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In Vitro Evaluation of Plant Extracts and Bio- Agents Against Alternaria

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In Vitro Evaluation of Plant Extracts and Bio- Agents Against Alternaria International Journal of Chemical Studies 2018; 6(2): 504-507 P-ISSN: 2349–8528 E-ISSN: 2321–4902 IJCS 2018; 6(2): 504-507 In vitro evaluation of plant extracts and bio- © 2018 IJCS Received: 04-01-2018 agents against Alternaria tenuissima (Fr.) keissl Accepted: 05-02-2018 causing leaf blight of kodo millet K Hariprasad Department of Plant Pathology, UAS, GKVK, Bengaluru - 65 K Hariprasad, A Nagaraja, Suresh Patil and Rakesha *Project Coordinating Unit (Small Millets), ICAR, GKVK, Abstract Bengaluru, Karnataka, India Kodo millet (Paspalum scrobiculatum L.) is nutritionally important millet. Leaf blight has been a major production constraint and fungicidal sprays for the management of any disease on this crop may not be A Nagaraja economically viable and feasible as the farmers cultivating the crop are resource poor and the crop is less Department of Plant Pathology, remunerative, but is important in tribal and rainfed agriculture. Hence, botanicals and bio-agents were UAS, GKVK, Bengaluru - 65 *Project Coordinating Unit evaluated in vitro against Alternaria tenuissima the cause of leaf blight. Ten plant extracts and 14 bio- (Small Millets), ICAR, GKVK, agents were evaluated following dual culture. The results revealed 100 per cent inhibition of the mycelial Bengaluru, Karnataka, India growth of A. tenuissima by Eucalyptus sp. and Clerodendron infortunatum at 7.5 and 10.0 per cent concentrations. Among the fungal bio control agents tested, maximum inhibition (100 %) was recorded Suresh Patil in Trichoderma harzianum (NBAIR), followed by T. viride (81.38 %); whereas the bacterial bio agent Department of Plant Pathology, Bacillus amyloliquefaciens (P-42) showed only 77.40 % inhibition of mycelial growth revealing that UAS, GKVK, Bengaluru - 65 fungal antagonists were more effective than the bacterial antagonists. *Project Coordinating Unit (Small Millets), ICAR, GKVK, Keywords: Alternaria tenuissima, Plant extracts, Bio agents, In vitro evaluation, Leaf blight, kodo millet Bengaluru, Karnataka, India Rakesha Introduction Department of Plant Pathology, Millets are unique due to their short growing season. They are highly nutritious, non-glutinous UAS, GKVK, Bengaluru - 65 and not acid forming foods. Among the millets, kodo millet (Paspalum scrobiculatum L.) is *Project Coordinating Unit one of the oldest cultivated cereals. The crop has rich medicinal values and provides protein, (Small Millets), ICAR, GKVK, carbohydrates and fibre for the body growth (Nagaraja et al., 2007). Leaf blight is one of the Bengaluru, Karnataka, India most destructive diseases after head smut, which if occurs at early stages of crop growth may [5] reduce the crop yield drastically (Nagaraja et al., 2016). As kodo is normally grown by the resource poor farmers under rainfed ecosystems with minimal investment on disease management, botanicals which are renewable, inexhaustible, indigenously available, easily accessible, non-phytotoxic, ephemeral and readily biodegradable, with relatively low cost may [4] be utilized in plant protection (Khadar, 1999). Also, biological control assumes special significance as it is ecologically conscious and cost-effective alternative strategy and presently has vital role in the organic farming system. Hence, the present study was taken up to screen in vitro both the plant products andbio-agents for their antifungal activity against A. tenuissima the causal fungus of kodo blight. Materials and Methods Plant extracts The present investigation was carried out to evaluate different plant species for the possible presence of fungi toxicant properties by poisoned food technique. The list of botanicals used in the study is presented in Table 1. Hundred grams of fresh leaf material was taken and cut into small pieces, 20 ml of 5 per cent acetone was added and the samples were ground thoroughly. Different plant extracts of varyingconcentrations i.e. 5.0, 7.5 and 10.0 percent were tested in three replications. In vitro evaluations of leaf extracts was done against A. tenuissima using Potato dextrose agar medium. Correspondence The leaf extracts were mixed to the medium by proper stirring and poured to Petri plates and Suresh Patil Department of Plant Pathology, allowed for solidification. Seven mm disc from twelve days actively growing culture was UAS, GKVK, Bengaluru - 65 transferred aseptically using cork borer to the Petri plates containing leaf extracts. The PDA *Project Coordinating Unit plates without any plant extracts served as control. The plates were incubated at 27±1°C for 10 (Small Millets), ICAR, GKVK, days and the colony diameter was recorded. Per cent inhibition was worked out according to Bengaluru, Karnataka, India the equation of Vincent (1947). ~ 504 ~ International Journal of Chemical Studies C – T Where, I = Per cent inhibition of the mycelium I = x 