摘要
为了研究甜叶菊中主要甜菊醇糖苷积累特点与其生物合成关键基因表达量的关系,本研究选取3个品种甜叶菊(普赛科3号、中山2号、同心)为试验材料,定期在幼苗期、营养生长期、花蕾期采收植株的上位3对叶片,采用液相色谱-质谱法(LC-MS)检测9种甜菊醇糖苷,即莱宝迪苷A、B、C、D、E、F(RA、RB、RC、RD、RE、RF)、甜菊糖苷(ST)、甜茶糖苷(RBS)和杜尔可苷A(DA)含量,同时利用实时荧光定量PCR(qRT-PCR)分析糖苷生物合成的3个UDP-糖基转移酶(UGTs)关键基因,即SrUGT91D2e、SrUGT74G1和SrUGT76G1的表达水平。结果表明,甜味糖苷RA在普塞科3号营养生长期含量最高(1.60 mg·g^(-1)),RD在普塞科3号的花蕾期含量最高(1.58 mg·g^(-1));苦味糖苷ST、RC、DA在普塞科3号中的积累量显著低于中山2号和同心(普塞科3号ST为0.50 mg·g^(-1)、RC为0.26 mg·g^(-1)、DA为0.01 mg·g^(-1)),因此普塞科3号可作为甜味糖苷高产选育品种。对甜菊醇糖苷生物合成关键基因表达量与糖苷含量进行相关性分析,甜味糖苷RD和RA的含量与其关键影响因子SrUGT91D2e和SrUGT76G1的表达量显著相关,苦味糖苷RC、ST、DA的含量与其关键影响因子SrUGT74G1的表达量显著相关。本试验为甜叶菊不同甜度糖苷的品种选育及采收时间的确定提供了依据。
In order to study the correlation between the accumulations of main steviol glycosides(SGs)and the expressions of the key genes that are involved in their biosynthesis in Stevia rebaudiana,three cultivars(Pusaike3,Zhongshan2,and Tongxin)were selected as materials.The upper three layer leaves were harvested at 3 growth stages,including the young seedling stage,vegetative stage,and flower budding stage.The contents of 9 SGs,such as rebaudioside A,B,C,D,E,F(RA,RB,RC,RD,RE,RF),stevioside(ST),rubusoside(RBS),dulcoside A(DA),were test by liquid chromatography mass spectrometry(LC-MS),and the expression levels of 3 key genes encoding uridine diphosphate-glycosyltransferases(UGTs),SrUGT91D2e,SrUGT74G1 and SrUGT76G1,were analyzed by quantitative real-time PCR.The results showed that the contents of the sweet SGs,RA was the highest at vegetative stage(1.60 mg·g^(-1)),and RD was the highest at flower budding stage(1.58 mg·g^(-1))in Pusaike3;the contents of bitter SGs,St(0.50 mg·g^(-1)),RC(0.26 mg·g^(-1))and DA(0.01 mg·g^(-1))were significant lower in Pusaike3 than those in Zhongshan 2 or Tongxin,thus Pusaike3 was considered as a candidate cultivar for high yield of sweet SGs.The correlation analysis between the contents of SGs and the expressions levels of the key genes involved in their biosynthesis showed that the contents of sweet SGs,RD and RA were remarkably correlated with the expression levels of SrUGT91D2e and SrUGT76G1,while the contents of bitter SGs,RC,ST,DA were remarkably correlated with the expression level of SrUGT74G1.This research provides a scientific basis for Stevia rebaudiana breeding and suitable harvest time selection.
作者
张虹
路国栋
袁春春
郎思睿
陈任
ZHANG Hong;LU Guodong;YUAN Chunchun;LANG Sirui;CHEN Ren(School of Life Science,NingxiaUniversity/Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China/Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia,Yinchuan,Ningxia 750021)
出处
《核农学报》
CAS
CSCD
北大核心
2022年第1期75-82,共8页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金资助项目(31860065)
宁夏自然科学基金项目(2020AAC03106)。
关键词
甜叶菊
甜菊醇糖苷
基因表达
相关性分析
Stevia rebaudiana
steviol glycosides
gene expression
correlation analysis
