摘要
为了揭示竹叶花椒萜类代谢的分子机理及嫁接对其风味的影响,该文依据转录组数据设计特异性引物,采用RT-PCR方法从竹叶花椒(Zanthoxylum armatum)中克隆得到一个全新的牻牛儿基牻牛儿基焦磷酸合成酶(GGPPS)基因的全长cDNA序列,命名为ZaGGPPS,并利用NCBI、ProParam、SignalP 4.1 server、DNAMAN和MEGA 7.0软件对ZaGGPPS基因进行生物信息学分析,并比较其在嫁接树和实生树中的表达量。结果表明:ZaGGPPS包含完整的cDNA开放阅读框(OFR),由1086 bp组成,编码361个氨基酸。其蛋白的相对分子量为39079.14 Da,理论等电点pI为6.38。Blast比对结果显示该蛋白质属于GGPPS家族蛋白,含有2个GGPPS蛋白特有的天冬氨酸富集基序,分别是“DDXXXXD”和“DDXXD”,以及5个特征性功能结构域。系统进化树结果显示竹叶花椒与芸香科植物甜橙(Citrus sinensis)、克里曼丁桔(C.clementina)、柚子(C.maxima)等亲缘关系较近。荧光定量PCR检测显示,ZaGGPPS基因在竹叶花椒中的表达量从高到低分别为实生树的叶、嫁接树的叶、实生树的茎、嫁接树的茎。牻牛儿基牻牛儿基焦磷酸合成酶是竹叶花椒萜类化合物生物合成途径中的关键酶,通过嫁接可影响ZaGGPPS基因在叶和茎中的表达量。该文对竹叶花椒ZaGGPPS基因进行了克隆与分析,为后续深入研究竹叶花椒香气形成的分子机理及利用分子生物学手段选育优良品种提供理论依据。
In order to reveal the molecular mechanism of terpenoid metabolism in Zanthoxylum armatum and the effect of grafting on its flavor,we designed specific primers based on transcriptome data and cloned a novel full-length cDNA sequence of geranylgeranyl pyrophosphate synthase(GGPPS)gene from Z.armatum by RT-PCR,and named ZaGGPPS.We analyzed the ZaGGPPS gene by using NCBI,ProParam,SignalP 4.1 server,DNAMAN and MEGA 7.0 softwares.The expression of ZaGGPPS gene in grafted and seedling trees were compared.The results were as follows:ZaGGPPS contained a complete open reading frame(OFR),consisting of 1086 bp,encoding 361 amino acids.The relative molecular weight of the protein was 39079.14 Da and the theoretical isoelectric point pI was 6.38.Blast comparison results showed that the protein belonged to the GGPPS family and contains two specific aspartic acid enrichment motifs,namely“DDXXXXD”and“DDXXD”,and five characteristic functional domains.Phylogenetic tree results showed that Z.armatum had close relationships with sweet orange(Citrus sinensis),clementine mandarin(C.clementina)and pomelo(C.maxima).Fluorescence quantitative PCR showed that the expression level of ZaGGPPS gene in Zanthoxylum armatum ranged from high to low as follows:leaf of seedling tree,leaf of grafted tree,stem of seedling tree and stem of grafted tree.Geranylgeranyl pyrophosphate synthase was a key enzyme in terpenoid biosynthesis pathway of Z.armatum,and grafting can affect the expression of ZaGGPPS gene in leaves and stems.The cloning and analysis of ZaGGPPS gene of Z.armatum provides theoretical basis for further study on the molecular mechanism of aroma formation of Z.armatum and selection of excellent varieties by molecular biological means.
作者
关淑文
王毅
郝佳波
原晓龙
陆斌
李贤忠
GUAN Shuwen;WANG Yi;HAO Jiabo;YUAN Xiaolong;LU Bin;LI Xianzhong(College of Forestry,Southwest Forestry University,Kunming 650224,China;Yunnan Key Laboratory of Forestry Plant Cultivation and Utilization,State Forestry and Grassland Administration Key Laboratory of Yunnan Rare and Endangered Species Conservation and Propagation,Yunnan Academy of Forestry and Grassland,Kunming 650201,China)
出处
《广西植物》
CAS
CSCD
北大核心
2021年第4期598-605,共8页
Guihaia
基金
国家自然科学基金(31860177)
云南特色经济林新种质创新与利用(2018BB008)。
