-Guerrero et al., 2010). Segers et al. (1996) showed that the enzyme VCP1, an alkaline serine protease produced by P. chlamydosporia, can break down the protein layer in the Meloidogyne egg shell and expose the inner chitin layer. Siddiqui and Shaukat (2003) reported that production of the metabolite 2,4- diacetylphloroglucinol (2,4-DAPG) by Pseudomonas fluorescens strain CHA0 inhibited egg hatching and induced mortality in J2 root-knot nematodes. Similarly Al Kader (2008) and Sun et al. (2006) revealed that Purpureocillium lilacinum caused 77% eggs infection after 4 days of incubation and high in vitro parasitism rate of PL strain YESX-2-14 on M. hapla egg respectively. However, Morgan-Jone (1984) and Holland et al. (1999) claimed that their PL strain infected eggs at all developing stages. Fungal bioagents can directly parasitize nematodes and/or secrete nematicidal metabolites and enzymes that affect nematode viability (Meyer and Chitwood 1999). The activity of P. lilacinus against root knot nematode is based on its ability to destroy eggs (Dube and Smart, 1987; Holland et al., 1999; Jatala, 1986). Loganathan et al. (2010) observed that in vitro studies the Tvc1, Tvc2 and the isolates of Trichoderma effectively inhibited the egg hatchability of M. incognita. The secondary metabolites includes chitinase enzyme are produced by the fungus that help in degrading the chitin composed the outer shell of nematode’s eggs (Haggag and Amin 2001; Jin et al., 2005). El Hamshary et al. (2004) who reported that P. fluorescens affected M. incognita juveniles survival in vitro and the mortality percentages of nematode were dependent on bacterial concentration and exposure time. The findings suggested that J2 mortality of M. incognita was directly proportional to concentrations and duration of exposure. Our findings are in conformity with the previous studies (Ayatollahy and Fatemy, 2010; Goswami and Mittal, 2002; Satyandra and Mathur, 2010).
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Discussion
Effect of bare-root dipping penetration
The bare-root dip treatments of tomato seedlings with the extracts of different plants, oil cakes, bioagents and biochar provide defense against M. incognita infection. In the initial experiment, use of plants extract as bare- root dip treatment to the tomato seedling prevent root- knot nematode penetration thereby protect from nematode infection. Our results are in confirmation with the findings of Mojumder and Gowswami (1987), where Calotropis gigantea extracts treated to eggmasses of M incognita reduced the juvenile penetration into tomato roots. Akhtar and Mahmood (1994) and Aziz et al. (1995) reported that the extract showed prophylactic and therapeutic effect against the nematode on tomato. Tariq and Siddiqui (2005) evaluated that bare-root dip in tomato seedlings in extract of neem oil cake +carbofuran for 120 min caused the highest inhibition of root-knot larvae penetration. Maximum percent inhibition in nematode penetration was observed in Mexican poppy extract at 120 minutes of the dip duration. This might be due to the chemical present in the extract absorbed by the root that may induce resistance and may cause toxicity against the nematode.
In another experiment on the same procedure, use of oil cakes viz., castor, cotton cake, mahua cake, mustard cake and soybean cake as bare-root dip treatment of tomato seedlings results in rendering defense thereby inhibited larvae penetration. Our results are in agreements with Khan and Saxena (1980) and Battacharya and Gowsami (1988) who suggested that neem cake and groundnut cake alone or in combination with aldicarb significantly reduced nematode penetration compared with the control. Proct and kornprobst (1986) found that penetration of tomato seedlings with neem seed extract inhibited the penetration of M.incognita juveniles into its roots. Goswami and Meshram (1990) observed 50%reduction in root penetration of juveniles of M.incognita with the cake of mustard and karanj. Application of various dilutions of biochar as bare-root dip treatment inhibited the juveniles penetration in the roots of tomato ultimately produced the defense mechanism in the roots against the juveniles. Inhibition in the juvenile varied with the concentrations of the extract used. Concentration and time exposure worked as dependent factors for the inhibition of juveniles penetration. Huang et al. (2015) stated that at 7 dpi, most of the nematodes developed to third-stage juveniles (J3). The mean number of nematodes inside the roots and their development was not different between the biochar exudates treated 149
Discussion and control nematodes. At 14 dpi, most of the nematodes had developed into adult females. Again, significant differences were not observed in the number of adult females or the total number of nematodes in the biochar exudates-treated and water- treated nematodes. The inhibitory effect on juvenile penetration in the root of tomato might me due to the release of toxic chemicals from the biochar dissolution in the water. All the treatments showed significant inhibition of juvenile penetration in the root. This might be the results of absorbed chemicals by the roots that act as defense activator thereby prevents the penetration of juveniles in the root. Beside this, the exudates release from the biochar might formed a covering or coating over the root surface that may prevent juvenile penetration in the root.
Bare-root dip in different treatments of bioagents viz., Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum and Trichoderma viride inhibited the juvenile penetration in the seedlings of tomato root. Percent inhibition of juvenile penetration in the root was found directly proportional to the concentration of the extract and duration of the exposure. Our results are in accordance with the work of Hallmann and Sikora (1994); De Freitas et al., (1997); Kokalis-Burelle, et al. (2002). Siddiqui et al. (2007) observed that the isolates having aggressive root colonization and inhibitory effect on hatching and penetration of nematodes are known to suppress diseases. Kerry (2000) reported that the ability of Pasteuria penetrans to inhibit the penetration of juveniles of root-knot nematode into the root is influenced by the nematode host plant involved. Secondary metabolites from fungi also contain compounds that can be toxic to plant parasitic nematodes
(Dababat and Sikora, 2007). Mechanism of percent inhibition in the penetration may be due to the release of repellent substance by the bioagents in nearby root area and or conversation of plants exudates by the antagonistic bioagents. In our results the statistically significant repellence and antagonistic behaviour of M incognita against the bioagents was observed.
