Tsuboyama Shoko, Kodama Yutaka
Center for Bioscience Research and Education, Utsunomiya University, Tochigi 321-8505, Japan.
Plant Biotechnol (Tokyo). 2018 Dec 25;35(4):399-403. doi: 10.5511/plantbiotechnology.18.0917a.
The liverwort L. is an important model species for investigating land plant evolution. Effective genetic transformation techniques are crucial for plant molecular biology and simplified or improved techniques for specific cultivars or strains can accelerate research. Over the past several years, we developed a simple -mediated transformation technique for named AgarTrap (Agar-utilized transformation with pouring solutions). AgarTrap is an easy technique that involves pouring the appropriate solutions onto plant materials on a single solid plate of medium. We recently improved AgarTrap using gemmalings (G-AgarTrap) of the female model strain BC3-38 and achieved a transformation efficiency of nearly 100%. Based on this improved technique, in the current study, we adopted two factors (sealing the Petri dish with Parafilm and dark treatment during co-cultivation) and optimized two factors ( strain and pre-culture period) of the improved G-AgarTrap for other model strains of , the male strain Takaragaike-1 (Tak-1) and the female strain Takaragaike-2 (Tak-2). After optimization, the transformation efficiency of Tak-1 using G-AgarTrap was as high as 55% compared to approximately 30% using the previous protocol. Furthermore, using Tak-2, we achieved a transformation efficiency of nearly 100%. Our improved G-AgarTrap technique for Tak-1 and Tak-2 represents a promising tool for promoting the study of .
苔类植物L.是研究陆地植物进化的重要模式物种。有效的遗传转化技术对植物分子生物学至关重要，针对特定品种或菌株简化或改进技术可以加速研究进程。在过去几年中，我们开发了一种针对苔类植物的简单介导转化技术，命名为琼脂陷阱法（Agar-utilized transformation with pouring solutions，即通过倾注溶液利用琼脂进行转化）。琼脂陷阱法是一种简便的技术，只需将适当的溶液倾注到单个固体培养基平板上的植物材料上即可。我们最近利用苔类植物雌性模式菌株BC3-38的幼芽（G-琼脂陷阱法）改进了琼脂陷阱法，并实现了近100%的转化效率。基于这项改进技术，在本研究中，我们针对苔类植物的其他模式菌株，即雄性菌株高贺川池-1（Tak-1）和雌性菌株高贺川池-2（Tak-2），采用了两个因素（用Parafilm密封培养皿和共培养期间进行暗处理）并优化了改进后的G-琼脂陷阱法的两个因素（菌株和预培养期）。优化后，使用G-琼脂陷阱法的Tak-1的转化效率高达55%，而使用先前方案时约为30%。此外，对于Tak-2，我们实现了近100%的转化效率。我们针对Tak-1和Tak-2改进的G-琼脂陷阱法是促进苔类植物研究的一种很有前景的工具。
