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Brief Communication Enhancing the accumulation of eicosapentaenoic acid and docosahexaenoic acid in transgenic Camelina through the CRISPR-Cas9 inactivation of the competing FAE1 pathway

Phosphatidylcholine:diacylglycerol cholinephosphotransferases (PDCT) regulate the fatty acid composition of seed oil (triacylglycerol, TAG) by interconversion of diacylglycerols (DAG) and phosphatidylcholine (PtdCho). PtdCho is the substrate for polyunsaturated fatty acid biosynthesis, as well as for a number of unusual fatty acids. By the action of PDCT, these fatty acids can be transferred into the DAG pool to be utilized in TAG biosynthesis by the action of acyl-CoA:DAG and phospholipid:diacylglycerol acyltransferases. Despite its importance in regulating seed oil composition, biochemical characterization of PDCT enzymes has been lacking. We characterized Camelina sativa PDCT in microsomal preparations of a yeast strain expressing Camelina PDCT and lacking the capacity of producing TAG. Camelina PDCT was specific for PtdCho and the sn-1,2 enantiomer of DAG and could not utilize ceramide. The interconversion reaches equilibrium within 15 min of incubation, indicating that only distinct pools of DAG and PtdCho were available for exchange. However, the pool sizes of DAG and PtdCho involved in the exchange were not fixed but increased with the amount of exogenous DAG or PtdCho added. Camelina PDCT showed about the same selectivity for di-oleoyl, di-linoleoyl, and di-linolenoyl species in both PtdCho and DAG substrates, suggesting that no unidirectional transfer of particular unsaturated substrates occurred. Camelina PDCT had a good activity with erucoyl-DAG as a substrate despite low erucic acid levels in PtdCho in plant species accumulating a high amount of this fatty acid in the seed oil.

Cutworms (Lepidoptera: Noctuidae) are economically important pests of multiple field crops, but are difficult to control with insecticides. Biological control with native parasitoids may offer a sustainable alternative for pest management. Cover crops improve the availability of resources required by parasitoids to increase their effectiveness as biocontrol agents. Copidosoma spp. (Hymenoptera: Encyrtidae) have the potential to control several species of cutworms in the Canadian Prairies. However, their foraging preferences for non‐host food items are unknown. In this study, we tested flower color and odor preferences of Copidosoma bakeri (Howard) across nine potential cover crop species and investigated how these resources affect survival and longevity of the female wasps. Results indicated that the wasps were most attracted to plants with yellow flowers in the family Brassicaceae (i.e., camelina, mustard, and canola). In multiple‐choice tests, the wasps preferred camelina over all other flower species, and showed an intermediate preference for mustard and canola. In dual‐choice tests with color paper, the wasps preferred yellow over white, and the light yellow of camelina over the bright yellow of canola and mustard, whereas they showed a preference for the odors of mustard and canola. Life expectancy was increased when the wasps were provided floral resources compared to the absence of a food source. Later planting dates and mixed‐species plantings of cover crops resulted in flowering times that better coincided with native wasp emergence. Camelina is known to suppress weeds and may be used as a habitat management tool for enhancing parasitism of cutworms by C. bakeri in the Canadian Prairies. However, further testing is needed to achieve late flowering well into August and early September and to ensure planting additional Brassica spp. does not attract more late‐season pests to cash crops such as canola.

Flowers of field pennycress (Thlaspi arvsense L.) and winter camelina (Camelina sativa (L.) Crantz.) produce abundant pollen and nectar in early spring and thereby may be valuable for pollinators. Insects observed in field plots of these flowers were classified into seven easily identifiable groups (bumblebee, honeybee, solitary bee, butterfly/moth, beetle, fly and other) and monitored for 2 years at three sites in the Upper Midwest region of the USA. Average seasonal observations across years and sites varied from 1.6 to 5.3 total insects/min for field pennycress and 1.4 to 4.5 insects/min for winter camelina. Lowest visitation rates occurred in central Iowa and highest rates in south‐eastern Minnesota for both crops. Multiple regressions showed that visitation rates for specific insect groups were correlated poorly but significantly (p

