Top–down control analysis (TDCA) is a useful tool for quantifying constraints on metabolic pathways that might be overcome by biotechnological approaches. Previous studies on lipid accumulation in oilseed rape have suggested that diacylglycerol acyltransferase (DGAT), which catalyses the final step in seed oil biosynthesis, might be an effective target for enhancing seed oil content. Here, increased seed oil content, increased DGAT activity, and reduced substrate:product ratio are demonstrated, as well as reduced flux control by complex lipid assembly, as determined by TDCA in Brassica napus (canola) lines which overexpress the gene encoding type-1 DGAT. Lines overexpressing DGAT1 also exhibited considerably enhanced seed oil content under drought conditions. These results support the use of TDCA in guiding the rational selection of molecular targets for oilseed modification. The most effective lines had a seed oil increase of 14%. Moreover, overexpression of DGAT1 under drought conditions reduced this environmental penalty on seed oil content.
Hybridizations among Brassica napm, B. rapa and B.juncea and their two weedy relatives B, nigra and Sinapis arvensis under open pollination conditions in the field
AbstractThe rate of natural crosses occurring among the cultivated Brassica species B. napus, B. rapa, and B. juncea and their two weedy relatives B. nigra and Sinapis arvensis was studied in co-cultivation experiments under field conditions in Saskatchewan, Canada, with special reference to evaluation of the possibility of transgene escape from the cultivated to the weedy species. Natural crosses occurred among B. napus, B, rapa, and B.juncea, indicating that hybridizations among these three species do occur under field conditions. On the other hand, no natural crosses occurred between the cultivated species and B. nigra or S, arvensis. It is concluded that the crosses between the cultivated and weedy species are practically impossible under field conditions in Saskatchewan, and that the escape of transgenes from transgenic cultivars of B. napus, B. rapa and B. juncea into B. nigra and 5, arvensis is basically zero in this region.
We report the tagging of genes involved in blackleg resistance, present in the French cultivar Crésor of B. napus, with RFLP markers. A total of 218 cDNA probes were tested on the parental cultivars Crésor (resistant) and Westar (susceptible), and 141 polymorphic markers were used in a segregating population composed of 98 doubled-haploid lines (DH). A genetic map from this cross was constructed with 175 RFLP markers and allowed us to scan for specific chromosomal associations between response to blackleg infection and RFLP markers. Canola residues infested with virulent strains of Leptosphaeria maculans were used as inoculum and a suspension of pycnidiospores from cultures of L. maculans, including the highly virulent isolate Leroy, was sprayed to increase disease pressure. QTL mapping suggested that a single chromosomal region was responsible for resistance in each of the four environments tested. This QTL accounted for a high proportion of the variation of blackleg reaction in each of the assays. A second QTL, responsible for a small proportion of the variation of blackleg reaction, was present in one of four year-site assays. A Mendelian approach, using blackleg disease ratings for classifying DH lines as resistant or susceptible, also allowed us to map resistance in the region of the highly significant LOD scores observed in each environment by interval mapping. Results strongly support the presence of a single major gene, named LmFr 1 controlling adult plant resistance to blackleg in spring oil-seed rape cultivar Crésor. Several RFLP markers were found associated with LmFr 1.
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