[关键词]
[摘要]
目的 探讨不同来源的酸枣Ziziphus jujuba var. spinosa果实及酸枣仁外观形态差异的分子机制及筛选影响其形态发育的核心基因。方法 以野生酸枣及嫁接酸枣的果实、酸枣仁为材料,利用表型组进行外观形态差异分析;利用Illumina HiSeq平台进行转录组测序,经过数据质控、组装及基因注释后对差异表达基因(differentially expressed genes,DEGs)进行分析及筛选;利用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)分析核心基因在不同组织部位,赤霉素(gibberellin,GA3)及干旱胁迫处理后的表达水平。结果 嫁接酸枣果实的横径、纵径及质量,酸枣仁纵径、仁厚及质量均分别显著高于野生来源酸枣果实及酸枣仁;转录组分析揭示嫁接酸枣与野生酸枣果实及酸枣仁之间的DEGs与对应激的反应、转录调控、植物激素信号转导及苯丙烷类物质生物合成有关。通过qRT-PCR筛选出WRKY53、WRKY40、TIFY9、NCED3、TSJT1、NRT3.1、BZIP9、TEN1、BXL1和LRX4等为影响酸枣果实和酸枣仁形态发育的核心DEGs。150 mg/L GA3处理后WRKY53、LRX4、NRT3.1及TIFY9显著上调,而BZIP9及NCED3显著下调;PEG-6000干旱胁迫下,WRKY53、WRKY40、TEN1、TSJT1及TIFY9基因表达显著上调,BZIP9及LRX4显著下调。结论 初步明确了嫁接酸枣与野生酸枣果实及酸枣仁之间形态差异的分子机制,并筛选出影响酸枣果实和酸枣仁形态发育的关键候选基因及其在不同胁迫条件下的表达模式,为酸枣的分子育种、环境适应性研究及资源定向改良提供了理论依据和关键靶点。
[Key word]
[Abstract]
Objective To explore the molecular mechanisms underlying the morphological differences in fruits and seeds of Ziziphus jujuba var. spinosa from different sources and to identify key genes influencing the morphological variation in Z. jujuba var. spinosa fruits and seeds. Methods Wild and grafted Z. jujuba var. spinosa fruits and seeds were used as materials. Phenotypic differences in their appearance were analyzed using a phenotypic group approach. Transcriptome sequencing was performed using the Illumina HiSeq platform. After data quality control, assembly, and gene annotation, differentially expressed genes (DEGs) were analyzed and screened. Quantitative real-time PCR (qRT-PCR) was used to assess the expression levels of key genes in different tissues, under gibberellin (GA3) and drought stress treatments. Results The transverse and longitudinal diameters, as well as the weight of grafted Z. jujuba var. spinosa fruits, were significantly larger than those of wild Z. jujuba var. spinosa fruits. Similarly, the longitudinal diameter, thickness, and weight of grafted Z. jujuba var. spinosa seeds were significantly higher than those of wild Z. jujuba var. spinosa seeds. Transcriptome sequencing revealed that DEGs between grafted and wild Z. jujuba var. spinosa fruits and seeds were related to stress response, transcriptional regulation, plant hormone signal transduction, and phenylpropanoid biosynthesis. Through qRT-PCR, core DEGs influencing the morphological development of Z. jujuba var. spinosa fruits and seeds were identified, including WRKY53, WRKY40, TIFY9, NCED3, TSJT1, NRT3.1, BZIP9, TEN1, BXL1, and LRX4. After treatment with 150 mg/L GA3, the expression levels of WRKY53, LRX4, NRT3.1, and TIFY9 were significantly upregulated, whereas BZIP9 and NCED3 were significantly downregulated. Under PEG-6000 drought stress, the expression levels of WRKY53, WRKY40, TEN1, TSJT1, and TIFY9 were significantly upregulated, while those of BZIP9 and LRX4 were significantly down-regulated. Conclusion This study preliminarily elucidated the molecular mechanisms underlying the morphological differences between grafted and wild Z. jujuba var. spinosa fruits and seeds. It also identified key candidate genes influencing the morphological development of fruits and seeds and their expression patterns under different stress conditions. These findings provide a theoretical basis and key targets for molecular breeding, environmental adaptability studies, and resource improvement of Z. jujuba var. spinosa.
[中图分类号]
R282.12
[基金项目]
中央本级重大增减支项目名贵中药资源可持续利用能力建设项目(2060302);陕西省重点研发计划(2024CY-JJQ-41);现代农业产业技术体系建设专项(CARS-21);咸阳市重点研发计划项目(L2024-ZDYF-ZDYF-SF-0041,L2024-ZDYF-ZDYF-SF-0035)