Enhancing efficiency of the gene transform system via Agrobacterium tumefaciens mediated in apple

摘要

A simple and high-efficient genetic transform system via Agrobacterium tumefaciens mediated by using leaf disc regeneration was developed by analyzing the factors that affect the transformation rate, suitable selecting pressure and methods for the main conventional apple cultivars in Northern China. The leaves selected from in vitro cultured plantlets were soaked in Agrobacterium solution (OD=0.6-0.8) about 10-20 min, co-cultured for 3-4d, sterilized on the medium amended with cefotaxime (Cef) for 2d, then selected at the selecting pressure of kanamycin (Km) 30-40 mg/L for 15-20d, and finally, transferred onto the Km-free medium to induce Km-resistant adventious shoots. Transgenic plants were acquired by subculture of shoots on selecting medium containing Km 50mg/L. Southern blotting confirmed that CpTI gene was successfully inserted into 'Fuji', 'Jonagold', 'Orin' and 'Royal Gala' apples.