Analysis of the endophytic actinobacterial population in the roots of wheat (Triticum aestivum L.) by Terminal Restriction Fragment Length Polymorphism (T-RFLP) and sequencing of 16S rRNA clones

dc.contributor.authorConn, V.
dc.contributor.authorFranco, C.
dc.contributor.organisationAustralian Centre for Plant Functional Genomics (ACPFG)
dc.date.issued2004
dc.description.abstractThe endophytic actinobacterial population in the roots of wheat grown in three different soils obtained from the southeast part of South Australia was investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis of the amplified 16S rRNA genes. A new, validated approach was applied to the T-RFLP analysis in order to estimate, to the genus level, the actinobacterial population that was identified. Actinobacterium-biased primers were used together with three restriction enzymes to obtain terminal restriction fragments (TRFs). The TRFs were matched to bacterial genera by the T-RFLP Analysis Program, and the data were analyzed to validate and semiquantify the genera present within the plant roots. The highest diversity and level of endophytic colonization were found in the roots of wheat grown in a dark loam from Swedes Flat, and the lowest were found in water-repellent sand from Western Flat. This molecular approach detected a greater diversity of actinobacteria than did previous culture-dependent methods, with the predominant genera being Mycobacterium (21.02%) in Swedes Flat, Streptomyces (14.35%) in Red Loam, and Kitasatospora (15.02%) in Western Flat. This study indicates that the soil that supported a higher number of indigenous organisms resulted in wheat roots with higher actinobacterial diversity and levels of colonization within the plant tissue. Sequencing of 16S rRNA clones, obtained using the same actinobacterium-biased PCR primers that were used in the T-RFLP analysis, confirmed the presence of the actinobacterial diversity and identified a number of Mycobacterium and Streptomyces species.
dc.identifier.citationApplied and Environmental Microbiology, 2004; 70(3):1787-1794
dc.identifier.doi10.1128/AEM.70.3.1787-1794.2004
dc.identifier.issn0099-2240
dc.identifier.issn1098-5336
dc.identifier.urihttp://hdl.handle.net/2440/44699
dc.language.isoen
dc.publisherAmer Soc Microbiology
dc.source.urihttps://doi.org/10.1128/aem.70.3.1787-1794.2004
dc.titleAnalysis of the endophytic actinobacterial population in the roots of wheat (Triticum aestivum L.) by Terminal Restriction Fragment Length Polymorphism (T-RFLP) and sequencing of 16S rRNA clones
dc.typeJournal article
pubs.publication-statusPublished

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