葡萄VvZF-HD11基因克隆及表达特性分析  

Cloning and Expression Characteristics Analysis of VvZF-HD11 Gene in Grape(Vitis vinifera)

作　　者：张瑶瑶 吕晓元 赵方贵[1] 车永梅[1] 刘新[1] 侯丽霞[1] ZHANG Yao-Yao;LYU Xiao-Yuan;ZHAO Fang-Gui;CHE Yong-Mei;LIU Xin;HOU Li-Xia(Key Lab of Plant Biotechnology in Universities of Shandong Province,College of Life Science,Qingdao Agricultural University,Qingdao 266109,China)

机构地区：[1]青岛农业大学生命科学学院山东省高校植物生物技术重点实验室,青岛266109

出　　处：《农业生物技术学报》2025年第3期547-560,共14页Journal of Agricultural Biotechnology

基　　金：山东省农业良种工程项目(2022LZGC0Y019);国家自然科学基金面上项目(31872082);山东省自然科学基金面上项目(ZR2023MC074);青岛市自然科学基金原创探索项目(23-2-1-178-zyyd-jch)。

摘　　要：锌指同源域(zinc finger-homeodomain, ZF-HD)蛋白是植物特有的一类转录因子,主要包括锌指结构域(zinc finger, ZF)和同源结构域(homeodomain, HD)。ZF-HD家族可通过与DNA结合调节靶基因的表达模式,从而影响植物的生长发育过程。本研究以葡萄(Vitis vinifera)品种'左优红'叶片为材料,克隆得到VvZF-HD11基因(GenBank No. PP735488),其编码区为969 bp,编码322个氨基酸。该基因编码的蛋白保守结构域为N端的ZF (72~127 aa)和位于C端的HD (202~258 aa)。氨基酸序列比对和系统发育树分析表明,VvZF-HD11与野生烟草(Nicotiana tabacum) NaZF-HD9同源性最高,亲缘关系最近。进一步通过qPCR分析了VvZF-HD11在多种非生物胁迫下的表达特性,结果显示,VvZF-HD11基因受多种逆境胁迫因子和信号分子的诱导表达。其中,低温和水杨酸(salicylic acid, SA)处理下VvZF-HD11基因表达量在3h最高,高温和H2O2处理后表达量在6 h变化最显著,盐胁迫和H2S、脱落酸(abscisic acid, ABA)处理后表达量12 h最高,干旱胁迫处理后9 h达到峰值。将VvZF-HD11瞬时转化葡萄叶片,经45℃高温处理4 h,发现过表达VvZF-HD11提高了葡萄叶片抵御高温的能力。检测生理指标和高温胁迫相关基因的表达量发现,VvZF-HD11能够通过提高葡萄叶片的抗氧化酶活性,减轻细胞膜氧化损伤,促进高温胁迫相关基因的表达,提高葡萄的抗高温能力。综合以上结果推测,VvZF-HD11可能参与调控葡萄响应逆境胁迫的过程。本研究为葡萄耐高温品种的遗传改良提供了理论基础。Zinc finger-homeodomain(ZF-HD)proteins are plant-specific transcription factors composed of a zinc finger(ZF)domain and a homeodomain(HD).The ZF-HD family plays a crucial role in plant growth and development by regulating the expression of target genes through binding to their promoters.In this study,the VvZF-HD11 gene was isolated from the leaves of the grape(Vitis vinifera)variety'Zuoyouhong'and assigned the GenBank accession number PP735488.The full length of the coding region of this gene was 969 bp,encoding 322 amino acids.The conserved domains of the VvZF-HD11 protein were the zinc finger domain ZF at the N-terminus(72~127 aa)and the homeodomain domain HD at the C-terminus(202~258 aa).Comparison of the amino acid sequence and phylogenetic analysis indicated that VvZF-HD11 shares the highest homology with wild tobacco(Nicotiana tabacum)NaZF-HD9.Further analysis through qPCR showed that the expression of the VvZF-HD11 gene was induced by various abiotic stress factors and signaling molecules.Specifically,VvZF-HD11 expression peaked under specific conditions,such as low temperature and salicylic acid(SA)induction at 3 h,heat stress and H2O2 treatment at 6 h,salt stress,H2S and abscisic acid(ABA)treatment at 12 h,and drought stress at 9 h.Moreover,transient transformation of VvZF-HD11 into grape leaves followed by heat stress at 45℃for 4 h demonstrated that overexpression of VvZF-HD11 enhanced the leaves'ability to resist heat stress.By detecting physiological indicators and the expression levels of genes related to heat stress,it was found that VvZF-HD11 could enhance the antioxidant enzyme activity of grape leaves,reduce membrane oxidative damage,and promote the expression of genes related to heat stress,thereby improving the plant's ability to resist heat stress.Based on the above results,it was speculated that VvZF-HD11 might be involved in regulating the stress response of grape.This study establishes a foundational theoretical framework for the genetic enhancement of grape varieties exhibiting tolerance

关 键 词：葡萄 VvZF-HD11 基因克隆 表达特性 

分 类 号：S663.1[农业科学—果树学]