江西农业大学学报2025，Vol.47Issue(4)：865-877,13.DOI:10.3724/aauj.2025074
薏苡ClVQ12的表达模式、抗旱功能及互作蛋白分析
Expression pattern,drought-resistant function and analysis of interacting proteins of ClVQ12 in Coix lacryma-jobi L
摘要
Abstract
[Objective]Coix lacryma-jobi L.is an important crop with both medicinal and edible value.Drought and soil salinization pose serious threats to its yield and quality.The VQ gene family plays a crucial role in plant responses to external stresses and hormone induction.This study aims to clone ClVQ12,a member of the VQ gene family in Coix,and analyzes its expression patterns,drought resistance,and interacting proteins,thus laying the foundation for further investigation into the functions of VQ genes in Coix growth,development,and stress responses.[Method]Using the Coix variety"Wanyi No.2"as material,ClVQ12 was cloned and subjected to bioinformatics analysis.Subcellular localization was performed to determine its cellular distribution.qRT-PCR was used to analyze its expression patterns under methyl jasmonate(MEJA)and abscisic acid(ABA)induction,as well as under high-temperature,salt,and drought stress.A yeast expression vector carrying ClVQ12 was constructed and transformed into yeast to compare the growth differences between transgenic and control yeast under drought stress.Yeast two-hybrid(Y2H)assays were conducted to verify the interactions between ClVQ12 and other ClVQs or ClWRKYs.[Result]ClVQ12 was successfully cloned,with an open reading frame(ORF)of 579 bp encoding 192 amino acids.The protein had an isoelectric point(pI)of 5.06,an average hydrophilicity index of-0.52,and was classified as an unstable hydrophilic protein.Subcellular localization revealed that ClVQ12 was localized in the cell membrane and nucleus.It lacked transmembrane domains and signal peptides but contained 18 phosphorylation sites.Secondary structure prediction indicated that the protein consisted mainly of random coils(61.46%),α-helices(32.29%),extended strands(4.17%),and β-sheets(2.08%).Promoter cis-acting element analysis identified multiple hormone-responsive and stress-related elements.Expression analysis showed that ClVQ12 was significantly induced by MEJA,ABA,and drought stress but suppressed under high-temperature and salt stress.Compared with the pYES2 empty vector control,ClVQ12-transformed yeast exhibited higher survival rates and better growth under mannitol-induced drought stress.Y2H assays confirmed that ClVQ12 interacted with ClVQ4,ClVQ11,ClVQ26,and ClWRKY51 but not with ClWRKY5.[Conclusion]ClVQ12 exhibits typical characteristics of the VQ gene family,localizes to the cell membrane and nucleus,and responds to multiple hormones and abiotic stresses.It interacts with ClVQs and ClWRKY51 and enhances drought tolerance in transgenic yeast.关键词
薏苡/ClVQ12/VQ基因家族/生物信息学分析/抗旱验证/蛋白互作Key words
Coix lacryma-jobi L./ClVQ12/VQ gene family/bioinformatics analysis/drought tolerance validation/protein interactions分类
农业科技引用本文复制引用
王永乐,路献勇,王红娟,王玉娇,朱加保..薏苡ClVQ12的表达模式、抗旱功能及互作蛋白分析[J].江西农业大学学报,2025,47(4):865-877,13.基金项目
国家科学自然基金青年项目(3240151950) Project supported by the Youth Fund of the National Natural Science Foundation of China(3240151950) 安徽省农业科学院人才引进项目(XXBS-202309)同时对本研究给予了资助,谨致谢意! （3240151950）
