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<p><i>Euphorbia lathyris</i> L. is an important medicinal and fuel plant due to its abundant flavonoids, diterpenoids, and fatty acids. However, the lack of a reliable genetic transformation system in <i>E. lathyris</i> currently hinders the discovery and functional verification of genes related to its valuable secondary metabolites biosynthesis. In this study, we established and optimized tobacco rattle virus (TRV) based virus-induced gene silencing (VIGS) system in <i>E. lathyris</i> using magnesium chelatase I subunit (<i>ElCHLI</i>) as the reporter gene to expedite the gene verification process. The silencing efficiency reached the highest when the two-leaf stage seedlings were infiltrated at the concentration of <i>Agrobacterium</i> OD<sub>600</sub> = 1.0 under the vacuum pressure of 6 kPa. <i>E. lathyris ACT</i> was validated as the reference gene for qRT-PCR experiments for TRV-infected plants. Based on the optimal conditions, the gene encoding casbene synthase (CBS) <i>in E. lathyris</i> was successfully silenced and the formation of ingenol in <i>E. lathyris</i> was significantly inhibited, indicating that CBS is a key enzyme for ingenol biosynthesis. These results demonstrate that TRV-mediated VIGS system provides an efficient approach for the gene functional verification in <i>E. lathyris</i>.</p>

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Establishment of virus-induced gene silencing in Euphorbia lathyris L. and its application in casbene synthase (CBS) gene functional analysis

  • 期刊论文

  • 发表日期：

    2025年5月13日

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  • Wei Lyu,
  • Mei Tian,
  • Long Huang,
  • Bi Wang,
  • Pirui Li,
  • Wanli Zhao,
  • Linwei Li,
  • Xu Feng,
  • Shu Xu,
  • Yu Chen

摘要

Euphorbia lathyris L. is an important medicinal and fuel plant due to its abundant flavonoids, diterpenoids, and fatty acids. However, the lack of a reliable genetic transformation system in E. lathyris currently hinders the discovery and functional verification of genes related to its valuable secondary metabolites biosynthesis. In this study, we established and optimized tobacco rattle virus (TRV) based virus-induced gene silencing (VIGS) system in E. lathyris using magnesium chelatase I subunit (ElCHLI) as the reporter gene to expedite the gene verification process. The silencing efficiency reached the highest when the two-leaf stage seedlings were infiltrated at the concentration of Agrobacterium OD600 = 1.0 under the vacuum pressure of 6 kPa. E. lathyris ACT was validated as the reference gene for qRT-PCR experiments for TRV-infected plants. Based on the optimal conditions, the gene encoding casbene synthase (CBS) in E. lathyris was successfully silenced and the formation of ingenol in E. lathyris was significantly inhibited, indicating that CBS is a key enzyme for ingenol biosynthesis. These results demonstrate that TRV-mediated VIGS system provides an efficient approach for the gene functional verification in E. lathyris.