Effects of bare-root dip treatments
The bare-root dip treatment of tomato seedlings with the aqueous extract of leaves and biocontrol agents worked as protectant against the root-knot disease. Leaf extract of different plants viz., Indian mallow, Mexican poppy, Ivy gourd, Trailing eclipta, Wild eggplant and Black pig weed in various concentrations of S, S/2, S/10
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Discussion and S/100 showed significant suppression in the nematode multiplication and improvement in plant growth characters of tomato cv.K-21. The S concentration of the extract at 90 minutes of the dip duration showed highest inhibition in root-knot, nematode infestation. Our observations are in conformity of Tiyagi et al. (2009) who found that leaf extracts of C. procera and T. peruviana was effective in enhancing plant growth parameters when used as a bare-root dip treatment for the management of root-knot nematode, M. incognita infecting tomato plants. Taba et al. (2008) reported that the repellent action and chemotaxis responses after treatment of natural plant extracts. They stated that these two protective activities could be antagonistic if low concentration repels the nematodes sufficiently. Natarajan et al. (2006) suggested that the aqueous extracts of T.erecta reduced the galling index of tomato roots caused by M.incognita. The root-knot development caused by M. incognita significantly reduced by bare-root dip treatment (Ajaz and Tiyagi, 2003; Usman and Siddiqui, 2011a, b; Khan et al., 2011; Meena et al., 2013).
Reduction in the root-knot nematode development after the bare root dip treatment of the seedling might be the results of the absorption of the chemicals by the root worked as defense molecule and repelled the juvenile, beside this it may form a layer or coating on the surface of the root and don’t allow to penetrate into the root for further infection. Undipped plants showed significant reduction in plant growth parameters due to M.incognita inoculation. But the use of leaf extract as bare root dip treatment suppressed the root knot nematode infection thereby manages the reduction. This can be best represented by the toxic chemicals present in the leaf extract showing pathogenic effect to the root -not nematode. The chemicals associated with the roots during root-dip treatment may stimulate activity of biological control agents towards the nematodes (Mc Sorley and Gallaher, 1995). The reduction may be due to root dip treatment causing some chemical reactions triggered by these chemicals (Bunt, 1975).It may also cause resistance of cells against the invasion and development of pathogens (Bell, 1981; Giebel, 1982; Tiyagi and Alam, 1995).
In another experiment, bare root dip treatment of tomato seedling with the biocontrol agents viz., Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum and Trichoderma viride caused significant reduction in root-knot nematode infestation. Hence they enhance the plant growth parameters. Ability of pronounced colonization by the bioagents in the rhizosphere of the plant is 151
Discussion one of the most important factors for the reduction of pathogenic effect of the pest. Aqueous extract of fungal spores and bacterial suspension at S concentration and 90 minute of the dip duration showed most toxic effect for the reduction in root knot nematode incidence. The efficacy of the bioagents was found directly proportional to the concentration of the extract and root dip exposure. The perusal of the results demonstrated that the culture filtrate of P. fluorescens alone and in combination with leaf extract was the most effective followed by P. chlamydosporia, P. lilacinum and T. viride extract. Sahebani and Hadavi (2008) recorded that bare-root-dip treatment of tomato plants treated with filamentous fungi, Trichoderma harzianum caused minimum number of galls/plant over the control. Rao et al. (1999) observed that the combined effect of the leaf extract of Calotropis and castor along with the P.lilacinus affected M.incognita and significantly reduced nematode population. Zaki and Bhatti (1990) reported that the soil amendment with castor leaves and P. lilacinus was effective for the management of M. javanica on tomato. Similar results were also demonstrated by Rao et al. (1997) who reported the successful management of M. incognita on eggplant with the root dip treatment of neem leaf extract mixed with P. lilacinus spores. The adverse effects on penetration and development of nematodes on plants inoculated with different fungi tested may be due to antagonistic fungal metabolites (Frank and de Boer 1959, Mankau 1969, Tripathi 1973, Reddy et al. 1975, Agarwal and Bisen 1984). Suggested mechanisms include the production of metabolites which reduce hatch and attraction and/or degradation of specific root exudates which control nematode behavior (Wheeler 1975; Ciancio et al. 1988; Sikora and Hoffmann-Hergarten, 1993). Root exudates affect hatching, attraction, repellence and invasion of nematodes (Perry and Gaur, 1996).This reduction in root knot infestation might be due to the poor invasion of juveniles or may be due to the formation of the strong and prominent colonization around the rhizosphere of the plant. From the study it was revealed that the individual application of bioagents is effective up to some extent. However, the combined use of leaf extract along with the biocontrol agent enhances and improves the effect by suppressing the nematode infestation. This can be explained by the integrative effect of significant increase in the colonization by the Pseudomonas fluorescens on the root. Leaf extract alone and in combination with biocontrol agents showed variation in their efficacy because of the varying degree of interactive effect in between the leaf extracts and biocontrol
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Discussion agents. Finding of the study clearly revealed this interaction in the form of additive effect in all the treatment. Among all the treatment highest efficacy of additive property was observed in P. fluorescens combined with water hyacinth leaf extract.
