ABSTRACT
The molecular and physiological mechanisms behind the maturation and maintenance of N(2)-fixing nodules during development of symbiosis between rhizobia and legumes still remain unclear, although the early events of symbiosis are relatively well understood. Azorhizobium caulinodans ORS571 is a microsymbiont of the tropical legume Sesbania rostrata, forming N(2)-fixing nodules not only on the roots but also on the stems. In this study, 10,080 transposon-inserted mutants of A. caulinodans ORS571 were individually inoculated onto the stems of S. rostrata, and those mutants that induced ineffective stem nodules, as displayed by halted development at various stages, were selected. From repeated observations on stem nodulation, 108 Tn5 mutants were selected and categorized into seven nodulation types based on size and N(2) fixation activity. Tn5 insertions of some mutants were found in the well-known nodulation, nitrogen fixation, and symbiosis-related genes, such as nod, nif, and fix, respectively, lipopolysaccharide synthesis-related genes, C(4) metabolism-related genes, and so on. However, other genes have not been reported to have roles in legume-rhizobium symbiosis. The list of newly identified symbiosis-related genes will present clues to aid in understanding the maturation and maintenance mechanisms of nodules.
Subject(s)
Azorhizobium/genetics , Bacterial Proteins/metabolism , Fabaceae/microbiology , Nitrogen Fixation , Plant Stems/microbiology , Symbiosis , Azorhizobium/growth & development , Azorhizobium/metabolism , Bacterial Proteins/genetics , DNA Transposable Elements , Fabaceae/growth & development , Molecular Sequence Data , Mutagenesis, Insertional , Mutation , Plant Stems/metabolism , Sequence Analysis, DNAABSTRACT
There has always been a great need for simple and accurate bioassays for evaluating nutrient limitation in aquatic ecosystems. Whereas organic carbon is usually considered to be the limiting nutrient for microbial growth in many aquatic ecosystems, there are, however, many water sources that are limited by phosphorus or nitrogen. A method named "nitrogen fixing bacterial growth potential" (NFBGP) test, which is based on pre-culturing of autochthonous (target) microorganisms was described. The method was applied to evaluate phosphorus or nitrogen nutrient limitation in lake and sewage water samples using an isolate of the nitrogen fixing bacterium, Azorhizobium sp. WS6. The results corresponded well to those from the traditional algal growth potential (AGP) test and the bacterial regrowth potential (BRP) test, suggesting that the NFBGP test is a useful supplementary method for evaluating the limiting nutrient, especially phosphorus, in an aquatic environment.
