摘要
【目的】CYP707A是细胞色素P450家族的亚家族成员,所编码的ABA8'-羟化酶是ABA分解代谢的关键酶,对植物生长发育起着重要作用。鉴定葡萄CYP707A基因家族,可为进一步研究葡萄果实成熟机理和生物育种技术提供理论依据。【方法】利用生物学信息方法分析葡萄中CYP707A家族基因的基因结构、蛋白基序、染色体定位、共线性、系统进化关系及启动子区顺式作用元件等,并利用RT-q PCR分析VvCYP707A基因在葡萄不同品种(早熟和中熟)的时空表达模式、组织表达模式以及对外源ABA处理的响应模式,并利用果实瞬时表达技术对Vitvi07g01751进行功能验证。【结果】葡萄基因组鉴定出5个VvCYP707As基因,分布于5条染色体,其中Vitvi02g01269、Vitvi07g01751和Vitvi18g00792为片段复制基因。CYP707A基因家族高度保守,其中,基因结构和蛋白基序相似,亚细胞定位均位于内质网;VvCYP707A启动子区域富含与生长发育和激素响应相关的顺式作用元件,均含有ABA激素响应元件;通过RT-qPCR检测基因的表达模式发现,VvCYP707A基因具有时空特异性和组织特异性,主要在果实的幼果期与膨大期和茎、叶组织中高表达,其中Vitvi07g01751基因在早熟品种‘甬早红’膨大期果实中特异性高表达,且在两个品种间的表达模式一致,随着果实的成熟其表达先升高后降低。外源ABA处理下VvCYP707A分为两种表达模式,在浆果肉中外源ABA可诱导5个VvCYP707A基因高表达,在浆果皮中会显著抑制VvCYP707A基因的表达;在葡萄果实中瞬时表达Vitvi07g01751,葡萄的糖度积累缓慢,显著增加编码ABA的受体VvPYL4基因表达量,推测了Vitvi07g01751介导ABA调控葡萄果实成熟的机制。【结论】研究结果揭示了葡萄Vitvi07g01751在果实成熟中起着重要作用。
【Objective】CYP707A is a subfamily member of cytochrome P450 family,encoding ABA8'-hydroxylase,which is a key enzyme in ABA catabolism and plays an important role in plant growth and development.Identification of grape(Vitis vinifera L.)CYP707A gene family may provide theoretical basis for further study of grape fruit ripening mechanism and biological breeding techniques.【Method】Bioinformatics methods were used to analyze the gene structure,protein motif,chromosome localization,collinearity,phylogenetic relationship and cis-acting elements of promoter region of CYP707A family gene in the grape.RT-qPCR was used to analyze the spatiotemporal expression patterns of VvCYP707As gene in tissues,different grape varieties(early and medium maturity variety)and in response to the exogenous ABA treatment were analyzed by.Fruit transient transfection was applied to verify the function of Vitvi07g01751.【Result】There were five VvCYP707As genes identified in grape genome,located on five chromosomes,among which Vitvi02g01269,Vitvi07g01751 and Vitvi18g00792 were fragment replicators.The CYP707A gene family had a high degree of conservation,sharing similarities in gene structure and protein motifs,and all subcellular localization were predominantly localized to the endoplasmic reticulum.The cis-acting elements of growth,development and hormone response were significantly enriched in the promoter regions of the CYP707A gene family,containing ABA hormone-corresponding elements.Furthermore,the RT-qPCR analysis demonstrated that VvCYP707A genes demonstrated both spatiotemporal-and tissue-specific expression patterns,with elevated expression levels during the early and expansion stages of fruit development as well as in stem and leaf tissues.Notably,Vitvi07g01751 gene was highly expressed in the fruits of the early maturity variety‘Yong Zaohong’,and the expression pattern was consistent between these two varieties.The expression pattern of Vitvi07g01751first increased and then decreased with fruit development.Under exogenous ABA treatment,VvCYP707A showed two distinct expression patterns:it induced the high expression of 5 VvCYP707A genes in the grape flesh while significantly suppressing the expressions of these genes in the grape pericarp.Furthermore,upon transient expression of Vitvi07g01751 in the grape flesh,there was a slow accumulation of sugar content,accompanied by a significant upregulation of the ABA receptor VvPYL4 gene,suggesting that Vitvi07g01751 mediated ABA regulation of grape fruit ripens.【Cconclusion】Vitvi07g01751 plays an important role in fruit ripening of grape.
作者
龚丽丽
余花
杨杰
陈天池
赵双滢
吴月燕
GONG Li-li;YU Hua;YANG Jie;CHEN Tian-chi;ZHAO Shuang-ying;WU Yue-yan(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 200120;College of Biology and Environment,Zhejiang Wanli University,Ningbo 315100)
出处
《生物技术通报》
CAS
CSCD
北大核心
2024年第2期160-171,共12页
Biotechnology Bulletin
基金
宁波市科技创新2025现代种业重大专项(2019B10015)
浙江省重点研发计划(2021C02053)
生物工程一流学科创新项目(CX2021015)。
