International Journal of Agriculture and Biosciences www.ijagbio.com P-ISSN: 2305-6622 E-ISSN: 2306-3599 [email protected]

RESEARCH ARTICLE

Effect of Leachates of Alien Weeds on Seed Germination, Seedling Growth and Physiology in Mungbean

NA Ghayal1, MV Biware2 and KN Dhumal3

1Department of Botany; 2Department of Chemistry, Abasaheb Garware College, Pune 411004, M.S., India 3Department of Botany, University of Pune, Pune – 411007

ARTICLE INFO ABSTRACT

The invasive weeds like Cassia uniflora Mill. non Spreng and Synedrella Received: March 10, 2014 nodiflora (L) Gaertn, are showing dominance in Pune University campus. They Revised: May 12, 2014 are observed in the form of pure stands reducing the phytodiversity in the areas Accepted: July 15, 2014 studied. The allelopathic potential of invasive weeds was screened by using

Key words: their aqueous leaf leachates for seed germination bioassay which indicated that Allelobiogenesis the seed germination, root and shoot length, root: shoot ratio and vigour index Cassia uniflora were significantly inhibited in mungbean at all concentrations. The results of germination studies seed germination bioassay were in conformity with that of seedling physiology. Synedrella nodiflora The effect of Synedrella leachates was more pronounced than that of Cassia. It was confirmed that both the weeds had strong allelopathic or phytotoxic *Corresponding Address: influence due to presence of different allelochemicals. [email protected]

Cite This Article as: Ghayal NA, MV Biware and KN Dhumal, 2014. Effect of leachates of alien weeds on seed germination, Seedling growth and physiology in mungbean. Inter J Agri Biosci, 3(4): 141-148. www.ijagbio.com

INTRODUCTION adaptability to diverse habitat, deep root system, faster growth rate, most effective and efficient pollination and Presently the scientists working in the field of weed seed formation ability, seed polymorphism and its biology are developing an integrated approach towards effective dispersal mechanisms, biotic and abiotic stress weed management for their long term control (Rao, 2000), tolerance along with synthesis of some novel and as they cause heavy losses in crop yield, than from the defensive allelochemicals. agricultural pests. At present the crop fields and forests It has been claimed (Inderjit and Weston, 2000) that are being invaded by many exotic weeds like Cassia, detailed allelopathic research on weed – weed and weed – Synedrella, Parthenium, Chromolaena, Lantana etc. crop interactions is lacking in India and many other which are causing a serious problem. These invasive developing countries. The present floristic studies have exotic species greatly influence the phytodiversity and revealed that plant diversity had been reduced by invasive replace the native species (Lee and Klasing, 2004). alien weed species like Alternanthera, Cassia, Synedrella, Regardless of several allelopathic studies on these weeds, Parthenium, and Acalypha etc. in the campus of Pune their mechanism of invasion are not yet fully understood University (Almeida, 2003; Varadpande, 1972). Hence to (Sutherland, 2004). explore the mechanism of successful invasion of Cassia Cassia uniflora Mill.non Spreng. (Syn. Cassia and Synedrella in the University campus, this study was sericea Sw.- Caesalpinaceae) having worldwide undertaken. The allelopathic potential of both these alien distribution. It is an annual, erect herb with stipulate, weeds was also investigated using seed germination glandular, paripinnate leaves. Flowers are yellow with bioassay technique on sensitive test crops like mungbean. clustered pods. If plants get water, they keep on The physiological and biochemical changes in leaf flowering throughout the year. This weed grows leachates treated seedlings of mungbean along with luxuriantly if it gets full sunlight and high soil moisture. control were analyzed at seventh day. It has been recorded at many other places like the highways, railway tracks, barren lands and sometimes MATERIALS AND METHODS even in agricultural lands. Synedrella nodiflora (L.) Gaertn. (family- Preparation of leachates Asteraceae) originated in tropical South America is an The leaves of Cassia and Synedrella were collected annual, herb and is now distributed all over India during the flowering stage and brought to laboratory, (Almeida, 2003). Its dominance is attributed to wide cleaned with distilled water and spread on filter paper for

141 142 Inter J Agri Biosci, 2014, 3(4): 141-148. shade drying. The dry leaves were ground in Wiley Mill fast and repeatable tools for investigating the potential for to pass two mm sieve. 100g powdered leaves were soaked different types of interactions. To prove the hypothesis of in 1000 ml distilled water for 24 h at 250C and the allelopathic interactions between test crop like mungbean leachate was filtered through Buchner funnel. It was and invasive weeds like Cassia and Synedrella, present stored in refrigerator in amber coloured bottle (to avoid investigation was focused on seed germination bioassay degradation). and analysis of the seedlings.

