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ORIGINAL ARTICLE
Year : 2012  |  Volume : 8  |  Issue : 30  |  Page : 93-97

Identification and elimination of bacterial contamination during in vitro propagation of Guadua angustifolia Kunth


1 Department of Biotechnology and Environmental Sciences, Thapar University, Patiala, Punjab; Division of Biotechnology, CSIR- Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India
2 Division of Biotechnology, CSIR- Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India
3 Department of Biotechnology and Environmental Sciences, Thapar University, Patiala, Punjab, India

Correspondence Address:
Anil Sood
Scientist G and Head, Division of Biotechnology, CSIR- Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh - 176 061
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0973-1296.96547

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Background: Guadua angustifolia Kunth is a very important bamboo species with significant utility in pharmaceutical, paper, charcoal, and construction industries. Microbial contamination is a major problem encountered during establishment of in vitro cultures of Guadua. Objective: This study has been designed to analyze the identity of contaminating bacteria and to develop the strategy to eliminate them during micropropagation of Guadua. Materials and Methods: We isolated and consequently analyzed partial sequence analysis of the 16S rRNA gene to identify two contaminating bacteria as (1) Pantoea agglomerans and (2) Pantoea ananatis. In addition, we also­ performed antibiotic sensitivity testing on these bacterial isolates. Results: We identified kanamycin and streptomycin sulfate as potentially useful antibiotics in eliminating the contaminating bacteria. We grew shoots on multiplication medium containing BAP (2 mg/l) and adenine sulfate (10 mg/l) supplemented with kanamycin (10 μg/ml) for 10 days and transferred them to fresh medium without antibiotics and found that bacterial growth was inhibited. Moreover, we observed intensive formation of high-quality shoots. Streptomycin sulfate also inhibited bacterial growth but at higher concentration. We also demonstrated that shoots grown in streptomycin sulfate tended to be shorter and had yellow leaves. Conclusion: Thus, we have developed a novel strategy to identify and inhibit intriguing microbial contaminations of (1) Pantoea agglomerans and (2) Pantoea ananatis during establishment of in vitro cultures of Guadua. This would improve in vitro establishment of an important bamboo, Guadua angustifolia Kunth for large scale propagation.


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