Scholarlink

<p>L-Dopa, the frontline drug for Parkinson’s disease (PD), is naturally synthesized by <i>Hybanthus enneaspermus</i> (L.) F. Muell., highlighting its pharmacological significance. Multi-walled carbon nanotubes (MWCNTs) function as promising nano-elicitor for enhancing and industrially scaling metabolites in plants. On this basis, the present investigation targets in vitro L-Dopa elicitation using MWCNTs feeding, employing a transgenic hairy root culture system of <i>H. enneaspermus</i>. The transgenic hairy root lines were successfully and consistently induced from callus cultures of <i>H. enneaspermus</i> using an optimized <i>Agrobacterium rhizogenes</i>-mediated transformation protocol. Synergistic effects were observed when acetosyringone, sonication, and vacuum infiltration were precisely optimized to 150 µM, 3&#xa0;min, and 90&#xa0;s, respectively. When applied together, these conditions led to the highest transformation efficiency (24%) after 3 days of co-cultivation with strain <i>Agrobacterium rhizogenes</i> (ATCC 15834). Additionally, PCR and RT-PCR amplification of the <i>rolA</i> gene provided clear evidence of the transgenic status of the induced hairy roots. Furthermore, feeding <i>H. enneaspermus</i> hairy root cultures with 60&#xa0;mg L⁻¹ MWCNTs for 24&#xa0;h led to the highest L-Dopa yield of 34.56&#xa0;mg/g DW. This marks a 10.97-fold improvement relative to hairy roots not elicited with the MWCNTs. The results support the application of MWCNTs as sustainable platform for enhancing L-Dopa yields, thereby reducing dependence on synthetic production. The CRISPR/Cas9 tool can be effectively used in upcoming research to fine-tune gene expression within the hairy root system for improved L-Dopa synthesis.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Multi-walled carbon nanotubes (MWCNTs) elicitation enhances L-Dopa biosynthesis in transgenic hairy root cultures of Hybanthus enneaspermus (L.) F. Muell.

  • 期刊论文

  • 发表日期：

    2025年10月2日

查看全文
  • Shanthi Pandurengan Parthasarathy,
  • Ananthakumar Archana,
  • Markandan Manickavasagam

摘要

L-Dopa, the frontline drug for Parkinson’s disease (PD), is naturally synthesized by Hybanthus enneaspermus (L.) F. Muell., highlighting its pharmacological significance. Multi-walled carbon nanotubes (MWCNTs) function as promising nano-elicitor for enhancing and industrially scaling metabolites in plants. On this basis, the present investigation targets in vitro L-Dopa elicitation using MWCNTs feeding, employing a transgenic hairy root culture system of H. enneaspermus. The transgenic hairy root lines were successfully and consistently induced from callus cultures of H. enneaspermus using an optimized Agrobacterium rhizogenes-mediated transformation protocol. Synergistic effects were observed when acetosyringone, sonication, and vacuum infiltration were precisely optimized to 150 µM, 3 min, and 90 s, respectively. When applied together, these conditions led to the highest transformation efficiency (24%) after 3 days of co-cultivation with strain Agrobacterium rhizogenes (ATCC 15834). Additionally, PCR and RT-PCR amplification of the rolA gene provided clear evidence of the transgenic status of the induced hairy roots. Furthermore, feeding H. enneaspermus hairy root cultures with 60 mg L⁻¹ MWCNTs for 24 h led to the highest L-Dopa yield of 34.56 mg/g DW. This marks a 10.97-fold improvement relative to hairy roots not elicited with the MWCNTs. The results support the application of MWCNTs as sustainable platform for enhancing L-Dopa yields, thereby reducing dependence on synthetic production. The CRISPR/Cas9 tool can be effectively used in upcoming research to fine-tune gene expression within the hairy root system for improved L-Dopa synthesis.