FAS Abstracts 2004 Meeting Page

Agricultural Sciences (AGR): AGR-2

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Development of an alternate genetic transformation system for Vitis.  M. DUTT, Z. LI, and D.J. GRAY.  Mid Florida Research and Education Center, Univ. of Florida, Apopka, FL, 32703. Agrobacterium mediated transformation in Vitis sp. (grape) is routinely carried out using somatic embryos derived from proembryogenic masses (PEM). However initiation and proliferation of PEM is laborious and genotype dependent in that somatic embryogenesis cannot be obtained from all Vitis species and cultivars. Among those genotypes that produce PEM, proliferation and culture maintenance is often difficult-to-impossible.  An alternate transformation method utilizing in vitro shoot (microprop) cultures is being explored.  Such cultures are easier to maintain when compared to embryogenic cultures and there is less genotypic variation in response.  Genetic constructs containing neomycin phosphotransferase (nptII) and green fluorescent protein (GFP) genes are being utilized to select transgenic tissues from  Agrobacterium infected microprop cultures.  The nptII gene allows selection of transgenic tissue using antibiotics such as kanamycin and GFP provides non-destructive visualization of transformed cells and tissues.  To date, stable transgenic shoots were produced within two months after transformation.  Procedures to optimize transformation of microprop cultures from a range of Vitis species and cultivars are being developed.