半夏珠芽发育相关PteAUX1基因的克隆与表达分析  

Cloning and Expressional Analysis of PteAUX1 Gene Related to Bulbil Development of Pinellia ternata

作　　者：罗睿 田晨 夏丽莎 陈海丽 杜声珊 罗须 Luo Rui;Tian Chen;Xia Lisha;Chen Haili;Du Shengshan;Luo Xu(College of Life Sciences,Institute of Agro-Bioengineering,Collaborative Innovation Center for Mountain Ecology&Agro-Bioengineering(CICMEAB),Guizhou University,Guiyang,550025)

机构地区：[1]贵州大学生命科学学院/农业生物工程研究院(山地植物资源保护与保护种质创新教育部重点实验室),山地生态与农业生物工程协同创新中心,贵阳550025

出　　处：《分子植物育种》2024年第18期6000-6009,共10页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31560081);贵州大学研究生创新基金项目(研理工2017027)共同资助。

摘　　要：生长素在中药植物半夏的珠芽发育中可能具有重要调节作用,但生长素调控珠芽发育的分子机制尚不清楚。本研究利用半夏及近缘植物滴水珠的RNA-seq结果筛选珠芽发育过程中影响内源生长素浓度的共差异表达基因,对筛选序列RACE克隆获得全长CDS或测序验证后进行序列分析,通过定量PCR对该基因在半夏珠芽发育过程中的时空表达特性进行分析。研究结果表明,珠芽发生过程中生长素合成、失活和运输环节都有差异表达基因,2个物种珠芽发生过程中共差异表达生长素输入蛋白AUX1/LAX家族成员(PteAUX1和PcoLAX2);序列分析发现编码蛋白二级结构都具有11个跨膜结构域,功能相关重要位点保守;PteAUX1在半夏叶柄的珠芽发生部位上/下随珠芽发生而差异表达。上述结果表明生长素输入蛋白AUX1/LAX家族成员在半夏叶柄珠芽发生部位上/下不同时期的差异表达可能导致该部位的内源生长素积累从而调节珠芽的发生。研究结果可为进一步探讨半夏珠芽发育调节的信号转导途径提供理论基础。Auxin may play an important regulatory role in bulbil development of Pinellia ternata(Thunb.)Breit.,but the molecular mechanism of auxin regulating the development of the bulbil is still unclear.In this study,RNA-seq results of P.ternata and P.cordata,which are relatives,were used to screen candidate unigenes that are involved in regulation of auxin concentration during bulbil development.Full CDS sequences were obtained by RACE cloning or were confirmed by sequencing.Furthermore,quantitative real-time PCR was used to analyze the spatio-temporal expression model of candidate genes during bulbil development of P.ternata.The results showed that there were differentially expressed genes in auxin synthesis,inactivation and transport during bud formation,and members of the AUX1/LAX family of auxin input proteins(PteAUX1 and PcoLAX2)were differentially expressed in the two species during bud formation;sequence analysis showed that the secondary structure of encoded proteins had 11 transmembrane domains,and functionally related important sites were conserved;PteAUX1 is differentially expressed on the upper/lower parts of P.ternata petiole with the occurrence of bead buds.These results indicated that differential expression of auxin influx carrier AUX1/LAX family members in the upper/lower regions of P.ternata petioles during different stages of bulbil formation may lead to localized accumulation of endogenous auxin,thereby regulating bulbil initiation.The research results can provide a theoretical basis for further exploring the signal transduction pathways of bulbil development regulation of P.ternata.

关 键 词：半夏(Pinellia ternata) 滴水珠(Pinellia cordata) 生长素 AUX1/LAX 珠芽 

分 类 号：S567.239[农业科学—中草药栽培]