Molecular diversity of root and nodule endophytic bacteria from field pea (Pisum sativum L.)

Endophytic bacteria from root and nodules of field pea being grown in CCS Haryana Agricultural University farm Hisar were isolated. A total of 15 endophytic bacteria from surface sterilized roots and 60 from the nodules of field pea were isolated. Out of 75 isolates, 67% in roots and 68% in nodules were gram positive. In roots 70% and in nodules 90% of the gram positive isolates was spore formers. Selected 9 endophytes from roots and 48 from nodules were used to determine molecular diversity by amplication of 16S rDNA by PCR, followed by RFLP by three restriction endonucleases Hae III, RsaI & Hinf I. Wide diversity among field pea bacterial endophytes was observed and considering each cluster as one genotype at 80% level of similarity coefficient, thereby in CCS Haryana Agricultural University soils, 3 genotypes were present in field pea roots while 17 genotypes were present in the nodules. A combined dandrogram showed that entirely a separate cluster was formed by the root endophytes as compared to nodules endophytes, which indicates that the number and type of bacterial genera enters depending upon the plant tissue and environmental conditions in field pea, besides the microbial population present in the soil.