The transgene‐directed accumulation of non‐native omega‐3 long chain polyunsaturated fatty acids in the seed oil of Camelina sativa (Camelina) was evaluated in the field, in distinct geographical and regulatory locations. A construct, DHA2015.1, containing an optimal combination of biosynthetic genes, was selected for experimental field release in the UK, USA and Canada, and the accumulation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) determined. The occurrence of these fatty acids in different triacylglycerol species was monitored and found to follow a broad trend irrespective of the agricultural environment. This is a clear demonstration of the stability and robust nature of the transgenic trait for omega‐3 long chain polyunsaturated fatty acids in Camelina. Examination of non‐seed tissues for the unintended accumulation of EPA and DHA failed to identify their presence in leaf, stem, flower, anther or capsule shell material, confirming the seed‐specific accumulation of these novel fatty acids. Collectively, these data confirm the promise of GM plant‐based sources of so‐called omega‐3 fish oils as a sustainable replacement for oceanically derived oils.

Plant oils are valuable commodities for food, feed, renewable industrial feedstocks and biofuels. To increase vegetable oil production, here we show that the nonspecific phospholipase C6 (NPC6) promotes seed oil production in the Brassicaceae seed oil species Arabidopsis, Camelina and oilseed rape. Overexpression of NPC6 increased seed oil content, seed weight and oil yield both in Arabidopsis and Camelina, whereas knockout of NPC6 decreased seed oil content and seed size. NPC6 is associated with the chloroplasts and microsomal membranes, and hydrolyzes phosphatidylcholine and galactolipids to produce diacylglycerol. Knockout and overexpression of NPC6 decreased and increased, respectively, the flux of fatty acids from phospholipids and galactolipids into triacylglycerol production. Candidate-gene association study in oilseed rape indicates that only BnNPC6.C01 of the four homeologues NPC6s is associated with seed oil content and yield. Haplotypic analysis indicates that the BnNPC6.C01 favorable haplotype can increase both seed oil content and seed yield. These results indicate that NPC6 promotes membrane glycerolipid turnover to accumulate TAG production in oil seeds and that NPC6 has a great application potential for oil yield improvement.

Camelina sativa(L.) Crantz is an oilseed crop with favorable potentials for bio-diesel production, such as the high plant yield, high oil content in the seed, high net energy ratio, and low oil production cost. This review paper deals with the present state and perspectives of biodiesel production from camelina oil. First, important issues of camelina seed pretreatment and biodiesel production are reviewed. Emphasis is given to different biodiesel technologies that have been used so far worldwide, the economic assessment of the camelina oil biodiesel (COB) production, the camelina- based biorefineries for the integrated biodiesel production, the COB life cycle analysis, and impact human health and ecosystem. Finally, the perspectives of COB production from the techno- economic and especially genetic engineering points of view are discussed.

Erucic (22:1, cisΔ13) and gondoic acids (20:1, cisΔ11) are building blocks obtained from renewable sources for the oleochemical industry. Different biocatalytic strategies for the enrichment of these compounds with high recovery yields were developed in our group. Geotrichum candidum lipases (GCL) strongly discriminate against fatty acids longer than 18 carbon atoms. Thus, GCL‐I and ‐II were investigated using hydrolysis or ethanolysis reactions with Crambe and Camelina oils. Hydrolysis was also studied using fatty acid ethyl esters (FAEE) derived from the corresponding oil. Both isoforms were highly selective; however, interesting differences were observed. Although it has been reported that GCL‐I displays a higher preference toward 18 cisΔ9, which is present in the studied oils at high levels, GCL‐II showed higher enrichment values during hydrolysis independent of the substrate used. Hence, enrichments of 87% (Crambe oil) and 82% (Crambe FAEE) for erucic acid and 50% (Camelina oil) and 45% (Camelina FAEE) for gondoic acid, with recovery values between 89% and 99%, were achieved. On the contrary, the best enzyme for ethanolysis was GCL‐I (82% and 41% for erucic and gondoic acid, respectively). In this case, although GCL‐II also displayed good enrichment and recovery levels (77% and 28%, respectively), they were lower compared to the former reactions. In both ethanolysis reactions, the FAEE fraction contained between 92% and 97% of 18 unsaturated fatty acids.

