Regeneration Of Strawberry And Genetic Transformation By AFP Gene

The effect of culture media supplemented with different hormones on the rapid propagation and plantlet regeneration was compared using creeping stem tip, leaf disc, petiole and its lower part of strawberry as explants. The results show that for shoot inducing of the creeping stem tip the suitable culture medium is MS + 6-BA 1.0 mg/I + NAA 0. 01 mg/I, for rooting is MS + IBA 0. 5 mg/I. The different genotype of strawberry and hormone supplement are remarkably affected on the adventitious bud inducing of leaf disc, petiole and its lower part of strawberry. The genotype of "Allstar" is easy to regenerate shoots, but most varieties of strawberry in medium MS+B₅+6-BA 2. 3 mg/I + IAA 1.8 mg/I can produce shoots with high inducing frequency. its lower part is the most suitable explant for plantlet regeneration and genetic transformation. The leaf disc and lower part of petiole of strawberry were transformed by the pre, pro, and mature antifreeze protein genes mediated by Agrobacterium tumefaciens carried a binary vector pPCV002. An efficient transformation system was established. The total DNA of transgenic plants was isolated. PCR amplification showed that the foreign AFP genes encoding pre and pro were integrated independently into the genome of transgenic strawberry. The assay of resistance of detached leaves to cold treatment indicated that about 50% detached leaves of transgenic plants were survived under -8 —10℃ for 2 hours while all untransgenic detached leaves were died.