普通小麥中一氧化氮相關因子(TaNOA)編碼基因的克隆和分子生物學分析

郝麗芳，余春梅，李斌，王道文

1 中國科學院遺傳與發育生物學研究所，北京 100101 2 中國科學院研究生院，北京 100049 3 南通大學生命科學學院，南通 226007

普通小麥中一氧化氮相關因子(TaNOA)編碼基因的克隆和分子生物學分析

郝麗芳1,2*，余春梅1,3*，李斌1,2，王道文1

1 中國科學院遺傳與發育生物學研究所，北京 100101 2 中國科學院研究生院，北京 100049 3 南通大學生命科學學院，南通 226007

一氧化氮是動植物體內重要的信號分子。本研究利用同源克隆技術從六倍體普通小麥中獲得一個一氧化氮相關因子(TaNOA)編碼基因的全長基因組和cDNA克隆。該基因具有13個外顯子和12個內含子，與擬南芥以及水稻中同源基因結構相似。根據cDNA推導的氨基酸序列與擬南芥AtNOA1的序列一致性達60%以上，具備P-環GTPase G4-G5-G1-G2-G3的排列特征和保守的序列。對其中2個內含子的測序分析表明在六倍體小麥中TaNOA至少有3個成員。進一步用中國春小麥缺體-四體材料將這3個TaNOA基因成員分別定位在第六同源群的6A、6B和6D染色體上，本研究中獲得的成員定位于6B染色體上，因此將其命名為TaNOA-B1。原生質體表達實驗表明，TaNOA-B1可能定位在線粒體中。TaNOA基因在小麥根、葉片中表達較高，在幼穗和小花中有少量表達，莖中幾乎檢測不到表達。TaNOA的轉錄本水平還因脫落酸或鹽處理而上升，表明它可能參與小麥對非生物脅迫的反應。本研究為進一步克隆六倍體小麥中TaNOA的其他成員及研究該基因在小麥中的功能奠定了基礎。

普通小麥，一氧化氮相關因子(NOA)，TaNOA1-B1，防御反應

Abstract:Nitric oxide(NO)is an important signaling molecule with diverse physiological functions in both animal and plant cells.In this work, we isolated the full-length cDNA and genomic DNA sequences of TaNOA-B1 encoding a putative NO associated(NOA)factor in common wheat.Bioinformatic analysis showed that TaNOA-B1 possessed a similar intron/exon structure as its orthologous genes in Arabidopsis and rice.The amino acid sequence deduced from TaNOA-B1 was more than 60% identitical to those of Arabidopsis and rice NOA1 proteins.The primary structure of TaNOA-B1 contained the zinc finger and P-loop GTPase motifsconserved in Arabidopsis and rice NOA1 proteins.There existed at least three NOA gene members in common wheat, which were mapped to homoeologous group six chromosomes 6A, 6B and 6D, respectively.TaNOA-B1 investigated in this work was located on chromosome 6B.The transcripts of TaNOA members were found mainly in leaves.TaNOA-B1-GFP fusion protein may be located in mitochondria.TaNOA transcript level was up-regulated by abscisic acid(ABA)or NaCl treatments, indicating that TaNOA might be involved in wheat responses to abiotic stresses.

Keywords:common wheat, nitric oxide associated(NOA), TaNOA-B1, abiotic stress response

一氧化氮(Nitrite oxide，NO)作為重要的信號分子參與動物的神經傳導、肌肉伸縮、激素分泌、細胞凋亡、離子通路活性以及免疫反應等重要生理過程[1]。……