Sequencing of 16S rDNA gene: A rapid tool for ...
1
BIOTECHNOLOGY-2011 342 21-23 March 2011 World Congress on Biotechnology E ndophytic bacteria reside within plant hosts without causing disease symptoms. ese bacteria generally colonize the intercellular spaces, and they have been isolated from all plant parts. One endophytic bacterium, named as KAU-EC1 was isolated from shoot cultures derived from field plants as well as in vitro seedlings of cashew (Anacardium occidentale L.). Isolation of endophytic bacteria on nutrient agar media yielded large mucoid, opaque, circular convex colonies on nutrient agar medium. Characteristics of this bacterium were studied by cultural and morphological tests. For bacterial identification, 16S rDNA sequencing is particularly important in the case of bacteria with unusual phenotypic profiles, rare bacteria, slow-growing bacteria, uncultivable bacteria and culture-negative infections. 16S rRNA molecules contain both highly conserved regions and variable regions. e highly conserved regions provide priming sites suitable for polymerase chain reaction and sequencing applications. PCR was performed for the amplification of 16S rDNA gene. 16S rDNA gene was amplified using two universal bacterial primers: 16S 43-63 and 16S 1404-1387 . e PCR product when checked on agarose gel indicated the presence of band 1.3 Kbp. e 16S rDNA gene from KAU-EC1 was cloned in pGEMT vector, sequenced and analysed aſter vector and adapter editing. In silico analysis using bioinformatics tools revealed that sequence of KAU-EC1 showed 99 per cent homology with Klebsiella pneumoniae strain SA-D6-7 16S ribosomal RNA gene. is is the first study reporting endophytic bacteria in tissue cultured cashew plants. e possible role for this bacterium in the biology of cashew plant remains unknown. Sequencing of 16S rDNA gene: A rapid tool for identification of endophytic bacteria from In vitro cultures of cashew Jusna Mariya P L, Keshavachandran R, Nazeem P A, Girija D and Surendragopal K Centre for Plant Biotechnology and Molecular Biology, India Biography Jusna Mariya P.L. has completed her graduation and post graduation from Kerala Agricultural University, Thrissur and currently she is doing Ph. D. in Agri. Plant Biotechnology at Kerala Agricultural University, Thrissur and pursuing her research work. doi:10.4172/jpb.1000240
Transcript of Sequencing of 16S rDNA gene: A rapid tool for ...
BIOTECHNOLOGY-2011
342
21-2
3 M
arch
2011
World Congress on
Biotechnology
Endophytic bacteria reside within plant hosts without causing disease symptoms. These bacteria generally
colonize the intercellular spaces, and they have been isolated from all plant parts. One endophytic bacterium, named as KAU-EC1 was isolated from shoot cultures derived from field plants as well as in vitro seedlings of cashew (Anacardium occidentale L.). Isolation of endophytic bacteria on nutrient agar media yielded large mucoid, opaque, circular convex colonies on nutrient agar medium. Characteristics of this bacterium were studied by cultural and morphological tests. For bacterial identification, 16S rDNA sequencing is particularly important in the case of bacteria with unusual phenotypic profiles, rare bacteria, slow-growing bacteria, uncultivable bacteria and culture-negative infections. 16S rRNA molecules contain both highly conserved regions and variable regions. The highly
conserved regions provide priming sites suitable for polymerase chain reaction and sequencing applications. PCR was performed for the amplification of 16S rDNA gene. 16S rDNA gene was amplified using two universal bacterial primers: 16S43-63 and 16S1404-1387. The PCR product when checked on agarose gel indicated the presence of band 1.3 Kbp. The 16S rDNA gene from KAU-EC1 was cloned in pGEMT vector, sequenced and analysed after vector and adapter editing. In silico analysis using bioinformatics tools revealed that sequence of KAU-EC1 showed 99 per cent homology with Klebsiella pneumoniae strain SA-D6-7 16S ribosomal RNA gene. This is the first study reporting endophytic bacteria in tissue cultured cashew plants. The possible role for this bacterium in the biology of cashew plant remains unknown.
Sequencing of 16S rDNA gene: A rapid tool for identification of endophytic bacteria from In vitro cultures of cashewJusna Mariya P L, Keshavachandran R, Nazeem P A, Girija D and Surendragopal KCentre for Plant Biotechnology and Molecular Biology, India
BiographyJusna Mariya P.L. has completed her graduation and post graduation from Kerala
Agricultural University, Thrissur and currently she is doing Ph. D. in Agri. Plant Biotechnology at Kerala Agricultural University, Thrissur and pursuing her research work.
doi:10.4172/jpb.1000240
![static-curis.ku.dk · [Pasteurella ] haemolytica or [ P.] anatis by DNA-DNA hybridization, 16S rRNA and 16S-23S ITS sequencing, AFLP, PFGE, plasmid profiling and phenotypic characterization.](https://static.fdocuments.us/doc/165x107/5ceed30088c99330508cabea/static-curiskudk-pasteurella-haemolytica-or-p-anatis-by-dna-dna-hybridization.jpg)
![16S rRNA Amplicon Sequencing for Epidemiological Surveys ... · sequencing] or 16S rRNA amplicon sequencing) of the bacterial DNA or RNA in a sample, followed by comparison to a reference](https://static.fdocuments.us/doc/165x107/602453d03df69944be4d3def/16s-rrna-amplicon-sequencing-for-epidemiological-surveys-sequencing-or-16s.jpg)
