‘三红蜜柚’CmCAD基因的克隆与表达分析
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引用本文:黄汉唐,徐 玉,佘文琴,潘东明,潘腾飞.‘三红蜜柚’CmCAD基因的克隆与表达分析[J].西北植物学报,2019,39(12):2161~2169
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作者单位
黄汉唐1,徐 玉1,佘文琴1,2,潘东明1,2,潘腾飞1,2* (1 福建农林大学 园艺学院福州 350002 2 园艺产品贮藏保鲜研究所福州350002) 
基金项目:福建省科技重大专项(2013NZ0002 1B)
中文摘要:为了探究CmCAD基因在‘三红蜜柚’果实发育过程中的作用与表达规律,该研究以‘三红蜜柚’果实为材料,利用反转录PCR技术克隆获得3个‘三红蜜柚’CmCAD基因,分别命名为CmCAD1、CmCAD3和CmCAD4。对这3个基因所编码的蛋白进行了生物信息学分析,研究了它们在不同品种蜜柚及‘三红蜜柚’果实生长发育阶段的表达模式。结果表明:克隆得到的3个CmCAD基因cDNA长度分别为1 032 bp、1 080 bp和1 071 bp,编码344、360和357个氨基酸;生物信息分析发现3个CmCAD基因编码的蛋白含有相同的保守结构域,均有跨膜螺旋结构与磷酸化位点;系统进化树分析表明,CmCAD1、CmCAD3和CmCAD4分别与甜橙CsCAD1、拟南芥AtCAD9和甜橙CsCAD4的亲缘关系最近。实时定量PCR分析结果显示,3个CmCAD基因在不同品种蜜柚果实生长过程中表达水平存在差异,且3个CmCAD基因在‘三红蜜柚’果实发育过程中的表达规律也有所不同。研究推测,CmCAD4具有一定的潜在功能,可能参与调控‘三红蜜柚’果实中木质素的合成,但基因的具体功能与调控机理还需进一步探索。
中文关键词:‘三红蜜柚’  肉桂醇脱氢酶  基因克隆  表达分析
 
Cloning and Expression Analysis of CmCAD Genes in ‘Sanhongmiyou’ Pummelo
Abstract:To reveal the function and expression patterns of CmCAD genes in ‘Sanhongmiyou’ pummelo fruit during its fruit development stages, we cloned three CmCAD genes named CmCAD1, CmCAD3 and CmCAD4 by reverse transcription PCR. The proteins encoded by these genes were analyzed by series of bioinformatic methods, the expression patterns of CmCAD genes in ‘Sanhongmiyou’ pummelo and its bud mutation varieties at different development stages were analyzed either. The results showed that the cDNA length of the three CmCAD genes were 1 032 bp, 1 080 bp and 1 071 bp respectively, encoding 344, 360 and 357 amino acids. Bioinformatic analysis showed that proteins encoded by threse CmCAD genes contained same conserved domains. All of them had transmembrane spiral structure and phosphorylation sites. Phylogenetic analysis revealed that CmCAD1, CmCAD3 and CmCAD4 were closely related to Citrus sinensis CsCAD1, Arabidopsis thaliana AtCAD9 and Citrus sinensis CsCAD4, respectively. The results of real time quantitative PCR showed that the expression levels of CmCAD gene members in different pummelo varieties had significant differences at fruit development stages. Meanwhile, during ‘Sanhongmiyou’ fruit development stages, the expression patterns of CmCAD gene members were different either. We speculated that CmCAD4 gene has some potential functions and might involved the regulation of lignin synthesis in ‘Sanhongmiyou’ fruit, but the specific function and regulatory mechanism of the gene need to be further explored.
keywords:Citrus grandis ‘Sanhongmiyou’  cinnamyl alcohol dehydrogenase  gene cloning  expression analysis
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