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Analysis of the Involvement of Hydroxyanthranilate Hydroxycinnamoyltransferase and Caffeoyl-CoA 3-O-Methyltransferase in Phytoalexin Biosynthesis in Oat

January 2004 , Volume 17 , Number  1
Pages  81 - 89

Qian Yang , 1 Hoat Xuan Trinh , 1 Satoshi Imai , 1 Atsushi Ishihara , 2 Liqun Zhang , 1 Hitoshi Nakayashiki , 1 Yukio Tosa 1 and Shigeyuki Mayama 1

1Laboratory of Plant Pathology, Graduate School of Science and Technology, Kobe University, Kobe 657-8501, Japan; 2Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan

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Accepted 6 August 2003.

Two oat genes encoding hydroxycinnamoyl-CoA:hydroxyanthranilate N-hydroxycinnamoyltransferase (HHT) and S-adenosyl-l-methionine:trans-caffeoyl-CoA 3-O-methyltransferase (CCoAOMT), both of which are possibly involved in the biosynthesis of oat avenanthramide phytoalexins, were cloned and their expression profiles in response to biological stress were studied. Four distinct cDNAs of oat HHT (AsHHT1-4) were isolated with the degenerative polymerase chain reaction method. The enzymatic activity of AsHHT1 expressed in E. coli was found using hydroxyanthranilate and hydroxycinnamoyl-CoAs as cosubstrates. Cloned oat CCoAOMT (AsCCoAOMT) encoded a polypeptide of 130 amino acid residues with 77.7 to 80.8% identities to the CCoAOMT sequences from other plant species. The accumulation of AsHHT1 and AsCCoAOMT transcripts increased concomitantly with phytoalexin accumulation by the treatment of victorin, a specific elicitor in oat lines carrying the Pc-2/Vb gene. Pharmacological approaches indicated the involvement of Ca2+, NO, and protein kinases in the signaling pathways of AsHHT1 and AsCCoAOMT mRNA induction. When oat leaves were inoculated with Puccinia coronata, the mRNA expression of AsHHT1 and AsCCOAOMT increased in both incompatible and compatible interactions but more rapidly in incompatible interaction. Interestingly, however, significant phytoalexin accumulation was observed only in incompatible interaction during the experimental period, suggesting that phytoalexin accumulation may be inhibited in one or more posttranscriptional processes in the compatible interaction.

© 2004 The American Phytopathological Society