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<p>The North-eastern India is a hotspot of genetic biodiversity of flora and fauna. Among rice landraces, indigenous black rice varieties such as <i>Chakhao Amubi</i> and <i>Chakhao Poireton</i> have gained attention due to their superior grain quality and nutritional attributes. These genotypes are recalcitrant to <i>Agrobacterium</i> <i>tumefaciens</i> mediated genetic transformation. Although transformation protocols have been reported for these landraces, but their transformation and regeneration efficiencies remain moderate. We developed and optimized a tissue culture-based <i>Agrobacterium</i>-mediated transformation protocol for the black rice landraces <i>Chakhao Amubi</i> and <i>Chakhao Poireton</i> and optimized the callus induction and shoot regeneration media for above genotypes. Incorporation of a short resting phase following <i>Agrobacterium</i> infection significantly improved callus recovery and shoot regeneration. The hygromycin concentration previously reported for black rice (50&#xa0;mg L⁻¹ during regeneration) was excessively stringent, resulting in poor regeneration and delayed shoot development, indicating higher hygromycin sensitivity compared with japonica varieties. Evaluation of lower selection pressures identified 25&#xa0;mg L⁻¹ as the optimal concentration, enabling efficient regeneration of positive transformants. Using these genotype-specific protocols, transgenic rice could be successfully regenerated within 90–100 days, with efficiencies of 48% in <i>Chakhao Amubi</i> and 45% in <i>Chakhao Poireton</i>. This reproducible transformation system offers an opportunity for functional genomics studies and genome editing applications in black rice landraces.</p> Graphical Abstract <p></p>

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Efficient Agrobacterium-mediated genetic transformation for Indica black rice cvs. Chakhao Amubi and Chakhao Poireton

  • 期刊论文

  • 发表日期：

    2026年3月16日

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  • Bharti Yadav,
  • Aditi Arya,
  • Pragya Yadav,
  • V. V. Santosh Kumar,
  • Archana Watts,
  • Viswanathan Chinnusamy,
  • Shivani Nagar

摘要

The North-eastern India is a hotspot of genetic biodiversity of flora and fauna. Among rice landraces, indigenous black rice varieties such as Chakhao Amubi and Chakhao Poireton have gained attention due to their superior grain quality and nutritional attributes. These genotypes are recalcitrant to Agrobacterium tumefaciens mediated genetic transformation. Although transformation protocols have been reported for these landraces, but their transformation and regeneration efficiencies remain moderate. We developed and optimized a tissue culture-based Agrobacterium-mediated transformation protocol for the black rice landraces Chakhao Amubi and Chakhao Poireton and optimized the callus induction and shoot regeneration media for above genotypes. Incorporation of a short resting phase following Agrobacterium infection significantly improved callus recovery and shoot regeneration. The hygromycin concentration previously reported for black rice (50 mg L⁻¹ during regeneration) was excessively stringent, resulting in poor regeneration and delayed shoot development, indicating higher hygromycin sensitivity compared with japonica varieties. Evaluation of lower selection pressures identified 25 mg L⁻¹ as the optimal concentration, enabling efficient regeneration of positive transformants. Using these genotype-specific protocols, transgenic rice could be successfully regenerated within 90–100 days, with efficiencies of 48% in Chakhao Amubi and 45% in Chakhao Poireton. This reproducible transformation system offers an opportunity for functional genomics studies and genome editing applications in black rice landraces.

Graphical Abstract