Genetic transformation of Brassica nigra by agrobacterium based vector and direct plasmid uptake
Genetic transformation of Brassica nigra by agrobacterium based vector and direct plasmid uptake
复制标题
通过基于农杆菌的载体和直接质粒摄取对黑芥进行遗传转化
作者：
Vibha Gupta;G. Sita;M. Shaila;V. Jagannathan
建立了黑菜遗传转化体系。IC257)，通过使用农杆菌双载体以及通过直接DNA摄取质粒载体。两种载体均携带nptII基因和gus基因。农杆菌介导的转化采用下胚轴组织外植体，高达33%的外植体在选择培养基上产生了愈伤组织。组织化学染色均表达β-葡萄糖苷酸酶基因。从幼苗下胚轴组织中分离出的原生质体可以通过FEG介导的载体DNA被转化到载体DNA中。用这两种方法获得了一批可育的卡那霉素抗性植株，并通过Southern印迹分析和GUS组织化学染色证实了它们的转化性质。这些转化植株的回交和自交后代表现出npt和gus基因的存在。
SummaryGenetic transformation systems have been established for Brassica nigra (cv. IC 257) by using an Agrobacterium binary vector as well as by direct DNA uptake of a plasmid vector. Both the type of vectors carried nptII gene and gus gene. For Agrobacterium mediated transformation, hypocotyl tissue explants were used, and up to 33% of the explants produced calli on selection medium. All of these expressed B-glucuronidase gene on histochemical staining. Protoplasts isolated from hypocotyl tissues of seedlings could be transformed with a plasmid vector by FEG mediated uptake of vector DNA. A number of fertile kanamycin resistant plants were obtained using both the methods, and their transformed nature was confirmed by Southern blot analysis and histochemical staining for GUS. Backcrossed and selfed progenies of these transformed plants showed the presence of npt and gus genes.