Transformation of poplar (Populus alba) plastids and expression of foreign proteins in tree chloroplasts
Transformation of poplar (Populus alba) plastids and expression of foreign proteins in tree chloroplasts
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DOI：
10.1007/s11248-006-9009-3
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发表时间：
2006-10-01
影响因子：
3
通讯作者：
Tomizawa, Ken-Ichi
中科院分区：
文献类型：
--
作者：
Okumura, Satoru;Sawada, Machiko;Tomizawa, Ken-Ichi
与核基因组转化相比，叶绿体转化具有一些独特的优点，如在叶绿体中高水平的转基因表达，多个转基因作为操纵子表达，由于定点转基因整合而缺乏位置效应，以及减少由于叶绿体基因组的母系遗传而通过花粉进行基因流动的风险。叶绿体转化已应用于几种草本植物，但尚未应用于树种。我们在这里报道了第一个成功的树种--白杨的叶绿体转化。构建了以壮观霉素抗性基因和绿色荧光蛋白基因为标记基因的杨树叶绿体转化载体。在再生芽中，证实了外源基因的定点整合和高度同源体态的建立。免疫印迹分析和组织学观察证实了绿色荧光蛋白在叶绿体中的积累。杨树叶绿体转化体系的建立为树木生物技术提供了一种新的工具。
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.