阳桃木质素合成相关基因AcaCAD2克隆与表达分析  

Cloning and expression analysis of AcaCAD2 gene related to lignin biosynthesis inAverrhoa carambola

作　　者：赵亚梅 陈露[1] 李青女 肖云 翟俊文[1] 任惠[3] 吴沙沙[1] ZHAO Yamei;CHEN Lu;LI Qingnu;XIAO Yun;ZHAI Junwen;REN Hui;WU Shasha(Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization,College of Landscape Architecture and Art,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Forestry Bureau of Lanshan County,Yongzhou,Hunan 425800,China;Horticulture Research Insistute of Guangxi Academy of Agricultural Science,Nanning 530007,China)

机构地区：[1]福建农林大学风景园林与艺术学院,兰科植物保护与利用国家林业和草原局重点实验室,福州350002 [2]蓝山县林业局,湖南永州425800 [3]广西壮族自治区农业科学院园艺研究所,南宁530007

出　　处：《植物生理学报》2023年第7期1351-1360,共10页Plant Physiology Journal

基　　金：广西重点研发项目(桂科AB16380134);福建农林大学艺术学院、园林学院学科专业建设项目(YSYL-bdpy-1和YSYL-bdpy-2)。

摘　　要：肉桂醇脱氢酶(Cinnamyl alcohol dehydrogenase,CAD,EC1.1.1.195)是植物木质素合成途径的关键酶,参与多种生物和非生物胁迫的过程,与植物的生长发育密切相关。为探究AcaCAD2基因在阳桃木质素合成过程中的作用及表达规律,本研究通过克隆得到阳桃AcaCAD2基因,并对其进行生物信息学和表达模式分析。结果表明,AcaCAD2含有1074 bp的开放阅读框,编码368个氨基酸,属于稳定亲水性蛋白,不含跨膜结构和信号肽。结构域分析发现该蛋白具有典型的CAD结构域,包含2个Zn2+结合位点和1个NADP+辅酶结合位点。系统发育分析表明AcaCAD2与拟南芥At CAD5和At CAD4亲缘关系较近,亚细胞定位分析发现AcaCAD2蛋白定位在细胞膜中。q RT-PCR结果显示该基因在阳桃小枝发育的过程中呈现先增加后降低的趋势,在阳桃叶、花、果、叶芽中表达相对较弱,呈现出明显的组织特异性。本研究结果为进一步探究AcaCAD2对阳桃木质素合成的影响提供了理论参考。Cinnamyl alcohol dehydrogenase(CAD,EC1.1.1.195)is a key enzyme in the lignin biosynthesis pathway,which is involved in a variety of biotic and abiotic stress processes,closely related to plant growth and development.To reveal the function and expression patterns of AcaCAD2 gene in Averrhoa carambola during lignin biosynthesis,the AcaCAD2 gene was cloned,analyzed by bioinformatics and expression patterns.The results showed that AcaCAD2 contained 1074 bp open reading frame that encoding 368 amino acids,the protein was stable hydrophilic protein without signal peptide and transmembrane structure.Domain analysis revealed that the protein had a typical CAD domain,including two Zn^(2+)binding sites and one NADp*coenzyme binding site.The phylogenetic tree analysis revealed the AcaCAD2 was most closely related to AtCAD5 and AtCAD4;and subcellular localization analysis revealed that AcaCAD2 was localized in cell membrane.The result of qRT-PCR showed that the expression levels of AcaCAD2 increased first and then decreased during the development of twigs,and it was weakly expressed in leaves,flowers,fruits,and vegetative bud,showing obvious tissue specificity.The results about AcaCAD2 provide a theoretical basis for further research on the effect of lignin biosynthesis in A.carambola.

关 键 词：阳桃 木质素 肉桂醇脱氢酶 克隆 表达分析 

分 类 号：Q943.2[生物学—植物学]