Isolation of Glucose Fermenting Nitrogen Fixing Gene (nifH) Containing Endophytic Bacteria from Seed and Roots of Wheat (Triticum aestivation l.)

Abstract

The present study was conducted to study the nifH gene containing endophytic bacterial frequency in wheat seed and roots obtained from the soil of Chitwan and Kaski. One hundred and four isolates were studied for the presence of the nifH gene. There was a diversity in isolate characters obtained from root sample Root (R), direct seed sample (ds), and plant sample (P). None of the isolates from any sources showed indole-producing ability. About 18 isolates (15% of the total) contained the nifH gene through amplification of the gene by universal primers PolF and PolR. About 6 isolates from seed sample ds and 12 isolates from root sample R contained nifH gene. None of the isolates from root sample P manifested the presence of the nifH gene. Among 18 nifH-containing isolates, only 6 isolates manifested presence of cel3 gene of 400 bp, whereas, 11 isolates showed cel3 gene of 200 bp. All nifH gene containing isolates were confirmed to be bacteria by PCR amplification of 16s rRNA gene by universal primer 27F and 1492R and visualization of agarose gel matrix with bp range of approximately 530 under UV ray. Further research scope exists to use these microbes as a bio fertilizer in plant growth promotion studies by inoculation.