Work had been done with the purpose to illustrate the regulation mechanism and function of auxin receptors and their signaling pathway in cucumber(Cucumis sativus L.),and also with the wish to find new approaches for enhancing production.Compared with the sequences of auxin receptor gene TIR1/AFBs in Arabidopsis,two highly conserved sequences were selected by screening the cucumber genome database,which were named Cs TIR and CsAFB.cDNAs of both CsTIR and CsAFB genes were cloned and verified by sequencing.Overexpression vectors were constructed for further reverse genetic research in cucumber.Their sequences were submitted to GenBank and recruited with the accession of KC414935.1 and KC414934.1,respectively.The basic physical and chemical properties,as well as functional protein domains,of CsTIR and CsAFB were predicted by bioinformatics analysis.Phylogenetic and conservation analysis were taken between sequences from different plants,which made these two genes putative auxin receptors in cucumber.The expression patterns of the two genes were analyzed by semi-q RT-PCR.CsAFB and CsTIR both accumulated more in roots and leaves than in stems in 10-day-old plants while CsTIR was undetectable in stems.Expressions of both genes were detected in every tissue from 70-day-old flowering plants and showed no tissue specificity.This work provides new evidence of auxin receptors in cucumber,and will contribute to the further study of their functions and regulation mechanisms.%为了解生长素受体及其信号通路在黄瓜(Cucumis sativus L.)中的调控机制和生物学功能,寻找提高产量的新途径并指导农业生产,我们以拟南芥(Aabidopsis)生长素受体家族AtTIR1/AFBs基因序列为参照进行比对,从黄瓜全基因组数据库中筛选出2条具有很高同源性的基因序列,分别命名为Cs TIR和CsAFB.克隆和测定了这2个基因的cDNA序列,构建了过表达载体,拟在黄瓜中开展反向遗传学研究.基因序列提交至GenBank并得到收录,其中CsTIR的登录号KC414935.1,CsAFB登录号KC414934.1.采用生物信息学方法对CsTIR和CsAFB蛋白的基本理化性质、功能结构域进行初步预测,对其在物种间的保守性进行进化分析,推测它们可能是黄瓜的生长素受体.采用semi-q RT-PCR分析了这两个基因的时空表达模式,发现10d黄瓜幼苗中CsAFB基因的表达量在根和叶中比在茎中高,CsTIR基因在茎中几乎检测不到表达;70d成年黄瓜各组织都有CsTIR和CsAFB表达,无明显组织特异性.本研究为寻找黄瓜生长素受体提供了实验依据,为进一步研究生长素信号通路在黄瓜中的功能及调控机理提供了基础资料.
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