Seed Germination Bioassay Seed germination bioassay The seeds of mungbean (Vigna radiata var. Vaibhav) % Seed germination were obtained from College of Agriculture, Pune. Healthy In case of mungbean the effect of leaf leachates of seeds were used for bioassay study using sterilized Cassia and Synedrella the seed germination showed petriplates (9 cm diameter) lined with germination papers. inhibition as compared to control. The results were The seeds of radish and mustard were surface sterilized statistically significant at p = 0.001 (Tables 1, 2). with 0.02% aqueous HgCl2 for two minutes. Then the seeds were thoroughly washed with distilled water. Seed Root and shoot length germination papers were thoroughly moistened (5ml.) In mungbean (Tables 1, 2) both the leaf leachates with respective concentrations of leaf leachates of caused decrease in root length, except at 1.5% Synedrella (1.5% - 10% v/v) which were prepared by concentrations of Synedrella leaf leachates. At 2.5% dilutions with distilled water. The seeds kept in distilled concentration of leaf leachates it was at par with control. water were considered as control. Twenty seeds of radish The leaf leachates of Cassia and Synedrella caused and forty seeds of mustard were uniformly kept in each increase in shoot length at 1.5% and 2.5% concentration petri plate. The 2.0 ml leachates of respective over control respectively. The results were statistically concentration were added in each petri plate on third day. significant at p = 0.001. The petri plates were irrigated with the leachates only once. The treatments were replicated thrice. Seed Root: Shoot ratio germination percentage, root and shoot length, total In mungbean there was increase in R: S ratio as seedling height and vigour index (Gupta, 1996) were compared to control. The results were non-significant for recorded on 7th day. mungbean (Tables 1, 2).

Biochemical analysis of seedlings Vigour Index (VI) Randomly selected 0.1g material of seedlings (control The results of vigour index in mungbean (Tables 1, 2) and leachates treated) was taken for analysis of different were corroborating with that of % seed germination for biochemical parameters. Total sugars were estimated by both Cassia and Synedrella leaf leachates. the method of Hodge and Hofreiter (1962). Starch content was estimated from 1g of leaf samples (shade dried) by Seed germination using Anthrone reagent as per the method described by Emergence of root and shoot is a primary phase in Thayumanavan and Sadasivam (1984). Protein estimation seed germination, which determines the future plant was done according to Lowry et al. (1951) method. Total growth. Many workers have reported positive as well as phenolics in the leaf samples were estimated as per the negative impact of aqueous leachates on seed germination method of Farkas and Kiraly (1962). and development of root and shoot. In the present investigation, the lower and higher concentration Assay of antioxidant enzymes treatments of Cassia and Synedrella were inhibitory as The Peroxidase [EC 1.11.1.7] and Polyphenol compared to control for mungbean (Tables 1, 2) oxidase [EC 1.14.18.1] enzyme assay was carried out by The different types of allelochemicals present in using Vidyasekharan and Durairaj (1973) method. While leachates of Cassia and Synedrella might have caused the the Superoxide dismutase [EC 1.15.1.1] enzyme assay retardation in seed germination percentage by acting was carried out by using Dhindsa et al. (1981) method. negatively on the process. With increasing concentrations of leachates the allelochemicals present in them might Statistical analysis have acted more negatively, leading to complete The data were summarized as the pooled means of inhibition of seed germination of mung bean. The three replicates each over three years with standard favourable or adverse impact of allelochemicals on seed deviation as the measures of variability. One-way germination is concentration/ dose dependant. An (2005) ANOVA was used to compare the mean values as affected had described this phenomenon as “Hormesis”. by different leachate concentrations. Duncan’s Multiple The delay in seed germination can have important Range Test (DMRT) was applied at p<0.05 to compare biological and ecological implications, because it can the mean differences. The data were analyzed by affect the ability of the seedlings to establish in natural SYSTAT (ver.11) and Microsoft excel 2000. conditions (Chaves et al., 2001). The observations of the present investigation on seed germination and seedling RESULTS AND DISCUSSION growth are in agreement with Baber et al. (2009), Wakjira (2009) and Dave and Jain (2009). They noted significant According to Inderjit and Weston (2000) and Inderjit increase in seed germination of different crops due to and Nilsen (2003) laboratory bioassays and field studies lower concentration treatments of leaf extracts and are integral part of allelopathy research, because they are leachates of Ageratum, Anagallis, Parthenium etc. The 143 Inter J Agri Biosci, 2014, 3(4): 141-148.

