芍药远缘杂交不亲和PlABCG15基因的克隆及表达分析  

Cloning and expression analysis of distant hybridization incompatibility PlABCG15 gene from Paeonia lactiflora

作　　者：贺丹[1,2,3] 曹健康 何松林 张明星 华超 张佼蕊 刘艺平 HE Dan;CAO Jiankang;HE Songlin;ZHANG Mingxing;HUA Chao;ZHANG Jiaorui;LIU Yiping(College of Landscape Architecture and Art, Henan Agricultural University, Zhengzhou 450002, China;College of Horticulture and Landscape Architecture, Henan Institute of Science and Technology, Xinxiang 453000, China;Henan Province Quality Flower and Vegetable Seedling Engineering Research Center, Zhengzhou 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：《河南农业大学学报》2021年第6期1074-1080,1096,共8页Journal of Henan Agricultural University

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：为研究芍药ABCG家族基因在芍药远缘杂交不亲和中的作用,以芍药‘粉玉奴’与牡丹‘凤丹白’杂交24 h柱头为材料,利用PCR扩增技术克隆得到一个芍药PlABCG15基因。该基因CDS区域全长1098 bp,共编码366个氨基酸;蛋白共有6个跨膜区域,以不规则卷曲为主。qRT-PCR分析显示,PlABCG15在茎中的表达水平最高;在自、杂交不同时期的柱头中,PlABCG15基因的相对表达水平总体呈上升趋势,且自交高于杂交(除第4、11、12天外)。自、杂交不同时期柱头的ABA含量总体呈下降趋势,且基因相对表达量与ABA含量呈负相关关系(自交Pearson相关系数为-0.155,杂交Pearson相关系数为-0.474(P<0.01))。亚细胞定位表明,PlABCG15定位在质膜上,进一步验证了该基因可能具有转运功能。In order to elucidate the molecular mechanism that controls the incompatibility of distant hybridization in Paeonialactiflora,the stigma of the hybridization between Paeonia lactiflora‘Fenyunu’×Paeonia suffruticosa‘Fengdanbai’after 24 h of pollination was used as a template.A Paeonia lactiflora PlABCG15 gene was cloned by PCR amplification technology.The gene,with a total length of 1098 bp,encoded 366 amino acids;The protein had 6 transmembrane regions,mainly random coil.qRT-PCR analysis showed,PlABCG15 gene had the highest expression in the stem.The relative expression level of stigma at different stages of self-cross and hybridization showed an overall upward trend,and it was higher in self-cross than in the hybridization of the same periods(except for the 4h,11h,and 12h).The ABA content of the stigmas at different stages of self-cross and hybridization declined gradually.The relative expression of PlABCG15 was negatively correlated with the ABA content(The inbred Pearson correlation coefficient was-0.155,and the hybrid Pearson correlation coefficient was-0.474(P<0.01)).The subcellular location indicated that PlABCG15 was located on the plasma membrane,which further verified that the gene might have a transport function.

关 键 词：芍药 杂交不亲和 ABCG转运蛋白 基因克隆 亚细胞定位 

分 类 号：S682.1[农业科学—观赏园艺]