The diversity of stem-associated bacteria of non-nodulated (Nod À ), wild-type nodulated (Nod þ ) and hypernodulated (Nod þ þ ) soybeans were evaluated by clone library analyses of the 16S ribosomal RNA gene. Soybeans were dressed with standard nitrogen (SN) fertilization (15 kg N ha -1 ) and heavy nitrogen (HN) fertilization (615 kg N ha -1 ). The relative abundance of Alphaproteobacteria in Nod þ soybeans (66%) was smaller than that in Nod À and Nod þ þ soybeans (75-76%) under SN fertilization, whereas that of Gammaproteobacteria showed the opposite pattern (23% in Nod þ and 12-16% in Nod À and Nod þ þ soybeans). Principal coordinate analysis showed that the bacterial communities of Nod À and Nod þ þ soybeans were more similar to each other than to that of Nod þ soybeans under SN fertilization. HN fertilization increased the relative abundance of Gammaproteobacteria in all nodulation phenotypes (33-57%) and caused drastic shifts of the bacterial community. The clustering analyses identified a subset of operational taxonomic units (OTUs) at the species level in Alpha-and Gammaproteobacteria responding to both the nodulation phenotypes and nitrogen fertilization levels. Meanwhile, the abundance of Betaproteobacteria was relatively constant in all libraries constructed under these environmental conditions. The relative abundances of two OTUs in Alphaproteobacteria (Aurantimonas sp. and Methylobacterium sp.) were especially sensitive to nodulation phenotype and were drastically decreased under HN fertilization. These results suggested that a subpopulation of proteobacteria in soybeans is controlled in a similar manner through both the regulation systems of plant-rhizobia symbiosis and the nitrogen signaling pathway in plants.
Acid sulfate soil (ASS) has an extremely low pH (3.0) and a high capacity to fix phosphate; symptoms of phosphate deficiency are commonly observed in many crop plants. Arbuscular mycorrhizal (AM) fungi form mutualistic relationships with plant roots, and improve uptake of phosphate from soil. However, there is little information on the actual situation of AM fungi in ASS in Thailand. The purpose of the present study is to determine the indigenous AM fungal density and species in ASS in Thailand. AM fungal spores were retrieved and identified by molecular approaches from ASS field at the central plain of Thailand. This study showed that AM fungal spore density in ASS was 0.232 spores per g dry soil. Among the plant species growing in the natural ASS, there was no AM fungal colonization in the roots of four plant species, i.e. Digitaria sp., Fimbristylis sp., Mimosa pudica L., and Sesbania sp.; however, AM colonization was found in Wedelia roots. Using phylogenetic analysis, four operational taxonomic units (OTUs), i.e., one Glomus, one Entrophospora, one Paraglomus and one unknown species were identified from the AM fungal spores. Five OTUs, i.e., two Glomus, one Acaulospora, one Entrophospora and one unknown Glomeromycota were indentified from Wedelia roots. To our knowledge, this is the first report of the actual situation of AM fungi in ASS in Thailand determined by using molecular approaches.
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