扁桃SBP基因家族鉴定及PsdSBP1克隆表达分析  

Identification of SBP Gene Family and Expression Analysis of PsdSBP1 Clone in Almond

作　　者：张冬冬 杨佳惠 曾斌[1] 余镇藩 高雯雯 马昕彤 Zhang Dongdong;Yang Jiahui;Zeng Bin;Yu Zhenfan;Gao Wenwen;Ma Xintong(College of Forestry and Horticulture,Xinjiang Agricultural University,Urumqi,830000)

机构地区：[1]新疆农业大学园艺学院,乌鲁木齐830000

出　　处：《分子植物育种》2025年第2期415-427,共13页Molecular Plant Breeding

基　　金：新疆维吾尔自治区自然科学基金项目(2019D01A56);国家自然科学基金项目(31660557);新疆维吾尔自治区园艺学重点学科项目(2016-10758-3)共同资助。

摘　　要：S-RNase binding protein是与S-RNase蛋白质相互作用的一类蛋白,而SBP1在配子体植物中属于自交不亲和性基因,起SSK1和Rbx1作用,并形成一个SCF^(SLF/SFB)复合体,用于非自身S-RNase的泛素化进程,调控配子体植物自交不亲和反应进程。本研究利用生物信息学技术筛选扁桃基因组中SBP家族成员并进行表达模式分析,同时克隆PsdSBP1基因进行RT-qPCR分析。从扁桃中筛选到13个PsdSBPs家族蛋白成员,均为亲水性蛋白;系统进化树划分4个亚簇,分别具有2对片段复制和串联重复基因。PsdSBPs大部分成员主要在扁桃花组织中表达显著,而受冷冻胁迫后在花药和子房中的表达被抑制。克隆得到PsdSBP1和PsdSBP-4序列一致,扁桃自交异交授粉表达模式表明PsdSBP-4在异交中的表达量比花粉和自交高出一倍,表达活性增强,并且荧光定量结果表明PsdSBP1在花粉和花柱中均有表达活性,进而可以说明PsdSBP1在异交授粉后活性会增强。蛋白质关联网络结果表明PsdSBP-4(PsdSBP1)与CUL1蛋白具有互作关系,参与SCF复合体。综上所述,扁桃中存在SBP1基因并与CUL1互作,极大可能参与扁桃自交不亲和反应进程。本次研究为深入探究SBP1基因在扁桃中调控自交不亲和性提供理论依据。S-RNase binding pro tein is a class of proteins that interact with S-RNase protein,while SBP1 is a self-incompatibility gene in gametophytic plants,acting as SSK1 and Rbx1,and forming an SCFSLF/SFBcomplex for the ubiquitination process of non-self S-RNase.Coordinating the process of self-incompatibility in gametophyte plants.In this study,the SBP family members in the almond(Prunus dulcis)genome were screened by bioinformatics technology and the expression pattern was assessed.PsdSBP1 gene was cloned and assessed by RT-qPCR.Thirteen PsdSBPs family proteins were screened from slmond,all of which were hydrophilic proteins.The phylogenetic tree was divided into four clusters with two pairs of fragment duplication and tandem duplication genes respectively.Most members of PsdSBPs were notably expressed in slmond tissue but inhibited in anther and ovary under freezing stress.The sequence of PsdSBP1 and PsdSBP-4 was consistent.The expression pattern of self-pollinati on in almond showed that the expression level of PsdSBP-4 in the crossing was twice as high as that in pollen and self-pollination,and the expression activity was improved.The fluorescence quantitative results indicated that PsdSBP1 had expression activity in pollen and style.Consequently,the activity of PsdSBP1 could be improved after outcross pollination.The results of the protein association network showed that PsdSBP-4(PsdSBP1)interacted with CUL1protein and participated in the SCF complex.In conclusion,the SBP1 gene exists in the almond and interacts with CUL1,which is likely to participate in the process of self-incompatibility response in the almond.This study provided a theoretical basis for further research on SBP1 gene regulation of self-incompatibility in the almond.

关 键 词：扁桃 S-RNASE S-RNase binding protein 1 生物信息学分析 克隆表达分析 

分 类 号：S662.9[农业科学—果树学]