This project will use FISH (fluorescence in situ hybridization) technology to define the centromere region of the Brassica genome. Confirmed centromere clones will be used to construct Brassica mini-chromosomes that will remain distinct from the host genome and allow the delivery of very large DNA segments, including those that encode all of the genes in a complex pathway. In Phase I, DNA from Brassica seedlings will be harvested and DNA fragments containing centromere-specific modifications or centromere-specific repeats will be purified. These fragments will be labelled, then used as probes for hybridization to Brassica chromosomes. Confirmed centromere clones will be sequenced and used in Phase II to construct a Brassica oleracea specific mini-chromosome. Commercial Applications And Other Benefits as described by awardee: This project should impact agricultural biotechnology by enabling the transfer of multiple genes into a plant. Future applications include synthesis of biomaterials and pharmaceuticals, plants with improved yield and nutritional value, and environmentally friendly specialty chemicals.