Table 1: Effects of Cassia leaf leachates on seed germination and seedling growth in mungbean Conc. % % Germination Root length Shoot length Root: Shoot ratio Vigour index Control 99 a±3.96 6.23 a±0.37 10.33 a±0.85 0.60 bc±6.76E-02 930.00 a±76.54 1.5 95.46 a±4.77 5.73 ab±0.20 7.73 b±1.85 0.76 ab±0.15 696.00 b±167.24 2.5 85 b±5.95 5.70 ab±0.30 10.56 a±0.95 0.54 c±2.05E-02 845.33 ab±76.03 5.0 70 c±4.2 5.50 b±0.30 6.90 bc±0.96 0.80 a±0.12 483.00 c±67.50 7.5 55 d±2.2 4.53 c±0.15 5.86 bc±0.15 0.77 ab±3.46E-02 352.00 cd±9.16 10.0 34 e±1.02 4.10 c±0.36 5.00 c±0.50 0.82 a±8.81E-02 200.00 d±20.00 p = <0.001 <0.001 <0.001 0.014 <0.001

Table 2: Effects of Synedrella leaf leachates on seed germination and seedling growth in mungbean Conc. % % Germination Root length Shoot length Root: Shoot ratio Vigour index Control 99 a±3.96 6.23 b±0.37 10.33 a± 0.85 0.60±6.765E-02 930.00 a±76.54 1.5 90.46 b±4.52 7.20 a±0.75 9.63 b±0.80 0.74±9.769E-02 871.12 a±80.82 2.5 85 b±5.95 6.33 ab±0.61 7.26 c±0.47 0.87±0.13 654.00 b±42.53 5 74 c±2.22 4.00 c±0.60 5.20 d±0.30 0.76±7.128E-02 312.00 c±18.00 7.5 54 d±3.24 3. d 00±0.20 3.60 e±1.05 0.88±0.3062 180.00 d±52.67 10 36 e±2.16 1.23 e±0.25 1.56 f±0.32 0.79±7.358E-02 62.66 e±12.85 p = <0.001 <0.001 <0.001 0.286 <0.001 # Data is the pooled means of three estimates each over two years ±standard deviation. ‘p-value’ denotes the significance of difference between the means by one way ANOVA statistics. a The values followed by different letters differ significantly by Duncan’s multiple range test at p=0.05.

Table 3: Effects of Cassia uniflora leaf leachates on organic constituents in seedlings of mungbean Conc. % Total sugars mg/g Starch mg/g Protein mg/g Phenolsmg/g Control 64.63 b±1.94 10.41 d±0.78 81.66 a±7.74 8.55 b±0.64 1.5 65.62 b±3.94 16.04 c±1.04 111.66 b±7.24 9.53 ab±0.62 2.5 67.66 b±4.74 18.87 c±1.03 122.5 ab±12.24 9.17 b±0.50 5 69.98 ab±2.80 22.45 b±2.13 126.66 ab±6.96 9.78 ab±0.92 7.5 72.08 ab±3.60 25.70 b±2.57 127.5 ab±9.56 9.90 ab±0.99 10 75.29 a±5.27 31.67 a±2.53 131.66 a±10.52 10.88 a±0.87 p = 0.043 <0.001 <0.001 <0.001

