摘要
为了探究芍药属不同种间远缘杂交不亲和的分子作用机制,以粉玉奴芍药自交授粉后24 h、36 h和粉玉奴芍药×凤丹白牡丹杂交授粉后24 h、36 h的柱头为材料,根据柱头转录组差异基因序列,采用逆转录(RT)-PCR技术,克隆得到SPL3基因的cDNA序列,将其命名为PlSPL3(GenBank登录号:MN842720),随后对其进行生物信息学分析和表达特性分析。结果表明,PlSPL3基因的编码区(Coding sequence,CDS)全长1305 bp,共编码434个氨基酸。分析结果显示,PlSPL3蛋白是一种带负电荷的不稳定的亲水性蛋白质,无跨膜结构。蛋白质进化树显示,芍药PlSPL3蛋白的氨基酸序列与拟南芥AtSPL7蛋白的氨基酸序列具有较高的同源性,同时芍药PlSPL3蛋白与木瓜、向日葵和无花果SPL3蛋白的氨基酸序列同源性也较高。实时荧光定量PCR(RT-qPCR)结果显示,在自交、杂交授粉后不同时期的柱头中,PlSPL3基因在杂交授粉后36 h的相对表达量最高。研究结果为进一步阐明PlSPL3基因在芍药与牡丹远缘杂交不亲和中的生物学功能提供了理论依据。
In order to explore the molecular mechanism of distant hybridization incompatibility in Paeonia,the stigmas from P.lactiflora Fenyunu×P.lactiflora Fenyunu and P.lactiflora Fenyunu×P.ostii Fengdanbai were harvested as the materials after 24 h and 36 h of pollination.The cDNA sequence of SPL3 gene was cloned by reverse transcription(RT)-PCR technique based on differential gene sequence of transcriptome in stigma,and was named as PlSPL3(GenBank accession No:MN842720),the bioinformatics and expression characteristics of PlSPL3 were then analyzed.The results showed that 434 amino acids were encoded by the coding sequence of PlSPL3 gene with a length of 1305 bp.Analysis results showed that PlSPL3 protein was a kind of unstable,hydrophilic protein with negative charges and without transmembrane structure.The results of phylogenetic tree indicated that the amino acid sequence of PlSPL3 protein from P.lactiflora had high homology with that of AtSPL7 protein from Arabidopsis thaliana,and the amino acid sequences of PlSPL3 protein in P.lactiflora were also highly homologous with those of SPL3 protein in Chaenomeles sinensis,Helianthus annuus and Ficus carica.Real-time quantitative PCR(RT-qPCR)results indicated that the relative expression level of PlSPL3 gene was the highest in stigmas at 36 h after hybridization.These results provide a theoretical basis in further elucidating the biological functions of PlSPL3 gene in distant hybridization incompatibility between P.lactiflora and P.ostii.
作者
张佼蕊
贺丹
何松林
谢栋博
李朝梅
王政
刘艺平
ZHANG Jiao-rui;HE Dan;HE Song-lin;XIE Dong-bo;LI Chao-mei;WANG Zheng;LIU Yi-ping(College of Landscape Architecture and Art,Henan Agricultural University,Zhengzhou 450002,China;School of Horticulture and Landscape Architecture,Henan Institute of Science and Technology,Xinxiang 453000,China)
出处
《江苏农业学报》
CSCD
北大核心
2020年第6期1537-1542,共6页
Jiangsu Journal of Agricultural Sciences
基金
国家自然科学基金项目(31600568、31870698)
河南省科技攻关项目(202102110234)
河南农业大学科技创新基金项目(KJCX2015A03)。
关键词
牡丹
芍药
PlSPL3基因
基因克隆
基因表达
Paeonia lactiflora
Paeonia ostii
PlSPL3 gene
gene cloning
gene expression
