Evaluation of Fungicides, Botanicals, Neem products and Bio-agents against Wilt of Pigeonpea caused by Fusarium udam.butler
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1 Vol.2 No. 2, (2013) Received: March.2013; Accepted Oct Evaluation of Fungicides, Botanicals, Neem products and Bio-agents against Wilt of Pigeonpea caused by Fusarium udam.butler K.Chaudhary 1, S.R Singh 2, A.K.Singh 3 and S.Mala 4 1 Departmentf of Plant Pathology, CSAU&T, Kanpur, 2, 3 Krishi Vigyan Kendra, Hathras (U.P.) and 4 G.M.Aligarh (U.P.) Abstract A study was conducted to identify the effect of fungicides, botanicals, neem products and biocontrol agents to control wilt of Pigeonpea. Mancozeb 80 WP (200) ppm completely inhibited the growth of Fusarium udam. In botanical s Makoy (10%) found most effective against F.udam. marigold (10%), Ashoka (10%), Parthenium (10%), ginger (10%) were also found effective to check the fungal growth. Achook (5%) has given good inhibition zone on fungus. out of 8 tested bio-agents the maximum (69%) colony growth inhibition of F.udam was recorded in the P.citrinum isolate of lucknow followed by A.niger (Delhi),G.virens (Pantnagar),T.harzianum (Pantnagar),A.flavus (Pantnagar),T.viride (Coimbatore), T.harzianum (Kanpur) and T.viride(Kanpur) showing 67.5,60.0, 59.0,56.8,54.5,50.0 and 47.7 % colony growth inhibition of Fusarium udam in vitro,respectively. Keywords: Fungicides, botanicals, neem products, bio-agents, pigeonpea, wilt, fusarium udam agent colonizes the rhizosphere, the site Introduction requiring protection and leaves no toxic residues as opposed to chemicals. Pulses are important sources of protein for India s large and growing population. Arhar (Cajanus cajan L. Millsp) also known as Pigeonpea, red gram is one of the most important leguminous crops and constitutes chief source of protein for majority of population of India dependening totally on vegetarian diet. Low yield of Pigeonpea attributed to its susceptibility to several fungal, bacterial and viral diseases [3]. In recent times, there has been a worldwide swing to the use of eco-friendly methods of protecting the crops from pests and diseases. The use of potential harmful chemical sprays is viewed with dissatisfaction in many countries. As such in the present context, biological control of wilt with bioagents offers a great promise. A biological control 1 Material and methods Laboratory bioassay of fungicides Relative efficacy of eight fungicides at the recommended doses was tested against the pathogen under laboratory. The required quantities of the fungicides were incorporated into 100 ml of 2% sterilized unsolidified potato dextrose agar and shaken well to make it homogenous. Medium was then poured in 90 mm. sterilized petri-dishes with the three replications of each treatment and allowed to solidify. These dishes were than inoculated from 7 days old culture and one disc was placed in the center of each petri
2 dish in such a way that the fungus may contact with the medium. The medium without any fungicides poured and inoculated similarly, served as control. The petri-dishes were incubated at 28 0c for 7 days. Measuring the radial growth of the fungal colony in mm assessed the efficacy of fungicides. The observations were taken after 24, 48, 72 and 92 hours. In vitro evaluation of Plant extract Relative efficacies of eleven plant extracts belonging to different families were tested under laboratory conditions. Standard dual culture technique [6] was followed for bioassay of bio-pesticides. The efficacy was judges by extant of their inhibitory effect on growth of pathogen on PDA. For this purpose the fresh plant leaves were separately homogenized with sterile distilled water at 1:1 w/v in a pestle and mortar and filtered through muslin cloth. This makes 100 per cent extract solution. The plant extract so prepared were heated at 65 0c for 15 minute to avoid contamination [4]. Medium was then poured in 90 cm sterilized petri-dishes with three replications each treatment and allowed to solidify. In each petri -dish, 4 wells of 6 mm dug at 4 corner at 5 cm apart from each other than 2 ml of the each plant extract was added to these wells using