• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

基于第三代纳米孔测序技术的茶树全长转录组分析

Full-length Transcriptomes of Tea Obtained by Oxford Nanopore Technology

  • 摘要:
      目的  研究茶树的全长转录组测序,以期为解析控制茶叶质量形成的分子机制提供理论依据。
      方法  以贵州都匀地区福鼎大白茶叶、根和茎为材料,采用牛津大学的纳米孔技术(Oxford Nanopore Technology, ONT)进行全长转录组测序;将茶树叶、根和茎进行基因表达量分析;为了验证转录组测序结果的可靠性,随机抽取4个基因进行实时荧光定量PCR(qRT-PCR)。
      结果  (1)获得69 379条全长转录本、93 102个SSR、45 852个ORF、6 335个转录因子和2 229个lncRNA;(2)获得叶与根、叶与茎以及茎与根的差异转录本,叶与根的差异转录本注释到GO、KEGG和KOG数据库分别有9 646、2 825和7 349个,其中参与茶叶品质形成中茶叶滋味相关代谢途径的有各种氨基酸代谢、类黄酮生物合成和苯丙素生物合成等,与香气相关的代谢途径有泛醌和其他萜烯醌生物合成等;qRT-PCR结果显示其表达量与转录组相一致。
      结论  筛选到2个差异基因(ONT.24127.2和TEA003892.1)与谷氨酸合成酶有关,在根部合成茶氨酸后再运输到叶部,进而影响茶叶的品质。研究结果为保障茶叶产量和质量打下良好基础,为后续良种选育和引种及开展后续分子生物学研究提供重要的数据支持。

     

    Abstract:
      Objective  The full-length transcriptomes of tissues of the famed Camellia sinensis (L.) O. Kuntze cv. Fuding Dabaicha were obtained to facilitate cultivar selection in upgrading the Duyun Maojian tea industry and increasing farmers’ income in Guizhou province.
      Method  The Oxford nanopore technology (ONT) was applied to secure the full-length transcriptomes in the leaves, roots, and stems of the tea plant. Gene expressions in the tissues were analyzed. For verification of the ONT results, expressions of 4 randomly selected genes were tested by qRT-PCR.
      Result  The ONT sequencing rendered 69,379 full-length transcripts with approximately 93,102 SSRs, 45,852 ORFs, 6,335 transcription factors, and 2,229 lncRNAs. The differential expression transcripts (DETs) among the leaves, roots, and stems were identified. Between the leaves and roots, there were 9,646 DETs annotated to GO database, 2,825 to KEGG, and 7,348 to KOG. The metabolisms of various amino acids, flavonoid biosynthesis, and phenylpropanol biosynthesis were related to the taste that affects the quality of a tea. Whereas the metabolic pathways involving aroma formation might include biosynthesis of ubiquinone and other terpene quinone. The ONT transcriptomes agreed with the qRT-PCR test results.
      Conclusion  Two differentially expressed genes, ONT.24127.2 and TEA003892.1, were identified to be related to glutamate synthase. Theanine, which is closely associated with tea quality, was synthesized in the roots, then transported to the leaves. The full-length transcriptomes on Fuding Dabaicha demonstrated the practical application of the information for tea cultivar selection.

     

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