Biotechnology Research Centre, Jilin Academy of Agricultural Sciences, 1363, Caiyu Street, Changchun 130033, People's Republic of China,
Biotechnol Lett. 2011 Dec;33(12):2487-94. doi: 10.1007/s10529-011-0709-2. Epub 2011 Jul 23.
The ability to transform chloroplasts in multiple species is important for improving agricultural traits. Chloroplast transformation of alfalfa (Medicago sativa L.), a useful forage plant with high market value, was achieved using a vector carrying aadA and gfp genes being introduced into the chloroplasts of alfalfa via particle bombardment using leaves and calli as explants. Resistant somatic embryos were generated and developed into plantlets from explants. The transformation efficiency was 1.3% for callus explants and 2.7% for leaf explants. PCR and Southern blotting analyses revealed that the foreign genes were integrated into the transformed chloroplast genome. The occurrence of GFP was further confirmed by fluorescence microscopy. Expression of foreign genes in alfalfa chloroplasts is therefore possible, and provides a novel means for genetic improvement of agronomically important traits and production of value-added proteins.
在多个物种中转化叶绿体的能力对于改善农业性状很重要。通过利用携带 aadA 和 gfp 基因的载体，通过粒子轰击将其导入紫花苜蓿（Medicago sativa L.）叶绿体中，实现了紫花苜蓿这种具有高市场价值的有用饲料植物的叶绿体转化。利用叶片和愈伤组织作为外植体，生成了抗性体细胞胚，并从外植体中发育成了植株。转化效率在愈伤组织外植体中为 1.3%，在叶片外植体中为 2.7%。PCR 和 Southern 印迹分析表明，外源基因已整合到转化的叶绿体基因组中。通过荧光显微镜进一步证实了 GFP 的存在。因此，外源基因在紫花苜蓿叶绿体中的表达是可能的，为遗传改良重要农艺性状和生产增值蛋白提供了新的手段。
