Graduate School of Informatics, Nagoya University, Nagoya, Japan.
Methods Mol Biol. 2021;2317:321-331. doi: 10.1007/978-1-0716-1472-3_19.
The moss Physcomitrium (Physcomitrella) patens performs efficient homologous recombination in both the nucleus and plastid enabling the study of individual gene function by generating precise inactivation or modification of genes. Polyethylene glycol (PEG)-mediated transformation of protoplasts is routinely used to study the nuclear gene function of P. patens. PEG-mediated protoplast transformation is also applied for plastid transformation of this moss. The efficiency of plastid transformation is quite reliable and one or two homoplasmic transplastomic lines are obtained in a plastid transformation experiment (5 × 10 protoplasts) by selection for spectinomycin resistance.
藓类植物 Physcomitrium(泡叶藻)patens 在细胞核和质体中都能进行高效的同源重组,这使其能够通过精确灭活或修饰基因来研究单个基因的功能。聚乙二醇(PEG)介导的原生质体转化通常用于研究 P. patens 的核基因功能。PEG 介导的原生质体转化也应用于该藓类植物的质体转化。质体转化的效率相当可靠,通过壮观霉素抗性选择,在一个质体转化实验(5×10 个原生质体)中可以获得一到两个同质质体转化系。