枣ZjMADS1-box基因克隆与表达分析  

Cloning and Expression Analysis of ZjMADS1-box Gene Related to Floral Organ Development in Chinese Jujube(Ziziphus jujuba Mill.)

作　　者：段经华[1,2] 张琳[3] 李芳东[2] 杜红岩[2] 袁德义[3] 马履一[1] 

机构地区：[1]北京林业大学,北京100083 [2]中国林业科学研究院经济林研究开发中心,郑州450003 [3]中南林业科技大学林学院,长沙410004

出　　处：《西北植物学报》2012年第12期2374-2382,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家林业局948项目(2012-4-42);国家林业公益性行业科研专项(201204407)

摘　　要：以‘金丝4号’枣花蕾和叶片为材料,通过抑制消减杂交文库(SSH文库)的构建、RACE技术和半定量RT-PCR等方法,克隆了枣花发育基因并研究了其时空表达模式,为枣花发育机理研究奠定基础。结果表明:(1)从枣花SSH文库中鉴定出476bp的花发育B类PISTILLATA(PI)基因的EST序列,命名为ZjMADS1-box;通过RACE技术获得了枣ZjMADS1-box基因的全长cDNA序列。(2)枣ZjMADS1-box基因表现出PI基因的序列结构特征。其全长cDNA长度为957bp,编码220个氨基酸残基;在氨基酸序列水平上,该基因与巨桉EgM2(Eu-calyptus grandis M2)和白千层MqPI(Melaleuca quinquenervia PI)的相似性最高为72.6%,与辣椒CaPI(Capsi-cum annuum PI)的相似性最低为61.8%;系统进化树分析显示,枣ZjMADS1-box与MqPI和EgM2基因聚在一起;枣ZjMADS1-box基因编码蛋白的等电点为9.75,分子量为25 587.5Da,属亲水性的非分泌性蛋白。(3)Zj-MADS1-box基因在花器官中特异表达,在嫩茎、成熟叶、腋芽等营养器官中不表达;在4月30日和5月8日花蕾中的表达量略高于5月4日、5月12日、5月16日的表达量,但差异不显著。With developing floral buds and leaves as materials, we studied ZjMADSI-box cloning and its ex- pression profiles by construction of a suppression subtractlve hybridization （SSH） library,RACE technolo- gy,and semi-quantitative RT-PCR. The results are as follows： （1）A 476 bp EST was identified as PISTIL- LATA （PI） gene related to floral development that was denominated as ZjMADSI-box. Based on the 476 bp sequence information, the full-length cDNA of ZjMADSI-box was successfully obtained by RACE from floral buds of the cultivar ＇Jinsi No. 4＇. （2）The ZjMADSI-box displays structural characters of PI homo- logs. The full-length eDNA of ZjMADS1-box is 957 bp,encoding 220 amino acids. On the amino acid se- quence level,this gene shows the highest similarity （72.6%） with EgM2 gene of Eucalyptus grandis and MqPI gene of Melaleuca quinquenervia ,and the lowest （61.8%） with CaPI gene of Capsicum annuum. ZjMADSI-boa： clusters with EgM2 and MqPI in phylogenetic tree. The predicted isoelectric point （pI）and molecular weight （Mw） of ZjMADSI-box is 9.75 and 25 587. 5 Da, respectively. ZjMADSI-box is predicted to be a kind of hydrophilic and non-secreted proteins. （3）ZjMADSI-box is specifically expressed in floral organs. It is not expressed in vegetative tissues （organs） including tender stems,mature leaves,and axillary buds. In floral buds,expression level of ZjMADSI-box on April 30 and May 8 is higher than that on May 4 ,May 12 ,and May 16. The expression level,however,has no remarkable difference between differ- ent developing stages. This study will provide a scientific basis for fully clarifying the mechanism of floral development of Z. jujuba.

关 键 词：枣 MADS-box基因家族 PI基因 抑制消减杂交文库 快速扩增CDNA末端 

分 类 号：Q786[生物学—分子生物学]