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<p>The present study illustrates the development of reliable and reproducible plant regeneration protocol for endangered plant species <i>Lindernia minima</i> using leaf derived callus. Leaf explants were cultured on quarter strength MS (Murashige and Skoog) medium augmented with different concentrations (0.5 to 3.0&#xa0;mg L<sup>−1</sup>) of 2,4-D and NAA individually was tested. The study achieved 95.6% of yellowish-green compact nodular callus from the leaf explants on quarter-strength MS medium fortified with 2.0&#xa0;mg L<sup>−1</sup> of 2, 4-D. Further, organogenic calli were cultured on quarter strength MS medium containing various concentrations of (0.5 to 3.0&#xa0;mg L<sup>−1</sup>) cytokinins (BA, KIN, and TDZ) with various natural growth stimulants like purified vermicast extract (VCE), VCE and Coconut Water mixture (Vermi-Co) for shoot regeneration. The highest shoot response (98%) with maximum mean shoot number (27.3 ± 0.16 shoots per explants) and shoot length (9.8 ± 0.28&#xa0;cm) were achieved on quarter strength MS medium containing 2.0&#xa0;mg L<sup>−1</sup> of BA and 15% of Vermi-Co mixture. The well elongated healthy shoots were cultured on various concentrations of IBA and IAA for root induction. The maximum rooting response (97%) was observed on quarter-strength MS medium supplemented with 2.0&#xa0;mg L<sup>−1</sup> of IBA. After root induction, the well rooted plantlets were successfully acclimatized with a 93% survival rate. The genetic purity of regenerated plantlets and mother plant assessed by the DNA based ISSR molecular marker showed no somoclonal variations. The described <i>in vitro</i> plant regeneration protocol can be more reliable for rapid mass multiplication of <i>L. minima.</i></p>

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Growth-promoting effects of natural biostimulants on Lindernia minima regeneration and genetic stability assessment

  • 期刊论文

  • 发表日期：

    2025年3月25日

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  • H. David Raja,
  • R. Tamilvanan,
  • P. Srinivasan,
  • S. Soosai Raj,
  • D. Jeferson,
  • A. Vijay,
  • S. Vigneshwaran

摘要

The present study illustrates the development of reliable and reproducible plant regeneration protocol for endangered plant species Lindernia minima using leaf derived callus. Leaf explants were cultured on quarter strength MS (Murashige and Skoog) medium augmented with different concentrations (0.5 to 3.0 mg L−1) of 2,4-D and NAA individually was tested. The study achieved 95.6% of yellowish-green compact nodular callus from the leaf explants on quarter-strength MS medium fortified with 2.0 mg L−1 of 2, 4-D. Further, organogenic calli were cultured on quarter strength MS medium containing various concentrations of (0.5 to 3.0 mg L−1) cytokinins (BA, KIN, and TDZ) with various natural growth stimulants like purified vermicast extract (VCE), VCE and Coconut Water mixture (Vermi-Co) for shoot regeneration. The highest shoot response (98%) with maximum mean shoot number (27.3 ± 0.16 shoots per explants) and shoot length (9.8 ± 0.28 cm) were achieved on quarter strength MS medium containing 2.0 mg L−1 of BA and 15% of Vermi-Co mixture. The well elongated healthy shoots were cultured on various concentrations of IBA and IAA for root induction. The maximum rooting response (97%) was observed on quarter-strength MS medium supplemented with 2.0 mg L−1 of IBA. After root induction, the well rooted plantlets were successfully acclimatized with a 93% survival rate. The genetic purity of regenerated plantlets and mother plant assessed by the DNA based ISSR molecular marker showed no somoclonal variations. The described in vitro plant regeneration protocol can be more reliable for rapid mass multiplication of L. minima.