青杄PwRhomboid基因表达特性及其抗旱作用  

The expression characteristics of PwRhomboid gene in Picea wilsonii and its effects on drought resistance

作　　者：杨沛欣 刘奎 李兴芬 汪晴晴 曹一博 张凌云[1] YANG Peixin;LIU Kui;LI Xingfen;WANG Qingqing;CAO Yibo;ZHANG Lingyun(School of Forestry,Beijing Forestry University,National Key Laboratory of Efficient Production of Forest Resources,State Key Laboratory of Forest Cultivation and Protection of Ministry of Education,Beijing 100083,China)

机构地区：[1]北京林业大学林学院,林木资源高效生产全国重点实验室,森林培育与保护教育部重点实验室,北京100083

出　　处：《陕西师范大学学报（自然科学版）》2024年第5期1-11,共11页Journal of Shaanxi Normal University：Natural Science Edition

基　　金：北京市自然科学基金(6222041);北京林业大学“5·5工程”科研创新团队项目(BLRC2023B08);北京林业大学大学生创新创业项目(X202310022018)。

摘　　要：通过对青杄转录组测序分析,筛选出干旱响应基因PwRhomboid,分析该基因在耐旱方面的功能特性,为揭示青杄的耐旱机理提供理论基础。采用生物信息学对PwRhomboid的氨基酸序列和同源基因进行比对;利用RT-qPCR技术分析PwRhomboid对不同非生物胁迫和激素的响应情况;通过瞬时转化烟草叶片检测PwRhomboid蛋白的亚细胞定位;获得拟南芥和马铃薯的PwRhomboid转基因株系并进行干旱表型验证。研究结果显示,PwRhomboid的氨基酸序列在N端与其同源物种的氨基酸序列具有较大差异,在C端相似度较高,保守性较好;其蛋白主要定位于细胞膜。PwRhomboid的表达量经干旱、低温和脱落酸诱导后发生有显著变化,在成熟叶中表达量最高,其能够提高转基因拟南芥和马铃薯的耐旱性。经干旱处理后,与对照组拟南芥野生型和pCM1205株系相比,PwRhomboid过表达株系PwRhomboid-L1和PwRhomboid-L2的存活率更高,叶绿素荧光最大光化学效率和实际光化学效率更高;在经聚乙二醇模拟干旱处理后,与野生型相比,过表达株系PwRhomboid-L3、PwRhomboid-L4的马铃薯苗高更高。结果表明PwRhomboid的表达量会受到干旱等逆境和激素的影响,过表达PwRhomboid能够提高拟南芥和马铃薯的耐旱性。By sequencing and analyzing the transcriptome of Picea wilsonii,the potential drought-related gene PwRhomboid was screened out,and its functional characteristics of drought resistance in Picea wilsonii were analyzed.Through bioinformatics analysis,the amino acid sequence and homologous gene of PwRhomboid were compared.The responses of PwRhomboid to different abiotic stresses and hormones were analyzed by RT-qPCR.Instantaneous transformation of tobacco leaves was conducted to detect the subcellular localization of PwRhomboid protein.PwRhomboid transgenic strains of Arabidopsis and potato were obtained and their drought phenotypes were verified.The results showed that the amino acid sequence of PwRhomboid has significant difference with those of its homologous species at N-terminus,but shows high similarity and good conservation at C-terminus.PwRhomboid protein is mainly localized on cell membrane.The expression level of PwRhomboid was significantly induced by drought,low temperature and abscisic acid,with the highest expression level observed in mature leaves.PwRhomboid can improve the drought resistance in transgenic Arabidopsis and potato.After drought treatment,compared with the control group Arabidopsis WT and pCM1205 strains,PwRhomboid overexpression strains PwRhomboid-L1 and PwRhomboid-L2 had higher survival rates,chlorophyll fluorescence F v/F m andΦPSⅡ.After polyethyleneglycol simulated drought treatment,the overexpression strains PwRhomboid-L3 and PwRhomboid-L4 resulted in higher potato seedling heights compared to the wild-type.The results showed that the expression level of PwRhomboid is affected by drought and hormones,and overexpression of PwRhomboid could improve the drought resistance of Arabidopsis and potato.

关 键 词：青杄 Rhomboid蛋白 耐旱性 基因表达 

分 类 号：S791.18[农业科学—林木遗传育种]