| 摘要: |
| 【目的】明确余甘子转录组SSR位点的分布特征，开发余甘子SSR分子标记技术，为余甘子的亲缘关系鉴定、遗传育种和创新利用提供理论依据。【方法】以6份表型差异较大的余甘子叶片为材料，通过高通量测序技术获得余甘子叶非参转录组数据，分析其转录组和SSR位点分布特征，设计SSR引物并进行有效性和多态性筛选，最终开发余甘子SSR分子标记技术。【结果】余甘子叶转录组测序共获得65 074条基因，平均长度为1 064 bp，71.77%的基因成功获得功能注释；所有基因中共获得18 245个SSR位点，发生频率为21.68%，含有192种重复基元，其中A/T重复基元数目最多；多态性SSR位点共2 714个，占比为14.88%，表明SSR位点可用于遗传多样性分析的潜力较大。对18 245个含有SSR位点的基因进行SSR引物设计,共设计出12 488对SSR引物，成功率为73.88%，其中7 602对引物的重复基元为单核苷酸，占比为60.87%。随机挑选154对引物进行SSR位点有效性和多态性分析，筛选出10对扩增条带清晰且多态性较好的SSR引物，这10对SSR引物重复基元分别为二核苷酸7个，三核苷酸1个和五核苷酸2个，扩增产物大小为127~270 bp，平均为220.5 bp。【结论】多态性SSR位点用于余甘子遗传多样性分析的潜力较大，筛选出10对扩增条带清晰且多态性较好的SSR引物。 |
| 关键词: 余甘子 转录组 SSR 引物筛选 分子标记 |
| DOI：10.13207/j.jnwafu.2026.03.004 |
| 分类号: |
| 基金项目:国家热带植物种质资源库 余甘子种质资源分库项目（NTPGRC2024-011）；福建省属公益类科研院所专项（2022R1028004，2024R1027002）；福建省人民政府与中国农业科学院农业高质量发展超越“5511”协同创新工程项目（XTCXGC2021019-GSS01） |
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| Analysis of Phyllanthus emblica transcriptome and SSR information and molecular marker development |
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WANG Jianchao, ZHANG Xiaoyan, XIE Lixue, ZHANG Lijie, LI Tao
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Fruit Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350013,China
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| Abstract: |
| 【Objective】This research aims to clarify the distribution characteristics of SSR loci in the transcriptome of Phyllanthus emblica and develop the SSR molecular marker technology of P.emblica,to provide a theoretical basis for the identification of genetic relationships,genetic breeding,and the innovative utilization of P.emblica.【Method】Six leaf samples of P.emblica exhibiting significant phenotypic differences were utilized as materials.Non-parametric transcriptome data for these leaves were obtained through high-throughput sequencing technology,followed by an analysis of the transcriptome and SSR locus characteristics.SSR primers were then designed and evaluated for their effectiveness and polymorphism.【Result】The results indicated that a total of 65 074 genes were identified through transcriptome sequencing of P.emblica leaves,with an average length of 1 064 bp.Among them,71.77% of the genes were successfully annotated.Additionally,a total of 18 245 SSR loci were identified across all genes,showing a frequency of 21.68%,which included 192 repeat motifs,with A/T repeat motifs being the most abundant.Among all loci,2 714 polymorphic SSRs were detected,accounting for 14.88%,suggesting that SSR loci had great potential for genetic diversity analysis.A total of 12 488 pairs of SSR primers were successfully designed for the 18 245 genes containing SSR loci,with a success rate of 73.88%.Among all the primers,7 602 pairs of primers had the repeat motif of mononucleotide,accounting for 60.87%.Subsequently,154 pairs of primers were randomly selected for validation and polymorphism analysis.Ultimately,10 pairs of SSR primers exhibiting clear amplified bands and good polymorphism were identified.The repeat motifs of these 10 pairs consisted of 7 dinucleotides,1 trinucleotide,and 2 pentanucleotides.The size of the amplified products ranged from 127 to 270 bp,with an average of 220.5 bp.【Conclusion】Polymorphic SSR loci hold significant potential for genetic diversity.10 pairs of SSR primers,which exhibited clear amplified bands and substantial polymorphism were selected. |
| Key words: Phyllanthus emblica transcriptome SSR primers screening molecular marker |
