Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (5): 1412-1423.DOI: 10.1016/j.jia.2023.03.005

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西番莲中参与黄酮类和萜类生物合成的关键基因鉴定和三萜类生物合成的途径分析

  

  • 收稿日期:2022-09-16 接受日期:2023-01-09 出版日期:2023-05-20 发布日期:2023-01-09

Identification of key genes involved in flavonoid and terpenoid biosynthesis and the pathway of triterpenoid biosynthesis in Passiflora edulis

XU Yi1, 2*#, HUANG Dong-mei1*, MA Fu-ning1, 2, YANG Liu3, WU Bin1, XING Wen-ting1, SUN Pei-guang1, 2, CHEN Di1, XU Bing-qiang1, SONG Shun1, 2#   

  1. 1 National Key Laboratory for Tropical Crop Breeding/Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Genetic Improvement of Bananas, Sanya Research Institute of Chinese Academy of Tropical Agricultural Sciences, Sanya 572000, P.R.China

    2 Hainan Yazhou Bay Seed Laboratory, Sanya 572000, P.R.China

    3 Guangxi Crop Genetic Improvement and Biotechnology Laboratory, Guangxi Academy of Agricultural Sciences, Nanning 530007, P.R.China

  • Received:2022-09-16 Accepted:2023-01-09 Online:2023-05-20 Published:2023-01-09
  • About author:#Correspondence SONG Shun, E-mail: songs@catas.cn; XU Yi, E-mail: xuyi@catas.cn * These authors contributed equally to this study.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (32260737), the Sanya Yazhou Bay Science and Technology City (SCKJ-JYRC-2022-84, SCKJ-JYRC-2022-93), and Natural Science Foundation of Hainan Province Hainan Provincial Natural Science Foundation of China (320QN305, 321MS091, 320RC686).

摘要:

西番莲(Passiflora edulis Sims)也称百香果,是西番莲科西番莲属的藤本植物。其全株提取的黄酮类和萜类对人体有良好的抗焦虑和抗炎作用,西番莲除了鲜食之外还可药用。在这项工作中,我们利用RNA-Seq分析了紫果品种的四个组织的转录表达,注释了大量基因功能。西番莲叶片中的黄酮类和萜类化合物主要是木犀草素、芹菜素、环状三萜皂苷和其他活性物质的衍生物。利用基于同源BLAST和系统发育分析,筛选了转录组数据中可能参与黄酮类和萜类合成途径的一系列候选单基因。结果显示,西番莲的三萜类化合物的生物合成来自甲羟戊酸(MVA)和2-C-甲基-D-赤藓糖醇-4-磷酸/1-脱氧-D-木糖醇-5-磷酸(MEP/DOXP)途径,这与其他果树通过MVA途径为主的生物合成不同。大多数候选基因在叶和/或花中高表达。8个关键基因的定量实时PCR(qRT-PCR)验证,证实了RNA-Seq数据的可靠性,并获得了其在西番莲属八个种(species)和其中一个种(栽培种)的四个组织中的表达规律。这些工作对分析关键基因在西番莲黄酮类化合物和萜类化合物的生物合成提供基础。

Abstract:

Passion fruit (Passiflora edulis Sims) is a vine of the Passiflora genus in the Passifloraceae family.  The extracted components include flavonoids and terpenoids, which have good anti-anxiety and anti-inflammatory effects in humans.  In this study, we analyzed the transcriptomes of four tissues of the ‘Zixiang’ cultivar using RNA-Seq, which provided a dataset for functional gene mining.  The de novo assembly of these reads generated 96 883 unigenes, among which 61 022 unigenes were annotated (62.99% yield).  In addition to its edible value, another important application of passion fruit is its medicinal value.  The flavonoids and terpenoids are mainly derivatives of luteolin, apigenin, cycloartane triterpenoid saponins and other active substances in leaf extracts.  A series of candidate unigenes in the transcriptome data that are potentially involved in the flavonoid and terpenoid synthesis pathways were screened using homology-based BLAST and phylogenetic analysis.  The results showed that the biosynthesis of triterpenoids in passion fruit comes from the branches of the mevalonate (MVA) and 2-C-methyl-D-erythritol 4-phosphate/1-deoxy-D-xylulose 5-phosphate (MEP/DOXP) pathways, which is different from the MVA pathway that is used in other fruit trees.  Most of the candidate genes were found to be highly expressed in the leaves and/or flowers.  Quantitative real-time PCR (qRT-PCR) verification was carried out and confirmed the reliability of the RNA-Seq data.  Further amplification and functional analysis of these putative unigenes will provide additional insight into the biosynthesis of flavonoids and terpenoids in passion fruit.

Key words: passion fruit , RNA-Seq ,  gene mining ,  flavonoids ,  terpenoids