薄壳山核桃MADS-box基因保守片段的克隆与序列分析  

Cloning and sequence analysis of MADS-box gene conservative fragment from pecan (Carya illinoensis)

作　　者：莫正海[1] 张计育[1] 翟敏 贾晓东[1] 宣继萍[1] 李永荣 郭忠仁[1] 

机构地区：[1]中国科学院植物研究所,南京210014 [2]南京绿宙薄壳山核桃科技有限公司,南京210072

出　　处：《南方农业学报》2013年第5期730-734,共5页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31200502);江苏省科技支撑计划项目(BE2012338);江苏省农业科技自主创新资金项目[CX(12)2012];南京市科技计划项目(201101025)

摘　　要：【目的】克隆薄壳山核桃MADS-box基因的保守片段,进行系统发育分析,为研究薄壳山核桃花发育相关MADS-box家族基因及其发育的分子机理奠定基础。【方法】以薄壳山核桃品种‘马罕’雄花花序为材料,提取总RNA反转录cDNA,采用RT-PCR克隆MADS-box基因的保守片段,并将其推导氨基酸序列与已知拟南芥的MADS-box家族基因进行系统发育分析。【结果】分离获得28条MADS-box基因的cDNA片段,片段长度均为137bp,包含基因起始密码子,核苷酸序列同源性为65.7%～98.5%,其推导氨基酸序列中有11个存在差异。系统发育树分析结果表明,这些基因片段分别归入拟南芥MADS-box基因不同亚家族中,包含ABCDE模型中的各类基因。【结论】薄壳山核桃中存在多种MADS-box家族基因,克隆的片段包含ABCDE模型中的各类花发育基因。[Objective]The conservative fragment sequence of MADS-box genes from pecan was cloned and their phy- logeny was assayed in order to provide references for cloning pecan new MADS-box family genes related to flower development and exploring molecular mechanism of pecan flower development. [Method]The total RNA was extracted from male flower of pecan variety ＇Mahan＇ and cDNA was synthesized using inverse transcription. The conservative fragment of MADS-box genes was cloned using RT-PCR, and the phylogenetic analysis was conducted using the deduced amino acid sequences with known Arabidopsis MADS-box gene subfamily. [ResultlTwenty-eight cDNA fragments of MADS-box gene were obtained by RT-PCR, and the length of these fragments was 137 bp including the initiation codon of genes. The homology of these nu- cleotide sequences was between 65.7% and 98.5%. The 11 deduced amino acid sequences were different. According to phyloge- netic tree analysis, all these gene fragments involving different types of gene in ABCDE model were divided into the Arabidopsis MADS-box gene subfamily. [Conclusion]There were many MADS-box genes in pecan. These gene fragments included all types of floral development genes in ABCDE model.

关 键 词：薄壳山核桃 MADS-BOX基因 RT-PCR 克隆 序列分析 

分 类 号：S664.1[农业科学—果树学]