瘤野螟抗性水稻品種清流之蛋白質體分析

Abstract

水稻為亞洲主要的糧食作物，瘤野螟為危害台灣水稻的主要害蟲之一。在先前的研究顯示，水稻品種清流對瘤野螟具有抗蟲特性。在本實驗中， 以蛋白質體學的層面比較抗蟲品種清流與感蟲品種台中在來一號在瘤野螟幼蟲咬食葉片處理 0、 6、 24、72 小時後葉片可溶性蛋白組成與含量。透過非標定定量技術 (sequential windowacquisition of all theoretical fragment ion spectra； SWATH)分析，進行水稻葉片蛋白質鑑定及定量分析。經質譜分析以及統計方法計算並比對資料庫以及去除未知蛋白後可鑑定到 505 個蛋白質。 經 agriGO 分析， 比較不同時間點清流與 TN1 葉片蛋白質表現量與趨勢， 發現清流在抗蟲相關的黃酮類生合成途徑與茉莉酸(JA)相關途徑皆有特殊的表現狀況。 在未經咬食與面對瘤野螟幼蟲咬食的清流與 TN1 相比phenylalanine ammonia lyase 皆會大量表現，並且快速的表現黃酮類生合成途徑下游的 chalcone synthase、速率決定步驟酵素 chalcone--flavonone isomerase，推斷清流能快速地啟動黃酮類合成反應。 茉莉酸訊號傳遞路徑的關鍵酵素 lipoxygenase 與12-oxo-phytodienoic acid reductase 在清流表現量與趨勢也都顯示清流具有比 TN1更快速表現茉莉酸相關防禦反應的能力。這兩點都是能提升植物抗蟲能力的重要反應。其他生理表現如光合作用、氮的分配也都會和植物的耐受性有關，結果顯示光合作用相當複雜，但是仍有特殊的趨勢表現可以在未來深入探討，氮分配的部分則發現清流大量表現在二次代謝物生合成的反應，在解毒相關的半胱胺酸合成酶也有特殊的表現情，抗氧化逆境相關蛋白 Glutathione (GSH)的表現趨勢也顯示清流可能有更高效率的抗氧化能力。透過本研究可以更全面了解清流對瘤野螟的抗性，並可在未來將其特性應用於害物整合管理上。

Rice (Oryza sativa L.) is a major crop in Asia. Rice leaffolder (Cnaphalocrocis medinalis Guenée) is one of serious rice pests caused severe damage in Taiwan. Our previous study showed that rice variety QingLiu has C. medinalis–resistant ability. In this study, we compared protein expression pattern on C. medinalis–resistant rice variety, QingLiu, with susceptible check, Taichung Native 1 (TN1) under C. medinalis infestation. We collected damaged leaves and further extracted proteins after four time points (0, 6, 24, 72 hour infestation). The soluble proteins were further analyzed by sequential window acquisition of all theoretical fragment ion spectra (SWATH) to identify and quantify proteins in treatments. The identified proteins were further analyzed by agriGO to understand the protein gene ontology. After filtering out unknown proteins, there was 505 proteins identified. After compared protein expression pattern, we found proteins in flavonoid biosynthesis process (phenylalanine ammonia-lyase (PAL), chalcone synthase and rate limited step chalcone--flavonone isomerase) and jasmonic acid biosynthesis pathway (lipoxygenase and 12-oxo-phytodienoic acid reductase) were highly expressed in QingLiu. Based on our finding, QingLiu would have better defense response ability against C. medinalis. In addition, we found that proteins involved in hotosynthesis and nitrogen resource distribution would have unique expression pattern after C. medinalis infestation. Furthermore, detoxification-related cysteine synthase and ROS-related protein GSH had higher expression pattern in QingLiu. Through our study, we could have better understanding on the resistant mechanism of QingLiu against C. medinalis.