马尾松PmCBL3基因的克隆及其表达分析

发布时间：2018-04-12 07:32

本文选题：马尾松 + PmCBL　；参考：《南京林业大学学报(自然科学版)》2017年04期

【摘要】：【目的】为明确马尾松(Pinus massoniana)CBL基因结构特征及干旱胁迫下的表达特性,探讨该基因在马尾松抗旱调控过程中的生物学功能。【方法】以马尾松优良家系为试材,采用RT-PCR和RACE方法克隆马尾松CBL基因全长cDNA,运用生物信息学方法,分析其蛋白质性质和亲缘关系,利用荧光定量PCR分析干旱胁迫下的基因表达特性。【结果】获得马尾松的一个CBL同源基因,命名为PmCBL3,该基因cDNA序列全长1 035 bp,包括681 bp的完整开放阅读框,编码226个氨基酸。PmCBL3蛋白含4个EF-hand功能域,且相邻EF-hand功能域之间的氨基酸数目非常保守,属于EFh家族蛋白。系统进化分析表明:该蛋白与其他植物CBL同源性达62%~98%,其中与北美云杉(Picea sitchensis)CBL亲缘关系最近。qRT-PCR结果显示,PmCBL3在马尾松根中表达量最高,茎和叶次之;干旱第15天表达量最大,随干旱胁迫加剧,表达量逐渐降低。【结论】PmCBL3基因属于CBLs家族,与北美云杉亲缘关系最近。该基因响应干旱胁迫,推测其可能参与马尾松干旱逆境的应答调控。
[Abstract]:[objective] to clarify the structural characteristics of Pinus massoniana)CBL gene and its expression under drought stress, and to explore the biological function of Pinus massoniana)CBL gene during drought resistance regulation of Pinus massoniana. [methods] A good family of Pinus massoniana was used as the test material.The full-length cDNAs of CBL gene of Pinus massoniana were cloned by RT-PCR and RACE, and their protein properties and genetic relationships were analyzed by bioinformatics.Fluorescence quantitative PCR was used to analyze the gene expression characteristics under drought stress. [results] A CBL homologous gene of Pinus massoniana, named PmCBL3, was obtained. The cDNA sequence of the gene was 1 035 BP, including a complete open reading frame of 681 BP.The protein encoding 226 amino acids. PmCBL3 contains 4 EF-hand functional domains, and the number of amino acids between the adjacent EF-hand functional domains is very conserved, which belongs to the EFh family.Phylogenetic analysis showed that the homology of PmCBL3 with other plants was 62kyun98, among which, the expression of PmCBL3 was the highest in roots of Pinus massoniana, followed by stem and leaf, and the highest on the 15th day of drought, especially in the roots of Pinus massoniana, and at the 15th day of drought, the expression of PmCBL3 was the highest in the roots of Pinus massoniana, and in the 15th day of drought.[conclusion] the PmCBL3 gene belongs to the CBLs family and is closely related to Picea spruce.In response to drought stress, the gene may be involved in the regulation of response to drought stress of Pinus massoniana.
【作者单位】：贵州省森林资源与环境研究中心贵州大学林学院;
【基金】：国家自然科学基金项目(31260183) “十三五”国家科技支撑计划(2015BAD09B0102) 贵州省重大专项项目(黔科合重大专项字[2012]6001号) 贵州省人才基地建设项目(黔人领发[2009]9号)
【分类号】：S791.248

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