越南油茶CdMYC2基因克隆、生物信息学分析与表达模式研究

Cloning, bioinformatics analysis and expression pattern of CdMYC2 gene in Camellia drupifera

摘要

MYC2 是一种重要的转录因子，可以调控植物萜类次生代谢产物的合成。然而，关于越南油茶（Camellia drupifera Lour.）中的 CdMYC2 转录因子知之甚少。本研究基于越南油茶的转录组注释数据获取了 CdMYC2 基因的相关序列信息。首次从越南油茶叶片中成功分离并克隆了一个 CdMYC2 基因，其 CDS 序列长度为 1 515 bp，编码 504 个氨基酸，具有典型的 bHLH-MYC_N super family 和 bHLH_SF super family 保守结构域，通过预测该基因位于细胞核中。多重序列比对结果进一步证实了扩增的 CdMYC2 属于 MYC2 家族转录因子。系统进化树显示，CdMYC2 与同科同属植物茶树（Camellia sinensis）聚为一支，而与壳斗科栎属植物白栎（Quercus lobata）、夏栎（Q. robur）的分支距离远。RT-qPCR 结果表明，CdMYC2 基因在叶片中的表达水平高于其他组织部位；而在果实的不同发育阶段中，9 月份果实的 CdMYC2 表达量达到最高。本研究为后续探究 MYC2 转录因子在调控越南油茶三萜皂苷生物合成途径中的功能提供了重要的参考依据。

Abstract

MYC2 is an important transcription factor that regulates the synthesis of terpenoid secondary metabolites in plants. However, little is known about the CdMYC2 transcription factor in C. drupifera. In this study, the related sequence information of CdMYC2 gene was obtained based on the transcriptome annotation data of C. drupifera. A CdMYC2 gene was successfully isolated and cloned from the leaves of C. drupifera for the first time. Its CDS sequence length was 1 515 bp, encoding 504 amino acids, with typical bHLH-MYC_N super family and bHLH_SF super family conserved domains. This gene was predicted to be located in the nucleus. Multiple sequence alignment further confirmed that the amplified CdMYC2 belongs to the MYC2 family transcription factor. The phylogenetic tree analysis showed thatCdMYC2 was clustered into a clade with Camellia sinensis, a plant belonging to the same family and genus, while it was far away from the clades of Quercus lobata and Q. robur. RT-qPCR results showed that the expression level of CdMYC2 gene was higher in the leaves than in other tissues. Among the different developmental stages of the fruit, the expression of CdMYC2 in the fruit reached the highest in September. This study provides an important reference for the subsequent exploration of the function of MYC2 transcription factor in the regulation of triterpenoid saponin biosynthesis pathway in C. drupifera.

关键词

越南油茶 / CdMYC2 / 基因克隆 / 生物信息学 / 表达分析

Key words

Camellia drupifera / CdMYC2 / gene cloning / bioinformatics / expression analysis

参考文献

[1] Wasternack C, Hause B. Jasmonates: biosynthesis, perception, signal transduction and action in plant stress response, growth and development. An update to the 2007 review in Annals of Botany [J]. Annals of Botany, 2013, 111(6): 1021-1058. https://doi.org/10.1093/aob/mct067

[2] 郑嘉瑞, 杨晓燕, 叶家保, 等. MYC2 转录因子在植物中的功能研究进展[J]. 园艺学报, 2023, 50(4): 896-908. https://doi.org/10.16420/j.issn.0513-353x.2022-0102

[3] Kazan K, Manners J M. MYC2: the master in action[J]. Molecular Plant, 2013, 6(3): 686-703. https://doi.org/10.1093/mp/sss128

[4] Hong G J, Xue X Y, Mao Y B, et al. Arabidopsis MYC2 interacts with DELLA proteins in regulating sesquiterpene synthase gene expression [J]. The Plant Cell, 2012, 24(6): 2635-2648. https://doi.org/10.1105/tpc.112.098749

[5] Liu T, Liao J, Shi M, et al. A jasmonate-responsive bHLH transcription factor TaMYC2 positively regulates triterpenes biosynthesis in Taraxacum antungense Kitag [J]. Plant Science, 2023, 326: 111506. https://doi.org/10.1016/j.plantsci.2022.111506

[6] 刘进平, 周扬, 胡海燕, 等. 热带油茶种质资源综述[J]. 热带林业, 2022, 50(4): 70-76. https://doi.org/10.3969/j.issn.1672-0938.2022.04.015

[7] 代佳妮. 越南油茶茶皂素合成关键酶基因 CvSQS、CvSQE 的克隆与功能分析[D]. 海口: 海南大学, 2021. https://doi.org/10.27073/d.cnki.ghadu.2021.000501

[8] 刘运锋, 邱兆志, 申新刚, 等. 茶油的化学成分和药理作用研究进展[J]. 南方农业, 2020, 14(9): 132-134. https://doi.org/10.19415/j.cnki.1673-890x.2020.09.067

[9] 黄运安. 油茶中三萜皂苷及其生物活性研究进展[J]. 食品安全质量检测学报, 2024, 15(13): 59-67. https://doi.org/10.19812/j.cnki.jfsq11-5956/ts.20240312008

[10] Li Y, Wang J, Li L, et al. Natural products of pentacyclic triterpenoids: from discovery to heterologous biosynthesis[J]. Natural Product Reports, 2023, 40(8): 1303-1353. https://doi.org/10.1039/D2NP00063F

[11] Zheng J, Liao Y, Ye J, et al. The transcription factor MYC2 positively regulates terpene trilactone biosynthesis through activating GbGGPPS expression in Ginkgo biloba [J]. Horticulture Research, 2024, 11(10): uhae228. https://doi.org/10.1093/hr/uhae228

[12] 王晨, 安立成, 李剑超, 等. 北柴胡 MYC2 转录因子的克隆及茉莉酸诱导的调控分析[J]. 植物生理学报, 2021, 57(2): 439-450. https://doi.org/10.13592/j.cnki.ppj.2020.0381

[13] Alix K, Gérard P R, Schwarzacher T, et al. Polyploidy and interspecific hybridization: partners for adaptation, speciation and evolution in plants[J]. Annals of Botany, 2017, 120(2): 183-194. https://doi.org/10.1093/aob/mcx079

[14] 焦晓林, 高微微. 环境因子对药用植物三萜皂苷合成影响的研究进展[J]. 中草药, 2011, 42(2): 398-402.

[15] Dellas N, Thomas S T, Manning G, et al. Discovery of a metabolic alternative to the classical mevalonate pathway[J]. eLife, 2013, 2: e00672. https://doi.org/10.7554/eLife.00672

[16] 郑蔚, 代佳妮, 严和琴, 等. 越南油茶 HMGCR 基因的克隆及表达分析[J]. 分子植物育种, 2022, 20(13): 4296-4304. https://doi.org/10.13271/j.mpb.020.004296

[17] 李好, 方学智, 钟海雁, 等. 油茶果成熟过程茶皂素含量分布状况[J]. 粮食与油脂, 2013, 26(9): 24-26. https://doi.org/10.3969/j.issn.1008-9578.2013.09.007

基金资助

“崖州湾”菁英人才项目(SKJC-JYRC-2024-30)

海南省重点研发计划项目(ZDYF2022SHFZ020)