Effect of seed dressing
Seed dressing or seed treatment is one of the beneficial and favourable methods for introducing biocontrol agents into the root environment since it protects the seed from soil and seed-borne pathogens and promote the seedling germination (Chang and kommedhal, 1968). Beside this, other beneficial factors creating by this approach includes formation of antagonistic behaviour of the rhizopshere in the early stage of the infection and with simple application of very small amount of the inoculum. Hence, a study was carried out with the aim to appraise the effect of seed dressing of tomato cv.K-21 seeds with the selected bioagents on plant length (cm), fresh and dry weight (g), chlorophyll content (mg/g), carotenoid content (mg/g), yield and pollen fertility (%) and disease intensity in terms of number of eggmasses/root, eggs/eggmass, nematode population and root-knot indices. Results of the study revealed that all the bioagents in the form of seed dressing agents showed assurable results, howevever, P. chlamydosporia and P. fluorescens by using 2% cellulose @4g/pot were recorded most effective. It was followed by the P. lilacinum and T. viride. Moreover, among the tested sticky agents 2% cellulose was most effective as a sticker to adhere the bioinoculants. Our results are in accordance with previous work where potato-associated strains of Pseudomonas inhibited both the soil-borne fungal wilt pathogen and the root-knot nematode, M. incognita (Krechel et al., 2002). Rao et al. (2004) reported that the formulation of P. chlamydosporia was significantly more effective in reducing root galling index and the numbers of nematodes in roots and soil and increasing the percent parasitization of eggs and yield. Devi and Dutta (2002) suggested that higher reduction in M. incognita population and improvement in plant growth was observed with seed treatment of okra with P. fluorescens than with foliar spray. Goswami and Mittal (2002) reported greatest egg parasitizing efficiency of Purpureocillium lilacinum (80%) as compared to other fungi. In greenhouse studies rhizobacteria applied as a seed dressing can reduce nematode penetration of the root system of potato and sugar beet (Racke and Sikora, 1985; Oostendorp and Sikora, 1986). Nama et al. (2015) observed improved plant growth of chick pea and reduction in nematode infestation when T. harzianum was applied in 153
Discussion the form of soil and seed treatment. Similar results were observed by Munfie et al. (2001) who reported that endophytic bacterium Pseudomonas putida reduce M. incognita on tomato when applied as seed treatment. The effect of the production of metabolites (Siddiqui and Shaukat, 2003) reduced hatch and attraction/degradation of specific root exudates and control the nematode behaviour. Trichoderma species are reported to mineralize phosphorous in the soil and make it available to the soil (Sharma et al., 2012). Similar type of mechanism was observed in P. chlamydosporia and P. lilacinum combination which in enhance the plant growth parameters and improving the yield (Zvala-Gonzalez et al., 2015).
In another similar experiment, the effect of aqueous extracts of various oil cakes viz., castor cake, cotton cake, mahua cake, mustard cake and soybean cake as seed dressing @7, 14, 21% concentration (w/w) were tested for the management of root knot nematode, M. incognita tomato. Finding of the study suggested that all the treatments brought about significant improvement in plant growth characters and reduced root-knot nematode infestation. Among all the oil cakes screened castor cakes@21% (w/w) was found most efficacious in enhancing the growth and root knot index while cotton cakes results least potential for promoting plant growth activity and reduction in nematode infestation. Our results are in conformity of various workers (Dcka and Rahman, 1998; Ram and Babeti, 2003; Baheti, L.B., 2005; Yadav et al., 2006). Yang et al. (2015) showed that Camellia seed cake extracts suppressed M. javanica, and promoted the banana plant growth. Singh et al. (1980) reported that the tomato seeds treated with either oilcakes of castor, mustard, neem, mahua and groundnut (2 g oilcake/10 g seeds) (0.1 or 0.2 g nematicides/10 g seeds) and planted in soil infested with Meloidogyne incognita helped in improvement of plant growth and reduction in root knot index. Similar observation was also reported by Mojumder and Mishra (1991). Khan and Hussain (1988) revealed that neem cake @ 20 % as seed treatment reduced multiplication of M incognita and R. reniformis on cowpea. Large number of organic additives of plant origin, including oil-seed cakes, chopped plant parts and seed dressing with plant extracts has been used in the past as nematode control agents (Akhtar and Alam 1993; Tiyagi and Alam 1995; Panteado et al., 2005; Monfort et al., 2006).
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Effect of organic amendment
Integrated pest management (IPM) is a concept proposed as a strategy for sustainable and effective plant protection (Abrol and Shankar, 2012). The aim of this approach is to assign the ecological theory in a manner of management catalogue to evade the loss in yield of agriculture and horticulture system without and negative and repugnant impact on the environment. The approach is commonly implemented with the combination of different cultural practices. The effort of such combination practices is to diminish the utilization of synthetic pesticides which are dangerous to the flora and fauna of the environment. Strategies based on the IPM concept will be necessary in the development of future management programs for Meloidogyne (Stirling, 2014). Addition of organic amendment to the soil is a traditional cultural practice that enables to maintain the soil structure; texture and fertility of the soil. In addition to this it facilitate to promote the rhizospheric activity of soil thereby results in improvement of soil health and worked as a soil conditioner. Application of organic wastes viz., agricultural wastes, farm yard wastes, oil cakes, saw dust, cellulosic waste other organic products, which are easily accessible and available on large scale especially in the developing countries, furnishes a new channel for their exploration and safe disposal in eco-friendly manner for nematode management. Results presented in the study clearly indicated that all the treatments were found to be potent for the tomato plants as they promoted the growth thereby results in decreasing the nematode infestation. Among all the treatments Mexican poppy combined with wild spinach leaf powder recorded highest improvement in plant growth and maximum reduction in the nematode multiplication and root-knot indices. Other treatments viz., Trailing eclipta, Wild eggplant, Black pigweed, Indian mallow and Ivy gourd combined with spinach leaf powder also showed significant enhancement in plant growth of tomato and reduction in the population of root-knot nematode, Meloidogyne incognita. A beneficial relationship was detected in the combined treatment of fresh chopped leaves plus wild spinach leaf powder with the tomato plants in terms of morphological and biochemical parameters as compared to inoculated control. The potential of all treatments to inhibit nematode activity resulted in increased number of fruits and yield of tomato. This beneficial relationship and improvement in plant growth parameters may be due to release of allochemicals and nutrients on the decomposition of the amendment that enhance the beneficial
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Discussion microflora of the soil rhizosphere that act as promoter of growth activity. While the reduction in nematode infestation may be due to release of some toxic chemicals which either disrupt the life cycle of the nematode or lesser down the infectivity of the juvenile. Yujil et al. (2002) reported that incorporation of rock fleabane plant (Inula viscosa) powder at a concentration of 0.1 % in soil reduced second stage juveniles of root- knot nematodes (M. javanica). The mechanisms of plant extract action may include denaturing and degrading of proteins, inhibition of enzymes and interfering with the electron flow in respiratory chain or with ADP phosphorylation (Konstantopoulou et al., 1994). Sensitivity of plant-parasitic nematodes varies from plant to plant. Generally, J2s of Meloidogyne spp. are very sensitive to plant-derived nematicides (Oka et al., 2001).
