ISSN: 0973-7510

E-ISSN: 2581-690X

Ronak Prajapati , R.V. Vyas, H.N. Shelat and H.K. Patel
Department of Agricultural Microbiology and Biofertilizers Project, Anand Agricultural University, Anand – 388 110, India.
J. Pure Appl. Microbiol., 2016, 10 (2): 1655-1661
© The Author(s). 2016
Received: 13/01/2016 | Accepted: 19/04/2016 | Published: 30/06/2016
Abstract

The native diazotrophic bacterial isolates (ABA-1, ABA-10, ABA-14, ABA-2010, ACG-2 and ASA-1) and four reference strains (MTCC-446 and MTCC-124 (Azotobacter chroococcum), MTCC-1226 (Acetobacter diazotrophicus) and MTCC-2306(Azospirillum lipofarum) were used for the study. Bacterial culture were grow on their specific nitrogen free medium. Detection of nifH (Nitrogen fixation) gene by polymerase chain reaction (PCR) amplification using specific nifH primer and whole cell protein profiling by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Presence of nifH gene confirms the nitrogen fixing nature of native diazotrophic isolates thus native diazotrophic isolates and reference strains which have showed the best nitrogen fixation ability and further validation of the Nitrogenases enzyme (MoFe- 220 to 240 kDa or Fe- 60 kDa protein) which is responsible for biological nitrogen fixation in all native nitrogen fixing bacterial isolates as well as standard cultures viz., (MTCC-446 and 124), (MTCC-1226) and (MTCC-2306) at three different incubation hours 24, 48 and 96 hrs proving their diazotrophic nature. Detection of nifH gene and Fe- 60kDa protein validation, which gave authentication to all the native nitrogen fixer bacterial isolates belonging to Azotobacter, Acetobacter and Azospirillum.

Keywords

Diazotrophs, PCR, nifH gene, Nitrogenases, SDS-PAGE, Biological Nitrogen Fixation (BNF).

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© The Author(s) 2016. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.