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转录组分析揭示了参与小檗科鬼臼属植物体细胞胚胎发生和鬼臼毒素生物合成的差异表达基因。

Transcriptome analysis reveals differentially expressed genes involved in somatic embryogenesis and podophyllotoxin biosynthesis of Sinopodophyllum hexandrum (Royle) T. S. Ying.

机构信息

Research Center of Agricultural Biotechnology, Ningxia Academy of Agricultural and Forestry Sciences, Yinchuan, 750002, Gansu, China.

Ningxia Key Laboratory of Agricultural Biotechnology, Yinchuan, 750002, Gansu, China.

出版信息

Protoplasma. 2023 Jul;260(4):1221-1232. doi: 10.1007/s00709-023-01843-9. Epub 2023 Feb 25.

Abstract

Sinopodophyllum hexandrum (Royle) T. S. Ying, an important source of podophyllotoxin (PTOX), has become a rare and endangered plant because of over-harvesting. Somatic embryogenesis (SE) is the main way of seedling rapid propagation and germplasm enhancement, but the regeneration of S. hexandrum has not been well established, and the PTOX biosynthesis abilities at different SE stages remain unclear. Therefore, it is extremely important to elucidate the SE mechanism of S. hexandrum and clarify the biosynthesis variation of PTOX. In this study, the transcriptomes of S. hexandrum at different SE stages were sequenced, the contents of PTOX and 4'-demethylepipodophyllotoxin were assayed, and the transcript expression patterns were validated by qRT-PCR. The results revealed that plant hormone (such as auxins, abscisic acid, zeatin, and gibberellins) related pathways were significantly enriched among different SE stages, indicating these plant hormones play important roles in SE of S. hexandrum; the expression levels of a series of PTOX biosynthesis related genes as well as PTOX and 4'-demethylepipodophyllotoxin contents were much higher in embryogenic callus stage than in the other stages, suggesting embryogenic callus stage has the best PTOX biosynthesis ability among different SE stages. This study will contribute to germplasm conservation and fast propagation of S. hexandrum, and facilitate the production of PTOX.

摘要

六角莲(Sinopodophyllum hexandrum (Royle) T. S. Ying)是足叶草毒素(PTOX)的重要来源,由于过度采集,已成为一种稀有濒危植物。体细胞胚胎发生(SE)是幼苗快速繁殖和种质增强的主要途径,但六角莲的再生尚未得到很好的建立,不同 SE 阶段的 PTOX 生物合成能力也不清楚。因此,阐明六角莲 SE 的机制,阐明 PTOX 的生物合成变化极为重要。在这项研究中,对不同 SE 阶段的六角莲进行了转录组测序,测定了 PTOX 和 4'-去甲表鬼臼毒素的含量,并通过 qRT-PCR 验证了转录表达模式。结果表明,不同 SE 阶段的植物激素(如生长素、脱落酸、玉米素和赤霉素)相关途径显著富集,表明这些植物激素在六角莲 SE 中发挥重要作用;一系列 PTOX 生物合成相关基因的表达水平以及 PTOX 和 4'-去甲表鬼臼毒素的含量在胚性愈伤组织阶段均高于其他阶段,表明胚性愈伤组织阶段在不同 SE 阶段具有最佳的 PTOX 生物合成能力。本研究将有助于六角莲的种质保存和快速繁殖,并促进 PTOX 的生产。

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