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<p>A highly efficient plant regeneration protocol for medicinally important <i>Cardiospermum halicacabum</i> L. has been devised by preculturing cotyledonary node (CN) explant, significantly enhancing shoot multiplication and acclimatization process. Direct inoculation of 15&#xa0;days (d) old CN explants on Murashige and Skoog’s (MS) medium with 0.2 to 1.0&#xa0;µM thidiazuron (TDZ) resulted in 80% regeneration but with a low shoot formation yield (8.7 ± 1.8 per explant). To overcome this limitation, a two-step culture strategy was applied, where CN explants were first preconditioned for 4, 8, 12, and 16 d in liquid MS medium supplemented with higher concentrations of TDZ (05 to 25&#xa0;µM) followed by their transfer to hormone-free semi-solid MS medium. A five-fold increase in the shoot production rate was achieved when preconditioning was done for 8 d in liquid MS medium augmented with 15.0&#xa0;µM TDZ, differentiating a maximum of 47.7 ± 1.5 number of shoots with highest shoot length (4.3 ± 0.3&#xa0;cm) in about 90% of cotyledonary nodes after 8 wk of culture duration. Rooting was successfully induced by a 24-h pulse treatment with 200.0&#xa0;µM indole-3-acetic acid (IAA) for approximately 24&#xa0;h and then transferring it to half-strength MS medium yielding a maximum of 6.3 ± 0.8 roots in 86% of cultures. Acclimatization success reached with the production of 96% regenerated plantlets demonstrated enhanced net photosynthetic rate (P<sub>N</sub>) and photosynthetic pigments and increased antioxidant enzymes activities (Superoxide Dismutase (SOD) and Catalase (CAT)) showing the ability of regenerated plants to regulate oxidative stress. This protocol demonstrates a highly efficient and reproducible procedure for mass multiplication of <i>C. halicacabum</i> plants, paving a way for its conservation and commercialization.</p>

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Enhancing shoot multiplication in Cardiospermum halicacabum L. by using two-step in vitro strategy with thidiazuron

  • 期刊论文

  • 发表日期：

    2025年5月23日

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  • Iram Siddique,
  • Anushi Arjumend Jahan

摘要

A highly efficient plant regeneration protocol for medicinally important Cardiospermum halicacabum L. has been devised by preculturing cotyledonary node (CN) explant, significantly enhancing shoot multiplication and acclimatization process. Direct inoculation of 15 days (d) old CN explants on Murashige and Skoog’s (MS) medium with 0.2 to 1.0 µM thidiazuron (TDZ) resulted in 80% regeneration but with a low shoot formation yield (8.7 ± 1.8 per explant). To overcome this limitation, a two-step culture strategy was applied, where CN explants were first preconditioned for 4, 8, 12, and 16 d in liquid MS medium supplemented with higher concentrations of TDZ (05 to 25 µM) followed by their transfer to hormone-free semi-solid MS medium. A five-fold increase in the shoot production rate was achieved when preconditioning was done for 8 d in liquid MS medium augmented with 15.0 µM TDZ, differentiating a maximum of 47.7 ± 1.5 number of shoots with highest shoot length (4.3 ± 0.3 cm) in about 90% of cotyledonary nodes after 8 wk of culture duration. Rooting was successfully induced by a 24-h pulse treatment with 200.0 µM indole-3-acetic acid (IAA) for approximately 24 h and then transferring it to half-strength MS medium yielding a maximum of 6.3 ± 0.8 roots in 86% of cultures. Acclimatization success reached with the production of 96% regenerated plantlets demonstrated enhanced net photosynthetic rate (PN) and photosynthetic pigments and increased antioxidant enzymes activities (Superoxide Dismutase (SOD) and Catalase (CAT)) showing the ability of regenerated plants to regulate oxidative stress. This protocol demonstrates a highly efficient and reproducible procedure for mass multiplication of C. halicacabum plants, paving a way for its conservation and commercialization.