山葡萄VaSR基因家族的鉴定及VaSR1抗寒功能验证与互作蛋白筛选  

Identification of VaSR Gene Family in Vitis amurensis,Verification of Cold Resistance Function of VaSR1 and Screening of Interacting Proteins

作　　者：梁国平 曾宝珍 刘铭 边志远 陈佰鸿[1] 毛娟[1] LIANG Guoping;ZENG Baozhen;LIU Ming;BIAN Zhiyuan;CHEN Baihong;MAO Juan(College of Horticulture,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学园艺学院,兰州730070

出　　处：《园艺学报》2025年第1期37-50,共14页Acta Horticulturae Sinica

基　　金：甘肃农业大学公招博士科研启动项目(GAU-KYQD-2022-30);甘肃省自然科学基金项目(23JRRA1425);高校教师创新基金项目(2024B-073)。

摘　　要：为探究葡萄中丝氨酸/精氨酸富集剪接因子(SR)的抗寒功能,对山葡萄(Vitis amurensis Rupr.)低温休眠期不同温度时段处理下的枝条韧皮部转录组中筛选出的VaSR1进行克隆、抗寒功能鉴定与互作蛋白筛选。从山葡萄基因组数据库中鉴定到了13个VaSR家族成员,其保守功能结构域为RRM_SF superfamily。对其进行系统进化和共线性分析,结果显示VaSR的多数成员被聚类到SC亚族,且与苹果SR蛋白亲缘关系较近。对VaSR1进行抗寒功能验证,转基因拟南芥植株的冰冻存活率显著低于野生型;4℃低温胁迫后转基因植株叶片中可溶性糖含量和抗氧化酶及淀粉酶活性显著低于野生型,而淀粉含量显著增加;酵母双杂交显示VaU1SNRNP和VaCYP63是VaSR1的互作蛋白,具有促进前体mRNA的剪接和参与调节细胞中相关基因的聚腺苷酸化作用。综上,VaSR1过表达降低了转基因拟南芥中的淀粉酶活性,导致叶片中可溶性糖含量下降,减弱了植株的低温耐受性。此外,山葡萄中存在VaU1SNRNP和VaCYP63,能够与VaSR1发生互作。To investigate the cold-resistance function of serine/arginine-rich splicing factor(SR)in grapevine,the present study selected VaSR1 from the transcriptome data of the phloem of branches treated with various temperature periods during the low-temperature dormancy period of Vitis amurensis Rupr.,and performed cloning,identification of cold-resistant functions,and screening of interacting proteins.Thirteen VaSR family members with the conserved functional domain RRM_SF superfamily were identified from the genome database of V.amurensis.Phylogenetic and collinearity analysis revealed that the majority of the members of the VaSRs were clustered into the SC subfamily and were closely related to the SR proteins of apple.The cold tolerance function of VaSR1 was verified,and the freezing survival rate of transgenic Arabidopsis plants was significantly lower than that of the wild type;the soluble sugar content and the activities of antioxidant enzymes and amylase in the leaf of transgenic plants were significantly lower than those of the wild type after 4℃cold stress,while the starch content was significantly increased;the yeast two-hybridization elucidated that VaU1SNRNP and VaCYP63 were the intercalating proteins of VaSR1 with the ability to promote precursor mRNA splicing and involved in regulating polyadenylation of related genes in cells.Collectively,VaSR1 overexpression reduced amylase activity in transgenic plants,leading to a reduction in soluble sugar content in the leaves and attenuating the low-temperature tolerance of the plants.Furthermore,the presence of VaU1SNRNP and VaCYP63 in mountain grapes was capable of interacting with VaSR1.

关 键 词：山葡萄 转录因子 VaSR1 低温胁迫 酵母双杂交 

分 类 号：S663.1[农业科学—果树学]