This study investigated the optimal culture conditions of anther-derived callus induction according to heat shock treatment, culture medium, and the concentration of myo-inositol, AgNO3, and Fe-EDTA. We also explored the optimal culture conditions for the regeneration of haploid-derived plants, made by using anther-derived callus, according to the concentration of Murashige and Skoog (MS) medium and rhizosphere temperature. The highest callus induction rate by heat shock treatment was found when incubated at 32 °C for 24 h. Of the three kinds of culture medium used (MS, Lichter [NLN], and Gamborg B5), NLN showed the highest callus induction rate. The addition of 100 mg L−1 of myo-inositol showed a higher callus induction rate in all three media compared with treatments without myo-inositol addition. When AgNO3 was added, the highest callus induction rate was 71.4% at a concentration of 25 mg L−1. Fe-EDTA showed the highest callus induction rate of 51.0% when 100 mg L−1 was added. The 0.5-fold concentration of MS medium exhibited the highest regeneration rate (92.7%). To improve the acclimation efficiency and growth of the regenerated plants in vitro, the growth and survival rates of the regenerated plants were examined by adjusting the rhizosphere temperature to either 25 ± 2 °C or 18 ± 2 °C. The survival rate was 62.5% in both rhizosphere temperature treatments. However, the rhizosphere temperature of 25 ± 2 °C showed significantly higher shoot and root growth than treatment at 18 ± 2 °C. Summarizing the results of this study, embryogenic callus was most effective when cultured in a NLN medium supplemented with 100 mg L−1 Myo-inositol, 25 mg L−1 AgNO3, 75 mg L−1 Fe-EDTA for 24 h heat shock treatment at 32 °C. And the regeneration rate of anther-derived callus induced in the above conditions was highest when regenerated in × 0.5 MS medium. In addition, the regenerated plant had the highest growth and survival rate when acclimated in a growth chamber where the rhizosphere temperature was adjusted to 25 ± 2 °C.

中文翻译：

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草莓'Jukhyang'花药愈伤组织、胚及再生的最佳培养环境

本研究根据热休克处理、培养基和肌醇、AgNO3 和 Fe-EDTA 的浓度研究了花药衍生愈伤组织诱导的最佳培养条件。我们还根据 Murashige 和 Skoog (MS) 培养基的浓度和根际温度探索了使用花药衍生愈伤组织再生单倍体衍生植物的最佳培养条件。当在 32°C 下培养 24 小时时，发现通过热休克处理的最高愈伤组织诱导率。在使用的三种培养基（MS、Lichter [NLN] 和 Gamborg B5）中，NLN 显示出最高的愈伤组织诱导率。与不添加肌醇的处理相比，添加 100 mg L-1 的肌醇在所有三种培养基中均显示出更高的愈伤组织诱导率。当加入 AgNO3 时，在 25 mg L−1 的浓度下，最高的愈伤组织诱导率为 71.4%。当添加 100 mg L-1 时，Fe-EDTA 显示出最高的愈伤组织诱导率为 51.0%。0.5 倍浓度的 MS 培养基表现出最高的再生率 (92.7%)。为了提高体外再生植物的驯化效率和生长，通过将根际温度调整到 25 ± 2 °C 或 18 ± 2 °C 来检查再生植物的生长和存活率。两种根际温度处理的成活率为62.5%。然而，25 ± 2 °C 的根际温度显示出明显高于 18 ± 2 °C 处理的地上部和根系生长。总结这项研究的结果，胚胎发生愈伤组织在补充有 100 mg L-1 肌醇、25 mg L-1 AgNO3、75 mg L-1 Fe-EDTA 在 32 °C 下进行 24 小时热休克处理。在上述条件下诱导的花药衍生愈伤组织在×0.5 MS培养基中再生时再生率最高。此外，再生植物在根际温度调节为 25±2°C 的生长室中适应时具有最高的生长和成活率。