Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup, Republic of Korea.
Department of Biotechnology, Chonnam National University, Gwangju, Republic of Korea.
PLoS One. 2023 Feb 24;18(2):e0279627. doi: 10.1371/journal.pone.0279627. eCollection 2023.
Whole-plant regeneration via plant tissue culture is a complex process regulated by several genetic and environmental conditions in plant cell cultures. Recently, epigenetic regulation has been reported to play an important role in plant cell differentiation and establishment of pluripotency. Herein, we tested the effects of chemicals, which interfere with epigenetic regulation, on the plant regeneration from mesophyll protoplasts of lettuce. The used chemicals were histone deacetylase inhibitors trichostatin A (TSA) and sodium butyrate (NaB), and the DNA methyltransferase inhibitor azacytidine (Aza). All three chemicals increased cell division, micro-callus formation and callus proliferation in lettuce protoplasts. Cell division increased by more than 20% with an optimal treatment of the three chemicals. In addition, substantial increase in the callus proliferation rates was observed. In addition, TSA enhances cell division and adventitious shoot formation in the protoplast culture of Nicotiana benthamiana. The regenerated tobacco plants from TSA-treated protoplasts did not show morphological changes similar to the control. TSA increased histone H3 acetylation levels and affected the expression of CDK, CYCD3-1, and WUS in tobacco protoplasts. Thus, we investigated the effect of TSA, NaB, and Aza on Lactuca sativa L. protoplasts and the effect of TSA on cell division and callus formation in Nicotiana benthamiana protoplasts, which facilitates plant regeneration from mesophyll protoplasts. Furthermore, these chemicals can be directly applied as media additives for efficient plant regeneration and crop improvement in various plant species.
通过植物组织培养进行全植物再生是一个复杂的过程，受到植物细胞培养中几种遗传和环境条件的调节。最近，表观遗传调控被报道在植物细胞分化和多能性建立中发挥重要作用。在此，我们测试了干扰表观遗传调控的化学物质对生菜叶肉原生质体植物再生的影响。所用的化学物质是组蛋白去乙酰化酶抑制剂曲古抑菌素 A（TSA）和丁酸钠（NaB），以及 DNA 甲基转移酶抑制剂氮杂胞苷（Aza）。这三种化学物质都能增加生菜原生质体的细胞分裂、微愈伤组织形成和愈伤组织增殖。三种化学物质的最佳处理使细胞分裂增加了 20%以上。此外，还观察到愈伤组织增殖率的大幅度增加。此外，TSA 增强了烟草原生质体培养中的细胞分裂和不定芽形成。用 TSA 处理的原生质体再生的烟草植物没有表现出与对照相似的形态变化。TSA 增加了组蛋白 H3 乙酰化水平，并影响了烟草原生质体中 CDK、CYCD3-1 和 WUS 的表达。因此，我们研究了 TSA、NaB 和 Aza 对生菜原生质体的影响，以及 TSA 对烟草原生质体细胞分裂和愈伤组织形成的影响，这有助于从叶肉原生质体进行植物再生。此外，这些化学物质可以直接用作培养基添加剂，以提高各种植物物种的植物再生和作物改良效率。