sterile pipette to get 2 ml of the appropriate plant extract were added to these wells using sterile to get 10 per cent concentration of plant extract in medium. The fungal discs of were then inoculated from 7 days old culture and one discs was placed in the center of each petri-dish in such a way that the fungus may contact with medium. The medium without any bio-pesticides poured and inoculated similarly, served as control. The petri - dishes were incubated at 4 mm at 28 0c for 3 days. The efficacy of different plant extracts was determined after statistical analysis. In vitro evaluation of Neem products The extracts of neem leaves are prepared by the same procedure as followed for the other plant extracts. Neem seed kernel extract is prepared by crushing it and keeping in distilled water for overnight. Then it was filtered through muslin cloth and sterilized. The 5 ml neem product was incorporated in to two per cent PDA medium which was shaken well to make it homogeneous. The medium was then poured in to 90 cm petri- dishes with three replications of each treatment. A separate treatment as control was also maintained having no neem product, served as control. Circular discs were 4mm from 7 days old culture of the pathogen in petri dishes were cut and sterilized cork borer. One such disc was placed in centre of each petri dish containing solidified medim.after inoculation of petri dishes were incubated for 7 days at 28 0c. After incubation the final observations on radial growth were taken when petri dishes in control were fully covered with growth of the fungus. In vitro evaluation of Trichoderma spp. The isolates of Trichoderma viride, Trichoderma harzianum and Trichoderma virens were evaluated in laboratory by dual culture technique as [6] to screen out the most efficacious one. Petri dishes (90 mm) containing PDA were inoculated with 5 mm diameter mycelial disc of 7 days old culture of F. udam and Trichoderma spp. 2
3 at equal distance from the periphery. Inoculated plates were incubated at 25+1ºC in BOD incubator and the radial growth of F. udam was measured 2, 3 and 6 days after incubation. Controls without Trichoderma were maintained side by side. Three replications were maintained for each treatment. Percent inhibition of radial growth of F. udam was calculated using the formula the percent growth inhibition was calculated by the following formula C-T I = X 100 C I = Percent growth inhibition = Colony diameter of pathogen in control T = Colony diameter/radial growth of pathogen in treatment The percent inhibition data were transformed in Sin -1 percentage transformation and analyzed statistically in completely randomized design (CRD). From the zone of interaction between the antagonist and Fusarium udam in dual culture plate, the mycelial mats were gently lifted with a needle and put in a drop of cotton blue on a microscopic slide and spread with a needle and observed under microscope for hyphal interaction. Results and Discussion Laboratory bioassay of fungicides Eight fungicides were evaluated for their inhibitory effect against the pathogen in vitro. The results of the average radial diameter of the fungus colony after 7 days of incubation at 28 0c. Results from the Table 1 indicated that out of eight different fungicides tested in laboratory; Mancozeb 80 WP (200) ppm Table1 Evaluation of fungicides against F.udam Fungicides Dose (ppm) Av. Dia. of fungal colony(mm) Per cent inhibition Mancozeb 80 WP Thiram80 WP Carbendazim 75 WP Captan 50WP Topsin-M Ziram 80 WP Ridomil Vitavax Control C.D at 5% 3.30 SEM completely inhibited the growth o the fungus. Other fungicides which were also found effective to check the growth of fungus were thiram (200ppm), Ziram 80 3
