利用抑制差减杂交技术分离茄子单性结实相关ESTs  

Isolation of Parthenocarpy Related ESTs in Parthenocarpic Eggplant Using Suppression Subtraction Hybridization (SSH)

作　　者：周亚君[1] 陈钰辉[1] 刘富中[1] 张映[1,2] 连勇[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]中国农业大学农学与生物技术学院,北京100193

出　　处：《园艺学报》2010年第12期1944-1952,共9页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30771475);国家‘863’计划项目(2006AA10Z1A6);国家科技支撑计划项目(2006BAD13B06-3);中央级公益性科研院所基本科研业务费专项项目(2060302-2-10);农业部园艺作物遗传改良重点开放实验室项目

摘　　要：以茄子(Solanum melongena)单性结实品系D-10和非单性结实品系03-2的子房cDNA分别为试验组(Tester)和驱动组(Driver),利用抑制差减杂交(SSH)技术构建了正反两个SSH-cDNA文库,分别包含472和124个克隆。通过测序、序列拼接后,共获得384个Unique ESTs。将其与非冗余蛋白数据库进行BLASTx比对,在E值小于等于1e-10条件下,有257个ESTs能找到相匹配的蛋白质,其中121个ESTs与非冗余蛋白质数据库中已知功能的蛋白质具有高度的相似性。功能注释结果表明,具生物过程、分子功能和细胞组分的EST分别为107、102和119条。推测与单性结实相关的EST有5条:2条ESTs属于MADS-box转录因子家族,1条与生长素增长蛋白同源,1条属于P450还原酶系,1条属于MAPKK家族;另外136个ESTs代表了未知功能基因。A forward and a reverse suppression subtraction hybridization（SSH）cDNA library with 472 and 124 positive clones were constructed respectively with cDNA from eggplant（Solanum melongena）parthenocarpic line D-10 ovary as the tester and cDNA from un-parthenocarpic line 03-2 ovary as the driver. After removing repeat and redundancy sequences,384 expressed sequence tags （ESTs）were obtained. Protein homology search in non-redundant protein database revealed that,under E-value less than or equal to 1e-10,121 ESTs were highly homologous with known proteins,and 136 ESTs could be new genes. The results of gene ontology showed that 107,102,119 ESTs were involved in biological process,molecular function and cell component respectively. Five parthenocarpic related ESTs were obtained,including two MADS-box family,one auxin growth promoter protein,one cytochrome P450 NADPH-reductase and one MAPKK family.

关 键 词：茄子 单性结实 抑制差减杂交 

分 类 号：S641.1[农业科学—蔬菜学]