Okada Atsushi, Okada Kazunori, Miyamoto Koji, Koga Jinichiro, Shibuya Naoto, Nojiri Hideaki, Yamane Hisakazu
Biotechnology Research Center, The University of Tokyo, Tokyo 113-8657, Japan.
J Biol Chem. 2009 Sep 25;284(39):26510-8. doi: 10.1074/jbc.M109.036871. Epub 2009 Jul 27.
Production of major diterpenoid phytoalexins, momilactones and phytocassanes, is induced in rice upon recognition of pathogenic invasion as plant defense-related compounds. We recently showed that biosynthetic genes for momilactones are clustered on rice chromosome 4 and co-expressed after elicitation, mimicking pathogen attack. Because genes for most metabolic pathways in plants are not organized in gene clusters, examination of the mechanism(s) regulating the expression of such clustered genes is needed. Here, we report a chitin oligosaccharide elicitor-inducible basic leucine zipper transcription factor, OsTGAP1, which is essential for momilactone biosynthesis and regulates the expression of the five genes in the cluster. The knock-out mutant for OsTGAP1 had almost no expression of the five clustered genes (OsCPS4, OsKSL4, CYP99A2, CYP99A3, and OsMAS) or production of momilactones upon elicitor treatment. Inductive expression of OsKSL7 for phytocassane biosynthesis was also largely affected in the ostgap1 mutant, although phytocassane accumulation still occurred. Conversely, OsTGAP1-overexpressing lines exhibited enhanced expression of the clustered genes and hyperaccumulation of momilactones in response to the elicitor. Furthermore, enhanced expression of OsKSL7 and hyperaccumulation of phytocassanes was also observed. We also found that OsTGAP1 overexpression can influence transcriptional up-regulation of OsDXS3 in the methylerythritol phosphate pathway, eventually leading to inductive production of diterpenoid phytoalexins. These results indicate that OsTGAP1 functions as a key regulator of the coordinated transcription of genes involved in inductive diterpenoid phytoalexin production in rice and mainly exerts an essential role on expression of the clustered genes for momilactone biosynthesis.
在水稻中,当识别到病原体入侵时,会诱导产生主要的二萜类植保素——稻瘟菌素和植二苯,并将其作为与植物防御相关的化合物。我们最近发现,稻瘟菌素的生物合成基因聚集在水稻第4号染色体上,在模拟病原体攻击的诱导后共同表达。由于植物中大多数代谢途径的基因不是成簇排列的,因此需要研究调节此类成簇基因表达的机制。在这里,我们报道了一种几丁质寡糖诱导型碱性亮氨酸拉链转录因子OsTGAP1,它对稻瘟菌素的生物合成至关重要,并调节簇中五个基因的表达。OsTGAP1的敲除突变体在诱导处理后,这五个成簇基因(OsCPS4、OsKSL4、CYP99A2、CYP99A3和OsMAS)几乎没有表达,也不产生稻瘟菌素。尽管仍有植二苯积累,但在ostgap1突变体中,用于植二苯生物合成的OsKSL7的诱导表达也受到很大影响。相反,过表达OsTGAP1的株系在诱导剂处理下,簇基因的表达增强,稻瘟菌素超积累。此外,还观察到OsKSL7的表达增强和植二苯的超积累。我们还发现,OsTGAP1的过表达可以影响甲基赤藓糖醇磷酸途径中OsDXS3的转录上调,最终导致二萜类植保素的诱导产生。这些结果表明,OsTGAP1作为水稻中诱导二萜类植保素产生相关基因协同转录的关键调节因子,主要对稻瘟菌素生物合成的成簇基因的表达发挥重要作用。