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内生真菌 Pseudodidymocyrtis lobariellae KL27 促进了中国红豆杉中紫杉醇的生物合成和积累。

Endophytic fungus Pseudodidymocyrtis lobariellae KL27 promotes taxol biosynthesis and accumulation in Taxus chinensis.

机构信息

School of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu, 221116, People's Republic of China.

Laboratory of Medicinal Plant Biotechnology, College of Pharmacy, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People's Republic of China.

出版信息

BMC Plant Biol. 2022 Jan 3;22(1):12. doi: 10.1186/s12870-021-03396-6.

Abstract

BACKGROUND

Taxol from Taxus species is a precious drug used for the treatment of cancer and can effectively inhibit the proliferation of cancer cells. However, the growth of Taxus plants is very slow and the content of taxol is quite low. Therefore, it is of great significance to improve the yield of taxol by modern biotechnology without destroying the wild forest resources. Endophytic fungus which symbiosis with their host plants can promote the growth and secondary metabolism of medicinal plants.

RESULTS

Here, an endophytic fungus KL27 was isolated from T. chinensis, and identified as Pseudodidymocyrtis lobariellae. The fermentation broth of KL27 (KL27-FB) could significantly promote the accumulation of taxol in needles of T. chinensis, reaching 0.361 ± 0.082 mg/g·DW (dry weight) at 7 days after KL27-FB treatment, which is 3.26-fold increase as compared to the control. The RNA-seq and qRT-PCR showed that KL27-FB could significantly increase the expression of key genes involved in the upstream pathway of terpene synthesis (such as DXS and DXR) and those in the taxol biosynthesis pathway (such as GGPPS, TS, T5OH, TAT, T10OH, T14OH, T2OH, TBT, DBAT and PAM), especially at the early stage of the stimulation. Moreover, the activation of jasmonic acid (JA) biosynthesis and JA signal transduction, and its crosstalk with other hormones, such as gibberellin acid (GA), ethylene (ET) and salicylic acid (SA), explained the elevation of most of the differential expressed genes related to taxol biosynthesis pathway. Moreover, TF (transcriptional factor)-encoding genes, including MYBs, ethylene-responsive transcription factors (ERFs) and basic/helix-loop-helix (bHLH), were detected as differential expressed genes after KL27-FB treatment, further suggested that the regulation of hormone signaling on genes of taxol biosynthesis was mediated by TFs.

CONCLUSIONS

Our results indicated that fermentation broth of endophytic fungus KL27-FB could effectively enhance the accumulation of taxol in T. chinensis needles by regulating the phytohormone metabolism and signal transduction and further up-regulating the expression of multiple key genes involved in taxol biosynthesis. This study provides new insight into the regulatory mechanism of how endophytic fungus promotes the production and accumulation of taxol in Taxus sp.

摘要

背景

来自红豆杉属植物的紫杉醇是一种用于治疗癌症的珍贵药物,能有效抑制癌细胞的增殖。然而,红豆杉植物的生长非常缓慢,紫杉醇的含量也相当低。因此,通过现代生物技术在不破坏野生森林资源的情况下提高紫杉醇的产量具有重要意义。与宿主植物共生的内生真菌可以促进药用植物的生长和次生代谢。

结果

从中国红豆杉中分离得到一株内生真菌 KL27,并鉴定为拟珊瑚菌 Pseudodidymocyrtis lobariellae。KL27 的发酵液(KL27-FB)能显著促进中国红豆杉针叶中紫杉醇的积累,在 KL27-FB 处理后 7 天达到 0.361±0.082mg/g·DW(干重),是对照组的 3.26 倍。RNA-seq 和 qRT-PCR 结果表明,KL27-FB 能显著上调萜类合成上游途径(如 DXS 和 DXR)和紫杉醇生物合成途径(如 GGPPS、TS、T5OH、TAT、T10OH、T14OH、T2OH、TBT、DBAT 和 PAM)中关键基因的表达,特别是在刺激的早期阶段。此外,茉莉酸(JA)生物合成和 JA 信号转导的激活,以及与赤霉素酸(GA)、乙烯(ET)和水杨酸(SA)等其他激素的相互作用,解释了与紫杉醇生物合成途径相关的大多数差异表达基因的上调。此外,还检测到转录因子(TF)编码基因,包括 MYB、乙烯响应转录因子(ERF)和碱性/螺旋-环-螺旋(bHLH),作为 KL27-FB 处理后的差异表达基因,进一步表明激素信号对紫杉醇生物合成基因的调控是由 TF 介导的。

结论

我们的结果表明,内生真菌 KL27-FB 的发酵液能有效促进中国红豆杉针叶中紫杉醇的积累,通过调节植物激素代谢和信号转导,进一步上调参与紫杉醇生物合成的多个关键基因的表达。本研究为内生真菌促进红豆杉属植物紫杉醇产生和积累的调控机制提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d10/8722197/f5c4fa3a4902/12870_2021_3396_Fig1_HTML.jpg

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