基于转录组测序的红肉火龙果淀粉降解相关基因挖掘及表达分析  

Mining of starch-degrading related genes in red-fleshed dragon fruit and expression analysis based on transcriptome sequencing

作　　者：魏治远 晏霜 解璞[2] 郑乾明 WEI Zhi-yuan;YAN Shuang;XIE Pu;ZHENG Qian-ming(College of Life Sciences,Guizhou University/Institute of Agricultural Bioengineering/Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),Guiyang,Guizhou 550025,China;Institute of Pomology Science,Guizhou Academy of Agricultural Sciences,Guiyang,Guizhou 550006,China;Guizhou Academy of Agricultural Sciences/Key Laboratory of Crop Gene Resources and Germplasm Innovation in Karst Mountainous Area,Ministry of Agriculture and Rural Affairs,Guiyang,Guizhou 550006,China)

机构地区：[1]贵州大学生命科学学院/农业生物工程研究院/山地植物资源保护与种质创新教育部重点实验室,贵州贵阳550025 [2]贵州省农业科学院果树科学研究所,贵州贵阳550006 [3]贵州省农业科学院/农业农村部喀斯特山区作物基因资源与种质创新重点实验室,贵州贵阳550006

出　　处：《南方农业学报》2025年第2期553-562,共10页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(32060674);贵州省高层次创新型人才项目(黔科合平台人才-GCC〔2022〕025-1);贵州省科技计划项目(黔科合中引地〔2023〕033)。

摘　　要：【目的】挖掘红肉火龙果淀粉降解相关基因并分析其在果实不同发育时期的表达模式,为阐明红肉火龙果果实成熟过程中的淀粉降解途径提供理论依据。【方法】供试红肉火龙果品种为软枝大红,选取4个不同发育时期(花后19 d、花后22 d、花后25 d和花后28 d)果实进行转录组测序,以差异倍数(Fold Change)≥2且错误发现率(FDR)<0.01为标准,筛选不同发育时期果实间的差异表达基因(DEGs),进行GO功能注释和KEGG信号通路富集分析,通过实时荧光定量PCR验证转录组测序数据的可靠性并分析候选基因的表达模式。【结果】经转录组测序共获得22.84 Gb有效数据(Clean data),共获得76618317条有效序列(Clean reads),GC含量为44.78%~46.83%,Q20为98.89%~99.74%,Q30为96.73%~98.53%。与花后19 d相比,花后28 d共有4194个DEGs,其中1728个上调表达,2466个下调表达。GO功能注释分析结果表明,花后28 d与花后19 d比较组的DEGs注释到生物学过程、细胞成分和分子功能3个类别,在生物学过程中,涉及细胞过程的DEGs数量最多,其次是代谢过程。KEGG信号通路富集分析结果表明,花后28 d与花后19 d比较组的DEGs在淀粉和蔗糖代谢通路显著富集。筛选出红肉火龙果淀粉降解途径的10个相关基因,分别为葡聚糖水激酶(GWD)基因(HU02G00177)、磷酸葡聚糖水激酶(PWD)基因(HU09G02021)、磷酸葡聚糖磷酸酶(SEX)基因(HU07G01227)、β-淀粉酶(BAM)基因(HU10G00833、HU10G00777)、α-淀粉酶(AMY)基因(HU05G01323、HU02G00293、HU02G01323)、异淀粉酶(ISA)基因(HU11G01417)、歧化酶(DPE)基因(HU09G01260)。实时荧光定量PCR检测结果表明,10个淀粉降解基因的表达趋势与转录组测序数据一致。HU02G00177、HU07G01227、HU10G00833、HU10G00777、HU02G00293和HU11G01417基因相对表达量在果实发育后期均高于果实发育前期。【结论】红肉火龙果淀粉降解相关基因在果实不同发育时期的表达模式具有差异,其�【Objective】To explore genes related to starch degradation in red-fleshed dragon fruit and analyze their expression patterns at different developmental stages of the fruit,which could provide theoretical basis for elucidating the starch degradation pathway in the ripening process of red-fleshed dragon fruit.【Method】The tested red-fleshed dragon fruit variety was Ruanzhidahong.Four different developmental stages(19 d after flowering,22 d after flowering,25 d after flowering and 28 d after flowering)were selected for transcriptomic sequencing,and the Fold Change≥2 and error discovery rate(FDR)<0.01 were used as the standard.Differentially expressed genes(DEGs)in fruits at different developmental stages were screened for GO functional annotation and KEGG signaling pathway enrichment analysis.Realtime fluorescence quantitative PCR was used to verify the reliability of transcriptome sequencing data and analyze the expression patterns of candidate genes.【Result】A total of 22.84 Gb of clean data were obtained by transcriptome sequencing,with 76618317 clean reads,GC content of 44.78%-46.83%and Q20 ranging from 98.89%-99.74%,Q30 ranging from 96.73%to 98.53%.Compared with 19 d after flowering,there were 4194 DEGs on 28 d after flowering,of which 1728 were up-regulated and 2466 were down-regulated.The results of GO functional annotation analysis showed that the DEGs annotation in the comparison group at 28 d after flowering and 19 d after flowering were classified into 3 categories:biological processes,cell components and molecular functions.In biological processes,the number of DEGs involved in cellular processes was the largest,followed by metabolic processes.The results of KEGG signaling pathway enrichment analysis showed that DEGs of the comparison group at 28 d after flowering and 19 d after flowering were significantly enriched in starch and sucrose metabolic pathways.Ten genes related to starch and sucrose metabolic pathways were screened,including glucan water dikinase(GWD)gene(HU02G00177),phosphoglucan

关 键 词：红肉火龙果 淀粉降解 差异表达基因 转录组 

分 类 号：S667.9[农业科学—果树学]