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以矮牵牛(Petunia hybrida)花序cDNA为模板,利用EST数据库信息,克隆了一个MADS-box基因编码区序列,GenBank登录为PMADS20(GU129907)。进化树分析表明,PMADS20属StMADS11亚家族SVP分支,与番茄和辣椒的JOINTLESS基因氨基酸序列相似性最高。定量PCR检测结果表明,PMADS20在矮牵牛营养器官中的表达量高,在花中的表达量低。构建了PMADS20的超量表达载体(35S︰PMADS20)和CRES-T(Chimeric Repressor Gene Silencing Technology)载体(35S︰PMADS20-SRDX)。35S︰PMADS20和35S︰PMADS20-SRDX转基因矮牵牛的花器官表型变化相似,花萼增大,花瓣和雌蕊出现营养器官特征,但35S︰PMADS20-SRDX的变化更为明显。扫描电镜观察显示,35S︰PMADS20-SRDX转基因花瓣下表皮的表皮毛增多,并有气孔分布;子房和花柱表面为表皮毛覆盖,呈现超量表达UNHAVEN的矮牵牛心皮的特征,并且有气孔分布。35S︰PMADS20转基因植株株形无明显变化,35S︰PMADS20-SRDX植株花序的节间较野生型短。
Using Petunia hybrida cDNA as a template, a MADS-box gene coding region sequence was cloned using EST database. GenBank accession was PMADS20 (GU129907). Phylogenetic tree analysis showed that PMADS20 belongs to the StMADS11 subfamily SVP branch, and has the highest similarity with the JOINTLESS gene of tomato and pepper. Quantitative PCR results showed that PMADS20 expressed high in petunia vegetative organs and low in flowers. PMADS20 overexpression vector (35S: PMADS20) and CRES-T (Chimeric Repressor Gene Silencing Technology) vector (35S: PMADS20-SRDX) were constructed. 35S: PMADS20 and 35S: PMADS20-SRDX transgenic petunia flower organ phenotypic changes similar to the calyx, petals and pistillate vegetative organs, but 35S: PMADS20-SRDX changes more pronounced. Scanning electron microscopy showed that epidermal hair of the lower epidermis of 35S: PMADS20-SRDX transgene increased and had stomatal distribution; the surface of ovary and style was covered with epidermal hairs, showing the characteristics of petunia carpel of over-expressing UNHAVEN, and there were Stomatal distribution. 35S: PMADS20 transgenic plants without significant changes in plant shape, 35S: PMADS20-SRDX plant inflorescence internode shorter than the wild-type.