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<p>This study aimed to evaluate the response of different 2,4-D concentrations on callus induction and plant regeneration in <i>indica</i> and <i>japonica</i> rice genotypes, specifically the elite line CMG1590 and the model cultivar Nipponbare. Despite the importance of rice biotechnology, genotypic recalcitrance remains a major bottleneck for genome editing. Mature seeds were inoculated onto N6 medium supplemented with 2,4-D concentrations (0 to 3.0&#xa0;mg/L) targeting the scutellum as the primary explant. The results revealed a quadratic dose–response relationship: the absence of 2,4-D (T0) prevented callus formation, whereas the peak induction occurred at 2.5&#xa0;mg/L (T4), with 93.3% for Nipponbare and 80.0% for CMG1590. Mechanistically, Nipponbare’s superior performance is attributed to efficient auxin signaling via the SCFTIR1/OsAFB2 complex, whereas supra-optimal concentrations (3.0&#xa0;mg/L 2,4-D) were associated with reduced cellular performance, potentially reflecting oxidative stress responses and involvement of the OsMAPK6-ROS pathway. Regeneration on MS medium was most effective following induction at 2.0&#xa0;mg/L (T3). ANOVA confirmed a highly significant Genotype × Regulator interaction (p &lt; 0.01). In conclusion, this research validates a 2.0–2.5&#xa0;mg/L 2,4-D threshold as a strategic platform to overcome recalcitrance, providing a robust protocol for future CRISPR/Cas9-mediated improvement of local rice germplasm.</p>

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Analysis of callus induction and regeneration in rice genotypes under different concentrations of 2,4-D

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月15日

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  • Suamith Herminio Luís Leveque,
  • Alisson Willians Teixeira Silva,
  • Elisa Agilo,
  • Flávia Barbosa Silva Botelho

摘要

This study aimed to evaluate the response of different 2,4-D concentrations on callus induction and plant regeneration in indica and japonica rice genotypes, specifically the elite line CMG1590 and the model cultivar Nipponbare. Despite the importance of rice biotechnology, genotypic recalcitrance remains a major bottleneck for genome editing. Mature seeds were inoculated onto N6 medium supplemented with 2,4-D concentrations (0 to 3.0 mg/L) targeting the scutellum as the primary explant. The results revealed a quadratic dose–response relationship: the absence of 2,4-D (T0) prevented callus formation, whereas the peak induction occurred at 2.5 mg/L (T4), with 93.3% for Nipponbare and 80.0% for CMG1590. Mechanistically, Nipponbare’s superior performance is attributed to efficient auxin signaling via the SCFTIR1/OsAFB2 complex, whereas supra-optimal concentrations (3.0 mg/L 2,4-D) were associated with reduced cellular performance, potentially reflecting oxidative stress responses and involvement of the OsMAPK6-ROS pathway. Regeneration on MS medium was most effective following induction at 2.0 mg/L (T3). ANOVA confirmed a highly significant Genotype × Regulator interaction (p < 0.01). In conclusion, this research validates a 2.0–2.5 mg/L 2,4-D threshold as a strategic platform to overcome recalcitrance, providing a robust protocol for future CRISPR/Cas9-mediated improvement of local rice germplasm.