Application of a novel Paenibacillus-specific PCR-DGGE method and sequence analysis to assess the diversity of Paenibacillus spp. in the maize rhizosphere

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• 作者：Araú ; jo da Silva ; Katia Regina ; Salles ; Joana Falcã ; o ; Seldin ; Lucy ; van Elsas ; Jan Dirk
• 关键词：Diversity ; Maize rhizosphere ; Paenibacillus ; PCR-DGGE
• 刊名：Journal of Microbiological Methods
• 出版年：2003
• 出版时间：August, 2003
• 年：2003
• 卷：54
• 期：2
• 页码：213-231
• 全文大小：1.61 M

文摘

In this study, a Paenibacillus-specific PCR system, based on the specific primer PAEN515F in combination with bacterial primer R1401, was tested and used to amplify specific fragments of the 16S rRNA gene from rhizosphere DNA. The amplicons were used in a second (semi-nested) PCR for DGGE, in which bacterial primers F968GC and R1401 were used. The resulting products were separated into community fingerprints by DGGE. To assess the reliability of the method, the diversity of Paenibacillus species was evaluated on the basis of DNA extracted directly from the rhizospheres of four different cultivars of maize (Zea mays), i.e. CMS04, CMS11, CMS22 and CMS36, sown in two Brazilian field soils (Cerrado and Várzea). In addition, a clone library was generated from the PCR-generated 16S rDNA fragments, and selected clones were sequenced.