摘要
【目的】聚焦于森林草莓AHL基因家族,旨在分析其理化性质,基因启动子顺式元件及基因表达模式,并验证FveTEK(FveAHL16)的生物学功能。【方法】对森林草莓AHL与拟南芥AHL家族构建物种发育树,分析森林草莓AHL家族的理化性质,保守结构分析与启动子顺式元件,并构建FveTEK-SRDX转录抑制表达载体,稳定转化森林草莓稳定以验证FveTEK基因功能。【结果】森林草莓中共有12个AHL成员(FveAHL),分为2类Clade-A和Clade-B,不均匀分布在7条染色体上。保守基序和基因结构分析均表明,FveAHLs在进化过程中具有高度保守性。启动子顺式作用元件分析显示,FveAHL基因主要参与光响应、生长发育、非生物胁迫响应及植物激素调控等相关反应。克隆获得FveTEK基因长度为801 bp,编码266个氨基酸残基,在N端第89~202氨基酸残基处含有一个保守的DUF296结构域。亚细胞定位结果显示FveTEK蛋白定位于细胞核中。森林草莓遗传转化,获得FveTEK-SRDX转基因植株。qRT-PCR检测结果表明,FveTEK在FveTEK-SRDX株系中的表达均被下调,是WT的0.09~0.6倍。表型观察和花粉活力检测结果显示,FveTEK-SRDX株系的花粉活力降低,自花授粉的大部分果实不能正常发育膨大。【结论】本研究鉴定了森林草莓AHL基因家族,研究了FveAHLs基因启动子顺式元件及其在不同组织的表达,同时,验证了FveTEK抑制表达会导致果实败育。这些结果可为森林草莓雄性不育育种开发提供重要理论依据。
【Objective】This study focused on AHL gene family of Fragaria vesca and aimed to analyze its physicochemical properties,gene promoter cis-element and gene expression pattern analysis,and to validate the biological function of FveTEK(FveAHL16).【Method】A phylogenetic tree was constructed for the AHL gene family of both Fragaria vesca and Arabidopsis thaliana.The physicochemical properties,conserved structural analysis,and promoter cis-elements of the AHL gene in Fragaria vesca were examined.Additionally,an FveTEK-SRDX transcription repressor expression vector was constructed,and the functionality of the FveTEK gene was validated through stable genetic transformation in Fragaria vesca.【Result】In Fragaria vesca,there are a total of 12 AHL family members,categorized into two clades,Clade-A and Clade-B,unevenly distributed across seven chromosomes.Both conservative motif and gene structure analyses indicate that FveAHLs exhibit a high degree of conservation throughout evolution.Analysis of promoter cis-acting elements suggested that FveAHL genes are primarily involved in light response,growth and development,abiotic stress responses,and plant hormone regulation.The FveTEK gene spans 801 bp and encodes 266 amino acid residues,featuring a conserved DUF296 domain spanning amino acids 89-202 at the N-terminus.Subcellular localization results indicated that the FveTEK protein is localized within the nucleus.Through genetic transformation in Fragaria vesca,FveTEK-SRDX transgenic plants were obtained.Further qRT-PCR analysis revealed that the expression of FveTEK in the FveTEK-SRDX lines was downregulated,ranging from 0.09 to 0.6 times that of the wild type(WT).Phenotype observation and pollen viability testing of the FveTEK-SRDX lines showed decreased pollen viability,with most self-pollinated fruits failing to develop and expand normally.【Conclusion】This study identified the AHL gene family in Fragaria vesca,studied the ciselements of the FveAHLs gene promoter and its expression in different tissues,and verified that the inhibition of FveTEK expression leads to fruit abortion.These results can provide important theoretical basis for the development of male sterility breeding in Fragaria vesca.
作者
黄凯
刘和青
吴润琴
林丽旋
王菲
山室千鹤子
Huang Kai;Liu Heqing;Wu Runqin;Lin Lixuan;Wang Fei;Chizuko Yamamuro(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;College of Future Technology,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;College of JunCao Science and Ecology,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)
出处
《东南园艺》
2025年第2期128-140,共13页
Southeast Horticulture
作者简介
通讯作者:山室千鹤子(1973-),女,教授,博士,主要从事果树育种与生物技术研究,E-mail:c.yama⁃muro@fafu.edu.cn;黄凯(1994-),男,硕士研究生,主要从事果树育种与生物技术研究,E-mail:kevinwong1994@163.com。
