Studies On The Transformation Of B.campestris L.and B.napus L.with Fad2 Gene And Bar Gene

High oleic rape seed breeding play a important role in oilseeds breeding, and breeding herbicide-resistant crops is an available measure to agricultural labor force cost increasing and environment pollution. This study hopes to develop high oleic content, herbicide-resistant B.rape and B.napus breeding germ plasm through transfonnating bar and antisense fad2 fragment into XiangYou 15 and CangGubai 9-4 by using Agrobacterium-mediated genetic tansformation. The results were described as following:1. Introduction of bar cassettee (herbicide-resistant gene) into a plasmid pBI121.NF that antisenses express fade2 fragment (oleic acid desaturase gene), and constructed a new plant expression vector, named pBI121 .XG.2. Transformed bar and antisense fad2 fragment into XiangYou 15 and CangGubai 9-4 by using Agrobacterium-mediated genetic tansformation, and obtained 5 XiangYou 15 tansformants, 0.92% tansformation efficiency. Obtained 7 CangbaiGu 9-4 transformants, 2.52% tansformation efficiency.3. Established high efficiency transformation system of multiple shoot via Cotyledon nodes of rape seed.4. 3 methods: CTAB method, SDS method and ShanLichun method were used to extract genomic DNA from the seedling of XiangYou 15.The result DNA samples of these 3 methods were tested by using spectrophotometer, agarose gel electrophoresis. The difference in yield, quality of the obtained DNAs were compared, and ShanLichun methods as the best method in the test.5. The result of 5 pairs primers and 2 DNAploymerases in amplifying bar from rape seed genomic DNA were compared, and primer B441 and pfu DNAploymerases as the best for this PCR.