摘要
为了解XTH(木葡聚糖内糖基转移酶/水解酶)基因在芸芥自交亲和系突变体(self-compatibility,SC)和自交不亲和系(self-incompatibility,SI)育性调控中的作用,分别以芸芥野生型SI系和突变体SC为材料,取植株的全花cDNA为模板,采用mRNA差异显示技术(differential display reverse transcription PCR,DDRT-PCR)筛选差异片段,获得了芸芥XTH基因的全长cDNA(EsXTH1,1 074bp)。序列分析表明,EsXTH1的5'端和3'端非翻译区分别为36bp和180bp,包含1个858bp的开放阅读框,编码一条285个氨基酸残基组成的肽链;推导的EsXTH1蛋白序列含有XTH蛋白的活性部位DEIDFEFL;芸芥EsXTH1与其它植物的XTH蛋白具有较高的同源性,与拟南芥At-XTH6同源性最高(89.1%),与拟南芥AtXTH7、番茄SlXTH7及康乃馨DcXTH4的同源性分别为73%、73%及69%。分子进化树分析表明,EsXTH1与AtXTH7、SlXTH7、DcXTH4更近。EsXTH1具有木葡聚糖内糖基转移酶(xyloglucan endotran,XET)和水解酶(xyloglucan endohydrolase,XEH)的功能结构位点,N端有一个由23个氨基酸组成的信号肽。预测认为EsXTH1与蛋白质分泌相关,与芸芥自交亲和性的调控相关。
To understand the function of xyloglucan endotransglycosylase/hydrolase （XTH） genes in Eruca sativa selfing affinity regulation in Eruca sativa, SC （self-compatibility） mutant and SI （self-incompatibility） wild type of E. sativa were used to clone DDRT （Differential Display Reverse Transcription） cDNA fragment. An XTH gene cDNA sequence （EsXTH1, 1074bp） was cloned from the cDNA of the whole flowers. The lengths of EsXTH1 5＇ end and 3＇ end untranslated regions were 36bp and 180bp. The length of open reading frame including 858bp coding a 285-amino-acids peptide. Deduced EsXTH1 protein sequence contains XTH protein active site DEIDFEFL. Compared to other species, EsXTH1 had higher homology to Arabidopsis thaliana AtXTH6. The homology to AtXTH7, SlXTH7 （tomato） and DcXTH4 （carnation） were 73%, 73% and 69% respectively. Molecular evolution analysis showed that EsXTH1, as well as AtXTH7, SlXTH7 and DcXTH4 were in the same group. EsXTH1 had XET and XEH active motifs. Its N-end had one signal peptide of 23 amino acids. EsXTH1 was putatively related to protein secretion and E. sativa fertility regulation.
出处
《中国油料作物学报》
CAS
CSCD
北大核心
2013年第2期131-136,共6页
Chinese Journal of Oil Crop Sciences
基金
国家自然科学基金(0960199)
关键词
芸芥
自交亲和系
木葡聚糖内糖基转移酶
水解酶
基因克隆
功能预测
Eruca sativa
Self - compatibility
Xyloglucan endotransglycosylase/hydrolase
Gene cloning
Function prediction
