摘要
【目的】发掘调控黄瓜性别决定的关键基因及调控通路,为培育高产黄瓜品种提供理论依据,并为理解双子叶植物性别决定机制奠定基础。【方法】以黄瓜雌性系X8g(纯雌株)与普通系X8(雌雄株)近等基因系为材料,采用RNA-Seq技术对四叶一心期花芽分生组织进行转录组测序分析。通过差异表达分析、主成分分析和方差分解分析3种策略整合筛选关键基因。利用OrthoFinder鉴定黄瓜与拟南芥的直系同源基因,开展跨物种比较转录组分析,以探究调控网络的保守性与特异性。运用VIGS技术对核心候选基因CsACO2进行功能验证,采用qPCR检测基因沉默效率和特异性。使用DNA甲基化抑制剂5-氮杂胞苷处理雌性系植株验证表观遗传调控作用。【结果】在黄瓜雌性系与普通系之间共鉴定到197个差异表达基因,其中43个上调,154个下调,这些基因显著富集于花器官形态建成、α-亚麻酸(茉莉酸合成前体)代谢等通路。多维度分析共同确定了一些核心候选基因,包括已知的乙烯合成关键基因CsACO2和新发现的MADS-box家族转录因子AGL6、MADS4等。跨物种比较分析显示,黄瓜与拟南芥同源基因的表达模式总体相关性弱,但揭示了乙烯响应因子EIN3、NAC053转录因子靶基因集以及H3K27me3表观修饰在黄瓜性别调控中的潜在重要作用。功能验证实验证明,在雌性系中特异性沉默CsACO2能够诱导雄花和两性花的产生;而施用DNA甲基化抑制剂5-azacytidine同样能使雌性系植株开出雄花。【结论】系统揭示黄瓜性别决定是由乙烯信号通路主导,并可能由茉莉酸代谢、多类转录因子以及表观遗传协同作用的复杂分子网络。
【Objective】To identify the key genes and regulatory pathways that control sex determination in cucumber(Cucumis sativus L.),with the aim of providing a theoretical basis for breeding high-yield cucumber varieties and laying a foundation for understanding the mechanisms of sex determination in dicotyledonous plants.【Method】Near-isogenic lines of cucumber,the gynoecious line X8g and the monoecious line X8,were used as materials.Transcriptome sequencing(RNA-Seq)analysis was conducted on the apical meristems of flower buds at the four-leaf-one-heart stage.An integrated strategy combining differential expression analysis,principal component analysis(PCA),and variance decomposition analysis was employed to screen for key genes.OrthoFinder was used to identify orthologous genes between cucumber and Arabidopsis thaliana to perform a cross-species comparative transcriptomic analysis,investigating the conservation and specificity of the regulatory networks.The function of the core candidate gene,CsACO2,was validated using Virus-Induced Gene Silencing(VIGS),and its silencing efficiency and specificity were assessed by qPCR.The role of epigenetic regulation was verified by treating the gynoecious line with the DNA methylation inhibitor 5-azacytidine.This study investigated key genes and pathways in cucumber sex determination using RNA-Seq analysis of gynoecious and monoecious lines.【Result】A total of 197 differentially expressed genes were identified between the gynoecious and monoecious lines,with 43 being upregulated and 154 downregulated.These genes were significantly enriched in pathways such as floral organ morphogenesis andα-linolenic acid(a precursor for jasmonic acid synthesis)metabolism.The multi-dimensional analysis identified a set of core candidate genes,including the known key ethylene synthesis gene CsACO2 and newly discovered MADS-box transcription factors like AGL6 and MADS4.The cross-species comparative analysis showed a weak overall correlation in expression patterns of orthologous genes but revealed the potential importance of target gene sets of the ethylene response factor EIN3,the NAC053 transcription factor,and H3K27me3 epigenetic modification in regulating cucumber sex.Functional validation experiments confirmed that the specific silencing of CsACO2 induced the formation of male and bisexual flowers in the gynoecious line.Similarly,treatment with 5-azacytidine also caused the gynoecious line to produce male flowers.【Conclusion】This study systematically reveals that cucumber sex determination is governed by a complex molecular network dominated by the ethylene signaling pathway,potentially involving jasmonic acid metabolism,various transcription factors,and epigenetic modifications.
作者
杨宗辉
李利斌
孟昭娟
高天
祝利霞
杜海梅
董伟伟
曹齐卫
YANG Zong-hui;LI Li-bin;MENG Zhao-juan;GAO Tian;ZHU Li-xia;DU Hai-mei;DONG Wei-wei;CAO Qi-wei(Shandong Key Laboratory of Bulk Open-field Vegetable Breeding,Ministry of Agriculture and Rural Affairs Key Laboratory of Huang Huai Protected Horticulture Engineering,Institute of Vegetables,Shandong Academy of Agricultural Sciences,Jinan 250100;Chengdu Agricultural Technology Station,Chengdu 610041;School of Biological Science and Technology,University of Jinan,Jinan 250022;Agricultural Tech-nology Station,Fushan Town,Ningyang County,Tai’an City,Shandong Province,Tai’an 271408;Yinan County Agricultural Technology Center,Linyi City,Shandong Province,Linyi 276300)
出处
《生物技术通报》
北大核心
2025年第12期139-155,共17页
Biotechnology Bulletin
基金
国家自然科学基金项目(32200506,32402582)
国家重点研发计划项目(2023YFD1201503)
国家现代农业产业技术体系建设专项资金项目(CARS-23-G13)
泰安市农业良种工程项目(2024NYLZ13)
山东省科技型中小企业创新能力提升工程(2024TSGC0122)
山东省农业科学院农业创新工程(CXGC2025B08,CXGC2025C08)。
关键词
黄瓜
性别决定
比较转录组学
乙烯信号通路
CsACO2
表观修饰
病毒诱导的基因沉默
拟南芥
Cucumis sativus L.
sex determination
comparative transcriptomics
ethylene signaling pathway
CsACO2
epigenetic modification
virus-induced gene silencing(VIGS)
Arabidopsis thaliana
