Stable genetic transformation of garlic plants using particle bombardment.
The improvement of garlic plants (Allium sativum L.) via biotechnological approaches is currently limited by the lack of an applicable direct gene transfer system. In this paper, we present the development of a genetic transformation system using particle bombardment for gene delivery and immature clove-derived callus as the gene target. Plasmid DNA (pBI221.23), containing the selectable "hpt" gene for hygromycin resistance and the reporter "gus" gene, was delivered into callus tissue that had been previously treated with aurintricarboxylic acid as an endogenous nuclease inhibitor. The transformed calli were selected using hygromycin B, regenerated, and analysed at the molecular level using DNA hybridization, transgenome rescue and histochemical beta-glucuronidase assay. The results indicated that biolistic transformation can lead to the transfer, expression and stable integration of a DNA fragment into garlic chromosomal DNA. The relative simplicity of this system is a good recommendation for its future use in the production of genetically modified garlic plants.
Sawahel
Wagdy A
WA
Microbial Genetics Department, Genetic Engineering & Biotechnology Division, National Research Center, Dokki, Cairo, Egypt.
D019470:Biolistics / Q000379:methods*; D015139:Blotting, Southern; D002934:Cinnamates; D018744:DNA, Plant / Q000032:analysis; D004351:Drug Resistance; D005737:Garlic / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D006921:Hygromycin B / Q000031:analogs & derivatives*; D010641:Phenotype; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic
D002934:Cinnamates; D018744:DNA, Plant; D006921:Hygromycin B; C026273:hygromycin A