Organic farming is an area of great concern and interest for growing safe and healthy food in sustainable manner without disturbing the ecological diversity of the soil and maintaining the environmental and soil behaviour. The outcome of the study state that the application of Mexican poppy, Trailing eclipta and Wild eggplant in combination with wild spinach powder is quite efficient alternative of chemical nematicides for the nematode management and enhancing the yield. The leaf powder of I. viscosa, at a lower concentration, showed nematicidal activity against Pratylenchus mediterraneus, although this activity was much lower than that of M. javanica J2 (Oka et al., 2001). Saxena (1989) reported that neem seed cake improved growth of tomato and reduced the reproduction factor and root galling of nematodes. Some herbal powders and their aqueous extracts result in increasing plant growth and reduction in the infection rate (Moosavi, 2012).The observation of the findings are in conformity of Asif et al. (2013); Ganai et al. (2013); Rehman et al, (2015); Ansari et al. (2016). Addition of soil amendments results in a considerable increase in the liberation of CO through the saprophytic activities of soil saprophytes which can 2 suppress the activities of pathogens (Papavizas and Davey, 1992). The use of phytochemical compounds and natural plant extracts has substantially reduced the number of galls and the extent of nematode reproduction on tomato plants (Naz et al., 2013). More specifically, the decomposition of organic residues from some oily plant residues, such as cottonseed meal, or from meal from certain types of mustard have been reported as releasing toxic ammonia, organic acids and other compounds as a byproduct that can kill nematodes (Oka, 2010; McSorley, 2011; Thoden et al.,
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2011). The use of plants as nematicidal or nematostatic products has been regarded as effective, economical and eco-friendly by numerous researchers (Chitwood, 2002). Amendments not only change physical and chemical soil properties, but also support a wide variety of antagonistic microorganisms like fungi, bacteria, etc. (Jaffee et al., 1998; Timm et al., 2001). Application of A. indica (neem) caused maximum reductions in the number of galls, egg masses and reproduction factor (Rf) of the nematode and increases all growth parameters of okra (Usha and Sundararaj, 1995; Hussain et al., 2011). Exaggeration of the fertility of the soil and enhancement of microbial diversity of the rhizosphere may be the results of the suppression of root- knot nematode infestation due to the release of nematotoxic chemical constituents after decomposition thereby ameliorate the plant growth character of the crop. It can work for fulfill the demand of global food production through sustainable nematode management by the involvement of organic farming system. The plant parts/products are safer, friendly than synthetic nematicides and may be economic and easily available. Moreover, they are also responsible to increase in soil nutrient status, soil loosening and texture and to maintain the soil fertility. The present study also elucidate that the reduction in nematode population could be due to the decomposition of plant material in soil and release of secondary metabolites.
In another such experiment of the current study, biochar alone and in combination with saw dust of various plants viz., eucalyptus, lebbeck, jambul, mango, poplar and babool were assessed for the management of root-knot nematode, M. incognita on tomato cv.K-21. All the treatments either singly and in combination significantly ameliorate the plant growth parameters there by results in suppression of nematode infestation. Results of the study also state that the combined action of biochar and saw dust results in significantly pronounced enhancement in growth and reduction in pathogenic effect and nematode infestation although the alone placement of biochar was not found much efficient. Biochar and saw dust when applied together improved plant growth better and suppressed the nematode multiplication more than individual usage of biochar. Application of biochar strategies has little been studied. So far no available reports with respect to meticulous utilization of biochar integrated with sawdust to manage the root-knot nematode. The results attained by the single application of biochar are backed by several workers (Perry and Beane, 1988; Zhang et al., 2013; Al-Fraih, 2015). Biochar amendment directly into the rhizosphere is
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Discussion thought to be more beneficial for crop growth might be due to release of chemicals and enhancing the beneficial rhizospheric flora and improvement in soil health. Besides this, the positive effect of biochar might be due to the sorption capacity of toxic chemical compounds and hydrophobic organic compounds and high water and nutrient retention which favour the decomposition that may leads to improve the soil health and reduced the environmental risk. Vaccari et al. (2015) found that the use of biochar on tomatoes increased plant growth compared to that of non-amended control for processing tomatoes. Rahman (2014) has shown that the addition of biochar to a grape vineyard reduced the incidence of plant parasitic nematodes by a factor of eight compared to the control. Huang et al. (2016) observed activation of ethylene responses and H2O2 accumulation may contribute to the capacity of biochar to suppress nematode infection inside roots. Boerjan et al. (2003) observed that the increased production of H2O2 in plants can lead to the polymerization of monolignols by peroxides and the formation of lignin. The suppression of soil pathogens by biochar may be from several mechanisms, including improved nutrient solubilization and uptake, which helps enhance plant growth and resistance to the stresses of pathogens; microbe stimulation, which promotes direct competition or parasitism against pathogens; or induced plant defense mechanisms (Elad et al., 2011). This exogenous ethylene production from biochar-amended soil might induce resistance in plants to pathogens (Spokas et al., 2010; Lehmann et al., 2011). A number of explanations for these impacts have been offered, such as biochar sorption, including the presence of volatile organic compounds (VOCs) that can inhibit or stimulate microbial mineralization reactions or affect plant–microbial interactions (Graber et al., 2010; Spokas et al., 2010) and the availability of B and Mo, which are important cofactors in biological nitrogen fixation (Rondon et al., 2007). Maximum suppressive effect for the nematode infestation and ameliorating the growth attributes was observed when biochar was applied with Eucalyptus sawdust due to their highest additive or synergistic effect. The efficacy of Eucalyptus over all the oil cakes combined with biochar could be due to the amount of different chemicals released in the form of secondary metabolites and essential oils. Eucalyptus species contain some essential oils which are known to be toxic against bacteria, fungi, insects and nematodes (Batish et al., 2008). Rokiek and El-Nagdi (2011) indicate that aqueous extracts of E. citriodora leaves were effective in reducing galls and egg masses of M.
