芍药属远缘杂交不亲和PlABCF3基因克隆及表达分析

Cloning and Expression Analysis of Distant Hybridization Incompatibility PlABCF3 Gene from Paeonia

为进一步研究PlABCF3基因在芍药属远缘杂交不亲和中的作用机制,根据转录组测序结果,采用RT-PCR技术在芍药粉玉奴自交和芍药粉玉奴×牡丹凤丹白杂交24,36 h的柱头中克隆到1个ABCF3基因(PlABCF3)。生物信息学和表达特性分析表明,PlABCF3的CDS全长为2151 bp,编码716个氨基酸。芍药ABCF3的碱基序列与拟南芥ABCFs进行同源序列分析表明,其与AtABCF3具有较高的同源性。系统进化分析表明,PlABCF3与博落回的ABCF3同源性最高,PlABCF3氨基酸序列与博落回的ABCF3的相似性最高,故命名为PlABCF3(GenBank登录号:MT123594)。RT-qPCR分析表明,在不同时期的自交、杂交柱头中,PlABCF3基因在相同处理条件下,36 h的相对表达量均低于24 h,杂交36 h相对表达量最低。PlABCF3的相对表达量与脱落酸(ABA)、玉米素核苷(ZR)含量显著负相关(R分别为-0.837,-0.565,P<0.01)。说明PlABCF3基因在芍药属远缘杂交中与内源激素有着密切的关系。

To further study the mechanism of PlABCF3 gene in distant hybrid incompatibility of Paeonia lactiflora.The stigmas of combinations P.lactiflora Fenyunu×P.lactiflora Fenyunu and P.lactiflora Fenyunu×P.ostii Fengdanbai were harvested as the materials after 24,36 h of pollination and ABCF3 gene(PlABCF3)was cloned by RT-PCR according to transcriptome data.Bioinformatics and expression characteristics showed that the CDS of PlABCF3 was 2151 bp encoded 716 amino acids.Between PlABCF3 and ABCFs in Arabidopsis thaliana,phylogenetic analysis and homologous sequence analysis showed that there were high homology of base sequences between PlABCF3 and AtABCF3.Phylogenetic analysis showed that PlABCF3 had high identities with ABCF3 in Macleaya cordata,and the amino acid sequence of PlABCF3 had high similarity with ABCF3 in Macleaya cordata,so its name was designated PlABCF3(GenBank accession No.MT123594).RT-qPCR results indicated that during self-cross and hybridization of the stigmas,the PlABCF3 expression level was lower in 36 h at the same treatment and lowest in hybridization stigma in 36 h.PlABCF3 gene expression was negatively related with abscisic acid(ABA),and zeatin nucleoside(ZR)content in stigmas(R=-0.837,-0.565,P<0.01).It was speculated that PlABCF3 had a deep connection with endogenous hormones in the distant hybridization of Paeonia.