Abstract:

Using plantlet shoot sections near the shoot tip as explants, after studying the six factors to affect transformation, we have preliminarily determined an effective system for genetic transformation. The protocol was: 1～3 days for pre-culture, OD_{600 nm} =0.1～0.4, 10～15 minutes for infection, 3～5 days for co-culture, and acetosyringone (AS) 80 μmol·L^-1 as co-culture medium, and then lasting 3 days before selecting. And then 186 regenerated kanamycin(Km)-resistant buds were obtained from the first selection medium. 39 Km-resistant buds were obtained from the second selection medium. PCR analysis on 39 regenerated Km-resistant buds showed that 2 buds were PCR positive. The result indicated that gastrodia antifungal protein (GAFP) gene was integrated into the genome of the two Km-resistant buds.

Key words: Cunninghamia lanceolata, Agrobacterium tumefaciens mediated transformation, gastrodia antifungal protein gene