Authors
Jinjuan Bai, Feijie Wu, Yanfei Mao, and Yuke He
Summary
In planta transformation has been widely used for gene transfer in the model plant Arabidopsis. Unlike Arabidopsis, the genus Brassica that comprises many important vegetable, oil seed and weed crops are vernalization-required, cross-pollinated, and self-incompatible. The protocols for in planta transformation established for Arabidopsis are not directly applicable to these crops. We take advantage of Arabidopsis vacuum infiltration to establish vernalization-infiltration method for transformation of Brassica crops. The germinated seeds of Chinese cabbage and oilseed rape are exposed to cold treatment for a period of vernalization; the floral buds of the early inflorescences are opened by hand before Agrobacterium infection; Agrobacterium-infected inflorescences are infiltrated with vacuum; the transformed stigmas are pollinated with the non-infected pollens of siblings; and hygromycin resistance is used to select the transformants. Using the vernalization-infiltration method, we have identified 52 homozygous transgenic lines from the cultivars of Chinese cabbage and oilseed rape.Further details
The protocol was published on Protocol Exchange on 7 August 2013. To see the entire protocol, click on the source link.Advertisement
Stats
- Recommendations n/a n/a positive of 0 vote(s)
- Views 629
- Comments 0