Jeong Yeong Yeop, Noh Yoo-Sun, Kim Suk Weon, Seo Pil Joon
Department of Chemistry, Seoul National University, Seoul 08826, Korea.
Plant Genomics and Breeding Institute, Seoul National University, Seoul 08826, Korea.
Biol Methods Protoc. 2024 Feb 6;9(1):bpae008. doi: 10.1093/biomethods/bpae008. eCollection 2024.
Protoplast regeneration has become a key platform for genetic and genome engineering. However, we lack reliable and reproducible methods for efficient protoplast regeneration for tomato () cultivars. Here, we optimized cell and tissue culture methods for protoplast isolation, microcallus proliferation, shoot regeneration, and plantlet establishment of the tomato cultivar Micro-Tom. A thin layer of alginate was applied to protoplasts isolated from third to fourth true leaves and cultured at an optimal density of 1 × 10 protoplasts/ml. We determined the optimal culture media for protoplast proliferation, callus formation, shoot regeneration, and root regeneration. Regenerated plantlets exhibited morphologically normal growth and sexual reproduction. The entire regeneration process, from protoplasts to flowering plants, was accomplished within 5 months. The optimized protoplast regeneration platform enables biotechnological applications, such as genome engineering, as well as basic research on plant regeneration in species.
原生质体再生已成为遗传和基因组工程的关键平台。然而，对于番茄（）品种，我们缺乏可靠且可重复的高效原生质体再生方法。在此，我们优化了番茄品种Micro-Tom的原生质体分离、微愈伤组织增殖、芽再生和植株建立的细胞及组织培养方法。将一层薄薄的藻酸盐应用于从第三至第四片真叶分离的原生质体，并以1×10个原生质体/毫升的最佳密度进行培养。我们确定了原生质体增殖、愈伤组织形成、芽再生和根再生的最佳培养基。再生植株表现出形态正常的生长和有性繁殖。从原生质体到开花植物的整个再生过程在5个月内完成。优化后的原生质体再生平台可实现生物技术应用，如基因组工程，以及对物种植物再生的基础研究。
