Okumura Satoru, Sawada Machiko, Park Yong Woo, Hayashi Takahisa, Shimamura Masaki, Takase Hisabumi, Tomizawa Ken-Ichi
Plant Research Group, Research Institute of Innovative Technology for the Earth (RITE), 9-2 Kizugawadai, Soraku-gun, Kyoto 619-0292, Japan.
Transgenic Res. 2006 Oct;15(5):637-46. doi: 10.1007/s11248-006-9009-3. Epub 2006 Sep 2.
Plastid transformation offers several unique advantages compared with nuclear genome transformation, such as high level of transgene expression within plastids, expressing multiple transgenes as operons, lack of position effect due to site-specific transgene integration, and reducing risks of gene flow via pollen due to maternal inheritance of the plastid genome. Plastid transformation has been applied to several herbal species, but as yet there are no applications to tree species. We report here the first successful plastid transformation in a tree species, Populus alba. A vector for plastid transformation of poplar (Populus alba) was constructed, which carried the spectinomycin resistance gene and the green fluorescence protein gene as marker genes. In the regenerated shoots, the site-specific integration of foreign genes and the establishment of a high homoplastomic state were confirmed. Immunoblot analysis and histological observations corroborated the accumulation of green fluorescence protein in chloroplasts. The establishment of a plastid transformation system in poplar provides a novel tool for tree biotechnology.
与核基因组转化相比，质体转化具有几个独特的优势，例如质体内转基因表达水平高、作为操纵子表达多个转基因、由于转基因位点特异性整合而缺乏位置效应以及由于质体基因组的母系遗传而降低通过花粉进行基因流动的风险。质体转化已应用于几种草本植物，但尚未应用于树种。我们在此报告了树种银白杨中首次成功的质体转化。构建了杨树（银白杨）质体转化载体，其携带壮观霉素抗性基因和绿色荧光蛋白基因作为标记基因。在再生芽中，证实了外源基因的位点特异性整合和高同质化状态的建立。免疫印迹分析和组织学观察证实了叶绿体中绿色荧光蛋白的积累。杨树质体转化系统的建立为树木生物技术提供了一种新工具。
