Influence of enhanced malate dehydrogenase expression by alfalfa on diversity of rhizobacteria and soil nutrient availability

Transgenic alfalfa over-expressing a nodule-enhanced malate dehydrogenase (neMDH) cDNA and untransformed alfalfa plants were grown at the same field site and rhizosphere soils collected after 53 weeks of plant growth. These alfalfa lines differ in the amount and composition of root organic acids produced and exuded into the rhizosphere. Nucleotide sequencing of PCR-based 16S ribosomal DNA (rDNA) clone libraries and Biolog™ GN microtiter plates were employed to assess the activity of naturally occurring rhizobacteria in the two alfalfa rhizospheres. Selected macro- and micro-elements in the two alfalfa rhizosphere soils were also measured. Analysis of 240 16S rDNA clone sequences indicated the existence of about 11 bacterial phyla and their major subdivisions in the two alfalfa rhizosphere samples. There were qualitative changes in the abundance of bacterial phylogenetic groups between rhizosphere soils of transgenic and untransformed alfalfa. Carbon substrate utilization profiles suggested that rhizosphere samples from transgenic alfalfa had significantly greater microbial functional diversity compared with rhizosphere samples from untransformed alfalfa. The concentrations of nitric acid extractable P, K, Mn, Zn and Cu increased significantly in the transgenic alfalfa rhizosphere compared with the untransformed alfalfa rhizosphere. These observations indicate that organic acids produced by plant roots significantly influence rhizosphere microbial diversity and availability of macro- and micro-nutrients and demonstrate the utility of such trangenic plants as tools for studying the potential impact of plant root exudates on soil microbial ecosystems.