Wu Yuyong, Chang Ling, Jiang Chunmei, Xu Letian, Zhang Jiang
State Key Laboratory of Biocatalysis and Enzyme Engineering, College of Life Sciences, Hubei University, Wuhan, Hubei, China.
Methods Mol Biol. 2021;2317:257-265. doi: 10.1007/978-1-0716-1472-3_14.
Poplar (Populus) is an important forest tree and considered model for perennial trees. Here we describe a method for poplar plastid transformation, which involves preparation of explants, vector construction, biolistic bombardment, regeneration and selection of transplastomic poplar plants. The young leaves of 4-week-old poplar plants are used for biolistic bombardment and aadA gene as selectable marker. Homoplasmic transplastomic lines are obtained after regeneration and several rounds of selection with spectinomycin over 10 months. Homoplasmy is further confirmed by Southern blot. The establishment of a plastid transformation system for Populus is likely to make a significant contribution to tree genetic improvement.
杨树（Populus）是一种重要的林木，被视为多年生树木的模式植物。在此，我们描述了一种杨树质体转化方法，该方法包括外植体的制备、载体构建、生物枪轰击、再生以及转基因杨树植株的筛选。4周龄杨树植株的幼叶用于生物枪轰击，aadA基因作为选择标记。经过再生以及用壮观霉素进行10个月的多轮筛选后，获得了同质转基因株系。通过Southern杂交进一步确认了同质性。杨树质体转化系统的建立可能会对树木遗传改良做出重大贡献。
