Evaluation of leaflets from mature Zygotic Embryos as recipient Tissue for Biolistic gene transfer to peanut (Arachis hypogaea L.)
TE Clemente; D Robertson; MK Beute; TG Isleib; AK Weissinger
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Abstract: Leaflets from mature peanut embryos are a useful recipient tissue for biolistic DNA transfer. Fertile plants were regenerated from leaflets from genotypes representing all botanical types of peanut. Regeneration frequency was strongly influenced by genotype. NPT II ‘nd GUS chimaeric gene fusions, driven by the CaMV 35S promoter, were expressed transiently following biolistic delivery to unexpanded leaflets. Bombardment conditions affecting transient expression frequency were determined using a prototype of the Bio Rad PDS 1000/He helium-pcwered particle acceleration apparatus. Stably transformed calli were derived routinely from leaflet
tissue bombarded with the NPT II gene and subsequently cultured on kanamycin. Several plants have been regenerated from treated explants under kanamycin selection. Thus far, none of these has been stably transformed. The occurrence of escapes suggests that kanamycin is an inefficient selective agent for the recovery of transgenic peanuts from this explant. Experiments designed to regenerate plants using published regeneration
protocols from stably transformed calli, devoid of primary explant tissue, have been unsuccessful.