100 C = Growth of the mycelium in control C T = Growth of the mycelium in treatment. Table 1: List of plant extracts used Sl. No. Common name Scientific name Family 1 Eucalyptus Eucalyptu ssp. L. Her. Myrtaceae 2 Clerodendron Clerodendron infortunatum L. Verbenaceae 3 Nagadhale Ruta graveolens L. Rutaceae 4 Pongemia Pongamiapinata L. Leguminaceae 5 Lantana Lantana camera L. Verbenaceae 6 Cyprus Cyprus rotundus L. Cyperaceae 7 Neem Azadirachta indica A. Juss. Meliaceae 8 Cynodan Cynodon dactylon L. Poaceae 9 Parthenium Parthenium hysterophorus L. Asteraceae 10 Tulasi Ocimum tenuiflorum L. Lamiaceae B. Bio-agents streaked in the corner of the plate after which a fungal disc In vitro evaluation was carried out with selected bio-agents was placed at the opposite side. Each treatment was replicated listed in Table 2 employing dual culture technique. Twenty ml three times. After incubation for a required period (when of sterilized and cooled potato dextrose agar was poured into radial mycelial growth in control plate reached maximum), sterile Petri plates. Fungal antagonists were evaluated by the growth of the pathogen was measured. Per cent inhibition inoculating the pathogen on one side of the plate and the over control was worked out according to the equation of antagonist on the exact opposite side of the same plate by Vincent (1947) as shown above. leaving 3-4 cm gap between. Bacterial antagonist was Table 2: List of bio-agents used Sl No Bio-agent Source 1 Trichoderma asperullum Microbiology lab., UAS, GKVK, Bengaluru 2 Trichoderma viride ICAR-NBAIR, Bengaluru 3 Trichoderma koningii Microbiology lab., UAS, GKVK, Bengaluru 4 Trichoderma harzianum Microbiology lab., UAS, GKVK, Bengaluru 5 Trichoderma harzianum ICAR-NBAIR, Bengaluru 6 Lysini bacillus sphaericus Plant Pathology lab., UAS, GKVK, Bengaluru 7 Pseudomonas fluorescens Plant Pathology lab., UAS, GKVK, Bengaluru 8 Myroidesmarinus Plant Pathology lab., UAS, GKVK,Bengaluru 9 Bacillus subtilis Plant Pathology lab., UAS, GKVK,Bengaluru 10 Bacillus cereus Plant Pathology lab., UAS, GKVK, Bengaluru 11 Bacillus amyloliquefaciens Plant Pathology lab., UAS, GKVK, Bengaluru 12 Bacillus subtilis Plant Pathology lab., UAS, GKVK, Bengaluru 13 Paenibacillusbarcinonensis Plant Pathology lab., UAS, GKVK, Bengaluru 14 Bacillus thuriangiensis Plant Pathology lab., UAS, GKVK, Bengaluru Results and Discussion %) was noticed in 10.0 per cent concentration with the least Effect of plant extracts on thegrowth of A. tenuissima in vitro (67.14 %) at 5.0 per cent concentration. However, inhibition Significantly higher inhibition of mycelial growth was increased with increase in the concentration of extracts from observed in Eucalyptus sp. (95.30 %)followed by 5.0 to 10.0 per cent In the interactions, Eucalyptus sp. and C. Clerodendron (92.22 %); but least inhibition was noticed in infortunatum both at 7.5 and 10.0 per cent as well as R. case of O. tenuiflorum (51.60 %) (Plate 1). Among the graveolens at 10.0 per cent showed 100 per cent inhibition different concentrations, maximum mycelial inhibition (80.74 (Table 3). Table3: In vitro evaluation of plant extracts againstA. Tenuissima Botanicals Mycelial growth inhibition over control Mean Sl. No. Concentration 5.0 % 7.5 % 10.0 % Inhibition (%) 1. Eucalyptu ssp. 85.92(68.03) 100.00(90.00) 100.00(90.00) 95.30 2. Clerodendron infortunatum 76.66(61.11) 100.00(90.00) 100.00(90.00) 92.22 3. Ruta graveolens 68.51(55.87) 84.44(66.96) 100.00(90.00) 84.32 4. Pongemia pinnata 65.92(54.30) 70.00(56.79) 84.44(66.82) 73.45 5. Lantana camera 64.81(53.62) 70.00(56.79) 76.66(61.11) 70.49 6. Cyprus rotundus 61.48(51.65) 65.18(53.80) 81.48(64.63) 69.38 7. Azadirachta indica 69.62(56.56) 72.22(58.19) 76.29(60.87) 72.71 8. Cynodan dactylon 62.96(54.58) 60.00(50.77) 66.29(54.55) 63.08 9. Parthenium hysterophorus 66.29(54.58) 65.55(54.08) 70.00(56.80) 67.28 10. Ocimum tenuiflorum 49.25(44.57) 53.33(46.91) 52.22(46.27) 51.60 Mean 67.14 74.07 80.74 73.98 Extracts (E) Concentration(C) E × C S. Em± 0.35 0.64 1.10 CD (P 0.01) 1.32 2.41 4.17 Note: Figures in the parenthesis are arc sine transformed value ~ 505 ~ International Journal of Chemical Studies Plate 1: In vitro efficacy of plant extracts on A. tenuissima In the present investigation, the mycelial growth of the fungus Evaluation of bio-agents by dual culture method revealed was inhibited to a greater extent by Eucalyptus as the leaf significant difference between the bio-agents on percent extract contains 2',6'-dihydroxy-3'-methyl-4'-methoxy- inhibition of mycelial growth of A. tenuissima (Table 4 and dihydrochalcone, eucalyptin and 8-desmethyl-eucalyptin, 5). Maximum mycelial inhibition (100.0%) was noticed in T. which exhibits antimicrobial activities (Takahashi et al., harzianum (NBAIR)with minimum mycelial inhibition 2004). Similar results were obtained by Ramezani and (69.16%)noticed in T. harzianum (Pl. Path.) indicating strain Abdollahi (2015), [8] and Patni et al. (2005) [7]who found that differences in the efficacy(Table 4 and Plate 2a).
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