中国农业大学学报2025，Vol.30Issue(6)：47-56,10.DOI:10.11841/j.issn.1007-4333.2025.06.05
陆地棉GhASMT17基因的克隆及功能分析
Cloning and functional analysis of GhASMT17 gene in Gossypium hirsutum
摘要
Abstract
In order to explore the function of Gossypium hirsutum N-acetyl serotonin methyltransferase gene(GhASMT1 7)in cotton resistance to Verticillium wilt,the GhASMT17 gene was cloned from the upland cotton standard system'TM-1'and sequenced.The online bioinformatics analysis was adopted to predict its physical,chemical,structural,evolutionary characteristics and promoter elements of the encoded protein.qRT-PCR was used to detect its tissue expression specificity and expression pattern after Verticillium wilt infection,and virus-induced gene silencing(VIGS)technology was used to analyze the function of the GhASMT17 gene.The results show that:1)The length of the open reading frame(ORF)of the cloned cotton GhASMT17 gene is 726 bp,encoding 241 amino acids.Its molecular weight is 26 650.66 U,theoretical PI is 4.80,the instability index is 26.62,and average hydropathic index is 0.022,indicating that it is an acidic,stable and hydrophilic protein and may be localized in chloroplasts.2)The promoter sequence of GhASMT17 gene contains abscisic acid response element(ABRE)andlow temperature stress response element(LTR).3)The qRT-PCR results show that after verticillium wilt infection,compared with disease control(CK)the expression levels of GhASMT17 gene in plants inoculated with Verticillium wilt(V991)were significantly different at 24 and 48 h(P＜0.05).4)The VIGS results show that the resistance of GhASMT17-silenced plants to Verticillium wilt of cotton is significantly reduced(P＜0.05),the vascular bundles are severely browned,and in the stem segment recovery culture test,the number of stem segments growing fungal hyphae increased by 40％.In conclusion,the GhASMT17 plays a positive regulatory role in cotton resistance to Verticillium wilt.关键词
棉花/GhASMT17/基因克隆/表达模式分析/VIGSKey words
cotton/GhASMT17/gene cloning/expression pattern analysis/VIGS分类
生物科学引用本文复制引用
马鑫,郭云龙,侯奥成,陈梦,李月,代培红..陆地棉GhASMT17基因的克隆及功能分析[J].中国农业大学学报,2025,30(6):47-56,10.基金项目
新疆维吾尔自治区自然科学基金(2022D01A184,2023D01E03) （2022D01A184,2023D01E03）
新疆农业大学创新团队开放课题(ITB202106) （ITB202106）
新疆维吾尔自治区大学生创新项目(S202310758061) （S202310758061）
新疆维吾尔自治区"天山英才"培养计划(2023SNGGNT063) （2023SNGGNT063）
