水稻白葉枯病抗性突變體接種白葉枯病菌之轉錄體學研究

Transcriptomic Analysis of a Resistant Mutant in Response to ＂Xanthomonas oryzae＂ pv. ＂oryzae＂ Infection

黃巧宜(Chiao-Yi Huang) ； 周思儀(Szu-Yi Chou) ； 李雅琳(Ya-Lin Lee) ； 曾文彬(Wen-Bin Tseng) ； 陳純葳(Chun-Wei Chen) ； 王強生(Chang-Sheng Wang) ； 林大鈞(Da-Gin Lin)

《台灣農業研究》 64卷2期 (2015/06) Pp. 145-158

https://doi.org/10.6156/JTAR/2015.06402.07

摘要

白葉枯病是由＂Xanthomonas oryzae pv. oryzae＂所引起的水稻嚴重病害。本研究為了開發新的抗性基因，利用「台農67號」（'TNG67'）及具持久抗性之SA0423突變體為材料，接種台灣水稻白葉枯病抗性檢定圃專用菌株XF89b，並利用微陣列晶片分析接種後之轉錄體變化。所得結果經由One-way ANOVA及Student-Newman-Keuls （SNK）分析，發現在SA0423專一表現的轉錄子有2,727個，'TNG67'則有3,585個，而兩者皆有表現之轉錄子則高達18,432個。利用「代謝路徑的註解」、「比對已發表的抗性基因座」、「比對植物－病原菌交互作用代謝路徑」及「BioLayout Express 3D表現分群」等方法，進一步篩選出59個抗性相關基因，再經由real-time RT-PCR分析確認，結果發現17個基因的表現符合微晶片分析結果。最後利用文獻檢索，分析這17個基因與抗性的關聯性，結果發現它們可能參與「植物－病原菌交互作用」、「植物荷爾蒙生合成」、細胞自噬作用」及「免疫機制啟動的訊息傳遞」等與抗性相關的代謝路徑。這些結果除可作為探討SA0423的抗性機制外，亦可進一步選殖表現數量基因座（expression quantitative trait loci; eQTL）與分子標誌，應用於未來水稻白葉枯病抗性之分子育種。

關鍵字

水稻 ； 白葉枯病 ； 抗性 ； 突變體 ； 轉錄體

並列摘要

Bacterial blight disease, caused by ＂Xanthomonas oryzae pv. oryzae＂, is one of the most serious diseases in rice. To explore novel resistant genes in SA0423, harbored with broadly resistance against rice bacterial blight disease (RBBD), the transcriptomes of 'TNG67' and SA0423 were determined by microarray technologies with the RNA samples prepared from the leaves collected at 0, 0.5, 1, 2, and 6 h, respectively, after inoculation of XF89b, atypical isolate used in Taiwan nursery. One-way analysis of variance and Student-Newman-Keuls (SNK) test displayed that 2,727, 3,585, and 18,432 differentially displayed transcripts were found only in SA0423, 'TNG67', and both lines, respectively. Fifty-nine genes that might involve in providing the resistance in SA0423 were determined by bioinformatics analysis such as annotation of metabolic pathway, comparative mapping analysis with published resistance loci, ＂plant-pathogen interaction＂ pathway, and clustering with BioLayout Express3D. By confirming with real-time RT-PCR, 17 resistance gene candidates were selected in this work. Through bioinformatic analysis, results showed that they might be involved in the published ＂plant-pathogen interaction＂ pathway, biosynthetic pathway of plant hormones, autophagy and signal transduction before the plant immune system was induced. The function of these candidates will be further defined to understand the resistance mechanism of SA0423 and to develop eQTL and/or SSR markers to apply in marker-assisted selection for improving disease resistance.

並列關鍵字

Rice ； Bacterial blight disease ； Resistance ； Mutant ； Transcriptomes