Tsuyoshi Isawa

Publications of Tsuyoshi Isawa

• Complete genomic structure of the cultivated rice endophyte Azospirillum sp. B510.

  Authors: Takakazu Kaneko, Kiwamu Minamisawa, Tsuyoshi Isawa, Hiroki Nakatsukasa, Hisayuki Mitsui, Yasuyuki Kawaharada, Yasukazu Nakamura, Akiko Watanabe, Kumiko Kawashima, Akiko Ono [......] Chika Takahashi, Chiharu Minami, Tsunakazu Fujishiro, Mitsuyo Kohara, Midori Katoh, Naomi Nakazaki, Shinobu Nakayama, Manabu Yamada, Satoshi Tabata, Shusei Sato

  DNA research : an international journal for rapid publication of reports on genes and genomes. 02/2010; 17(1):37-50.

  We determined the nucleotide sequence of the entire genome of a diazotrophic endophyte, Azospirillum sp. B510. Strain B510 is an endophytic bacterium isolated from stems of rice plants (Oryza sativaWe determined the nucleotide sequence of the entire genome of a diazotrophic endophyte, Azospirillum sp. B510. Strain B510 is an endophytic bacterium isolated from stems of rice plants (Oryza sativa cv. Nipponbare). The genome of B510 consisted of a single chromosome (3,311,395 bp) and six plasmids, designated as pAB510a (1,455,109 bp), pAB510b (723,779 bp), pAB510c (681,723 bp), pAB510d (628,837 bp), pAB510e (537,299 bp), and pAB510f (261,596 bp). The chromosome bears 2893 potential protein-encoding genes, two sets of rRNA gene clusters (rrns), and 45 tRNA genes representing 37 tRNA species. The genomes of the six plasmids contained a total of 3416 protein-encoding genes, seven sets of rrns, and 34 tRNAs representing 19 tRNA species. Eight genes for plasmid-specific tRNA species are located on either pAB510a or pAB510d. Two out of eight genomic islands are inserted in the plasmids, pAB510b and pAB510e, and one of the islands is inserted into trnfM-CAU in the rrn located on pAB510e. Genes other than the nif gene cluster that are involved in N(2) fixation and are homologues of Bradyrhizobium japonicum USDA110 include fixABCX, fixNOQP, fixHIS, fixG, and fixLJK. Three putative plant hormone-related genes encoding tryptophan 2-monooxytenase (iaaM) and indole-3-acetaldehyde hydrolase (iaaH), which are involved in IAA biosynthesis, and ACC deaminase (acdS), which reduces ethylene levels, were identified. Multiple gene-clusters for tripartite ATP-independent periplasmic-transport systems and a diverse set of malic enzymes were identified, suggesting that B510 utilizes C(4)-dicarboxylate during its symbiotic relationship with the host plant.

• Azospirillum sp. strain B510 enhances rice growth and yield.

  Authors: Tsuyoshi Isawa, Michiko Yasuda, Hirotoshi Awazaki, Kiwamu Minamisawa, Satoshi Shinozaki, Hideo Nakashita

  Microbes and environments / JSME. 01/2010; 25(1):58-61.

  Inoculation experiments with the endophytic bacterium Azospirillum sp. strain B510, an isolate from surface-sterilized stems of field-grown rice, were conducted in pots in a greenhouse, and in paddyInoculation experiments with the endophytic bacterium Azospirillum sp. strain B510, an isolate from surface-sterilized stems of field-grown rice, were conducted in pots in a greenhouse, and in paddy fields in Hokkaido, Japan. B510 significantly enhanced the growth of newly generated leaves and shoot biomass under greenhouse conditions. When rice seedlings were treated with 1×10(8) CFU ml(-1), then transplanted to paddy fields, tiller numbers and seed yield significantly increased. Azospirillum sp. strain B510 is a promising bacterial inoculant for plant growth promotion and agricultural practices.

• Effects of Colonization of a Bacterial Endophyte, Azospirillum sp. B510, on Disease Resistance in Rice.

