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<p>Efficient and reproducible <i>in vitro</i> regeneration is essential for crop improvement strategies, including genetic transformation and genome editing. In this study, we developed and optimized a high-frequency shoot regeneration protocol using shoot apical meristem (SAM) explants of <i>Solanum lycopersicum</i> cv. <i>Pusa rubi, which is</i> an elite cultivar valued for its processing qualities and widespread cultivation in India. Explants were cultured on Murashige and Skoog (MS) medium supplemented with varying concentrations of cytokinins-zeatin (Zn), thidiazuron (TDZ), auxin indole-3-acetic acid (IAA) and the benzylaminopurine (BAP). Among all treatments, the combination of 1.0&#xa0;mg/L zeatin + 1.0&#xa0;mg/L TDZ + 1.5&#xa0;mg/L IAA + 0.1&#xa0;mg/L BAP achieved the highest regeneration efficiency of 92%, with an average of 46.0 ± 2.0 shoots per explant within 21–28&#xa0;days. Rooting was successfully induced on MS medium containing 0.6&#xa0;mg/L BAP and 0.1&#xa0;mg/L IAA, with a rooting efficiency of 94%. Acclimatised plantlets displayed normal phenotypes and high survival rates under greenhouse conditions. SAM explants facilitated direct organogenesis, minimizing somaclonal variation and improving protocol consistency. Overall, this study lays a strong foundation for developing a high-efficiency regeneration platform, serving as a crucial preparatory step toward future CRISPR/Cas9-based genome editing.</p> Graphical abstract

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Development of an efficient in vitro apical meristem-based regeneration system in tomato (Solanum lycopersicum L.)

  • 期刊论文

  • 发表日期：

    2026年2月20日

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  • Rashmi Kaul,
  • Arulprakash Thangaraj,
  • Jyotsna Bharti,
  • Rachana Verma,
  • Puja Chakraborty,
  • Isha Gupta,
  • R. Shubhra Maithreyi,
  • Sugyan Preet,
  • Shivani Sharda,
  • Tanushri Kaul

摘要

Efficient and reproducible in vitro regeneration is essential for crop improvement strategies, including genetic transformation and genome editing. In this study, we developed and optimized a high-frequency shoot regeneration protocol using shoot apical meristem (SAM) explants of Solanum lycopersicum cv. Pusa rubi, which is an elite cultivar valued for its processing qualities and widespread cultivation in India. Explants were cultured on Murashige and Skoog (MS) medium supplemented with varying concentrations of cytokinins-zeatin (Zn), thidiazuron (TDZ), auxin indole-3-acetic acid (IAA) and the benzylaminopurine (BAP). Among all treatments, the combination of 1.0 mg/L zeatin + 1.0 mg/L TDZ + 1.5 mg/L IAA + 0.1 mg/L BAP achieved the highest regeneration efficiency of 92%, with an average of 46.0 ± 2.0 shoots per explant within 21–28 days. Rooting was successfully induced on MS medium containing 0.6 mg/L BAP and 0.1 mg/L IAA, with a rooting efficiency of 94%. Acclimatised plantlets displayed normal phenotypes and high survival rates under greenhouse conditions. SAM explants facilitated direct organogenesis, minimizing somaclonal variation and improving protocol consistency. Overall, this study lays a strong foundation for developing a high-efficiency regeneration platform, serving as a crucial preparatory step toward future CRISPR/Cas9-based genome editing.

Graphical abstract