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標題: 水稻結鈣激活酵素基因群之表現分布 及其僅於花粉大量表達成員之功能探討
Expression analysis of CDPK gene family and functional characterization of pollen-predominant CDPKs in rice
作者: 林忠威
Lin, Chung-Wei
關鍵字: pollen;花粉;CDPK gene family;水稻結鈣激活酵素基因群
出版社: 生物科技學研究所
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對於植物的有性生殖而言,花粉管能否於雌蕊上伸長是對植物授粉成功與否的關鍵步驟。當花粉通過自交不親合的篩選後,萌發的花粉管會穿透柱頭並沿著花柱朝胚珠生長。花粉中許多無機離子的濃度,例如鈣離子和鉀離子,對於花粉的萌發甚或是花粉管的生長扮演著相當重要的角色。Calcium-dependent protein kinases(CDPKs)即是植物界當中普遍存在的一種鈣離子接受者,因此可能多方影響花粉之生物功能。植物的CDPK參與了許多鈣離子所調控的訊息傳導途徑,常專一表現於特定的組織或發育時期,並受到特定外來訊息的調控。目前的研究指出水稻中的CDPK共有29個,是一多基因家族。為能有效利用RNAi實驗探討花粉中CDPK的功能,則必須先了解有何CDPK參與花粉萌發生長的調控,以專一性引子進行RT-PCR實驗發現,OsCPK2, OsCPK14, OsCPK21, OsCPK22, OsCPK25/26, OsCPK28及OsCPK29會在成熟花粉中大量,甚或是專一表現,由此大致確認,參與花粉萌發生長的CDPK基因共有八個。為觀察此八個CDPK於花粉萌發時可能之功能及參與的調控機制,構築會大量表現CDPK融合螢光蛋白(ECFP)的質體,利用基因槍進行短暫轉殖,使CDPK於花粉萌發時大量表現,並觀察花粉所受到之影響,藉此推測此八個CDPK於花粉萌發延長中可能扮演的角色,目前結果發現,四個水稻CDPK基因,OsCPK2, OsCPK21, OsCPK26及OsCPK29會抑制花粉管的生長。然而對推測可能分為兩群的CDPK基因而言,大量表現CDPK基因對花粉的影響並無明顯的區別。

Pollen tube elongation in pistil is a crucial step in the sexual reproduction of plants. In a compatible pollination, pollen germinates to extrude a tube that penetrates the stigma and grows extracellularly through the stylar tissues to ovules. It is known that pollen germination and tube growth are significantly regulated by the transport of inorganic ions, such as Ca2+ and K+, across the plasma membranes of pollen and/or pollen tubes. Calcium dependent protein kinase (CDPK) is one kind of the calcium receptors prevalent in plant kingdom, therefore which may play critical roles in multiple aspects of pollen function. The expressions of CDPKs are finely tuned in different tissues and in response to specific signals. Twenty-nine CDPK genes have been identified in the rice genome through a complete search of genome and full-length cDNA databases. To clarify how many CDPK genes participate in control of pollen tube growth, RT-PCR was performed in this study to detect the expression patterns of the 29 CDPK genes. Eight CDPKs (OsCPK2, OsCPK14, OsCPK21, OsCPK22, OsCPK25/26, OsCPK28 and OsCPK29) are expressed predominantly (or specifically) in the mature pollen. Using transient overexpression of CDPK/CFP fusion proteins in the mature pollen of lily, four of them (OsCPK2, OsCPK21, OsCPK26 and OsCPK29) showed inhibition of pollen tube elongation in a concentration-dependent manner. However, no clear functional differences were observed in the putative two clades of 8 CDPKs that expressed in the mature pollen.
其他識別: U0005-2808200711195300
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