Camelina [Camelina sativa (L.) Crantz] of the Brassicaceae family is a short-season drought- and frost-tolerant oilseed. Camelina has been promoted as an alternative crop for the low-precipitation (

Interest is growing in camelina (Camelina sativa L. Crantz) as a biofuel feedstock. However, there has been little camelina research in irrigated arid systems. A 2-yr field study in Maricopa, AZ, under an overhead sprinkler irrigation system determined the effects of 10 water levels (irrigation fraction 0.5–1.1) and five N fertilizer rates (38–150 kg N ha–¹) on seed yield, seed oil content, and N use efficiency. Cultivar Robinson was planted in December 2012 and 2013. Nitrogen fertilizer (urea ammonium nitrate) was applied in three split applications. Irrigation amounts were from 125 to 380 mm, and in-season rain was 70 and 50 mm, in 2013 and 2014, respectively. Camelina seed yields were maximum at water level 7 (irrigation fraction 0.93) in 2013 at 1800 kg ha–¹. Maximum seed yields were 1600 kg ha–¹ at water level 6 (irrigation fraction 0.83) in 2014. These highest seed yields were achieved with 150 kg N ha–¹ in both years. Oil content (maximum 41%) decreased with N rate but increased with water level. Seed N increased with N rate but decreased with irrigation level. Recovery efficiency of N fertilizer by camelina ranged from 12 to 72%. The results indicate that good high-oil camelina yields can be produced in the southwestern United States with 320 to 380 mm irrigation plus rain and N fertilizer rates of 150 kg N ha–¹.

Winter cover crops might reduce nutrient loss to leaching in the Upper Midwest. New oilseed-bearing cash cover crops, such as winter camelina ( L.) and pennycress ( L.), may provide needed incentives. However, the abilities of these crops to sequester labile soil nutrients are unknown. To address this unknown, N in shoot biomass, plant-available N and P in soil, and NO-N and soluble reactive P in soil water collected from lysimeters placed at 30, 60, and 100 cm were measured in cover crop and fallow treatments established in spring wheat ( L.) stubble and followed through a cover crop-soybean [ (L.) Merr.] rotation. Five no-till cover treatments (forage radish [ L.], winter rye [ L.], field pennycress, and winter camelina) were compared with two fallow treatments (chisel till and no-till). Pennycress and winter camelina were harvested at maturity after relay sowing of soybean. Winter rye and radish sequestered more N in autumn shoot biomass, ranging from 26 to 38 kg N ha, but overwintering oilseeds matched or exceeded N uptake in spring, ranging 28 to 49 kg N ha before soybean planting. Nitrogen uptake was reflected by reductions in soil water NO-N during cover crop and intercropping phases for all cover treatments (mean = 4 mg L), compared with fallow treatments (mean = 31 mg L). Cash cover crops like pennycress and winter camelina provide both environmental and potential economic resources to growers. They are cash-generating crops able to sequester labile soil nutrients, which protects and promotes soil health from autumn through early summer.

Cover crops can serve as a valuable management tool for improving soil and water quality, but are an added expense for farmers. We evaluated the yields and economics of four cover crops and two winter fallow treatments in a spring wheat (Triticum aestivum L.)–soybean [Glycine max (L.) Merr.] rotation at three sites in Minnesota. The four cover crop treatments were winter rye (Secale cereal L.), forage radish (Raphanus sativus L.), winter camelina [Camelina sativa (L.) Crantz], and pennycress (Thlaspi arvense L.) planted into spring wheat stubble. The fallow treatments consisted of no-tilled and conventionally tilled soil. Radish winterkilled and rye was terminated chemically before planting soybean in early May. Soybean was inter-seeded between rows of camelina and pennycress at the same time it was planted in other treatments. Camelina and pennycress were harvested over soybean seedlings in late June. Camelina yields ranged from 600 to 1100 kg ha–¹, while pennycress ranged from 900 to 1550 kg ha–¹. Mono-cropped soybean averaged 1819, 3510, and 4180 kg ha–¹ in northern, central, and southern Minnesota, respectively. Soybean seedlings under oilseed cover crop canopies exhibited light-stress, which likely reduced soybean yield in these treatments by 22 to 30%. When oilseed and inter-seeded soybean yields were combined, total seed yields generally were equal to or exceeded those of mono-cropped soybean. In addition, net income for inter-seeded systems was typically equivalent to mono-cropped soybean. Improvements in net income are likely needed before the benefits of oilseed cover crops are fully realized.

