Wong-Villarreal, A. and Caballero-Mellado, J. (2010) Rapid identification of nitrogen-fixing and legume-nodulating Burkholderia species based on PCR 16S rRNA species-specific oligonucleotides. Systematic and Applied Microbiology, 33 (1). pp. 35-43.
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Abstract
Several novel N(2)-fixing Burkholderia species associated with plants, including legume-nodulating species, have recently been discovered. Presently, considerable interest exists in studying the diazotrophic Burkholderia species, both for their ecology and their great potential for agro-biotechnological applications. However, the available methods used in the identification of these Burkholderia species are time-consuming and expensive. In this study, PCR species-specific primers based on the 16S rRNA gene were designed, which allowed rapid, easy, and correct identification of most known N(2)-fixing Burkholderia. With this approach, type and reference strains of Burkholderia kururiensis, B. unamae, B. xenovorans, B. tropica, and B. silvatlantica, as well as the legume-nodulating B. phymatum, B. tuberum, B. mimosarum, and B. nodosa, were unambiguously identified. In addition, the PCR species-specific primers allowed the diversity of the diazotrophic Burkholderia associated with field-grown tomato and sorghum plants to be determined. B. tropica and B. xenovorans were the predominant species found in association with tomato, but the occurrence of B. tropica with sorghum plants was practically exclusive. The efficiency of the species-specific primers was validated with the detection of B. tropica and B. xenovorans from DNA directly recovered from tomato rhizosphere soil samples. Additionally, using PCR species-specific primers, all of the legume-nodulating Burkholderia were correctly identified, even from single nodules collected from inoculated common bean plants. These primers could contribute to rapid identification of the diazotrophic and nodulating Burkholderia species associated with important crop plants and legumes, as well as revealing their environmental distribution.
| Item Type: | Article |
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| Additional Information: | We thank L. Martı´nez-Aguilar for technical assistance, and A. Ramı´rez-Trujillo for his valuable advice on nodulation tests.We are grateful to Michael Dunn (CCG-UNAM) for reading the manuscript. We acknowledge Euan K. James (University of Dundee, U.K.) and W.-M Chen (National Kaohsiung University, Taiwan) for supplying B. mimosarum and B. nodosa strains. We also thank J.J. Pen˜a-Cabriales and J.A. Vera (CINVESTAV-Irapuato) for sorghum plant collection, and Jose´ Leyva for tomato collection.A. Wong-Villarreal is a Ph.D. student in the Programa de Doctorado en Biotecnologı´a-CEIB/UAEM, who acknowledges fellowships (Register No. 181684) from the Consejo Nacional de Ciencia y Tecnologı´a (CONACyT) Mexico. |
| Uncontrolled Keywords: | Diazotrophic Burkholderia, Nodulating Burkholderia, PCR species-specific primers, Nitrogen fixation, Sorghum, Tomato |
| Author Affiliation: | Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, México. |
| Subjects: | Crop Improvement |
| Divisions: | General |
| Depositing User: | Mr Arbind Seth |
| Date Deposited: | 12 Feb 2013 02:46 |
| Last Modified: | 12 Feb 2013 02:46 |
| Official URL: | http://dx.doi.org/10.1016/j.syapm.2009.10.004 |
| URI: | http://eprints.icrisat.ac.in/id/eprint/9601 |
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