Brassica carinata (A.) Braun and Camelina sativa (L.) Crantz are two re-emerging oilseed crops of the Brassicaceae family that are being adapted for cultivation in western Canada. Both seeds of these species reportedly accumulate considerable amounts of sulfur-containing secondary metabolites called glucosinolates. The purpose of the current work was to gain knowledge of the occurrence and distribution of glucosinolates during primary processing of these oilseeds, including during pressing and dehulling. In the first study, a reversed phase HPLC method was developed for the analysis of sinigrin, the major glucosinolate in B. carinata. Both C18 columns selected were able to separate the compound with an isocratic eluent containing 100% tetramethylammonium bromide (10 mM, pH 5) delivered at 1 mL/min at a column temperature of 25oC. These chromatographic conditions were applied and sinigrin concentration of whole B.carinata seed was estimated to be 29 μg/mg. Average matrix effect was estimated to be 104% that was caused by other components in the B. carinata seed matrix. In the second study, high concentrations of glucosinolates were detected and identified in fractions of C. sativa seeds using HPLC-ESI-MS. Methods for extraction, isolation, and purification of three individual glucosinolates from these fractions are reported. Quantitation of total glucosinolates was performed on proton NMR using DMF as an internal standard. Quantitation of individual glucosinolates was achieved by using MS extracted ion chromatogram data. Total glucosinolates were found in C. sativa whole seed at a concentration of 14 μg/mg, and glucocamelinin, the major glucosinolate, constituted 65% of the total amount. In addition, a dehulling treatment was applied to C. sativa seeds, from which both oil content and crude protein content increased after dehulling of the seeds.

Brassica carinata is being actively pursued as a new industrial oil crop platform for the Canadian Prairies. A genetic assessment of B. carinata was performed to elucidate its evolutionary origins and create a genetic map to assist in locating genes and traits of interest that would help in marker-assisted breeding. First, genetic analysis using simple sequence repeat (SSR) markers, previously tested on B. juncea and B. napus, was performed, to examine the genetic diversity of 37 B. carinata lines. SSR analysis revealed world accessions were more diverse than lines conditioned to grow in the prairies. Diversity analysis revealed that the parental lines of a double haploid (DH) population, 179 and 345, obtained from the John Innes Centre (JIC), were among the more genetically diverse lines, supporting the use of this population for linkage mapping. Genetic markers created from 3’ targeted SNP discovery between 179 and 345, were tested on the DH population resulting in the generation of a B. carinata genetic linkage map essentially with no prior sequence data knowledge. This genetic map contained 341 SNP and 86 SSR loci identifying eight linkage groups belonging to the B genome, nine belonging to the C genome and two unidentified groups spanning 2041 cM. Comparative mapping of polymorphic markers identified in the amphidiploid B. carinata indicated the orientation of B and C genomes coincide with that of other Brassica species, and the two genomes have remained essentially unaltered, with no major chromosomal rearrangements since the formation of B. carinata. A lesser number of polymorphic markers were detected in the C genome, which suggested the B genome is more genetically diverse in B. carinata. Limited field trials of the 179 x 345 DH population were performed during the 2011 and 2012 growing seasons. Preliminary quantitative trait loci (QTLs) for agronomic traits including flowering time (FT), plant height (PH), and seed quality were identified.

An efficient system for or 'Agrobacterium'-mediated transformation of <i>Brassica carinata</i> was used together with a promoterless <i> gus</i>::<i>nptII</i> gene fusion to isolate putative promoter sequences. Cotyledonary petioles were transformed using the promoterless gene fusion construct. Only transformation events in which the promoterless gene fusion had integrated downstream from plant regulatory sequences were expected to produce viable tissue under kanamycin selection. Forty-two transgenic plants were recovered. Transformation efficiency was approximately 0.6%. Regenerated plants were screened for GUS expression in different tissues and organs by histological and fluorometric assays. Tissue-specific GUS expression was detected (stigmas, seed coat, leaf edges and vascular tissue) in some plants, while strong constitutive GUS expression was detected in others (based on GUS histological assays). Using subgenomic libraries, putative promoter fragments were isolated from the plants which exhibited GUS expression in stigmas, leaf edges and constitutively. A putative promoter fragment from a plant which exhibited GUS expression only in the stigma was fused with the gus gene and reintroduced by <i>Agrobacterium </i> -mediated transformation into <i>B. napus, B. carinata, Arabidopsis' and tobacco </i>. GUS expression was observed in the stigma of <i>B. napus </i> but not in ' B. carinata'. In <i>Arabidopsis </i> and tobacco GUS expression. was not tissue specific (weakly constitutive or restricted to two or more tissues). The 3' DNA sequence (15 kb) flanking the <i> gus</i>::<i>nptII </i> insert in the plant with GUS expression in the stigma was also isolated using a subgenomic library. A gene for a cytochrome P450 like protein was discovered on the minus DNA strand of the 3' sequence with a start codon approximately 6.5 kb from the T-DNA left border.