水稻中受冷誘導的DEAD-box RNA helicase 42 參與 mRNA 的修飾作用;A cold-induced DEAD-box RNA helicase 42 participates in mRNA splicing in rice

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Title:	水稻中受冷誘導的DEAD-box RNA helicase 42 參與 mRNA 的修飾作用;A cold-induced DEAD-box RNA helicase 42 participates in mRNA splicing in rice
Authors:	黃文杉;Huang,Wen-Shan
Contributors:	生命科學系
Keywords:	冷逆境;水稻;mRNA修飾;DEAD-box RNA helicase;cold stress;rice (Oryza sativa);mRNA splicing
Date:	2016-07-26
Issue Date:	2016-10-13 13:03:03 (UTC+8)
Publisher:	國立中央大學
Abstract:	低溫和寒害產生的冷逆境影響全球農作物的產量，所以很多植物演化出抵抗冷逆境之分子機制包括維持RNA 分子的正常功能。RNA 解旋酶是一種以ATP 水解作為能量將 RNA 雙股解旋的酵素，它參與在與RNA 有關之機制，包括轉錄、核醣體生合成、 pre-mRNA 修飾、RNA 在細胞核與細胞質間運輸、轉譯、mRNA 降解和胞器中基因的表現。水稻為世界上重要糧食作物，至少含有62 種不同的基因編碼DEAD-box 蛋白，其中已知功能的水稻DEAD-box 蛋白質很少，本論文主要目的在探討水稻中受冷逆境誘導的DEAD-box 蛋白質OsRH42 基因為之生理功能，利用RNAi 轉殖株策略，發現三個 OsRH42 弱化表現之獨立轉殖株呈現對冷逆境極為敏感的性狀。OsRH42 位在細胞核中的splicing speckle。進一步的研究結果顯示OsRH42 功能為維持Pre-mRNA 的正確 splicing，一些對冷反應的基因， (OsSK12 (shaggy-related protein kinase) 、OsCIR1 (Circadian 1)、OsPRR95 (PSEUDO-RESPONSE REGULATOR)、OsPUB45 (PLANT U-BOX )、 OsEBF1 (EIN3-binding F-box protein 1 ) 、OsSPFH (SPFH/ PHB domain-containing membrane-associated protein))，在冷逆境下OsRH42-Ri 轉殖株會發生錯誤splicing 的現象。此外，在OsRH42 基因靜默轉殖株中CBF 基因表現增加。這些結果說明OsRH42 為水稻中表現受冷反應基因的正常splicing 不可或缺的RNA helicase，並且有助於水稻對抗冷逆境。;Chilling and freezing temperatures negatively affect global crop production. Plants have evolved several physiological and molecular adaptations to minimize damage from cold stress programs. RNA helicases are enzymes that are able to unwind RNA duplex in an ATP-dependent manner and play important roles in various aspects of RNA metabolism, including transcription, pre-mRNA splicing, ribosome biogenesis, mRNA transport, translation, and RNA decay. Rice, one of the most important crops in the world, contains at least 62 different genes encode DEAD-box proteins. Herein, we identified a rice DEAD-box RNA helicase gene, OsRH42, which expression was induced by cold stress. The OsRH42 knockdown transgenic plants were generated by RNAi approach and three independent lines of seedlings exhibited cold hypersensitive phenotype, regardless of whether or not they were cold acclimated. Subcellular localization indicated that the OsRH42 localized into splicing speckle. Cold-responsive genes, OsSK12 (shaggy-related protein kinase), OsCIR1 (Circadian 1), OsPRR95 (PSEUDO-RESPONSE REGULATOR), OsPUB45 (PLANT U-BOX ), OsEBF1 (EIN3-binding F-box protein 1 ), OsSPFH (SPFH/PHB domain-containing membrane-associated protein) were misspliced in OsRH42 knockdown transgenic plants under cold stress. Moreover, expression of cold response key regulators, CBF genes, were increased in these OsRH42 knowndown lines, as compared to wildtype. There results demonstrate that the cold-induced RNA helicase, OsRH42, is essential for splicing of cold-responsive gene thereby contribute to cold stress tolerance in rice.
Appears in Collections:	[生命科學研究所 ] 博碩士論文

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