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Discussion incognita on roots and increase in growth of sunflower plants. Effectiveness of sawdust against root- knot nematode on okra confirms the earlier findings of (Kokalis- Burelle et al., 1994; Imre et al., 2011; Obasi et al., 2013). Khan et al. (2004b) used Datura, Calotropis, Neem leaves and sawdust of Shisham alone and in combination reduced the number of galls of M. incognita on tomato and increased the growth significantly. Sitaramaiha et al. (1978) found salwood sawdust very effective against nematodes in increasing the growth of tomato. It may be because of their feudal effect. Increased microbial activity in amended soil is to bring about increased conversion of nitrogen to nitrate form, which results ultimately in increased metabolic activity of plants and plant growth (Tiyagi and Ajaz, 2004). The improvement in plant growth characters by the use of sawdust may be attributed due to the fact that these organic additives after their decomposition release some nutrients and serving as manure increase the nutrient status of the soil and improve physicochemical properties of soil (Huang and Huang, 1993), which promote rapid root development and finally overall plant growth. Hassan et al. (2010) have reported the growth promoting effects of sawdust on tomato plants. The inhibitory effect of sawdust on nematodes can be attributed to the formation of phenolic compounds by the decomposition of sawdust (Kokalis-Burelle et al., 1994). Other reason of their reduction was that of the development and colonization of nematode natural enemies (Oka, 2010; Thoden et al., 2011) in higher sawdust amended soil. Increased colonization and reproduction of the nematophagous fungi was reported in sawdust amended soil (Hassan et al., 2010). Although application of biochar as integrated management is quite effective and efficient method for nematode management and improvement in plant growth but exploration and further characterization is needed. Combined effect of biochar with the amendment results in positive and beneficial relationship in the form of synergistic effect that leads to enhance the plant growth characters and reduction of nematode infestation. Combined application of biochar with Eucalyptus results in greater root colonization although this synergism mechanism is not clear for disease management point of view.
In the next experiment of the current study the potential of biochar alone and in combination with various agriculture waste viz., tobacco, garlic waste, mentha waste, decomposed potato, black gram waste and bagasse were demonstrated for the management of root-knot nematode M.incognita and improvement of plant growth
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Discussion characters of tomato. Although all the treatments either singly and in combination substantially reduced the root knot nematode infestation but the integration of biochar along with the tobacco waste was found most momentous followed by garlic waste and mentha waste for escalating the plant growth characters and reduction in nematode population. Very little work has been done for the application of biochar for nematode management. Beside this no review literature is available for the management of nematode in integrated manner though the involvement of biochar with agriculture wastes. Utilization of agricultural wastes as an organic amendment alters the soil rhizospheric activity by enhancing and promoting the flora and fauna that are antagonistic to nematodes. The finding of various workers vindicates our results (McSorely and Gallaher, 1995b; Oka and Yerumiyahu, 2002; Ogwulumba et al., 2010; Hassan et al., 2010; Rashid et al., 2011). Eyal et al. (2006) reported that soil amended with spent tea, sugarcane bagasse, paddy husk, wheat straw, domestic garbage, saw dust and cellulosic waste significantly decreased the root-knot population. The use of composted dry cork, dry-grape (fruit residue after pressing) and mixture of dry-olive, dry-rice husk as an amendment to potting mixtures was assessed for the management of root-knot nematodes (Andres et al., 2012). Similar observation was elucidated by Gong et al. (2013) and Zasada et al. (2010) reported that garlic straw suppressed nematode population. Marrush (2007) observed that the amending mushroom compost into soil containing low organic matter resulted in poor performance in nematode suppression. Motha et al. (2010) demonstrated that addition of tobacco dust as soil amendment significantly reduced the nematode development and helps in improvement of plant growth. The positive and promoting effects of organic additive application have been generally considered to be due to increase in soil nutrients, improvement in physical and chemical properties of soil and direct or indirect stimulation of predators and parasites of plant parasitic nematodes (Kumar et al., 2005; Kumar and Jain, 2007) and release of toxic chemicals that act as natural nematicides (Olabiyi, 2004; Tobih et al., 2011; Kumar et al., 2011). Youssef and El- Nagdi (2010) revealed that the application of onion bulb, dry leaves of sugar beet, fleabane and jojoba, filter cake or mud as sugarcane industrial residue and nile fertile mineral bio-fertilisers enhanced the plant growth parameters and reduction of nematode infestation in banana. Some studies recorded promising results of biochar on several plant pathogens (Elad et al., 2010; Harel et al., 2012; De Tender et al.,
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2016) and pests (Elad et al., 2010). The amendment of poultry-litter biochar to the soil generally decreased the number of plant-parasitic nematodes while increasing the amount of free-living nematodes in the soil (Rahman et al., 2014). Elmer and Pignatello (2011) revealed that addition of biochar made by fast pyrolysis of wood powder results in decrease in Fusarium infection of Asparagus. Also, infection of tomato with another soil borne disease, bacterial wilt (Ralstonia solanacearum), was significantly reduced by adding wood biochar and biochar made from municipal biowaste (Nerome et al., 2005). Studies by Mian et al. (1982a) demonstrated that when organic amendments are added to soil at a rate of 1.0% (w/w) there is a direct relation between the N content, or inverse with the C: N ratio of the amendments and their effectiveness against phytonematodes. Although ammonia and organic amendments with low C: N ratios are nematicidal (Mian et al., 1982b and Rodriguez- Kabana, 1981). Haefele et al. (2011) observed yield increases in yield of rice from 16 to 35% with rice hull biochar Thus, straw biochar is a good soil conditioner in tobacco field, which can improve tobacco growth and nutrients adsorption at appropriate level through both directly and indirectly effect. Finding of the study concluded that integrated effect of soil organic amendment with biochar is one the potent and reliable method for the nematode management in a facile and ecofriendly manner without disturbing the equilibrium of the soil and environment. Suppression and reduction in nematode infestation due to integrated nematode management with biochar might be due to release of allochemicals by the enhancement microflora and fauna and that are produced due the degradation of the amendments in addition to the promoting a activity of antagonistic rhizosphere. Findings of my study also suggest that mixture of biochar with saw dust and agriculture waste may protect in a better from the nematode infestation.