  Authors: Michiko Yasuda, Tsuyoshi Isawa, Satoshi Shinozaki, Kiwamu Minamisawa, Hideo Nakashita

  Bioscience, biotechnology, and biochemistry. 12/2009;

  Agriculturally important grasses contain numerous diazotrophic bacteria, the interactions of which are speculated to have some other benefits to the host plants. In this study, we analyzed theAgriculturally important grasses contain numerous diazotrophic bacteria, the interactions of which are speculated to have some other benefits to the host plants. In this study, we analyzed the effects of a bacterial endophyte, Azospirillum sp. B510, on disease resistance in host rice plants. Rice plants (Oryza sativa cv. Nipponbare) were inoculated with B510 exhibited enhanced resistance against diseases caused by the virulent rice blast fungus Magnaporthe oryzae and by the virulent bacterial pathogen Xanthomonas oryzae. In the rice plants, neither salicylic acid (SA) accumulation nor expression of pathogenesis-related (PR) genes was induced by interaction with this bacterium, except for slight induction of PBZ1. These results indicate the possibility that strain B510 is able to induce disease resistance in rice by activating a novel type of resistance mechanism independent of SA-mediated defense signaling.

• Expression of the nifH gene of a Herbaspirillum endophyte in wild rice species: daily rhythm during the light-dark cycle.

  Authors: Mu You, Tomohiro Nishiguchi, Asami Saito, Tsuyoshi Isawa, Hisayuki Mitsui, Kiwamu Minamisawa

  Applied and environmental microbiology. 01/2006; 71(12):8183-90.

  The expression of nitrogenase genes of Herbaspirillum sp. B501 associated in shoot (leaf and stem) of wild rice, Oryza officinalis, was studied by means of reverse transcription-PCR (RT-PCR) targetedThe expression of nitrogenase genes of Herbaspirillum sp. B501 associated in shoot (leaf and stem) of wild rice, Oryza officinalis, was studied by means of reverse transcription-PCR (RT-PCR) targeted at the nifH gene. RT-PCR analyses indicate that nifH transcript was detected exclusively from nitrogen-fixing cells of gfp-tagged strain B501gfp1 in both free-living and endophytic states by using a constitutive gfp gene transcript as a positive control. Transcription of nifH and nitrogen fixation in free-living cells were induced maximally at a 2% O2 concentration and repressed in free air (21% O2). nifH transcription was monitored in the endophytic cells by using total RNA extracted from B501gfp1-inoculated wild rice plants during daily light-dark cycles. The level of nifH transcription in planta varied dramatically, with a maximum during the light period. Moreover, the light radiation enhanced nifH expression even in free-living cells grown in culture. These results suggest that in planta nitrogen fixation by the endophyte shows a daily rhythm determined by the plant's light environment.

• Phylogeny and distribution of extra-slow-growing Bradyrhizobium japonicum harboring high copy numbers of RSalpha, RSbeta and IS1631.

  Authors: Reiko Sameshima, Tsuyoshi Isawa, Michael J Sadowsky, Tohru Hamada, Hiroaki Kasai, Arawan Shutsrirung, Hisayuki Mitsui, Kiwamu Minamisawa

  FEMS microbiology ecology. 06/2003; 44(2):191-202.

  Abstract We previously reported that extra-slow-growing Bradyrhizobium japonicum isolates obtained from three field sites in Japan, designated as HRS (highly reiterated sequence-possessing) strains,Abstract We previously reported that extra-slow-growing Bradyrhizobium japonicum isolates obtained from three field sites in Japan, designated as HRS (highly reiterated sequence-possessing) strains, have high copy numbers of the insertion sequences RSalpha and RSbeta. When strain collections in the USA, Japan, Korea, Thailand and China were examined by Southern hybridization using RSalpha, RSbeta and IS1631 as probes, HRS strains were found in the Japanese, Chinese, and American collections, but not in the Korean and Thai ones. Copy number analyses of RSalpha and RSbeta, calibrated with the copy number of rrs (16S rRNA gene), indicated that the HRS stains can be divided into two major groups. Group A is comprised of members with a high copy number of RSalpha (mean+/-S.D., 139+/-27), a low number of RSbeta (mean+/-S.D., 30+/-13) sequences, and extremely slow growth rates (mean doubling time+/-S.D., 27+/-9 h). In contrast, group B is comprised of strains with a high copy number of RSbeta (mean+/-S.D., 93+/-6) and a lower number of RSalpha. These groupings of HRS strains were well correlated with phylogenetic clusters based on rrs, gyrB and serogroups (110/122 and 123/135). Growth rate of B. japonicum strains was also correlated exclusively with RSalpha copy number. The ecological, evolutionary and biotechnological implications of the findings are discussed.