ZHOU Xiaofeng,MO Zhenghai,SHANG Yangjuan,et al.Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis[J].Journal of Nanjing Forestry University(Natural Science Edition),2020,44(4):055-62.[doi:10.3969/j.issn.1000-2006.201905023]





Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis
周晓峰 莫正海 尚杨娟 骆敏 苏文川 谭鹏鹏 韩明慧 邓秋菊 田朝霞 彭方仁
作者单位:南京林业大学,南方现代林业协同创新中心,南京林业大学林学院, 江苏 南京 210037; 中国科学技术大学 生命科学学院 安徽 合肥 230027
ZHOU Xiaofeng1 MO Zhenghai1 SHANG Yangjuan1 LUO Min1 SU Wenchuan1 TAN Pengpeng1 HAN Minghui1 DENG Qiuju1 TIAN Zhaoxia2 PENG Fangren1
(1.Co-Innovation Center for the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China; 2.School of Life Sciences, University of Science and Technology of China, Hefei 230027,China)
薄壳山核桃 油脂合成 CiDGAT1 基因克隆 表达模式
lipids synthesis gene cloning gene expression patterns
目的 研究薄壳山核桃(Carya illinoinensisCiDGAT1基因的结构特征和表达模式,为深入分析CiDGAT1基因功能提供理论参考。 方法 以薄壳山核桃‘波尼’果实为材料,利用DGAT1基因保守片段设计特异引物,通过3’ RACE 和RT-PCR技术,克隆薄壳山核桃的CiDGAT1基因。运用生物信息学方法分析其蛋白性质、亲缘进化关系及基因结构。通过实时荧光定量PCR分析该基因在不同品种以及果实不同发育时期的表达模式。 结果 克隆到1条薄壳山核桃油脂合成相关基因CiDGAT1,该基因开放阅读框(ORF)1 617 bp,编码538个氨基酸,5’UTR区包含294个碱基,3’UTR区包含340个碱基。基因结构分析显示,CiDGAT1由15个外显子和 14个内含子组成。系统进化分析表明CiDGAT1蛋白聚在双子叶植物分支中,与栎树(Quercus suber)QsDGAT1和榛子(Corylus americana)CaDGAT1的亲缘关系最近。实时荧光定量PCR结果表明,在早熟品种‘波尼’和晚熟品种‘马罕’与‘金华’3个品种果实发育过程中,CiDGAT1在‘波尼’发育时表达量较高,在145 d成熟期时达到最高,而‘马罕’、‘金华’品种在生长过程中,CiDGAT1表达量一直缓慢增长,152 d后迅速升高,至最后成熟时达到最高值。 结论 CiDGAT1属于MBOAT家族,在薄壳山核桃果实发育过程中表达量呈上升趋势,是油脂合成过程中重要的调控因子。
Objective The purpose of the study was to elucidate the structure and expression patterns of the CiDGAT1 to provide a theoretical reference for a further analysis of the function of CiDGAT1 gene. Method A full-length CiDGAT1 gene was isolated from the fruit of Carya illinoinensis using rapid amplification of cDNA ends with specific primers according to the conserved DGAT1 fragment sequence. The genetic structure , protein character, and evolutionary relationship were analyzed using bioinformatics. In addition, the CiDGAT1 temporal expression pattern was analyzed in different varieties using quantitative real-time PCR. Result CiDGAT1 has an open reading frame with 1 617 bp and encodes 538 amino acids. The gene structure analysis showed that the CiDGAT1 gene consists of 15 exons and 14 introns. The phylogenetic analysis revealed that the CiDGAT1 protein is clustered in the dicotyledonous branch with a close relationship to oak DGAT1 and filbert DGAT1. The qRT-PCR analysis showed that the expression of CiDGAT1 is higher in the growth process of ‘Pawnee’ and peaks in the mature stage at 145 d. Whereas, CiDGAT1 expression is lower during the growth process of ‘Mahan’ and ‘Jinhua’ varieties; it rapidly increased after 152 d, reaching the highest level at final maturity. Conclusion CiDGAT1 belongs to the MBOAT gene family, and its expression tends to increase during the growth process of the fruit of C. illinoinensis, which is a key regulator in the process of oil synthesis in plants.


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更新日期/Last Update: 2020-08-13