华北农学报 ›› 2020, Vol. 35 ›› Issue (6): 81-89. doi: 10.7668/hbnxb.20191499

所属专题: 生物技术

• 作物遗传育种·种质资源·生物技术 • 上一篇    下一篇

芍药属远缘杂交不亲和PlABCF3基因克隆及表达分析

贺丹1,2, 张佼蕊1, 何松林2, 刘红利1, 王政1, 刘艺平1   

  1. 1. 河南农业大学 风景园林与艺术学院, 河南 郑州 450002;
    2. 河南科技学院 园艺园林学院, 河南 新乡 453000
  • 收稿日期:2020-08-19 出版日期:2020-12-28
  • 通讯作者: 何松林(1965-),男,河南郑州人,教授,博士,主要从事园林植物研究。
  • 作者简介:贺丹(1983-),女,河南新乡人,副教授,博士,主要从事园林植物育种研究。
  • 基金资助:
    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)

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

HE Dan1,2, ZHANG Jiaorui1, HE Songlin2, LIU Hongli1, WANG Zheng1, LIU Yiping1   

  1. 1. College of Landscape Architecture and Art, Henan Agricultural University, Zhengzhou 450002, China;
    2. College of Horticulture Landscape Architecture, Henan Institute of Science and Technology, Xinxiang 453000, China
  • Received:2020-08-19 Published:2020-12-28

摘要: 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2 151 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。

关键词: 牡丹, 芍药, PlABCF3基因, RT-PCR, 远缘杂交

Abstract: 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 2 151 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.

Key words: Paeonia ostii, Paeonia lactiflora, PlABCF3 gene, RT-PCR, Distant hybridization

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引用本文

贺丹, 张佼蕊, 何松林, 刘红利, 王政, 刘艺平. 芍药属远缘杂交不亲和PlABCF3基因克隆及表达分析[J]. 华北农学报, 2020, 35(6): 81-89. doi: 10.7668/hbnxb.20191499.

HE Dan, ZHANG Jiaorui, HE Songlin, LIU Hongli, WANG Zheng, LIU Yiping. Cloning and Expression Analysis of Distant Hybridization Incompatibility PlABCF3 Gene from Paeonia[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(6): 81-89. doi: 10.7668/hbnxb.20191499.

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