甘蔗光合系统Ⅰ psaD 基因的克隆和表达分析  

Cloning and expression analysis of sugarcane photosystem Ⅰ psaD gene

作　　者：牛俊奇[1,2] 王爱勤[1,2] 朱惠[1,2] 杨丽涛[1,2,3] 李杨瑞[2,3,4] 

机构地区：[1]广西大学农学院,南宁530005 [2]亚热带农业生物资源保护与利用国家重点实验室,南宁530005 [3]中国农业科学院甘蔗研究中心/广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,南宁530007 [4]广西作物遗传改良生物技术重点开放实验室,南宁530007

出　　处：《中国农业大学学报》2014年第1期43-50,共8页Journal of China Agricultural University

基　　金：863计划课题(2013AA102604);国家科技计划支撑项目(2007BAD30B00);国家国际合作项目(2013DFA31600);广西自然科学基金重点项目(2011GXNSFF018002、2013NXNSFAA019073);广西科学研究与技术开发计划项目(桂科产1123008-1,桂科攻1222009-1B,桂科合1347004-2);广西八桂学者/特聘专家专项经费

摘　　要：采用RACE-PCR方法,以甘蔗心叶为材料克隆了甘蔗PS Ⅰ反应中心亚基Ⅱ基因全长,命名为SopsaD,GenBank登录号JX866950.该基因cDNA序列全长为904 bp,编码200个氨基酸多肽,预测其分子质量和等电点分别为21.85 ku和10.5,其中N端的1～44 aa是叶绿体转运肽序列,62～199 aa是保守的Pfam:PasD结构域.SopsaD与玉米(EU953246)、水稻(AY224449)、大麦(M98254)和短柄草(XM003564060) psaD基因核苷酸同源性分别为85％、85％、84％和81％,氨基酸序列的同源性分别为92％、88％、87％和85％.荧光定量PCR表明甘蔗在叶、花序、芽、茎和根中都能检测到SopsaD基因表达,其中在叶中表达量最高,其次是花序中,而在根中的表达量最低.甘蔗在工艺成熟期和生理成熟期SopsaD基因整体上表现为功能叶中基因表达量高,而老叶中基因表达量低,证实该基因参与光合作用反应.In the present study,rapid amplification of cDNA ends (RACE PCR) was used to clone gene for PS Ⅰ reaction center subunit Ⅱ from the sugarcane leaf roll,which was named as SopsaD gene,with the GenBank accession number of JX866950.The full-length cDNA of SopsaD was 904 bp.It possessed the open reading frame of 603 bp encoding 200 amino acids with predicted molecular weight of 21.85 ku and an isoelectric point of 10.5.At the N-end of SoPsaD protein,1～44 aa was the chloroplast transit peptide sequence and 62～ 199 aa was the conserved Pfam:PasD domain.The similarity comparison revealed that the gene nucleotide sequence shared 85％,85％,84％ and 81％ homology with those of the psaD genes from Zea mays (EU953246),Oryza sativa (AY224449),Hordeum vulgare (M98254) and Brachypodium distachyon (XM003564060),while the similarity at the amino acid sequences with those species was 92％,88％,87％ and 85％ respectively.The results of real-time fluorescent quantitative PCR (qRT-PCR) showed expression of SopsaD could be detected in the leaves,flowers,buds,stalks and roots,among which the highest was found in leaves,followed by flowers,and the lowest in roots.Overall performance of SopsaD expression was high in the function leaves,but low at the old leaves during the periods of technical maturing and physiological maturing.

关 键 词：甘蔗 光合系统Ⅰ PSAD 克隆 基因表达 

分 类 号：S566.1[农业科学—作物学] Q516[生物学—生物化学]