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DNA-based stable isotope probing enables the identification of active bacterial endophytes in potatoes.
New Phytol. 181, 802-807 (2009)
* A (13)CO(2) (99 atom-%, 350 ppm) incubation experiment was performed to identify active bacterial endophytes in two cultivars of Solanum tuberosum, cultivars Desirée and Merkur. We showed that after the assimilation and photosynthetic transformation of (13)CO(2) into (13)C-labeled metabolites by the plant, the most directly active, cultivar-specific heterotrophic endophytic bacteria that consume these labeled metabolites can be identified by DNA stable isotope probing (DNA-SIP). * Density-resolved DNA fractions obtained from SIP were subjected to 16S rRNA gene-based community analysis using terminal restriction fragment length polymorphism analysis and sequencing of generated gene libraries. * Community profiling revealed community compositions that were dominated by plant chloroplast and mitochondrial 16S rRNA genes for the 'light' fractions of (13)CO(2)-incubated potato cultivars and of potato cultivars not incubated with (13)CO(2). In the 'heavy' fractions of the (13)CO(2)-incubated endophyte DNA, a bacterial 492-bp terminal restriction fragment became abundant, which could be clearly identified as Acinetobacter and Acidovorax spp. in cultivars Merkur and Desirée, respectively, indicating cultivar-dependent distinctions in (13)C-label flow. These two species represent two common potato endophytes with known plant-beneficial activities. * The approach demonstrated the successful detection of active bacterial endophytes in potato. DNA-SIP therefore offers new opportunities for exploring the complex nature of plant-microbe interactions and plant-dependent microbial metabolisms within the endosphere.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
C-13-DNA stable isotope probing; 16S rRNA gene-based community analysis; active bacterial endophytes; plant-microbe interaction; Solanum tuberosum (potato); soil type; rna; communities; plants; populations; diversity; microorganisms; tool
ISSN (print) / ISBN
0028-646X
e-ISSN
1469-8137
Journal
New Phytologist
Quellenangaben
Volume: 181,
Issue: 4,
Pages: 802-807
Publisher
Wiley
Reviewing status
Peer reviewed