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以黄瓜(Cucumis sativus L.)果实易发生弯曲的品种‘长春密刺’为试验材料,采用RT-PCR技术从果皮中分离并克隆了14-3-3蛋白基因,并将此基因命名为Cs14-3-3。Cs14-3-3基因cDNA全长792bp,编码263个氨基酸,分子量29589.287D,等电点为4.585。生物信息学分析表明此基因含有14-3-3蛋白基因的典型结构域,与其他物种14-3-3蛋白基因核苷酸序列同源性达到75%以上,编码的氨基酸序列同源性达到85%以上,属于14-3-3蛋白基因家族。同时,采用实时荧光定量PCR方法对顺直果实和弯曲果实腹部及脊部在果实开花的2、4、6、8、10、12d的基因表达量进行了研究。结果显示,在果实发育各个时期,Cs14-3-3的表达量均为在弯曲果实腹部>顺直果实>弯曲果实脊部的表达量;在各个部位,Cs14-3-3在果实开花2d的表达量明显高于开花后其他时期。试验结果表明,Cs14-3-3基因为黄瓜果实弯曲相关基因,在黄瓜果实开花早期发育过程中起重要作用。
The cucumber (Cucumis sativus L.) is susceptible to bending of the species ’Changchun secret thorn’ as the test material, using RT-PCR technology from the peel isolated and cloned 14-3-3 protein gene, and the gene was named Cs14 -3-3. The cDNA of Cs14-3-3 gene is 792bp in length and encodes 263 amino acids with a molecular weight of 29589.287D and an isoelectric point of 4.585. Bioinformatics analysis showed that this gene contains the typical domain of 14-3-3 protein gene, which has more than 75% nucleotide sequence homology with 14-3-3 protein of other species. The homology of the encoded amino acid sequence is up to More than 85% belongs to the 14-3-3 protein gene family. At the same time, real-time fluorescent quantitative PCR method was used to study the gene expression amount of 2,4 and 6, 8, 10 and 12 days of flowering in the abdomen and spine of the fruit and the curved fruit. The results showed that the expression of Cs14-3-3 in all stages of fruit development was expressed in the abdomen of Curcuma longa> straight fruit> curved ridges of fruit; at each site, The expression level was significantly higher than other periods after flowering. The results showed that the Cs14-3-3 gene was related to the bending of cucumber fruit and played an important role in the early flowering of cucumber fruit.