| 摘要: |
| 【目的】通过高通量RNA测序技术，初步探究云南松腋芽生长分子机制，为后续克隆云南松腋芽重要基因及其功能验证研究奠定基础。【方法】以无腋芽与有腋芽2种生长状态的云南松幼苗为研究材料，构建其茎部组织cDNA文库并进行转录组测序，从中筛选出差异表达基因（DEGs），利用生物信息学方法对DEGs进行GO和KEGG分析；从激素信号转导、淀粉和蔗糖代谢与糖酵解/糖异生代谢通路中选出6个与腋芽萌发和生长相关的基因，并采用RT-qPCR技术进行验证分析。【结果】比较无腋芽与有腋芽云南松的转录组文库，共鉴定到1 420个DEGs，其中上调基因与下调基因分别有524和896个。GO富集分析结果表明，在细胞组分类别中，差异基因数量最多的前3个亚类从多到少依次为胞外区、膜和细胞解剖实体部分；在分子功能中，注释为催化活性的差异基因数量最多，其次为电子传递活性、氧化还原酶和抗氧化活性；在生物学过程中，差异基因主要集中注释在细胞杀伤和刺激响应过程中。KEGG分析结果表明，云南松基因可能通过玉米素生物合成及糖酵解/糖异生显著富集通路参与对腋芽生长的响应。此外，在激素信号转导及淀粉和蔗糖代谢、糖酵解/糖异生代谢通路上共发现12个DEGs，从中筛选出6个候选基因，对其进行RT-qPCR验证，显示各基因表达趋势与转录组分析结果一致。【结论】通过分析植物激素代谢及糖代谢通路上相关基因表达的变化，筛选出6个与腋芽生长发育相关的基因，可用于进一步揭示云南松腋芽生长机制及云南松的遗传改良。 |
| 关键词: 云南松；腋芽 差异表达基因；转录组测序 |
| DOI：10.13207/j.cnki.jnwafu.2025.02.006 |
| 分类号: |
| 基金项目:云南省农业联合专项重点项目（202301BD070001-152）；云南省农业联合专项面上项目（202301BD070001-035）；国家自然科学基金项目(32360381) ；云南省万人计划青年拔尖人才项目（YNWR-QNBJ-2019-075）；云南省研究生导师团队建设项目(2022-97) |
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| Analysis of differentially expressed genes in the growth and development of Pinus yunnanensis axillary buds based on transcriptome sequencing |
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ZHOU Chiyu, KONG Di, XU Junfei, ZHENG Chaofan, SUN He, LI Ruilian, CHEN Lin, CAI Nianhui, XU Yulan, TANG Junrong
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Key Laboratory for Forest Resources Conservation and Utilization in the Southwest Mountains of China,Ministry of Education,Key Laboratory of Biodiversity Conservation in Southwest China,State Forestry Administration,Southwest Forestry University,Kunming,Yunnan 650224,China
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| Abstract: |
| 【Objective】This study employed high-throughput RNA sequencing technology to preliminarily explore the molecular mechanisms of axillary bud growth in Pinus yunnanensis for laying a foundation for subsequent cloning and functional validation of significant genes associated with the axillary buds of P.yunnanensis.【Method】Seedlings of P.yunnanensis with and without axillary buds were used as research materials.A cDNA library of their stem tissues was constructed and transcriptome sequencing was conducted.Differential Expression Genes (DEGs) were screened,and bioinformatics methods were applied for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of these DEGs.Six genes related to axillary bud germination and growth were selected from the hormone signaling,starch,sucrose metabolism and glycolysis/gluconeogenesis pathways,and their expression was validated using RT-qPCR technology.【Result】In comparing the transcriptome libraries of P.yunnanensis with and without axillary buds,a total of 1 420 DEGs were identified,among which 524 were upregulated and 896 were downregulated.GO enrichment analysis revealed that the top three subcategories with the most differentially expressed genes in the cellular component category were the extracellular region,membrane,and cellular anatomical entity from most to least.In terms of molecular function,differentially expressed genes annotated for catalytic activity was the most numerous,followed by electron transfer activity,oxidoreductases and antioxidant activities.In biological processes,the differentially expressed genes were primarily annotated in cell killing and response to stimulus processes.KEGG analysis suggested that the genes of P.yunnanensis may participate in axillary bud growth response through significantly enriched pathways of zeatin biosynthesis and glycolysis/gluconeogenesis.Moreover,twelve DEGs were identified in the hormone signaling pathway,starch,sucrose metabolism,and glycolysis/gluconeogenesis pathways,from which six candidate genes were selected for RT-qPCR validation,showing expression trends of each gene consistent with the transcriptome analysis results.【Conclusion】By analyzing the changes in the expression of genes related to plant hormone metabolism and carbohydrate metabolism pathways,six genes associated with the growth and development of axillary buds were identified.These genes may be used to further elucidate the growth mechanisms of axillary buds in P.yunnanensis and contribute to the genetic improvement of this species. |
| Key words: Pinus yunnannensis axillary bud differentially expressed genes (DEGs) transcriptome sequencing |
