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<p>Pomegranate (<i>Punica granatum</i> L.) is a high-value fruit, and efficient in vitro regeneration is crucial for the rapid propagation and conservation of elite cultivars. However, phenolic oxidation, particularly in the ‘Wonderful’ cultivar, remains a major obstacle during culture establishment. This study aimed to optimize early regeneration by reducing browning and improving the establishment of explants. The experiment comprised two integrated sections. The first section evaluated antioxidant-based anti-browning treatments (citric acid, ascorbic acid, and PVP). These treatments were applied at different sterilization stages under contrasting temperature and light conditions. Browning severity was monitored for 10 days using a standardized five-level index. Among the treatments, adding 1&#xa0;g L⁻¹ PVP to the medium and incubating explants in the dark at 4&#xa0;°C for three days consistently reduced browning the most. This treatment decreased browning index by 31–52% over the first eight days. In contrast, immersion in 0.05% citric acid and repeated 24-h subculturing had limited effects, and other treatments showed moderate reductions (1–22%). The second section investigated topophysis by comparing explants from apical, middle, and basal shoot positions. The study tested these explants under two temperature regimes (4&#xa0;°C and 25&#xa0;°C) and on MS medium with or without PVP. Explants from the middle and basal positions incubated at 4&#xa0;°C with PVP showed the lowest browning index (&lt; 2.0) throughout the 10-day period. In contrast, apical segments and higher temperatures resulted in greater browning. Overall, these results highlight the combined importance of anti-browning chemical treatments, explant position, and incubation temperature in reducing browning. The integration of PVP supplementation and low-temperature dark incubation provides a reliable and practical strategy for minimizing oxidative browning and enhancing early stage in vitro regeneration in high-value pomegranate cultivars.</p>

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Optimization of anti-browning treatments and explant position to reduce phenolic oxidation in ‘Wonderful’ pomegranate

  • 期刊论文

  • 发表日期：

    2026年4月22日

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  • Alireza Ghaderi,
  • Azam Jafari,
  • Mostafa Shirmardi,
  • Mohammadreza Vazifeshenas

摘要

Pomegranate (Punica granatum L.) is a high-value fruit, and efficient in vitro regeneration is crucial for the rapid propagation and conservation of elite cultivars. However, phenolic oxidation, particularly in the ‘Wonderful’ cultivar, remains a major obstacle during culture establishment. This study aimed to optimize early regeneration by reducing browning and improving the establishment of explants. The experiment comprised two integrated sections. The first section evaluated antioxidant-based anti-browning treatments (citric acid, ascorbic acid, and PVP). These treatments were applied at different sterilization stages under contrasting temperature and light conditions. Browning severity was monitored for 10 days using a standardized five-level index. Among the treatments, adding 1 g L⁻¹ PVP to the medium and incubating explants in the dark at 4 °C for three days consistently reduced browning the most. This treatment decreased browning index by 31–52% over the first eight days. In contrast, immersion in 0.05% citric acid and repeated 24-h subculturing had limited effects, and other treatments showed moderate reductions (1–22%). The second section investigated topophysis by comparing explants from apical, middle, and basal shoot positions. The study tested these explants under two temperature regimes (4 °C and 25 °C) and on MS medium with or without PVP. Explants from the middle and basal positions incubated at 4 °C with PVP showed the lowest browning index (< 2.0) throughout the 10-day period. In contrast, apical segments and higher temperatures resulted in greater browning. Overall, these results highlight the combined importance of anti-browning chemical treatments, explant position, and incubation temperature in reducing browning. The integration of PVP supplementation and low-temperature dark incubation provides a reliable and practical strategy for minimizing oxidative browning and enhancing early stage in vitro regeneration in high-value pomegranate cultivars.