Effect of biocontrol agents, agriculture waste and oilcakes Soil ecosystem is extremely a complex structure, involving determination of interactions between microbial, chemical, physical and plant host variables (Villenave and Duponnois, 2002). The first recorded case of interaction between nematodes and fungi was observed by Atkinson (1892). Interactions involving nematodes are important because they contribute substantially to variability in crop growth (Zadoks and Schein, 1979). Most of the soil-borne plant diseases are occasionally occurred due to the interaction of two or more pathogens of the same or different groups leads to
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Discussion the formation of disease complexes. Disease complexes are produced when synergistic interactions occur between organisms (Back et al., 2002). In another study, the inoculation of biocontrol agents viz., Pseudomonas fluorescens, Purpureocillium lilacinum, Pochonia chlamydosporia and Trichoderma viride were evaluated individually, concomitantly or sequentially with root-knot nematode, M.incognita in pots. All biocontrol agents tested during the experiment results in the significant enhancement in plant growth characters and physiological parameters (chlorophyll content, carotenoid content and nitrate reductase activity) as compared to untreated inoculated control. Alone inoculation of M.incognita showed highest nematode multiplication and most significant reduction in plant growth and physiological parameters followed by sequential inoculation of M.incognita prior to 15 days with fungus. Our results are also in conformity with other workers (Hasan, 2004; Vidyasagar et al., 2012). Ganaie and Khan (2010) found significant reduction in eggmass/root, eggs/eggmass and population of M. javanica on sequential inoculation of P. lilacinus 10 days prior to the inoculation of M. javanica. Lesser reduction comparatively, inoculated with biocontrol agents followed by M.incognita is acceptable as the plant applied with bioagents got sufficient to colonize the root tissue making it less susceptible to the nematode attack or toxic secretion of screened bioagents produced antagonistic effect on nematode also confirmed by Van and Gundy (1975), Sharma and Gill (1979) and Parveen et al. (2007). Moreover, in those treatments where M.incognita was inoculated prior to bioagents, the number of eggmasses/root, eggs/eggmass and M.incognita population was higher as compared to other combinations because nematode gets adequate time to multiply in the root system of the plants, as a consequence results in limiting the effect of latter inoculated bioagents. Our results are confirmed by the works of Ganai and khan (2010) and Vidyasagar et al. (2012). The findings of our results also revealed that P.fluorescens was the most effective followed by P.chlamydosporia and P.lilacinum. Although T.viride was found to be least efficient amongst all the tested biocontrol agents. Our results are in agreement with some others scientists (Kerry and Hirsh, 2005; Dong and Zhang, 2006; Wu et al., 2010). Esfahni and Pour (2006) observed that P.lilacinum an opportunistic biocontrol agent was most effective when the fungus and root knot nematode M. javanica were inoculated simultaneously or the fungus prior to the nematode in sequential inoculation on tomato in greenhouse conditions.
162
Discussion
In another experiment, the screened bioagents viz., Pseudomonas fluorescens, Purpureocillium lilacinum, Pochonia chlamydosporia and Trichoderma viride were integrated with various organic soil amendments viz., castor cake, press mud, mustard cake, soybean cake, mahua cake and cotton cake to test their potential and their impact on growth of tomato cv.K-21and nematode infestation. Although all the treatments considerably abate the root-knot nematode infestation but the alliance of bioagents with castor cake, press mud and mustard recorded to be most potent treatments for the improvement of growth and growth yielding attribute and reduction in root-knot nematode infestation. Oil cakes because of their high decomposition processes, low C/N ratio and release of toxic chemicals favours as one of the most important tactic for nematode management. They provide slow and steady nourishment and protection to host plant; create antagonistic micro flora and fauna for pathogens, including soil nematodes (Khan et al., 2011). The combined application of biocontrol agents and various oil cakes have been considered to be an efficient approach of the management of nematodes (Borah and Phukan, 2004; Zareena and Kumar, 2005; Sundraraju and Kiruthika, 2009; Parihar et al., 2015). Pandey (2005) reported that the application of oil seed cakes and T. harzianum significantly enhanced crop yield and significantly reduced the nematode populations and root-knot indices. Bhat et al. (1998) studied the combined application of P. lilacinus and oil cakes for protection of chickpea against M. incognita. The incorporation of neem cake along with P. lilacinus caused the reduction of root-knot nematode, the number of galls and egg masses (Sharma et al., 2007). Biocontrol agents such as P. chlamydosporia, P. lilacinus and T. harzianum generally colonize and establish themselves in soils rich in organic matter (Khan et al., 2012) and these agents usually offer improved biocontrol activity (Stirling, 1991). Pseudomonas species are aggressive colonizers of the rhizosphere of various crop plants (Schroth and Hancock, 1982) and have a broad spectrum antagonistic activity against plant pathogens. Wei et al. (1996) revealed that some species of Pseudomonas caused greater colonization as well as greater siderophore production. Due to this, P. fluorescens cause significant enhancement in plant growth characters of tomato and also reduced the root knot nematode infestation. Ahmad and Khan (2004) and Ahmad and Alam (1998) reported integrated approach of managing nematodes with P. lilacinus together with organic matters. The organic amendments also increased the parasitism of P. lilacinus and T. harzianum on M. incognita (Ahmad and Khan, 2004; Ashraf and Khan, 2008). 163
Discussion
Integrated effect of biocontrol agent and organic amendment were effectively used for nematode control (Tiyagi and Ajaz, 2004; Sharma and Pandey, 2009; Pandey et al., 2011). Many other researchers reported the nematicidal potential of T .harzianum and P .lilacinus alone and in combination with organic amendment and other biological control methods (Kiewnick and Sikora, 2006; Anastasiadis et al., 2008). Organic manuring caused a lot of advantageous role such as improvement of soil health and soil conditioner that favours for the plant growth. Nutrients availability after the decomposition may be helpful for the colonization of bioagents in the root. The improvement of plant growth characters and production of more biomass might be due enhancing the photosynthetic rate and mobilizations of growth promoting nutrients and suppression of nematode severity by the combined usage of bioagents and organic amendment.
