Identification of methanotrophs in the buffalo rumen by 16S rRNA sequence analysis

NISHA JHA; SACHINANDAN DE; S S KUNDU; CHANDER DATT

doi:10.56093/ijans.v83i12.35814

Authors

NISHA JHA Assistant Manager (Research and Development), Anmol Feeds Private Limited, Muzzarpur, Bihar
SACHINANDAN DE Senior Scientist, Animal Biotechnology Centre, ICAR-NDRI, Karnal, Haryana
S S KUNDU Principal Scientist and Head, National Dairy Research Institute, Karnal, Haryana 132 001 India
CHANDER DATT Senior Scientist, Dairy Cattle Nutrition Division, National Dairy Research Institute, Karnal, Haryana 132 001 India

https://doi.org/10.56093/ijans.v83i12.35814

Keywords:

16S rRNA gene, Buffalo, Rumen, Methanotrophs

Abstract

In the present study, methanotrophs inhabiting the rumen of Murrah buffalo were detected by molecular technique. The methanotrophs specific primer sets (type l and type ll methanotrophs) were used to amplify 16S rDNA sequence from buffalo rumen. The PCR product of an expected DNA length was observed in the 1% agarose gel, which was purified and then cloned. Different gene clones were screened for inserts using the respective PCR primers. Out of 15 colonies, 10 colonies were positive for inserts of 16S rRNA gene sequences. One recombinant plasmid DNA from each group was sequenced and the nucleotide sequence (592bp and 486bp) obtained with type I and type II primer sets were deposited in the GenBank database under the accession number HQ699778 and HQ699779, respectively. The sequences obtained in present study showed <83% sequence similarity with known methanotrophs and most closely (96 to 99%) related to the 16S rRNA gene of uncultured rumen bacteria. The findings lead to a supposition that the novel methanotrophs with 16S rRNA gene similar to uncultured rumen bacteria are present in the Murrah buffalo rumen.

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