山药DoSCF108基因的克隆及表达分析  

Cloning and Expression Analysis of Do SCF108 Gene in Chinese Yam(Dioscorea opposita)

作　　者：李钰婷 周芸伊 肖冬[1] 何龙飞[1] 詹洁[1] 王爱勤[1] Li Yuting;Zhou Yunyi;Xiao Dong;He Longfei;Zhan Jie;Wang Aiqin(College of Agriculture,Guangxi University,Nanning,530005)

机构地区：[1]广西大学农学院,南宁530005

出　　处：《分子植物育种》2023年第4期1084-1092,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(81860670);广西自然科学基金重点项目(2018GXNSFDA281029);广西薯类首席专家创新团队基金项目(nycytxgxcxtd-11-01)共同资助。

摘　　要：为研究F-box蛋白GID2在DELLA蛋白介导赤霉素调节山药块茎膨大发育的分子机理,本研究根据转录组序列,设计特异引物,利用RT-PCR技术克隆到一个山药F-box蛋白GID2基因,在利用生物信息学软件对其序列进行分析的同时通过RT-qPCR技术分析了赤霉素及其抑制剂多效唑(PP333)诱导下该基因在山药块茎膨大初期的表达动态。结果显示,从GH16山药块茎cDNA中克隆到一个长604 bp片段。该片段具有长522 bp的完整开放阅读框(ORF),编码173个氨基酸。预测其分子量为19.02 kD,等电点为9.23,属不稳定亲水蛋白,亚细胞定位在细胞核上,与DELLA蛋白互作系数最大。与已报道的48个物种F-box蛋白GID2氨基酸序列同源性在45.83%~93.64%之间,其中与薯蓣(XM_039286227)同源性最高,达93.64%。推测该基因属于F-box蛋白GID2家族,命名为DoSCF108(GenBank No.MW239683)。RT-qPCR结果表明,在山药块茎膨大初期,DoSCF108基因受外源GA3诱导,相对表达量显著提高,抑制剂PP333处理则相反,不同品种之间存在较大差异。综上所述,山药DoSCF108为F-box蛋白GID2基因,可能参与DELLA介导赤霉素调控山药块茎生长和发育。In order to study the molecular mechanism of the F-box protein GID2 in the DELLA protein-mediated gibberellin regulating the development of yam tubers,in this study,a pair of specific primers was designed by the transcriptome sequence and cloned the F-box protein GID2 of Chinese yam by RT-PCR.Bioinformatics software was used to analyze the gene and RT-qPCR was used to analyze the expression of gibberellin and PP333in the early stage of tuber expansion.The results showed that a 604 bp fragment was cloned from GH16 yam tuber cDNA,contained a complete ORF frame of 522 bp in length,the deduced encoding was 173 amino acids.The predicted molecular weight of the protein was 19.02 kD,theoretical isoelectric point was 9.23,and was unstable hydrophilic protein,subcellular localization in nucleus and maximum protein interaction coefficient with DELLA protein.NCBI-BLAST of amino acid sequences found that the DoSCF108 amino acid sequence homology with the reported F-box protein GID2 of 48 species was 45.83%~93.64%,and the homology with Dioscorea cayenensis(XM_039286227)was the highest,reaching 93.64%.It is speculated that this gene belongs to the F-box family of GID2 protein and is named DoSCF108(GenBank No.MW239683).The results of RT-qPCR showed that an upregulation of DoSCF108 under gibberellin treatment during the early expansion stage of yam tuber,while PP333treatment was opposite,and there were many differences between different varieties.In summary,the DoSCF108gene is the F-box protein GID2 gene,which may be involved in DELLA-mediated gibberellin to regulate the growth and development of yam tubers.

关 键 词：山药(Dioscorea opposite) F-box蛋白GID2 表达分析 

分 类 号：S632.1[农业科学—蔬菜学]