Technical Advance: Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker.
Technical Advance: Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker.
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发表时间：
1999
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通讯作者：
Sidorov;Kasten;Pang;Hajdukiewicz;Staub;Nehra
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作者：
Sidorov;Kasten;Pang;Hajdukiewicz;Staub;Nehra
我们在这里描述了马铃薯可再生质体转化系统的发展和均匀转化质体的植株再生。将两个不同的烟草特异性质体载体pZS197 （Prrn/aadA/TpsbA）和pMON30125 （Prrn/GFP/Trps16:PpsbA/aadA/TpsbA）分别用于整合到质体基因组的大单拷贝区和倒重复区，并将其轰击到马铃薯株系FL1607的叶片外植体中。在pZS197轰击的46个培养板中，共筛选到3个转基因品系；在pMON30125轰击的104个培养板中，共筛选到3个转基因品系。建立基于叶片的高频再生系统、严格的选择方案和优化的生物转化方案是恢复质体转化体的关键。利用质体表达的绿色荧光蛋白作为视觉标记，在选育前期和茎部再生阶段对质体转化体进行鉴定。马铃薯质体转化体系的建立，与常规的核转化相比，具有许多优点，为马铃薯遗传改良提供了新的可能性。
We describe here the development of a reproducible plastid transformation system for potato and regeneration of plants with uniformly transformed plastids. Two distinct tobacco-specific plastid vectors, pZS197 (Prrn/aadA/TpsbA) and pMON30125 (Prrn/GFP/Trps16:PpsbA/aadA/TpsbA), designed for integration into the large single copy and inverted repeat regions of the plastid genome, respectively, were bombarded into leaf explants of potato line FL1607. A total of three transgenic lines were selected out of 46 plates bombarded with pZS197 and three transgenic lines out of 104 plates were obtained with pMON30125. Development of a high frequency leaf-based regenera- tion system, a stringent selection scheme and optimization of biolistic transformation protocol were critical for recovery of plastid transformants. Plastid-expressed green fluorescent protein was used as a visual marker for identification of plastid transformants at the early stage of selection and shoot regeneration. The establishment of a plastid transformation system in potato, which has several advantages over routinely used nuclear transformation, offers new possibilities for genetic improvement of this crop.