金线莲中调控胚胎发育WRKY转录因子筛选及克隆分析  

Screening and cloning analysis of WRKY transcription factor regulating embryo development from Anoectochilus roxburghii

作　　者：邢丙聪 苏立样[1,2] 万思琦 邵清松 XING Bing-cong;SU Li-yang;Wan Si-qi;SHAO Qing-song(Zhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine,Hangzhou 311300,China;Department of Traditional Chinese Medicine,Zhejiang Agriculture and Forestry University,Hangzhou 311300,China)

机构地区：[1]浙江省特色中药资源保护与创新利用重点实验室,浙江杭州311300 [2]浙江农林大学中药学科,浙江杭州311300

出　　处：《中草药》2022年第12期3745-3754,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(82173916);浙江省农业新品种选育重大科技专项(2021C02074);浙江省特色中药资源保护与创新利用重点实验室(2021E10013)。

摘　　要：目的 探究金线莲Anoectochilus roxburghii中WRKY转录因子时空表达模式并筛选调控金线莲胚胎发育的Ar WRKY基因。方法 基于金线莲转录组数据库进行Ar WRKY转录因子筛选,利用软件进行蛋白质结构域预测、序列比对、理化性质预测与进化树构建等分析。采用实时荧光定量PCR(q RT-PCR)检测Ar WRKY基因时空表达模式。根据转录组数据Unigenes克隆Ar WRKY的CDS序列。结果 筛选得到金线莲中23条WRKY转录因子,分别命名为Ar WRKY1~Ar WRKY23;q RT-PCR结果表明Ar WRKY5与Ar WRKY20基因在金线莲叶芽期、花芽期及开花期的花中表达量都显著高于根、茎、叶;克隆得到Ar WRKY5的CDS为600 bp,Ar WRKY20的CDS为1149 bp。结论 筛选得到23个Ar WRKY转录因子并成功克隆获得Ar WRKY5与Ar WRKY20的CDS序列,对探究Ar WRKY转录因子调控金线莲生长发育及验证Ar WRKY5与Ar WRKY20基因功能奠定基础。Objective In order to lay a foundation for further study of EuTCP genes function, TCP(Teosinte branched 1/Cycloidea/Proliferating) gene family of Eucommia ulmoides(E. ulmoides) is screened and analyzed in this study. Method Based on the E. ulmoides genome database, the physicochemical properties, phylogeny, gene structure, promoter cis-acting elements and their expression levels in the leaf development and eu-rubber formation of the TCP gene family were analyzed by bioinformatics system. Result In this study, a total of 14 EuTCPs were identified from E. ulmoides genome, EuTCPs encoded 139—492 amino acids, and theoretical isoelectric points distribution were 5.54—9.72, molecular weight between 18 880 to 53 620 by physicochemical analysis, all of EuTCPs proteins were hydrophilic proteins and expressed in nucleus. EuTCP gene family was divided into three subfamilies: PCF, CIN and CYC/TB1, which contained 5, 6 and 3 EuTCPs proteins respectively. The expression pattern analysis showed that EuTCPs were significantly different at different developmental stages of E. ulmoides leaves, and the expression levels of EuTCPs were different at each developmental stage. EuTCPs positively regulated the formation of eu-rubber.Conclusion There are 14 members of TCP family identified in E. ulmoides, and their molecular characteristics and tissue expression specificity are different, which may be involved in the development of leaves and eu-rubber formation.

关 键 词：金线莲 胚胎发育 WRKY转录因子 生物信息学分析 表达模式 

分 类 号：R286.12[医药卫生—中药学]