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Nasushiobara, Japan

Oishi H.,Forage Crop Research Institute | Takahashi W.,Japan National Agriculture and Food Research Organization | Ebina M.,Japan National Agriculture and Food Research Organization | Takamizo T.,Japan National Agriculture and Food Research Organization
Breeding Science | Year: 2010

By data mining of EST databases and screening of a genomic library, the cold stimulated gene Lcs19 was isolated from Italian ryegrass (Lolium multiflorum Lam.). The amino acid sequence of the putative LCS19 was 74% identical to wheat (Triticum aestivum) WCS19. Transcripts of Lcs19 were detected after 8 h of cold treatment, and continued to increase up to 48 h after exposure. Lcs19 expression was induced only by cold temperature, and not by high salinity or exogenous abscisic acid. No Lcs19 expression was observed in seedlings that underwent the cold treatment in the dark. Comparison of the promoter of Lcs19 with that of homologous genes of wheat and barley (Hordeum vulgare) showed that regions that include the DRE/CRT cisacting element sequence were conserved. These results suggest that Lcs19 expression is regulated by signal pathway mediated by the DRE/CRT element and other signal pathways reflecting photosynthetic conditions such as a redox signal.


Takahashi W.,Japan National Agriculture and Food Research Organization | Oishi H.,Forage Crop Research Institute | Ebina M.,Japan National Agriculture and Food Research Organization | Komatsu T.,Japan National Agriculture and Food Research Organization | Takamizo T.,Japan National Agriculture and Food Research Organization
Breeding Science | Year: 2010

Transgenic Italian ryegrass plants expressing the betaine aldehyde dehydrogenase gene of zoysiagrass (ZBD1) were produced via particle bombardment of embryogenic calluses. Although the growth of both non-transgenic and transgenic plants expressing ZBD1 was inhibited under salt stress, isolated tillers of the transgenic plants showed better re-rooting ability in vitro in the presence of 300 mM NaCl. Chlorophyll fluorescence values of the transgenic plants declined under salt stress, but were significantly higher than those of non-transgenic plants under salt stress, indicating the improved salt stress tolerance of transgenic plants.


Matsukura K.,Japan National Agricultural Research Center | Shiba T.,National Institute of Livestock and Grassland Science | Shiba T.,Japan National Agricultural Research Center | Sasaki T.,Forage Crop Research Institute | Matsumura M.,Japan National Agricultural Research Center
Journal of Economic Entomology | Year: 2012

Particular alkaloids produced by Neotyphodium endophytes show toxicity to invertebrates. Italian ryegrass (Lolium multiflorum Lamarck) cultivars and strains that are symbiotic with Neotyphodium endophytes have been recently established in Japan. N. uncinatum-infected Italian ryegrass lines accumulate N-formylloline, a type of loline alkaloid (1-aminopyrrolizidine) showing neurotoxicity to herbivorous insects. This study investigated the toxicity of N-formylloline and resistance of N. uncinatum-infected Italian ryegrass to vascular-sap feeding Clypeorrhynchan pests. When four vascular-sap feeding insects: Laodelphax striatellus (Falln) (Homoptera: Delphacidae), Sogatella furcifera (Horvth) (Homoptera: Delphacidae), Cicadulina bipunctata (Melichar) (Homoptera: Cicadellidae), and Nephotettix cincticeps (Uhler) (Homoptera: Cicadellidae) fed on N. uncinatum-infected Italian ryegrass, significant decreases in survival rate were observed for three phloem-sap feeders but not for a xylem-sap feeder, N. cincticeps. This result suggests an uneven distribution of N-formylloline among plant tissues. A potency assay for N-formylloline using a Parafilm feeding sachet and a quantitative analysis of N-formylloline in plant showed a concentration-dependent lethal effect of N-formylloline on all four tested vascular-sap feeders. Our results strongly suggest that N. uncinatum-infected plants can control some Clypeorrhynchan pests in crop fields. © 2012 Entomological Society of America.


Matsukura K.,Japan National Agriculture and Food Research Organization | Shiba T.,Japan National Agriculture and Food Research Organization | Sasaki T.,Forage Crop Research Institute | Yoshida K.,Japan National Agriculture and Food Research Organization | Matsumura M.,Japan National Agriculture and Food Research Organization
Journal of Applied Microbiology | Year: 2014

Aims: A fungal endophyte, Neotyphodium uncinatum, accumulates N-formylloline, which is toxic to Hemipteran insects, in Italian ryegrass. This study aimed to clarify the dynamics of N. uncinatum and N-formylloline in Italian ryegrass, and their relationship to insect resistance. Methods and Results: Changes in the density and localization of N. uncinatum and N-formylloline in N. uncinatum-infected Italian ryegrass were examined by real-time PCR and gas chromatography, respectively. Neotyphodium uncinatum multiplied on pseudostems at the flowering stage, and then increased on inflorescences at the ripening stage. On the other hand, N-formylloline accumulated heavily in inflorescences and leaf blades, but lightly in pseudostems at the ripening stage. In field experiments, N. uncinatum-infected Italian ryegrass suppressed the occurrence of Stenotus rubrovittatus, which fed on the inflorescences, but was not effective to Laodelphax striatellus, which do not necessarily prefer a particular plant tissue. Conclusion: Localization of N. uncinatum and N-formylloline were discordant in Italian ryegrass. The N. uncinatum-infected Italian ryegrass was effective to only insects that prefer to feed on particular plant tissues containing N-formylloline. Significance and Impact of Study: Our data implies that the relationship between insect habits and the dynamics of alkaloids in plants is important for the effective use of endophyte-infected crops. © 2013 The Society for Applied Microbiology.


Kindiger B.,U.S. Department of Agriculture | Conley T.,Oklahoma City University | Keith-Stanley G.,Oklahoma City University | Cai H.,Forage Crop Research Institute
Grassland Science | Year: 2011

Microsatellite markers represent a proven class of genetic markers that have wide use in a variety of plant genetic studies and exhibit several advantages when compared to other DNA-based marker systems. Because of their successful use for genetic analysis in a number of plant species, their generation and availability in the genus Poa (Poaceae) would be advantageous for a multitude of Poa improvement, selection or marker-assisted breeding endeavors. However, due to the cost of their development, few microsatellite markers are available for Poa. In this study, we describe the isolation and characterization of 46 informative microsatellite primer pairs from an unenriched Poa arachnifera Torrey (Texas bluegrass) genomic library. Most of the microsatellite primer pairs amplify a single amplification product and were found to be useful across seven other Poa species. The availability of these informative microsatellite markers should serve a valuable role in the analysis and construction of genetic linkage maps and future marker assisted selection projects. © 2011 The Authors. Grassland Science © 2011 Japanese Society of Grassland Science.

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