小麦糖转运蛋白基因TaSWEET6的克隆与表达分析  

Cloning and Expression Analysis of a Sugar Transporter Protein Gene TaSWEET6 in Wheat

作　　者：徐磊[1] 王伟伟[2] 苏世超[2] 马锦绣[2] 孙辉[2] 房兆峰 高世庆[2] 唐益苗[2] 赵昌平[2] 蒋涛[2] 

机构地区：[1]首都师范大学生命科学学院,北京100048 [2]北京市农林科学院北京杂交小麦工程技术研究中心,北京100097

出　　处：《麦类作物学报》2016年第11期1411-1418,共8页Journal of Triticeae Crops

基　　金：国家转基因重大专项(2016ZX08002-003);北京市农林科学院科技创新能力建设专项(KJCX20140202);国家自然科学基金项目(31371699);北京自然科学基金项目(5131001);北京市科技计划项目(Z141100002314018)

摘　　要：植物SWEET基因家族是一类糖转运蛋白,参与生殖发育、衰老、逆境响应等多个生理过程。为研究小麦SWEET基因对逆境胁迫的响应及其在花药发育过程中的功能,采用同源克隆的方法从普通六倍体小麦中克隆出TaSWEET6基因(GenBank登录号为KU936097)后对其进行染色体定位、同源蛋白序列比对和系统进化树分析,同时分析TaSWEET6基因于普通六倍体小麦在正常生长情况下及非生物胁迫条件下不同组织器官中和光温敏雄性不育小麦BS366在不同发育时期的花药中的表达模式。序列分析结果表明,TaSWEET6基因包含1个732bp的完整开放阅读框,编码243个氨基酸。跨膜结构分析得知,TaSWEET6蛋白包含2个Mtn3_slv跨膜结构域和1个起连接作用的跨膜-螺旋。系统进化树分析表明,TaSWEET6蛋白与大麦处于同一分支,其亲缘关系最近。中国春缺四体定位表明,TaSWEET6基因位于7D染色体上。表达分析表明,TaSWEET6基因在小麦根、茎、叶、种子、小花(除去雄蕊)、各时期雄蕊中均有表达,小孢子时期雄蕊表达量最高;在低温、干旱、NaCl和ABA胁迫处理下,TaSWEET6基因表达均有上调;光温敏雄性不育系BS366在不育环境(低温短日照)下,TaSWEET6基因在雄蕊发育关键时期(二分体时期)高度表达。说明小麦TaSWET6基因可能参与了多种逆境应答反应,并在小麦雄蕊发育过程中发挥着重要作用。SWEET family is a sugar transporter protein family in plants,which are involved in multiple physiological processes, including reproductive development, senescence, and abiotic stress responses. Our objectives are to analyse the expression of TaSWEET6 under various abiotie stresses and how Ta- SWEET6 influences the development of anther. To understand the function of TaSWEET6,qRT-PCR assays were used to quantify the expression of TaSWEET6 within different wheat organs. The expres- sion of TaSWEET6 in anther of a photoperiod-thermos-sensitive genic male sterility line BS366 at dif- ferent developmental stages under various abiotic stresses was quantified by qRT-PCR. We cloned a full-length cDNA sequence of TaSWEET6 （GenBank Accession No. KU936097） from wheat by RT-PCR,which has 806 bp in length containing a 732 bp ORF （open reading frame）,encoding 243 amino acid residues. The phylogenetic analysis of TaSWEET6 indicated high similarity with the SWEET protein in barley. The chromosome localization of TaSWEET6 was confirmed by nullisomic-tetrasomic （NT） analysis, which was detected to be located on chromosome 7D in wheat. TaSWEET6 protein contains two Mnt3_slv transmembrane domains that are connected by an inversion linker helix using TMHMM tool. Expression of TaSWEET6 have been observed in different tissues of wheat organs such as root, stem, leaf, spikelet （except anther）, and anther at different stages, while TaSWEET6 preferentially expressed in the young anther at microspore stage. The qRT-PCR assays detected that TaSWEET6 was up-regulated by low temperature at 4 ℃, drought, NaCl and ABA treatments. The high expression of TaSWEET6 was detected in anther of BS366 at dyad stage under sterile condition （low temperature and short day）. These results provide basic information for the functional analysis of TaSWEET6 in response to various stresses,and also suggest that TaSWEET6 play a critical role in the process of wheat anther development.

关 键 词：小麦 糖转运蛋白 非生物胁迫 雄性不育 

分 类 号：S512.1[农业科学—作物学] S330