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Isolation and molecular characterization of a Spotted leaf 18 mutant by modified activation-tagging in rice

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Abstract

A lesion mimic mutant that we designated Spotted leaf 18 (Spl18) was isolated from 13,000 activation-tagging lines of rice produced by our modified activation-tagging vector and further characterized. Spl18 was dominant and its phenotype was linked to the T-DNA insertion. An ORF was located about 500 bp downstream of the inserted T-DNA, and the deduced protein, designated OsAT1, showed sequence similarity to an acyltransferase whose expression is induced by hypersensitive reaction in tobacco. The transcriptional level of OsAT1 was very low in the WT leaf blade but high in Spl18 leaf blade. In wild-type rice, OsAT1 was transcribed mainly in the young panicle, in the panicle just after heading, and in the leaf sheath. In addition, transcription of the genes for PR protein was upregulated in Spl18, accumulation of phytoalexins (both momilactone A and sakuranetin) was increased, and resistance to blast disease was improved. We then combined OsAT1 genomic DNA downstream of the modified 35S promoter and re-transformed it into rice. Lesion mimic and blast resistance phenotypes were detected in the transgenic lines produced, clearly indicating that overexpression of OsAT1 caused the Spl18 phenotypes. In addition, plants overexpressing OsAT1 showed resistance to bacterial blight.

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Acknowledgements

This work was supported in part by the Ministry of Agriculture, Forestry and Fisheries of Japan (Rice Genome Project nos. MP-1202 and 2142) and by a Grant-in-Aid for Scientific Research (C) (grant no. 17580087) from the Ministry of Education, Sports, Culture, Science and Technology of Japan. We thank Meiji Seika Co. (Tokyo, Japan) for kindly providing bialaphos and the PAT gene. We thank Elizabeth E. Hood for Agrobacterium strain EHA101. We thank Hiroaki Ichikawa for kindly providing pSMAB704. We thank Sei-ichi Toki, Hiroshi Tanaka, Haruko Onodera and Kazuko Ono for their technical advice on rice transformation and Nipponbare seeds, and Shin-ichiro Watanabe and Muneo Yamazaki for their technical advice on TAIL-PCR. We also thank Tomiko Senba, Lois Ishizaki, Toshiko Shibata, Noriko Nakajima, Hiromi Satoh, and Hisako Ooka for technical assistance.

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  1. National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

    Masaki Mori, Chikako Tomita, Kazuhiko Sugimoto, Nagao Hayashi, Joseph G. Dubouzet, Hirokazu Ochiai, Hirohiko Hirochika & Shoshi Kikuchi

  2. College of Agriculture, Ibaraki University, Ami, 300-0393, Japan

    Morifumi Hasegawa

  3. Faculty of Agriculture, Utsunomiya University, Utsunomiya, 321-8505, Japan

    Chikako Tomita & Hitoshi Sekimoto

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  1. Masaki Mori
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  2. Chikako Tomita
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  9. Hirohiko Hirochika
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Mori, M., Tomita, C., Sugimoto, K. et al. Isolation and molecular characterization of a Spotted leaf 18 mutant by modified activation-tagging in rice. Plant Mol Biol 63, 847–860 (2007). https://doi.org/10.1007/s11103-006-9130-y

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