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<p><i>Furcraea foetida</i> (Mauritius hemp), a drought-tolerant species of the Agavaceae family, holds economic importance due to its fiber, medicinal properties, and ornamental value. Despite its potential, there is limited progress in developing reproducible in vitro regeneration and genetic transformation systems for this species. This study presents a preliminary investigation into the establishment of an in vitro shoot and root regeneration system and explores the feasibility of <i>Agrobacterium</i>-mediated transient gene expression in <i>F. foetida</i>. Shoot regeneration was tested using Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinins (BAP, kinetin, and zeatin), while rooting was examined using auxins (IAA and IBA). The best shoot induction response was observed with 1.0&#xa0;mg/L BAP, and optimal rooting was achieved using 0.5&#xa0;mg/L IBA. Transient genetic transformation was performed using <i>Agrobacterium tumefaciens</i> strain LBA4404 harboring the binary vector pBI121 carrying the GUS and <i>nptII</i> genes. Factors influencing transformation, such as explant age, co-cultivation time, acetosyringone concentration, and kanamycin selection pressure, were evaluated. A maximum transient expression efficiency of 7% was recorded following 48&#xa0;h of co-cultivation with <i>Agrobacterium</i> in the presence of 250&#xa0;mM acetosyringone. GUS histochemical staining confirmed transient gene expression in transformed explants, and PCR analysis of in vitro regenerated leaf tissues verified the presence of both <i>gus</i> and <i>nptII</i> genes, indicating preliminary transgene integration. Although further validation of transgene stability, ex vitro survival, and true-to-type regeneration is needed, this study provides an essential baseline for future genetic improvement strategies in <i>F. foetida</i>, supporting its potential applications in agriculture, industry, and biotechnology.</p>

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Development of in vitro regeneration protocol and Agrobacterium-mediated transient transformation system in Furcraea foetida

  • 期刊论文

  • 发表日期：

    2025年6月12日

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  • Sheeba Naz,
  • Naheed Kauser,
  • Hammad Afzal Kayani,
  • Bushra Ahmad Saeed,
  • Ishrat Jamil,
  • Saddia Galani

摘要

Furcraea foetida (Mauritius hemp), a drought-tolerant species of the Agavaceae family, holds economic importance due to its fiber, medicinal properties, and ornamental value. Despite its potential, there is limited progress in developing reproducible in vitro regeneration and genetic transformation systems for this species. This study presents a preliminary investigation into the establishment of an in vitro shoot and root regeneration system and explores the feasibility of Agrobacterium-mediated transient gene expression in F. foetida. Shoot regeneration was tested using Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinins (BAP, kinetin, and zeatin), while rooting was examined using auxins (IAA and IBA). The best shoot induction response was observed with 1.0 mg/L BAP, and optimal rooting was achieved using 0.5 mg/L IBA. Transient genetic transformation was performed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pBI121 carrying the GUS and nptII genes. Factors influencing transformation, such as explant age, co-cultivation time, acetosyringone concentration, and kanamycin selection pressure, were evaluated. A maximum transient expression efficiency of 7% was recorded following 48 h of co-cultivation with Agrobacterium in the presence of 250 mM acetosyringone. GUS histochemical staining confirmed transient gene expression in transformed explants, and PCR analysis of in vitro regenerated leaf tissues verified the presence of both gus and nptII genes, indicating preliminary transgene integration. Although further validation of transgene stability, ex vitro survival, and true-to-type regeneration is needed, this study provides an essential baseline for future genetic improvement strategies in F. foetida, supporting its potential applications in agriculture, industry, and biotechnology.