摘要
【目的】鉴定葡萄(Vitis vinifera L.)VvOMTs基因家族启动子对激素和干旱信号的响应,阐明VvOMTs基因家族的启动子功能,为干旱条件下VvOMTs调控3-烷基-2-甲氧基吡嗪香气物质合成提供理论依据。【方法】采用生物信息学方法分析其蛋白理化性质、染色体定位、基因结构和启动子顺式作用元件;通过农杆菌介导的烟草瞬时转化体系验证VvOMT2和VvOMT3基因启动子对脱落酸(abscisic acid, ABA)、茉莉酸酸甲酯(methyl jasmonate, MeJA)和甘露醇的响应。【结果】葡萄VvOMTs基因家族编码序列CDS区介于1 062-1 110 bp,蛋白氨基酸数介于353-369 aa;4个VvOMTs在葡萄第3和第12号染色体上均匀分布;亚细胞定位预测结果表明,VvOMTs蛋白存在于细胞质;启动子顺式作用元件分析表明,VvOMTs基因可能受到光照、多种植物激素和干旱胁迫的诱导;瞬时转化本氏烟草GUS染色、GUS基因定量和酶活活性测定结果表明,在ABA、MeJA和甘露醇的诱导下,转化VvOMT2和VvOMT3启动子的烟草叶片均呈现蓝色反应;VvOMT2和VvOMT3的GUS基因表达水平呈现先升高后下降趋势,且显著上调16.80和77.73倍、14.09和34.15倍、30.77和36.40倍;VvOMT2和VvOMT3的GUS酶活性在ABA和MeJA处理下呈先升高后下降,且显著上调16.62和21.69倍、4.50和3.00倍,甘露醇处理下出现“双峰”变化趋势,显著上调15.13和18.64倍。【结论】从葡萄基因组中共鉴定出4个调控甲氧基吡嗪合成的甲氧基转移酶基因(VvOMT1-4),VvOMT2和VvOMT3基因启动子参与干旱、ABA和MeJA响应,并受其调控。
【Objective】To characterize the response of grape(Vitis vinifera L.)VvOMTs gene family promoters to hormone and drought signals,to elucidate the promoters functions of the VvOMTs gene family,and to provide a theoretical basis for the regulation of the synthesis of 3-alkyl-2-methoxypyrazine aroma substances by VvOMTs under drought conditions.【Method】Bioinformatics methods were used to analyze the physicochemical properties of the protein,chromosomal localization,gene structure,and promoters cis-acting elements;and the promoters of the VvOMT2/3 gene was verified by an Agrobacterium-mediated transient transformation system in tobacco in response to abscisic acid(ABA),methyl jasmonate(MeJA),and mannitol.【Result】The CDS region of the coding sequence of the grape VvOMTs gene family ranged from 1062‒1110 bp,and the number of protein amino acids ranged in353-369 aa;the four VvOMTs were evenly distributed on the two grape chromosome 3 and 12;subcellular localization predictions indicated that the VvOMTs proteins existed in the cytoplasm;the promoters cisacting element analysis indicated that the VvOMTs genes might be subjected to induction by light,multiple plant hormones and drought stress.The results of GUS staining,gene quantification and enzyme activity assay of transiently transformed tobacco showed that under the induction of ABA,MeJA and mannitol,the tobacco leaves transformed with VvOMT2/3 promoters showed a blue color reaction,and the expression of the GUS gene of VvOMT2/3 significantly increased and showed a tendency of increasing and then decreasing,and the GUS enzyme activity of VvOMT2/3 significantly increased.The GUS enzyme activity of VvOMT2/3 significantly increased and showed a trend of increasing and then decreasing under hormone treatment,and a bimodal trend under drought treatment.【Conclusion】A total of four methoxytransferase genes(VvOMT1-4)regulating methoxypyrazine synthesis are identified from the grape genome,and the VvOMT2 and VvOMT3 gene promoters are involved in and regulated by drought,ABA and MeJA responses.
作者
薛晓斌
宁琳
周鱼
刘虹君
高照祖
王振平
李栋梅
XUE Xiao-bin;NING Lin;ZHOU Yu;LIU Hong-jun;GAO Zhao-zu;WANG Zhen-ping;LI Dong-mei(School of Life Sciences,Ningxia University,Yinchuan 750021;School of Wine&Horticulture,Ningxia University,Yinchuan 750021)
出处
《生物技术通报》
北大核心
2025年第12期168-176,共9页
Biotechnology Bulletin
基金
宁夏重点研发(引才专项)(2023BSB03032)。
