苹果成花抑制蛋白SVP基因的克隆、表达及启动子活性分析  

Cloning,Expression Pattern and Promoter Activity Analysis of Flowering Regulatory Gene SVP in Apple(Malus×domestica)

作　　者：王世祥 左希亚 邢利博[1] 樊胜[1] 张东[1] 韩明玉[1] 张林森[1] WANG Shixiang;ZUO Xiya;XING Libo;FAN Sheng;ZHANG Dong;HAN Mingyu;ZHANG Linsen(College of Horticulture,Northwest Agriculture and Forestry University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学园艺学院

出　　处：《园艺学报》2019年第8期1445-1457,共13页Acta Horticulturae Sinica

基　　金：国家现代农业产业技术体系建设专项资金项目(CARS-27);陕西省科技统筹创新工程项目(2015KJZDNY02-03-02)

摘　　要：以‘长富2号’苹果短枝顶芽为材料,采用同源重组法克隆得到成花抑制蛋白SHORT VEGETATIVE PHASE(SVP)家族的1个关键基因MdMADS50,开放阅读框(ORF)长度为675 bp,编码224个氨基酸。序列分析发现,该基因含有1个保守的MADS-box结构域和1个K-box结构域,属于MIKC型。氨基酸多重序列比对和系统进化分析表明,MdMADS50蛋白序列与苹果MdSVP具有较高的同源性。qRT-PCR分析结果表明,MdMADS50具有组织表达特异性,在苹果芽和叶中表达量较高。外源GA3处理促进了MdMADS50的表达,且在难成花品种‘长富2号’苹果芽中的表达量显著高于其他苹果品种。另外,GUS活性检测结果表明MdMADS50基因启动子具有启动活性,且受外源GA3诱导后活性增强。综上,研究表明MdMADS50可能响应GA3处理对苹果成花诱导发挥抑制作用,并介导赤霉素信号参与苹果花芽孕育的调控。The key gene MdMADS50 of SHORT VEGETATIVE PHASE(SVP)was cloned by homologous recombination method using short shoot buds of‘Nagafu 2’apple.The open reading frame(ORF)of MdMADS50 was 675 bp,that encoded 224 amino acids.Protein sequence analysis revealed that this gene contains conserved MADS-box and K-box domains belonging to MIKC-type MADS-box genes.Phylogenetic analysis indicated that the MdMADS50 protein has high homology with MdSVP.By quantitative real-time PCR(qRT-PCR)tissue-specificity expression of MdMADS50 was identified,that displayed that MdMADS50 has higher expression in shoots and leaves.In addition,exogenous GA3 treatment promoted the expression of MdMADS50,and the expression level of cultivar‘Nagafu 2’in the initial stage of flower induction was significantly higher compared to other cultivars.Furthermore,the GUS activity assay showed that the MdMADS50 promoter had promoter activity and was enhanced by exogenous GA3.In conclusion,our study showed that MdMADS50 may significantly inhibit the flower induction of apple in response to GA3 treatment,mediating the regulation of gibberellin signaling in apple flower buds.

关 键 词：苹果 成花诱导 SVP GA 基因克隆 基因表达 GUS 

分 类 号：S661.1[农业科学—果树学]