Bansal Kailash C, Singh Ajay K
National Bureau of Plant Genetic Resources (ICAR), New Delhi, India.
Methods Mol Biol. 2014;1132:305-16. doi: 10.1007/978-1-62703-995-6_19.
Eggplant (Solanum melongena L.) is an important vegetable crop of tropical and temperate regions of the world. Here we describe a procedure for eggplant plastid transformation, which involves preparation of explants, biolistic delivery of plastid transformation vector into green stem segments, selection procedure, and identification of the transplastomic plants. Shoot buds appear from cut ends of the stem explants following 5-6 weeks of spectinomycin selection after bombardment with the plastid transformation vector containing aadA gene as selectable marker. Transplastomic lines are obtained after the regenerated shoots are subjected to several rounds of spectinomycin selection over a period of 9 weeks. Homoplasmic transplastomic lines are further confirmed by spectinomycin and streptomycin double selection. The transplastomic technology development in this plant species will open up exciting possibilities for improving crop performance, metabolic engineering, and the use of plants as factories for producing biopharmaceuticals.
茄子(Solanum melongena L.)是世界热带和温带地区的一种重要蔬菜作物。在此,我们描述了一种茄子质体转化方法,该方法包括外植体的制备、将质体转化载体通过基因枪导入绿色茎段、筛选过程以及对转质体植物的鉴定。用含有aadA基因作为选择标记的质体转化载体轰击后,经过5 - 6周的壮观霉素筛选,茎外植体的切口端会出现芽。再生芽经过9周的几轮壮观霉素筛选后获得转质体系。通过壮观霉素和链霉素双重筛选进一步确认同质性转质体系。该植物物种的转质体技术发展将为改善作物性能、代谢工程以及利用植物作为生产生物制药的工厂开辟令人兴奋的可能性。