In another experiment, tested biocontrol agents viz., Pseudomonas fluorescens, Purpureocillium lilacinum, Pochonia chlamydosporia and Trichoderma viride were integrated with different organic matters viz., straw of Pigeon pea, Pearl millet, Maize, Mustard, Sorghum and Potato waste and their effect on plant growth parameters of tomato cv.K-21 and root-knot nematode severity. All the treatments significantly improved the plant growth parameters but among all, P. fluorescens along with the mentha waste was found most inhibitory for the reduction in nematode multiplication and exaggerating the plant growth characters of tomato. This enhancement might be due to the better colonization of tomato root by the P.fluorescens in mentha waste amended soil which caused better protection of the plant with the nematode infection thereby improved the plant growth. Buena et al. (2007) stated the pepper crops residues showed inhibitory effect against nematode management. Sharma et al. (1997) observed that the application of water hyacinth compost, mustard straw, rice husk and asparagus compost reduced nematode multiplication and improved plant growth. Similar observations were also showed by Akthar (1993) who reported that dead vegetation, sugarcane trash, corn shucks, paddy straw and husk, press mud reduced the nematode multiplication and enhanced plant growth. Gupta and Kumar (1997) stated that level of reduction of Tylenchorhynchus spp. and Helicotylenchus spp. in soil increased at higher doses and the longer periods of treatment by compost, fenugreek and chickpea straw. Pearl millet increases the supply of the nutrients and provide favourable medium for plant growth (Southey, 1978) and inhibit pathogen
164
Discussion proliferation (Siddiqui, 2004). Rizvi et al. (2016) revealed that combined inoculation of Glomus fasciculatum + Avena sativa straw and Trichoderma harzianum was found to be the most efficient in improving the plant growth parameters and decreasing the root-knot development of M. incognita in chickpea plants. It has been considered that decomposition of organic amendment release chemicals that are toxic to the nematode (Sayre et al., 1964; Alam et al., 1979; Badra et al., 1979) and they dispersed within the soil pore spaces where most of the noxious nematodes occur. During watering, the soluble fractions and other products released into the pore space and kill the nematodes (Alam, 1979).
Finding of the study done in vitro and pots condition demonstrate that bioagents and organic amendment showed prominent effect on the reproductive potential of nematode and plant growth parameters of tomato. In future studies more detailed investigation is needed for the characterization, exploration and to develop the relationship for the biocontrol of plant disease. The present study also advocate to work on the integration of various bioagents with different organic amendment with other management practices for providing facile, ecofriendly, economically viable and environmentally adaptable and sustainable management practice.
165
Summary
SUMMARY
The present study was performed to cogitate the results of various experiments conducted under glasshouse conditions to find the resistance and susceptibility behaviour of different cultivars of tomato and to assess the efficacy of different organic soil amendments and biocontrol agents, viz., Pseudomonas fluorescens, Pochonia chlamydosporia, Trichoderma viride, and Purpureocillium lilacinum, applied either alone or in combinations against Meloidogyne incognita attacking Tomato, Solanum lycopersicum L cv. ‘K-21’. Different organic additives including various plant parts, organic cake of castor, mustard, mahua, soybean, cotton, press mud, biochar, sawdust and agriculture waste were used for the management of root knot nematode Meloidogyne incognita. Aqueous extracts of leaves of some selected plant species, oil cakes and biochar were tested for their antinemic properties in vitro. Similarly the culture filtrates of biocontrol agents were also examined for their nematostatic and nematicidal effect against eggs and juvenile of M.incognita. These extracts were also tested as bare-root dip penetration and root dip treatment evaluating their systemic activity against the M.incognita. In another study seed dressing of tomato cv. K-21 with bioagents, biochar and aqueous extracts of various oil cakes viz., castor, mustard, mahua, soybean and cotton were done to control root knot infestation caused by Meloidogyne incognita. The summary of the above experiments is as follows:
All the fourteen cultivars of tomato screened to test the resistance and susceptibility behaviour responded differently in terms of plant growth characters and root knot nematode infestation level. The overall reduction in plant growth characters was observed in all the tested cultivars of tomato which may be due the infestation of nematode. The cultivar H-88-78-1 showed highest resistance against the root knot nematode by the presence of number of eggmasses, eggs and root knot indices. The cultivar K-21 was found highly susceptible. On the basis of resistance, tomato cultivars can be arranged in the following descending order: VRT-101A> EC- 570018> GT-2 > PB Barkha bahar-2> FEB-02> CO-3> Kalyanpuri-T1> NDT-3> EC- 538380> GT-3> GT-1> S-22> K-21.
Aqueous dilutions of leaf extract of some selected plant species were found to be toxic to M.incognita. Similar results were received with the aqueous dilutions of
166
Summary oil cakes and biochar. The toxicity of different extracts and exudates was found to be concentration and time dependent. The aqueous leaves extract of Mexican poppy, wild eggplant and black pigweed showed maximum egghatching inhibitory effect and juvenile mortality of M.incognita. Similarly the aqueous extracts of castor cake and mustard cake produced the most stricking effects against the M. incognita juveniles. The inhibition in egg hatching was found directly proportional to the concentration of the extract and released exudates. In a related study the culture filtrates of biocontrol agents were tested for nematicidal and nemtostatic action under in vitro conditions. All the bioagents showed deleterious effect on the egg hatching and juvenile mortality of M. incognita juveniles. The inhibition in egg hatching was found directly dependent to the concentration of the extract whereas the mortality was inversely correlated to the dilution of the filtrate.
