The explosion in human population is a problem to the survival of human existence, so there is an urgent need to increase the production capacities of products, including vaccines, therapeutic molecules, antibodies, industrial enzymes, developing plant oil as well as biofuels and produce them at low-costs. Plant biotechnology based production was considered to be a solution for such a major global challenge. However, this approach is not yet exploited for the production of any recombinant protein for any major application so far. This was mainly due to lack substrates and/or high level expression of enzyme activity in a desired plant tissue making the whole process uneconomical. So, a combine of many molecular biology techniques, such as seeking synthetic or modified genes and use most sensible genetic engineering strategy can solve above problem. In this overview, I would like to focus just some topics which I had worked, and hope they may be support new products or increasing a product and produce at low-cost.

1. Boost  astaxanthin and EPA in soybean and camelina seeds

Astaxanthin accumulation turn soybean seed to orange (right) compare with non-transgenic seeds( left)

Camelina seeds look dark orange (top right and circle-bottom) compare with wild type seed (top left or center-bottom)

2. Redirection of Metabolic Flux for High Levels of Omega-7 Monounsaturated Fatty Acid Accumulation in Camelina seed

Seed oils enriched in omega-7 monounsaturated fatty acids, including palmitoleic acid (16:1D9) and cis-vaccenic acid (18:1D11), have nutraceutical and industrial value for polyethylene production and biofuels. Existing oilseed crops accumulate only small amounts (<2%) of these novel fatty acids in their seed oils. We demonstrate a strategy for enhanced production of omega-7 monounsaturated fatty acids in camelina (Camelina sativa) and soybean (Glycine max) that is dependent on redirection of metabolic flux from the typical delta 9 desaturation of stearoyl (18:0)-acyl carrier protein (ACP) to delta 9 desaturation of palmitoyl (16:0)-acyl carrier protein (ACP) and –coenzyme A (CoA). This was achieved by seed-specific co-expression of a mutant delta 9-acyl-ACP and an acyl-CoA desaturase with high specificity for 16:0-ACP and -CoA substrates, respectively. This strategy was most effective in camelina where seed oils with ~17% omega-7 monounsaturated fatty acids were obtained. Further increases in omega-7 monounsaturated fatty acid accumulation to ~ 65% of the total fatty acids in camelina seeds were achieved by inclusion of seed-specific suppression of 3-keto-acyl-ACP synthase II and the FatB 16:0-ACP thioesterase genes to increase substrate pool sizes of 16:0-ACP for the delta 9-acyl-ACP desaturase and by blocking C18 fatty acid elongation. Seeds from these lines also had total saturated fatty acids reduced to ~5% of the seed oil versus ~12% in seeds of non-transformed plants. Consistent with accumulation of triacylglycerol species with shorter fatty acid chain-lengths and increased monounsaturation, seed oils from engineered lines had marked shifts in thermotropic properties that may be of value for biofuel applications.

3. Camelina Seed Transcriptome: A Tool for Meal and Oil Improvement and Translational Research

Camelina (Camelina sativa), a Brassicaceae oilseed, has received intense interest as a biofuel crop and production platform for industrial oils. Limiting wider production of camelina for these uses is the need to improve the quality and content of the seed protein rich-meal and oil, which is enriched in oxidatively unstable polyunsaturated fatty acids that are deleterious for biodiesel. To identify candidate genes for meal and oil quality improvement, we built a transcriptome reference from 2,047 Sanger ESTs and over 2 million 454-derived sequence reads, representing genes expressed in developing camelina seeds. The transcriptome of ~60K transcripts from 22,597 putative genes includes camelina homologs of nearly all known seed-expressed genes, suggesting a high level of completeness and usefulness of the reference. These sequences included candidates for 12S (cruciferins) and 2S (napins) seed storage proteins (SSPs) and nearly all known lipid genes, which have been compiled into an accessible database. To demonstrate the utility of the transcriptome for seed quality modification, seed-specific RNAi lines deficient in napins were generated by targeting 2S SSP genes, and high oleic acid oil lines were obtained by targeting fatty acid desaturase 2 (FAD2) and fatty acid elongase 1 (FAE1). The high sequence identity between Arabidopsis and camelina genes was also exploited to engineer high oleic lines by RNAi with Arabidopsis FAD2 and FAE1 sequences. It is expected that this transcriptomic data will be useful for breeding and engineering of additional camelina seed traits and for translating findings from the model Arabidopsis thaliana to an oilseed crop.

4. Omega7, turn nothing into 70%

Gas chromatograph-mass spectrometer (GCMS) analysis fatty acid single seed of control (top) and transgenic (bottom) seeds

5. Engineering Plant Oils

Using genetic manipulation to modify the activity of a plant enzyme, we have converted an unsaturated oil in the seeds of a temperate plant to the more saturated kind usually found in tropical plants. The research had published by the Proceedings of the National Academy of Sciences (PNAS) with title “Modulating seed Beta-ketoacyl-acyl carrier protein synthase II level converts the composition of a temperate seed oil to that of a palm-like tropical oil”, vol.104, 2007.