4 WP (200 ppm) which appears promising in comparison to other fungicides i.e. Topsin M.Apron Mancozeb,Ridomil and Vitavax. The efficacy of fungicides was also evaluated against S.rolfssi in bioassay by several workers [7,8], and they found the same result as observed in present study. Vitavax were the best among the fungicides tested in vitro [2]. Agrosan GN were most promising against F.solani,R.bataticola and S.rolfsii isolated from C.arietinum [11]. Laboratory bioassay of plant extracts Table 2 Evaluation of plant extracts against F.udam Plant extract Dose (%) Av.diameter of fungal colony(mm) Per cent inhibition Makoy Marigold Ashoka Parthenium Ginger Bakayan Aak Dhatura Garlic Tulsi Control 87 - C.D.at 5% S.E The extracts of ten botanicals tested in laboratory inhibited the fungal growth to varying extent and were considered effective over control (table-2), the highest makoy (75.58%) inhibition and lowest in Laboratory bioassay of neem products Table 3 Evaluation of neem products against F.udam tulsi (4%) followed by marigold, Ashoka and parthenium. The remainining plant extract were also effective but noneffective in controlling disease [5,9]. Neem products Dose (%) Av.diameter of fungal colony(mm) Per inhibition Achook Nemark Neem gold Econim Neem Kernel Javan Nemata Control C.D.at 5% 7.94 S.E Seven neem products were evaluated for their inhibitory effect against the Fusarium cent udam in vitro.the results of average radial diameter of fungal colony after 7 days of 4
5 incubation at 28 0c. reveled the out of six neem products tested in the laboratory. Achook (5%) has given good inhibition zone on fungus. Rests are significantly effective [4]. Laboratory bioassay of bio-agent Table 4 Per cent colony growth inhibition of F.udam in vitro Bio-agents Colony growth(mm) % inhibition over control P.citrinum (L) (55.2) A.niger(L) (56.1) G.virens(P) (50.1) T.harzianum(P) (48.9) A.flavus(K) (43.6) T.viride(C) (50.7) T.harzianum(K) (45.0) T.viride(K) (47.5) Control(F.udam) C.D. at 5% It is evident from the results (Table4) that out of 8 tested bio-agents the maximum (69%) colony growth inhibition of F.udam was recorded in the P.citrinum isolate of lucknow followed by A.niger (D), G.virens(P),T.harzianum(P),A.flavus(P),T. viride(c), T.harzianum(K) and T.viride(k) showing 67.5,60.0,59.0,56.8,54.5,50.0 and 47.7 % colony growth inhibition of Fusarium udam in vitro, respectively. Microscopic examination of slides prepared from the junction points of dual culture on 6 th day revealed the entewining coiling and lysis of Fusarium udam mycelium by all the isolates of T.viride (C),T.harzianum(P) and G.virens (P),highest being with T,viride (K) isolate.a.flavus,a.niger(d) and P.citrinum(Lucknow)did not show much coiling but lysis of the mycelium through antibiosis [1,10]. Reference 1. Bulter, E.J. (1911). Fungi and Diseases in Plants. Bishan Singh, Mahendra Pal Singh, New Connaught Place, Dehradun and M/s Periodicals 42-D, Vivek Vihar, Delhi-32, pp. 547 (reprinted 1978). 2. Channamma,K.A.L., Hiremath, P.C. and Vishwanath, S. (1980).Efficacy of some fungicides in controlling foot of ragi caused by S.rolfsii caused by S.rolfsii, Phytopathology,9: Cook, R.J. (1996). Sustainable Agriculture: Introduction and Summary. Candian of Jounnal Plant Pathlogy 18: Jagnathan,R. and Narasimhan, V. (1988). Effect of plant extract/product on two fungal pathogens of finger millet. Indian Journal Mycology and Plant Pathology 18: Manian,S. And Udaiyan, K. (1992).Report on effect of plant extracts. Indian Phytopathology Supplementary issue of vol Morten,D.J. and Stroube,W.H. (1955). Antagonist and stimulatory effect of soil 5
6 microorganism.phytopathology,45: Shahid,M.A., Mukhtar, Khan A.M.A. and Ahmad, M. (1980). Chemical control of collar rot of lentil caused by Sclerotium rolfsii Sacc.Sarhad Journal of Agriculture, 6: Singh R.K., Dwivedi, R.S., Hasija, S.K., Rajak, R.C. and Singh, S.M. (1987). Studies on Some aspects of S.rolfsii causing foot root of Barley Perspec. Mycolgy Research1: Srikant, M.G., Kulkarni, Hedge, R.K. and Kulkarni, S. (1986). Studies on root rot of betal vine caused by S.rolfsii Sacc. In Karnatka.Plant Pathology Newsletter, 4: Van Emden, H.F., Ball, S.L. and Rao, M.R. (1988). Pest diseases and weed problems in pea lentil and faba bean and chickpea. In: R.J. Summerfield (ed.), World Crops: Cool season Food Legumes. ISBN Kluwer Academic Publishers. Dordrecht, The Netherlands. pp Vishwakarma,S.N. and Chaudhary, K.C.B. (1992).Laboratory evaluation of some fungicides against some root diseases pathogens of gram(c.arietinum l.)pesticides,16:
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