Aqueous leaf extract of some selected plants showed deleterious effect on the percent inhibition of juvenile penetration of M.incognita into the root of tomato cv.K- 21. Similar results were obtained with the aqueous extracts of oilcakes and biochar. The aqueous leaf extract of Mexican poppy, Wild eggplant and Trailing eclipta results in maximum inhibition in penetration, while in oil cakes aqueous extract of castor cake, mustard cake and soybean cake caused highest percent inhibition in penetration. Similarly the aqueous dilution of biochar at 5% of the concentration caused most pronounced effects against juvenile penetration. Penetration of juveniles significantly decreased with increase in the dip duration and differentially varied from treatment to treatment as compared to the undipped inoculated control. In the related study all the bioagents give rise to significant inhibition in the juvenile penetration. The percent inhibition in juvenile penetration was found directly proportional to the concentration of the extract.
The bare-root dip treatment of tomato cv. K-21 seedlings with aqueous dilutions of leaf extracts of Mexican poppy, Wild eggplant and Trailing eclipta conferred protection against root-knot nematode, M. incognita. The aqueous leaf extracts of Mexican poppy showed most promising results followed by Trailing eclipta and Wild eggplant in suppressing the root knot indices and provided curative therapeutic effect to seedlings. With increase in the concentration of the extracts and dip duration the lethality increases. Similarly the aqueous dilution of Pseudomonas fluorescens and Pochonia chlamydosporia attain the most encouraging results. In the 167
Summary allied study the combined utilization of bioagents P. fluorescens, P. lilacinum and P. chlamydosporia and extracts of water hyacinth also assessed as bare root dip treatment. Combined application of all the bioagents and plant extract significantly improved plant growth and check nematode infestation. P. fluorescens and P. chlamydosporia were found reliable in suppressing the root knot incidence and enhancement of the plant growth characters. As a consequence in the suppression of the root-knot development the plant growth characters increased.
The study was conducted under glasshouse conditions to appraise the nematicidal potential of chopped leaves of some selected plant species applied @ 50g/ pot in combination with Black nightshade (Seed powder@10g) against root knot infestation caused by Meloidogyne incognita on tomato cv. K-21. All the treatments were found to be effective in reducing the nematode population and root knot nematode multiplication in terms of number of eggmasses/plant, eggs/eggmass and nematode population/250g soil and thereby results in augmenting the various plant growth characters. Among all the applied treatments Mexican poppy + Black nightshade (Seed powder) brought about most pronounced effect followed by Trailing eclipta+ Black nightshade (Seed powder) Wild eggplant+ Black nightshade (Seed powder) in suppressing the nematode infestation and thus supporting the plant growth characters in terms of length, fresh and dry weight, pollen fertility and yield/plant.
In the related study exploration were done to investigate the potential of biochar alone and in combination with saw dust of different plants viz., eucalyptus, lebbeck, jambul, mango, poplar and babool against M.incognita under glass house conditions. The results of the study revealed that combined action of biochar with saw dusts significantly reduced the eggmasses/ root, eggs/eggmass, population of M.incognita and root knot indices and by virtue of this increased the plant growth parameters. Biochar alone caused antagonistic effect on nematode but the biochar along with the saw dust of eucalyptus saw dust showed maximum potential followed by biochar+ lebbeck and biochar +jambul in exaggerating the plant growth characters and reducing the nematode infestation. A similar study was conducted to examine the efficacy of biochar alone and in combination with the agriculture waste viz., tobacco, garlic, mentha, decomposed potato, black gram and bagasse waste against the root knot nematode, M.incognita under glasshouse conditions. It was observed that all the treatments either alone or in combination were effective in minimizing the root knot 168
Summary nematode infestation in terms of number of eggmasses/plant, eggs/eggmass and nematode population/250g soil and root knot indices or thus enable to improve the plant growth characters. The application of biochar in combination with tobacco was showed most pronounced effect in exaggerating the plant growth characters and to abating the nematode infestation. It was followed by biochar+ garlic waste and biochar+ mentha waste. Biochar alone also caused antagonistic effect in the suppression of nematode disease and improvement of plant growth characters but not too much effective.
The oil cakes were tested as seed dressing agents for tomato cv. K-21. Seed dressing with different oil cakes viz., castor cake, mustard cake, soybean cake, mahua and cotton cake rendering the safety against the root-knot nematode M.incognita infestation and also improved the plant growth characters, where as castor and mustard cake showed the most promising results. The efficacy of oil cake varied from treatment to treatment and dose dependent manner for nematode infestation and improvement of plant growth parameters. In the similar study seed treatment with biochar at various concentrations enhance the plant growth characters and suppression of nematode infestation. Among all concentration, 3.2 and 6.4% concentration of biochar showed most pronounced improvement in plant characters and suppression of root knot nematode development. In a related study seed treatment with culture filtrates of biocontrol agents, viz., P. fluorescens, P. chlamydosporia, P. lilacinum, and T. viride were investigated against the root knot nematode M.incognita infestation. All the bioagents significantly increased the plant growth characters and crop yields but also reduced the nematode severity. P. chlamydosporia and P. fluorescens caused most pronounced effect in arresting the root knot incidence and enhancing the plant growth characters.
Utilization of bioagents viz., Pochonia chlamydosporia, Pseudomonas fluorescens, Purpureocillium lilacinum and Trichoderma viride significantly reduced the root knot nematode development and multiplication when either applied individually or in combination with organic soil amendment. Alone application of bioagents and in combined use caused suppression in the nematode development in tomato cv.K-21 and enhancement of plant growth characters and yield. Among different treatments, P. fluorescens and P. chlamydosporia were found most effective whereas T. viride was least effective. Similarly the combined application of P. 169
Summary fluorescens + castor cake, P. fluorescens + mustard waste and P. chlamydosporia+ castor cake were observed most promising in reducing the nematode population and enhancing the plant growth and yield of test crops.
The integrated application of bioagents and organic soil amendments was very effective in managing the nematode development which improved the plant growth characters and crop yields. However, among the various treatments, the combined exploration of bioagents with castor cake brought about most significant suppression in nematode infestation and improved the growth and yield of test crops significantly.
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