National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692 Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
This research is aimed to optimize isolation processes and culturing the seed endophytic bacteria from wheat seed endosperm. Combination of 4% sodium hypochlorite, 75% ethanol was found effective for the surface sterilization of wheat seeds. Processing of seeds for removal of seed coat was to avoid the transient microbes interfering in the isolation of true bacterial endophytes from endosperm. Using plant based growth medium are more ideal for isolation of inherent microflora, hence a new growth medium with wheat flour as matrix (WMM) was developed to extract more number and diversity of endophytic bacteria and compared with the conventional media TSA & NA during the isolation. WMM medium directly delivered the lytic enzyme producing bacteria showing antagonism and prompt seed germination from placed endosperm on medium with highest numbers of cultivable bacteria, as well as the highest diversity compared with TSA and NA mediums. Based on endophyte growth outcome on WMM, TSA, NA medium during isolation, A total of 18 morphologically different endophytic bacteria have been isolated from seven wheat genotypes of Triticum aestivum and were tested for the production of hydrolytic enzymes. In preliminary screening 16 isolates were found gram-positive, 2 were gram-negative and 16 isolates showed positive results for endospore staining, out of which 7-subterminal spore formers, 8-terminal and 1-central spore former. Out of 18 isolates, NWPZ-9, NWPZ-10 isolates were positive for all lytic enzyme production except lipase, whereas NWPZ-60 were positive for all except esterase. Finally, maximum lytic enzyme producers could directly and indirectly promote the seed germination at initial seedling stage of wheat crop. Overall WMM medium has generated diverse type of core microbes with probability of finding endophyte producing different hydrolases during the isolation itself. These hydrolases coupled with antagonistic potential could have a major influence on the seed germination.
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