Table 4: Effects of Synedrella nodiflora leaf leachates on organic constituents in seedlings of mungbean Conc. % Total sugars mg/g Starch mg/g Protein mg/g Phenols mg/g Control 64.63 b±2.59 10.41 e±0.78 84.16 d±4.62 8.55 bc±0.64 1.5 67.36 ab±3.37 17.82 d±1.15 119.16 c±11.32 8.43 c±0.84 2.5 69.1 ab±4.84 30.15 c±1.65 127.5 c±9.56 8.80 bc±0.48 5 71.22 ab±2.14 32.65 bc±3.10 134.16 bc±10.7 8.92 ab±0.71 7.5 73.19 a±4.39 37.13 b±3.71 149.16 ab±9.68 9.90 ab±0.94 10 74.55 a±4.47 42.15 a±3.37 155.82 a±15.58 10.27 a±0.66 p = 0.06 <0.001 <0.001 <0.001 # Data is the pooled means of three estimates each over two years ±standard deviation. ‘p-value’ denotes the significance of difference between the means by one way ANOVA statistics. a The values followed by different letters differ significantly by Duncan’s multiple range test at p=0.05. leachates of Cassia and Synedrella, which might be allelochemicals and the species. As suggested by inducing the increase or decrease in membrane Einhellig (1995), Reigosa et al. (1999) the allelochemicals permeability, activities of metabolic enzymes and rate of present in the leachates might be responsible for the cell division in germinating seeds of mungbean. changes in water relations, membrane permeability and Oyun (2006) also reported that Acacia auriculiformis enzymatic activities of protein and carbohydrate and Gliricidia sepium leaf leachates inhibited both seed degradation during seed germination process, which germination and seedling growth of maize due to determined the success of seed germination. Same may be inhibition to water absorption in germinating seeds and the reason for the changes in the germination of mung that of nutrient uptake in the growing seedlings. bean due to leachates of invasive weeds. The negative influence of higher concentrations of The allelopathic impact of leachates or extracts is leachates and extracts of different allelopathic plant more harmful to radicle (Friedman, 1995). The species including weeds on seed germination was reported phytotoxicity was directly proportional to higher by Travlos and Paspatis (2008), Tanveer et al. (2008) and concentrations of leachates (Hashim Ali et al., 2005; Sharma et al. (2009). The stimulatory as well as inhibitory Wang et al., 2009). effects were concentration dependant. The results of the present investigation were in agreement with the above Root and shoot length studies. The roots are more sensitive to allelochemicals than According to Orr et al. (2005) potential allelopathic shoots (Rahman, 2006). Roots need to absorb water for impact of invasive species like Lolium and Elaeganus on growth of plants, which is done by the root cells that are native species like Acer, Populus and Platanus was due to directly exposed to the solution of leachates containing allelopathy, which was depending on the source of the allelochemicals which affect the growth of root and 144 Inter J Agri Biosci, 2014, 3(4): 141-148.

seedling growth of various crops had been well -1 1.2 a documented by several workers like Wu et al. (2008), min 1

-1 b 0.8 c a Cul Gatti et al. (2008) and many other allelopathic researchers 0.6 d b Snl explained that these effects were due to the 0.4 e e c d 0.2 e e allelochemicals existing in the leachates and extracts of OD µgOD protein ∆ 0 donor plants. Vasudevan et al. (2001) have also reported Control 1.50% 2.50% 5% 7.50% 10% allelopathic activity of Cassia sericea (Cassia uniflora) on Concentrations of leaf leachates germination and growth of bajra, horsegram and safflower. Fig. 1: Effect of leaf leachates of Cassia and Synedrella on activities of polyphenol oxidase in seedlings of mungbean Root: Shoot ratio

The root: shoot ratio is very confirmatory and reliable 1.2

-1 a 1 a indicator of morphological, physiological and biochemical min

-1 changes in germinating seeds and developing seedlings. In 0.8 c a d b Cul 0.6 e the present investigation it was found that all the f b Snl 0.4 b treatments slightly decreased the root: shoot ratio as c 0.2 c OD µg protein compared to control (Tables 1, 2). The decreased values ∆ 0 of R: S ratio indicated more shoot length as compared to Control 1.50% 2.50% 5% 7.50% 10% Concentrations of leaf leachates less root length. This may be due to direct contact of roots

