牡丹远缘杂交种子败育PsMTERF2基因的克隆、表达与生理机制分析  

Cloning,expression and physiological mechanism of Paeonia suffruticosa distant hybrid seeds abortion PsMTERF2 gene

作　　者：贺丹[1,2] 张明星 何松林 曹健康 华超 张佼蕊 刘艺平 HE Dan;ZHANG Mingxing;HE Songlin;CAO Jiankang;HUA Chao;ZHANG Jiaorui;LIU Yiping(College of Architecture and Art,Henan Agricultural University,Zhengzhou,Henan 450002,China;Colleg of Horticulture Landscape Architecture,Henan Institute of Science and Technology,Xinxiang,Henan 453003,China;Henan Province Quality Flower and Vegetable Seedling Engineering Research Center,Zhengzhou,Henan 450002,China)

机构地区：[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453003 [3]河南省优质花卉蔬菜种苗工程研究中心,河南郑州450002

出　　处：《西北农林科技大学学报（自然科学版）》2022年第12期108-116,共9页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(31600568,31870698);河南省科技攻关项目(202102110234)。

摘　　要：【目的】探究PsMTERF2基因在牡丹远缘杂交种子发育过程中的分子及生理机制,为克服牡丹远缘杂交种子败育的研究提供理论依据。【方法】采用RT-PCR技术克隆得到牡丹MTERF2基因,并对其进行生物信息学分析;采用实时荧光定量PCR(RT-qPCR)技术检测并比较PsMTERF2基因在牡丹种子不同发育时期(‘凤丹白’自交与‘凤丹白’ב粉玉奴’授粉后14,18和28 d的正常与败育种子)的表达与牡丹不同组织(根、茎、叶、花瓣、花药、鳞芽、柱头、种子)中的差异性表达;测定不同发育时期正常种子与败育种子中可溶性总糖、蔗糖、果糖、淀粉和可溶性蛋白的含量,并分析其与PsMTERF2基因表达量的相关性。【结果】牡丹MTERF2基因CDS全长972 bp,编码323个氨基酸,相对分子质量为3.77×10^(4) D,理论等电点为8.87,具有3个跨膜螺旋区,不具有信号肽切割位点;进化树及三级结构分析显示,牡丹MTERF2基因具有7个MTERF结构域,蛋白结构较为复杂,其碱基序列与AtMTERF2同源性较高,故命名为PsMTERF2,GenBank登录号为MZ505000。PsMTERF2基因在牡丹自交种子各发育时期的表达量均高于同一发育时期的杂交种子,且在种子发育后期二者差异更显著;PsMTERF2基因在牡丹不同组织中均有表达,但在种子、柱头和花药中的表达水平相对较高。授粉后14和28 d,可溶性总糖、蔗糖、果糖、淀粉和可溶性蛋白在自交正常种子中的含量均高于杂交败育种子,其中淀粉和可溶性蛋白含量与PsMTERF2基因相对表达量显著正相关,其余指标与PsMTERF2基因相对表达量正相关但相关性不显著(P>0.05)。【结论】PsMTERF2基因可能通过影响代谢产物的合成参与调控种子发育的后期进程。【Objective】This study explored the molecular mechanism of PsMTERF2 gene in seed development of Paeonia suffruticosa to provide basis for understanding seed abortion of distant hybridization.【Method】The bioinformatics and expression characteristics of the PsMTERF2 gene cloned by RT-PCR technology were analyzed.The quantitative Real-time PCR technology were applied to assess and compare the expression of PsMTERF2 gene in different developmental stages of peony seeds(14,18 and 28 d after pollination in P.ostii cv.‘Phoenix White’×P.ostii cv.‘Phoenix White’and P.ostii cv.‘Phoenix White’×P.lactiflora Fenyunu)and tissues of peony seeds(roots,stems,leaves,petals,anthers,scale buds,stigmas and mature seeds).Contents of total soluble sugar,sucrose,fructose,starch and soluble protein in normal seeds and aborted seeds at different developmental stages were determined for correlation analysis with PsMTERF2 gene expression.【Result】The CDS of MTERF2 was 972 bp in length,encoding 323 amino acids with the predicted molecular mass of 3.77×10^(4) D,and its theoretical isoelectric point(pI)was 8.87.It had three transmembrane helical regions,but no signal peptide cleavage site.The phylogenetic tree and three-dimensional structure analysis indicated that PsMTERF2 contained 7 MTERF domains and complicated protein structure.Phylogenetic analysis and homologous sequence analysis showed that there was high homology of base sequences between PsMTERF2 and AtMTERF2,so it was named PsMTERF2 with GenBank accession No.of MZ505000.The expression level of PsMTERF2 in all stages of self-pollinated seeds was higher than that of cross-pollinated seeds in the same period,especially in the later stage of seed development.Expression of PsMTERF2 was detected in different tissues with high level in seeds,anthers and stigmas.The contents of total soluble sugar,sucrose,fructose,starch and soluble protein in self-pollinated normal seeds were higher than those of cross-pollinated abortive seeds 14 and 28 days after pollination.Solu

关 键 词：牡丹育种 种子败育 远缘杂交 PsMTERF2基因 基因表达分析 

分 类 号：S685.11[农业科学—观赏园艺]