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I:\Ijta\Ijta No 4 Shimla Conf\I Population dynamics of microflora in the rhizosphere of Macrophomina phaseolina resistant... IJTA© Serials Publications Paritosh Kumar, V.K. Gaur, Anand Kumar Meena* Sunil Kumar Pipliwal and Ashok Kumar Meena ABSTRACT: Eighteen fungi and one bacterium showed different trends in population in initial infested soil and in rhizosphere at seedling and flowering stages of resistant MSJ-118 and susceptible RMG-62 varieties. The frequency and abundance (%) of microorganisms also showed different trends. Key words: Macrophomina phaseolina, Mungbean, rhizosphere, microflora. INTRODUCTION present study was carried out to compare the Mungbean [Vigna radiate (L.) Wilczek] is an important rhizosphere microflora of resistant MSJ-118 and pulse crop in Indian continents. Mungbean is being susceptible RMG-62 varieties of mungbean at infected by several fungal, bacterial and viral diseases different stages of plant growth. The extent of changes but dry root rot caused by Macrophomina phaseolina in population of micro organisms in rhizosphere and (Tassi) Goid. is considered as the most devasting in soil is to be determined by the enumeration of disease in all the mungbean growing areas of microflora. For soil-borne diseases the knowledge of Rajasthan.The disease is quite wide spread across the microbial activity in the root zone (rhizosphere) is of Rajasthan state due to congenial weather conditions prime importance. In rhizosphere the micro organism and causes considerable yield losses (Philip et al., 1969, are known to synthesize metabolites which in some Grewal, 1988). The pathogen may infect almost all cases may be antagonistic to root pathogens. The parts of plants i.e. root, stem, branches, petioles, leaves antagonists may be effective in reducing the and pods. Seed infection due to Macrophomina population of pathogen. phaseolina ranges from 2.2 to 15.7 per cent which may MATERIALS AND METHODS cause losses in grain yield to the extent of 10.8 per cent and protein content of 12.3 per cent (Kaushik et Soil from fields of Bikaner district of Rajasthan, where al., 1987). The infected s eeds act as an important mungbean is mainly grown was collected and to source of primary inoculum for new areas. Plant stand increase the bulk soil from Agronomy Farm, College is affected due to pre and post-emergence infection of Agriculture, Bikaner with farm yard manure was of the crop. In pre-emergence stage, the fungus causes mixed. The soil was filled in two wooden boxes which seed rot and mortality of germinating seedlings while were rinsed with formalin and dried. The highly in post emergence stage seedlings get blightened due pathogenic isolate Bikaner was multiplied on sand to soil and seed-borne infection. In later stages of crop maize flour medium and added to wooden box soil. growth decay of secondary roots and shredding of The infested soil was irrigated and left for ten days. the cortex region of the tap roots are commonly For sampling, surface soil about 1 cm was removed observed. Rhizosphere microflora plays an important and randomly three soil samples taken diagonally role in the manifestation of diseases, especially the from each wooden box. The soil samples were air soilborne ones. It is characterized by greater microbial dried in laboratory and passed through 2 mm sieve. activity which plays a role in biological control. The For enumeration of fungi ‘Soil plate method’ (Warcup, 1 Department of Plant Pathology, S. K. Rajasthan Agricultural University, Bikaner-334006, E-mail: anandraj.km@g mail.com Vol. 33, No. 4, October-December 2015 2537 Paritosh Kumar, V. K. Gaur, Anand Kumar Meena, Sunil Kumar Pipliwal and Ashok Kumar Meena 1950 and Parkinson, 1958) was followed. A lunatus, Fusarium oxysporum, Macrophomina phaseolina, microspatula was standardized for placing the Penicillium aurantiogriseum, Penicillium javanicum, required quantity of soil in Petri dishes (0.0035 g/ Rhizopus stolonifer, Trichoderma atroviride, T. harzianum microspatula). A drop of sterilized distilled water was and T. viride were present in initial infested soil and placed in the centre of Petri dish and on this drop soil rhizosphere soil of two stages of plant (seedling and with the help of microspatula of the sample was flowering) of resistant MSJ-118 variety of mungbean. placed to disperse it uniformly in the medium. The population of Aspergillus clavatus, A spergillus Martin’s peptone dextrose agar medium just above flavus, Fusarium oxysporum, Macrophomina phaseolina the solidifying temperature was poured in these and Rhizopus stolonifer decreased at flowerin g as dishes. Thornton’s standardized medium was used compared to seedling stage. Aspergillus niger, for the isolation of Bacillus bacterium following ‘The Cochliobolus lunatus, Penicillium aurantiogriseum, Soil Dilution and Plate Count Method’ (Waksman, Penicillium javanicum, Trichoderma harzianum, T. 1927). In each Petri dish 1 ml of prepared dilution atroviride and T. viride had increasing trend from was transferred aseptically to which 15-20 ml medium seedling to flowering stage. The population of just above the solidifying temperature was poured. Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, To disperse the soil or soil/water dilution uniformly, Chaetomium globosum, Cochliobolus lunatus, Fusarium the medium was rotated inside the Petri dish oxysporum, Rhizopus stolonifer, Trichoderma atroviride, clockwise as well as anti clockwise with swirling T. harzianum and T. viride increased at seedling stage motions of the dish. The poured Petri dishes were as compared to initial infested soil. Macrophomina incubated at 28 ± 1ºC for four to five days. The phaseolina, Penicillium aurantiogriseum and Penicillium recording of colonies for different microorganisms javanicum had decreasing trend from initial infested was continued for fifteen days or until new colonies soil to seedling stage. The population of Chaetomium ceased to develop. After studying soil samples from globosum increased from initi al infested soil to initial infested soil, seeds surface sterilized with 0.1% seedling stage but remains constant from seedling to mercuric chloride of dry root rot resistant MSJ-118 flowering stage. Similarly, Alternaria alternata, and susceptible RMG-62 varieties were sown in the Aspergillus flavus, Aspergillus niger, Chaetomium boxes separately for rhizosphere studies at seedling, globosum, Curvularia lunata, Fusarium oxysporum, flowering stages. The boxes were watered at intervals Macrophomina phaseolina, Penicillium javanicum, depending upon the dryness of soil and care was Rhizopus stolonifer, Tricoderma atroviride and taken to break the upper crust of soil. In order to Trichoderma harzianum were present in initial infested prevent lumping of soil about roots, water was not soil and rhizosphere soil at seedling and flowering given 24 hours prior to sampling. At seedling stage stages of susceptible RMG-62 variety of mungbean. three plants of each variety were removed from the The population of Alternaria alternata, Aspergillus niger, soil with roots intact, taking care that the roots were Penicillium javanicum, T. atroviride and T. harzianum not injured. The roots of these plants were shaken increased from seedling to flowering stage. The gently to dislodge the adhering soil which was population of Aspergillus flavus, Chaetomium globosum, collected in sterilized Petri dishes for further dispersal Curvularia lunata, Fusarium oxysporum and Rhizopus in different media. The same process was repeated at stolonifer decreased at flowering stage as c ompared flowering stage. For enumeration and isolation of to seedling stage. Population of Alternaria alternata, fungi and bacteria the techniques employed for initial Aspergillus flavus, Chaetomium globosum, Fusarium infested soil were followed. oxysporum, Penicillium javanicum and Rhizopus in which dishes Petri ofNumber ofNumber Petri dishes in which ofcolony a species appeared stolonifer increased from initial infested soil to seedling (%)Frequency � �100 dishes Petri ofnumber Total ofnumber Petri dishes stage. Population of Aspergillus niger, Curvularia lunata Total ofnumber colonies of speciesa theallin Petri dishes and Macrophomina phaseolina decreased at seedling (%)Abundance � �100 colonies of Total of colonies of all species allin Petri dishes stage as compared to initial infested soil.The population of Tricoderma atroviride and Trichoderma RESULTS AND DISCUSSION harzianum were constant at initial infested soil and Different trends were observed in population of each seedling stage but had increasing trend from seedling species estimated in 1 g rhizosphere soil of resistant to flowering stage. Bacillus subtilis appeared at all three MSJ-118 and susceptible RMG-62 varieties (Table 1 stages of resistant MSJ-118 as well as susceptible and 2). Aspergillus clavatus, Aspergillus flavus, RMG-62 variety of mungbean. The population of Aspergillus niger, Chaetomium globosum, Cochliobolus Bacillus subtilis was abundant in initial infested soil, 2538 International Journal of Tropical Agriculture © Serials Publications, ISSN: 0254-8755 Population dynamics of microflora in the rhizosphere of Macrophomina phaseolina resistant... decreased at seedling stage and further increased at infested soil to flowering stage of susceptible RMG- flowering stage in rhizosphere soil of resistant MSJ- 62 variety. Aspergillus niger, Cochliobolus lunatus, 118 variety. The population of Bacillus subtilis was Rhizopus stolonifer, Trichodrema harzianum, T. atroviride abundant in initial
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