Construction Of The RNAi Vectors Of Storage Protein Gene In Brassica Napus And Molecular Identification Of The Transformed Plants

Brassica napus is an important oil plant in the world. The oil content of the rapeseed influences the oil yield. Improving the oil content in oilseed, which could be resulted in a new species of Brassica napus with higher oil content, is a novel idea on development of Brassica napus genetic improvements.The seed of Brassica napus mainly contains fatty acid and storage protein. Previous studies demonstrated that the fatty acid and storage protein reserves simultaneously accumulate in the cotyledon during the seed development. Based on the TCA cycle, it was deduced that these two kinds of storage proteins may be produced from the same substrate and compete between each other. After analysis of the relationship between fatty acid and storage protein, it was estimated there is a negative correlation between each other.In order to down regulation the gene of the storage proteins NAPIN and Cruciferin in the seed of Brassica napus and get the higher oil content plant with lower storage protein, the RNAi tech was used to decrease the storage protein content in rapeseed. Then, the relationship between the overall oil content and the storage protein was evaluated by analysis of the oil content of oilseeds in different experiment groups.In this dissertation, several steps were taken to realize this idea:1.The storage protein gene sequence were collected from the database. Analysis the genesequence to confirm the conservative fragment.2.The conservative fragment used to construct the storage protein RNAi vectors. 2 vectorsnapin-pHELLSGATE and cruciferin-pHELLSGATE were constructed.3.This 2 vectors were transformated into the Brassica napus by Agrobacterium tumefaciems. There were 7 napin-pHELLSGATE transformation plants and 4 cruciferin-pHELLSGATE transformation plants maintained.4. Molecular identification, including PCR and southern bolt, were taken to make sure thegene from the vectors inserting into the transformation plants.