with allelopathic leachates with roots resulting in to Fig. 2: Effect of leaf leachates of Cassia and Synedrella on inhibition of root length. The rate of root growth might activities of peroxidase in seedlings of mungbean have retarded drastically due to their contact with allelochemicals, resulting in to reduced root length as 3.50E-02 compared to shoot length. The decreased values of root: a -1 3.00E-02 ab ab 2.50E-02 b shoot ratios were reported by Gogoi et al. (2002) in two c c min 2.00E-02 d Cul different varieties of rice due to aqueous extracts of -1 1.50E-02 d b a Snl 1.00E-02 Ageratum, Borreria. 5.00E-03 c b Units g 0.00E+00 The higher values of root: shoot ratios were reported by Bhalerao et al. (2000b) in seedlings of jowar and bajra 5% 10%

1.50% 2.50% 7.50% when treated with extracts of Aspidium rhizomes. But they Control Concentrations of leaf leachates have further reported decrease in root: shoot ratio in Jowar seedlings with higher concentrations. In the present study Fig. 3: Effect of leaf leachates of Cassia and Synedrella on the same trend was noticed in mungbean seedlings when superoxide dismutase activity in seedlings of mungbean treated with leachates of Cassia and Synedrella at higher # Data points are the pooled means of three replicates over two a concentrations. years with standard deviation as error bars. Different letters at The allelochemicals might be acting as inhibitors or the data points denote significant difference by Duncan’s multiple range test at p<0.05. stimulators on root and shoot length, resulting into increased or decreased values of root: shoot ratio. Dias et shoot. When the water is absorbed, a smaller fraction of al. (2004) have also reported the changes in root: shoot the solution is available to the cells of the stem and leaves ratio in pearlmillet. They have further explained that (Dana and Domingo, 2006). The more accentuated effect several physiological and metabolic alterations take place on the roots might be due to their closer contact with the due to allelochemicals, during seed germination, which leachates especially when maintained on germination affect the overall performance of seedlings. Thus, it can paper (Chung et al. 2001). This may explain the reason be stated that both the invasive weeds and their leachates for inhibition of seedlings of any test crop. The root and which are rich in allelochemicals, are responsible for shoot length is the important parameter of seedling inhibition of seed germination and growth of seedlings in growth, which determine the growth of future plant. Many both the test crops. workers have reported positive and negative impact of aqueous extracts, leachates, residues and different Vigour index allelochemicals existing in them on root and shoot length. The vigour index is a vital parameter to determine the In the present investigation the leachates of Cassia efficiency of any germinating seed to utilize and mobilize and Synedrella had caused significant inhibitory effect on the reserved food materials. VI indicates the allelopathic shoot and root length at almost all the concentration effects on seedling establishment. Jadhav (2006) obtained levels, and the reduction was dose/ concentration similar effect on seedling vigour in wheat, mungbean, dependent in both the test crops (Tables 1, 2). These chickpea and fenugreek with the higher concentrations of results corroborate with the findings of the allelopathy leachates of Pistia and Eichhornia. workers like Ghayal et al. (2007a, b) and Dave and Jain However, Yang et al. (2006) recorded the reduction (2009). They reported inhibitory effects on root and shoot in seedling vigor of rice due to higher concentrations of length due to treatments of leachates or extracts, but only Ageratina and Eucalyptus leaf leachates. They further at higher concentrations. claimed that the presence of terpenoids in Ageratina was The negative impact of higher concentrations of possible reason for reduction in seedling vigour at higher leachates of different weeds on seed germination and concentrations. 145 Inter J Agri Biosci, 2014, 3(4): 141-148.

