甜瓜自毒作用相关基因CmSCL13的克隆及表达分析

Cloning and Analysis of Autotoxicity Related Gene CmSCL13 in Melon

以甜瓜品种"新银辉"为试验材料,利用cDNA-AFLP技术分离并获取了甜瓜自毒作用相关scarecrow-like protein 13-like(SCL13)片段序列,利用RACE技术获得该基因全长cDNA序列,包含有1 623 bp的完整阅读框,编码541个氨基酸。在NCBI上通过Conserved Domain Search对该基因编码的氨基酸进行搜索,表明该基因具有GRAS家族特有的保守结构域,命名为:CmSCL13(GenBank登录号:KF134812)。Blastn分析显示该基因与黄瓜CsSCL13基因同源性较高;荧光定量PCR分析结果显示该基因随胁迫处理时间延长表达量呈上调趋势,该结果也与cDNA-AFLP的结果相一致。

The differential fragment of scarecrow-like protein 13-like （SCL13） was separated by the cD-NA-AFLP technique from the leaves of melon , which was associated with autotoxicity .‘New Silverlit ’ was cloned by RACE technology .The full-length cDNA sequence of CmSCL13 had an ORF of 1 623 bp and enco-ded 541 amino acids .Its amino acid was searched by the NCBI Conserved Domain Search ,which indicated that the gene had conserved domain of GRAS family .Therefore , the gene of melon could be named as CmSCL13 （ GenBank accession number：KF134812 ） .The result showed that this fragment was highly associated with SCL13 genes of cucumber .Fluorescence quantitative PCR method was used to study the expression characteris-tics of GRAS gene in melon under root exudates stress ,and the result showed that the gene expression changed after root exudates treatment ,which was consistent with the results of cDNA-AFLP .