水稻中五個DEAD-box RNA helicase - RH2、RH6、RH22、RH42和RH51基因之探討;Characterization of five DEAD-box RNA helicase genes, RH2, RH6, RH22, RH42 and RH51 in Oryza sativa

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题名:	水稻中五個DEAD-box RNA helicase - RH2、RH6、RH22、RH42和RH51基因之探討;Characterization of five DEAD-box RNA helicase genes, RH2, RH6, RH22, RH42 and RH51 in Oryza sativa
作者:	陳玉夫;Chen,Yu-fu
贡献者:	生命科學系
关键词:	RNA解旋?;RNA helicase
日期:	2013-01-21
上传时间:	2013-03-25 15:52:58 (UTC+8)
出版者:	國立中央大學
摘要:	陸生植物因無法移動，當遭受到生物逆境與非生物逆境時，必須透過改變分子、細胞和生理性機制來適應這些不同的逆境。在非生物逆境中，溫度是影響植物發育過程中重要因子之一。低溫會抑制種子發芽、莖部延長和花粉活力，進而使農作物的產量降低。Helicase是一種需要消耗ATP將雙股RNA與DNA解開的motor蛋白質。RNA helicase參與大部分的細胞生理反應的過程，包括RNA合成、修復、轉錄、重組、剪切和降解的表現。DEAD-box RNA helicase屬於 helicase superfamily 2，在motif II保留的區域中有一段胺基酸序列Asp-Glu-Ala-Asp，D-E-A-D，因而命名為DEAD。水稻是世界上重要的糧食作物之一，並已完成序列解序，可做為單子葉植物的模式植物。經由資料料庫比對，在水稻中找到5個和阿拉伯芥中會受低溫誘導的AtRH2、AtRH6、AtRH22、AtRH42和AtRH51相似的基因，分別命名為OsRH2、OsRH6、OsRH22、OsRH42和OsRH51。利用水稻懸浮系統進行初步分析這些基因在逆境下的表現量。結果顯示OsRH42和OsRH51會受溫度與高鹽影響，其餘的3個基因則未受逆境影響。為了更進一步瞭解這6個基因在水稻中的功能與扮演的角色，我分別建構這五個DEAD-box RNA helicase的RNAi knock-down轉殖株，並獲得T0轉殖株。在T0代轉植株生長的過程中發現，OsRH2 RNAi轉殖株株高較野生型水稻矮小，且節點間距也較短，種子的結實率也比野生型水稻低。用顯微鏡觀察可能影響結實率的花粉，發現OsRH2 RNAi轉殖株的花粉不健全率比野生型水稻高。綜合以上的結果，推測將OsRH2 RNAi knock-down後會造成植株成長減緩及花粉發育不健全。Land plants are immobile and therefore encounter for various biotic stresses and abiotic stresses. Plants have to change in molecular, cellular and physical mechanisms to adapt those stresses. Temperature is the one of important factor for all of developmental stage of plant, form germination to reproduction. Low temperature inhibits seed germination, shoot elongation and overall metabolism and therefore cause to decrease agricultural productivity reduction. RNA helicase is motor protein witch can unwind of duplex RNA/RNA in an ATP-dependent manner. RNA helicases are involved in most of the basic cellular progresses including nuclear transcription, ribosome biogenesis, pre-mRNA splicing, nucleocytoplasmic transport, translation, mRNA decay and organall gene expression. DEAD-box RNA helicases belong to the helicase superfamily 2. The name is derived from the amino-acid sequence D-E-A-D (Asp-Glu-Ala-Asp) in the conserved motif II of RNA helicase family. Rice is one of the most important crops and also is a model plant for monocots. Base on amino acid sequence analysis, found the five homologues DEAD-box RNA helicase genes, OsRH2, 6, 22, 42 and 51, in rice that are similar to cold-stress response genes in Arabidopsis. The expression of five genes, showed that OsRH42 and OsRH51 are induced of temperature and salt stress in WT suspension cell. But the OsRH2, 6, 22 are consistive expression in various stresses treated cells. We further generated the RNAi knock-down transgenic lines for those DEAD-box RNA helicase genes. OsRH2 RNAi transgenic plant present plant height and internode were shorter than WT, and seeds fertility are also less than WT. Microscope analysis showed that pollen aborted rate of OsRH2 RNAi transgenic plants were higher than WT. Our results suggest that OsRH2 may have function in plant growth and seed development.
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