The workers like Sharma et al. (2009) reported the Research workers like Maiti et al. (2008) reported effects of extracts of seven different weeds on seed accumulation of sugars in seedlings of crops due to germination and vigour index in different varieties of allelopathic impact of leaf extracts of weeds at higher wheat. They attributed the reduction in vigour index to the concentration treatments. Findings of present presence of phenolic compounds in the leachates of investigation also corroborate with above reports (Tables weeds. As stated above, in the present investigation also 3, 4). the presence of various allelochemicals from alien weeds The carbohydrates, proteins, lipids and fats stored in might be inhibiting the growth of seedlings, resulting in to the seeds are usually oxidized during seed germination, decreased vigour index, due to higher concentration for supplying the energy to the developing seedlings, as a treatments. The allelochemicals in the leachates might result of this the organic constituents are very much have altered the cellular activities such as cell division, reduced. However, in the present investigation there was mineral uptake and enzyme activities resulting into constant increase in total sugars, starch and proteins which reduction in root and shoot lengths as well as vigour might be due to their non-utilization, as there was very index. less or sometimes no seed germination in leachates’ treated seeds of mungbean. Effects of leaf leachates of Cassia and Synedrella on physiology of mungbean seedlings Proteins The physiological analysis of seedlings will illustrate The modification in protein metabolism caused by their metabolic status and capability to mobilize and treatments of leachates of different allelopathic plant effectively utilize the reserve food materials, resulting into species, plays an important role in seed germination and successful seedling development, growth and seedling growth, as it is the fundamental event in cell establishment. Hence investigation on physiology and metabolism governing growth and development. biochemistry of germinating seeds was under taken. Increase in protein contents due to higher concentration treatments of leachates of various weeds Total sugars were recorded by Pathipati Usha Rani (2008) in crop In mungbean (Tables 3, 4) both Cassia and seedlings. The influence of various bioactive Synedrella leaf leachates induced increase in total sugar allelochemicals (triterpenes, steroids, flavonoids etc.), contents over control with all the treatments ranging from organic and inorganic compounds present in Cassia and 1.5% to 10%. The increase in sugar content was more due Synedrella leaf leachates have inhibited the seed to Synedrella leaf leachates as compared to Cassia leaf germination and seedling growth leading to higher protein leachates. contents resulted from their non-utilization and immobilization. Starch The results of the present investigations on increase In mungbean seedlings, treated with the leaf leachates in protein contents during seed germination of mungbean of Cassia and Synedrella, similar steady and gradual due to lower as well as higher concentrations of leachates increase in starch content was observed (Tables 3, 4). All of both the invasive weeds are in agreement with the the results were statistically significant. above workers indicating induction of stress proteins because of the allelobiogenesis stress (Tables 3, 4). The Proteins allelochemicals in Cassia and Synedrella leaf leachates In mungbean both Cassia and Synedrella leaf might be responsible for increase in the protein contents in leachates induced similar increase in protein contents over the seedlings of test crops. The retarded mobilization and control with all the treatments ranging from 1.5% to 10%. utilization of proteins must have led to their increase. The increase in protein content was very high due to Alterations in over all protein metabolisms during seed Synedrella leaf leachates as compared to Cassia leaf germination may be the most appropriate reason for leachates (Tables 3, 4). explaining the increase in protein content in the leachates treated seeds of mungbean. Phenols In case of mungbean seedlings, the results (Tables 3, Phenols 4) on phenol contents showed that with the increasing The secondary metabolites like phenols indicate the concentrations (1.5% to 10%) of leaf leachates of Cassia intensity of stress on plants and also afford defense to (9.53 to 10.88 mg/g) and Synedrella (8.43 to 10.57 mg/g) them. They have a very profound effect on seed the amount of phenols also went on increasing as germination, growth, development and metabolic compared to control (8.55 mg/g). functioning of plants. It is for these reasons phenolics have been a major focus of chemical ecology also. Total sugars and starch Phenolic compounds are the major allelochemicals, Carbohydrates play very vital role(s) in plant’s through which higher plants interact with each other metabolism and their synthesis and accumulation govern (Waterman and Mole, 1994). To understand the influence the majority of cellular metabolic processes. The of phenolics present in the leachates of Cassia and fluctuations in the status of carbohydrates will give clear Synedrella on mungbean, efforts were made to study the indications of growth, development, flowering, fruiting process of seed germination. and yield of treated plants. With Synedrella leaf leachates However Batish et al. (2005) and Bagavathy et al. at 2.5% concentration only there was little decrease in (2007) recorded opposite trend. They noted increased total sugar contents than control. phenolic contents in crops due to higher concentrations of 146 Inter J Agri Biosci, 2014, 3(4): 141-148. fruit and leaf extracts as well as leachates of Parthenium The assessment of activities of various antioxidant and Eucalyptus respectively. They claimed that increase enzymes in leachate treated plants is very important to in phenolic contents caused decrease in seed germination know that whether the leachates generate any stress on the rate and seedling growth. treated plants. The stressful environment created by high The results of present study are in agreement with concentration of allelochemicals existing in the leachates above findings (Tables 3, 4). The accumulated phenolic will induce several metabolic alterations. contents might have caused adverse impact on cell The stimulation or inhibition of the activities of membrane permeability, water absorption and other antioxidant enzymes is helpful for predicting the metabolic functions in seeds leading to inhibition of seed sensitivity and resistance of test plants to allelobiogenesis. germination in mungbean and reduction in root as well as The intensity of stress and adaptability of plants to it, can shoot length. be judged on the basis of percent increase in their activities. Weir et al. (2004) reported increase as well Effect of leaf leachates of Cassia and Synedrella on decrease in SOD and POX activity in Arabidopsis enzymatic activities in seedlings of mungbean thaliana, Centaurea diffusa and Centaurea maculosa. Results They further explained that this may be due to the The plants exposed to abiotic or allelopathic stress different response of each of the affected process to a generate reactive oxygen species (ROS), responsible for particular concentration of a specific allelochemical. The inducing changes in biochemical and physiological findings of Moradshahi et al. (2003) indicated significant processes, leading to cellular damage (Cruz-Ortega et al., reduction in the activities of different antioxidant enzymes 2002). Hence, for self-protection the plants exposed to in various crops due to the treatments of aqueous stress conditions, highly stimulate the activities of extracts/leachates of different forest trees, weeds and fern different antioxidant enzymes like PPO, POX and SOD to species. scavenge the ROS. The leachate treated seedlings of Many researchers reported the stimulation of the mungbean might be responsible for generating some type activities of antioxidant enzymes in treated crop plants. of allelopathic stress on them. To find out this, the Geng et al. (2005) correlated the inhibition of seed activities of some antioxidant enzymes were assayed. germination in cucumber, watermelon, pepper, lettuce and tomato with the changes in activities of SOD, POX, and Polyphenol oxidase (PPO) [EC 1.14.18.1] CAT. They further noted that these changes were The results shown in Fig.1 revealed that the concentration dependant. The stimulation of antioxidant activity of PPO was comparatively less than control in enzymes in mustard and mungbean treated with leaf mungbean when treated with the leachates of Cassia and leachates of Cassia and Synedrella can be explained on Synedrella. But for remaining concentrations it showed the basis of above results (Figs. 1, 2, 3), as these leachates significant stimulation. All the results were significant at were containing different types of allelochemicals and the p= 0.001. stress tolerance or sensitiveness to such chemicals can be predicted from the level of stimulation or inhibition of Peroxidase (POX) [EC 1.11.1.7] such enzymes. The significant results on the activity of POX shown in Fig. 2 indicated that in mungbean, the activity of Conclusion POX was significantly stimulated at all the concentrations The inhibitory effect on seed germination, seedling of leachates used. growth, vigour index and seedling physiology including enzyme activities in seedlings have confirmed the Super oxide dismutase (SOD) [EC 1.15.1.1] allelopathic nature of Cassia uniflora and Synedrella The results illustrated in Fig. 3 on the changes in nodiflora which can be assigned to the complex mixture activity of SOD in mungbean seedlings, the SOD activity of different compounds in leaf leachates. The suppression had shown very less stimulation at 1.5% and 2.5% leaf and inhibition of different plant species naturally growing leachates treatments of Synedrella as compared to control in association with Cassia and Synedrella might be due to (Fig. 3). But it was at par with control with leaf leachate the same reason. treatment of Cassia (1.5%). However, at higher concentrations of both the leaf leachates the activity of Acknowledgements SOD was stimulated by manifold. One of the authors (Dr. Ghayal N.A.) is indebted to The stimulation or inhibition in the activities of UGC for providing financial assistance and awarding her antioxidant enzymes like POX, PPO and SOD indicate the a teacher fellowship. The authors are thankful also to the level of reactive oxygen species produced under the stress HOD, Department of Botany, University of Pune, M.S., conditions and interactions between plants. These India, for providing research facilities. enzymes scavenge the ROS and protect the plants during biotic, abiotic and allelopathic stress conditions (Yu et al., REFERENCES 2003). They further explained that exposure to allelopathic agents like exudates or extracts at higher Abdur Rahman, 2006. 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