西北植物学报2024，Vol.44Issue(3)：430-442,13.DOI:10.7606/j.issn.1000-4025.20230468
小桐子早期光诱导蛋白基因ELIP克隆及其与靶向miRNA的互作与低温下共表达分析
Cloning of early light-induced protein gene ELIP and its interaction and co-expression analysis with targeting miRNA in Jatropha curcas under low temperature
摘要
Abstract
[Objective]As a kind of thermophilic and chilling-sensitive plants,low temperature severely af-fected the growth,development,geographical distribution,and yield of Jatropha curcas L..Previous studies showed that chill-hardening at 12 ℃ significantly enhanced the chilling resistance of J.curcas.The early light-inducible protein(ELIP)gene in J.curcas was a highly responsive gene to low tempera-ture.The study aimed to explore the role of JcELIP in response to low temperature in J.curcas,to com-prehensively understand the structure,regulatory mechanisms,evolutionary relationships of JcELIP and its interaction with miRNAs,and to provide an important candidate gene resource for subsequent molecu-lar breeding of cold resistance J.curcas.[Methods]This study cloned the JcELIP gene from J.curcas by RT-PCR and conducted a comprehensive bioinformatics analysis.The expression of JcELIP gene in the roots,stems,and leaves,as well as during the chill-hardening at 12 ℃,were analyzed by RT-qPCR.The miRNAs interacting with JcELIP was identified,and a co-expression analysis was conducted during the chill-hardening at 12 ℃.[Results]The results showed that the complete open reading frame(ORF)of JcELIP was 585 bp,encoding 194 amino acids.The size of the protein was 2.04 kD with a theoretical iso-electric point of 9.59.It was a stable hydrophobic alkaline protein,with 3 hydrophobic transmembrane helices.The tertiary structure mainly consisted of α-helices and irregular coils and possessed chlorophyll a/b binding sites.Cis-acting element prediction showed that JcELIP had hormone response elements such as abscisic acid.Evolutionary analysis showed that the JcELIP from J.curcas had the highest homology with MeELIP from Manihot esculenta.RT-qPCR analysis revealed that under normal growth conditions,there was no significant difference in the expression of JcELIP in the roots,stems,and leaves of J.cur-cas.During chill-hardening at 12 ℃,the expression of JcELIP in the leaves was quickly up-regulated,reaching 64.8 times of the control at 48 h,indicating that JcELIP was involved in the response and adap-tation to cold stress.Based on the degradome data of J.curcas,eight miRNAs,including miR390-x,miR6476-x,and novel-m0090-3p,were identified as having regulatory effects on the expression of JcELIP.Co-expression analysis showed that the expression of JcELIP was significantly negatively regu-lated by miR390-x and novel-m0090-3p during the chill-hardening at 12 ℃.[Conclusion]JcELIP highly responded to low temperatures stress and interacted with miRNAs,showing that it played an important role in response to low temperature stress in J.curcas.关键词
小桐子/早期光诱导蛋白(ELIP)基因/冷锻炼/表达分析/miRNA互作Key words
Jatropha curcas/early light-inducible protein(ELIP)gene/chill-hardening/expression anal- ysis/miRNA interaction分类
农业科技引用本文复制引用
陈建南,朱晓琴,吕宇婧,龚明..小桐子早期光诱导蛋白基因ELIP克隆及其与靶向miRNA的互作与低温下共表达分析[J].西北植物学报,2024,44(3):430-442,13.基金项目
国家自然科学基金项目(31860062,31460059) （31860062,31460059）
