鐵皮石斛DoCDPK6基因及啟動子的克隆與表達分析
本文選題:鐵皮石斛 + 鐵皮石斛鈣依賴蛋白激酶基因(DoCDPK)。 參考:《農(nóng)業(yè)生物技術(shù)學(xué)報》2017年04期
【摘要】:鈣依賴蛋白激酶(calcium-dependent protein kinase,CDPK)是一類能響應(yīng)低溫等非生物脅迫的蛋白激酶,目前已從擬南芥(Arabidopsis thaliana)、水稻(Oryza sativa)和楊樹(Populus trichocarpa)等植物中分離出多個CDPK基因。為了研究鐵皮石斛(Dendrobium officinale)鈣依賴蛋白激酶的結(jié)構(gòu)和功能,探尋其在逆境脅迫下的作用,從鐵皮石斛中分離出1個DoCDPK6基因(Gen Bank No.KY682702)及其啟動子,通過生物信息學(xué)分析序列特征、結(jié)構(gòu)及進化關(guān)系,結(jié)果表明,DoCDPK6基因的cDNA全長1 976 bp,開放閱讀框1 725 bp,編碼574個氨基酸,分子式為C2845H4500N766U858S24,分子量為63.9 k D;具有CDPK典型的保守結(jié)構(gòu):可變區(qū)、蛋白激酶區(qū)、連接區(qū)和鈣調(diào)控區(qū);進化分析表明,DoCDPK6與同科植物蝴蝶蘭(Phalaenopsis amabilis)親緣關(guān)系最近,符合植物學(xué)分類地位;在DoCDPK6基因1 500 bp的上游序列含有核心啟動子元件TAAT-box和CAAT-box,并含有光、溫、水楊酸、脫落酸、生長素和赤霉素等逆境信號相關(guān)的響應(yīng)元件;通過q RT-PCR發(fā)現(xiàn),鐵皮石斛DoCDPK6主要在葉片中表達,其次是莖和根;在4℃低溫、脫落酸(abscisic acid,ABA)和Na Cl脅迫處理后,DoCDPK6基因表達量均有不同程度的上調(diào),因此可以推測,DoCDPK6基因在低溫等非生物脅迫中起到調(diào)控作用,該研究結(jié)果可為DoCDPK6基因及其啟動子在非生物脅迫中的調(diào)控機制和信號傳導(dǎo)途經(jīng)提供信息。
[Abstract]:Calcium-dependent protein kinase (CDPKK) is a kind of protein kinase that can respond to abiotic stress such as low temperature. At present, many CDPK genes have been isolated from Arabidopsis thaliana, Oryza sativa and Populus roseus. In order to study the structure and function of calcium-dependent protein kinase of Dendrobium officinaleum, and to explore its role in stress, a DoCDPK6 gene, Genin Bank No. KY682702) and its promoter were isolated from Dendrobium officinale. and the sequence characteristics were analyzed by bioinformatics. The results of structure and evolution showed that the cDNA length of DoddPK6 gene was 1 976 BP, open reading frame 1 725 BP, encoding 574 amino acids, the molecular formula was C2845H4500N766U858S24, and its molecular weight was 63.9 KD, which had the typical conserved structure of CDPK: variable region, protein kinase region. Phylogenetic analysis showed that DoCDPK6 was closely related to Phalaenopsis amabilis (Phalaenopsis amabilis) of the family Phalaenopsis. The upstream sequence of DoCDPK6 gene (1 500bp) contained TAAT-box and CAAT-box, which contained light and temperature. Salicylic acid, abscisic acid, auxin and gibberellin, and so on. By Q RT-PCR, it was found that DoCDPK6 was mainly expressed in leaves, followed by stems and roots. The expression of DoCDPK6 gene was up-regulated by abscisic acidad (Aba) and NaCl stress in different degrees, so it can be inferred that DoCDPK6 gene plays a regulatory role in abiotic stress such as hypothermia. The results may provide information for the regulation mechanism and signal transduction pathway of DoCDPK6 gene and its promoter in abiotic stress.
【作者單位】: 浙江農(nóng)林大學(xué)亞熱帶森林培育國家重點實驗室培育基地;
【基金】:國家自然科學(xué)基金(No.81603228)
【分類號】:S567.239
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