Stable transformation of petunia plastids
Stable transformation of petunia plastids
复制标题
DOI：
10.1007/s11248-004-2374-x
复制
发表时间：
2004-12
影响因子：
3
通讯作者：
M. Zubko;E. Zubko;K. V. Zuilen;P. Meyer;A. Day
中科院分区：
文献类型：
--
作者：
M. Zubko;E. Zubko;K. V. Zuilen;P. Meyer;A. Day
质体转化导致稳定表达的外源基因，对于大多数被子植物来说，这些外源基因在很大程度上被排除在精细胞之外，从而大大降低了外源基因通过花粉传播的风险。在此之前，可育质体转化体仅限于烟草、番茄和Lesquerella。质体工程在重要花卉产业中的应用需要在主要观赏植物如矮牵牛中开发稳定的质体转化。在这里，我们描述了在一个商业栽培品种的可育和稳定的质体转化体的成功分离。hybrida（var. Pink Wave）。利用烟草的质体靶向区域，将ad A和gusA整合到P.基因枪轰击后的杂种质体DNA。对于三个大观霉素和链霉素抗性品系，DNA印迹分析证实转基因整合到质体DNA和同质性。母本遗传和同质性导致自交后100个大观霉素抗性传递给后代。质体转化体表达thegusA基因均匀叶片内和可比的水平，在所有三个线。插入性状基因代替gusA编码序列使得我们的质体转化载体能够立即应用。建立了P. hybrida有助于安全可靠地使用这种重要的观赏植物进行研究和植物生物技术。
Plastid transformation results in stably expressed foreign genes, which for most Angiosperms are largely excluded from sperm cells, thereby greatly reducing the risk of foreign gene spread through pollen. Prior to this work, fertile plastid transformants were restricted to tobacco, tomato andLesquerella. Application of plastid engineering in the important floriculture industry requires the development of stable plastid transformation in a major ornamental plant species such asPetunia hybrida. Here we describe the successful isolation of fertile and stable plastid transformants in a commercial cultivar ofP. hybrida (var. Pink Wave). Plastid targeting regions from tobacco were used to integrateaad AandgusAbetween theacc Dandrbc Lgenes ofP. hybrida plastid DNA following particle bombardment of leaves. For three spectinomycin and streptomycin resistant lines, DNA blot analysis confirmed transgene integration into plastid DNA and homoplasmy. Maternal inheritance and homoplasmy resulted in 100 transmission of spectinomycin resistance to progeny after selfing. Plastid transformants expressed thegusAgene uniformly within leaves and to comparable levels in all three lines. Insertion of trait genes in place ofgusAcoding sequences enables immediate applications of our plastid transformation vector. Establishment of plastid transformation inP. hybrida facilitates a safe and reliable use of this important ornamental plant for research and plant biotechnology.