Hybridization between crops and their wild relatives has the potential to introduce novel variation into wild populations. Camelina (Camelina sativa) is a promising oilseed and cultivars with modified seed characteristics and herbicide resistance are in development, prompting a need to evaluate the potential for novel trait introgression into weedy relatives. Little‐podded false flax (littlepod; Camelina microcarpa) is a naturalized weed in Canada and the USA. Here we evaluated the hybridization rate between the three cytotypes of littlepod (♀) and camelina (♂), assessed characteristics of hybrids, and evaluated the fitness of hexaploid littlepod and camelina hybrids in the glasshouse and field. In total we conducted, 1,005 manual crosses with diploid littlepod, 1, 172 crosses with tetraploid littlepod, and 896 crosses with hexaploid littlepod. Hybrids were not produced by the diploids, but were produced by the tetraploids and hexaploids at rates of one hybrid for 2,000 ovules pollinated and 24 hybrids for 25 ovules pollinated, respectively. Hybrids between tetraploid littlepod and camelina showed low pollen fertility and produced a small number of seeds. In the glasshouse, hybrids between hexaploid littlepod and camelina also showed significantly lower pollen fertility and seed production than parental lines, but their seeds showed high viability. A similar pattern was observed in field trials, with hybrids showing earlier flowering, reduced biomass, seed production and seed weight. However, seed produced by the hybrids showed greater viability than that produced by hexaploid littlepod and is potentially the result of a shortened lifecycle. The introgression of lifecycle traits into littlepod populations may facilitate range expansion and contribute to crop gene persistence. Consequently, future work should evaluate the hybridization rate in the field, the fitness of advanced generation backcrosses, and the role of time to maturity in limiting hexaploid littlepod's distribution.

Biotechnological strategies using renewable materials as starting substrates are a promising alternative to traditional oleochemical processes for the isolation of different fatty acids. Among them, long chain monounsaturated fatty acids are especially interesting in industrial lipid modification, since they are precursors of several economically relevant products, including detergents, plastics and lubricants. Therefore, the aim of this study was to develop an enzymatic method in order to increase the percentage of long chain mono-unsaturated fatty acids fromCamelinaandCrambeoil ethyl ester derivatives, by using selective lipases. Specifically, the focus was on the enrichment of gondoic (C20:1cisΔ11) and erucic acid (C22:1cisΔ13) fromCamelinaandCrambeoil derivatives, respectively. The pursuit of this goal entailed several steps, including: (i) the choice of a suitable lipase scaffold to serve as a protein engineering template (Candida antarcticalipase A); (ii) the identification of potential amino acid targets to disrupt the binding tunnel at the adequate location; (iii) the design, creation and high-throughput screening of lipase mutant libraries; (iv) the study of the selectivity towards different chain lengthp-trophenyl fatty acid esters of the best hits found, as well as the analysis of the contribution of each amino acid change and the outcome of combining several of the aforementioned residue alterations and, finally, (v) the selection and application of the most promising candidates for the fatty acid enrichment biocatalysis. As a result, enrichment of C22:1 fromCrambeethyl esters was achieved either, in the free fatty acid fraction (wt, 78%) or in the esterified fraction (variants V1, 77%; V9, 78% and V19, 74%). Concerning the enrichment of C20:1 whenCamelinaoil ethyl esters were used as substrate, the best variant was the single mutant V290W, which doubled its content in the esterified fraction from approximately 15% to 34%. A moderately lower increase was achieved byV9 and its two derived triple mutant variants V19 and V20 (27%).

Camelina sativa seeds are rich in oil (30–49%) and protein (24–31%). They contain ω-3 acids, ω-6 acids, tocopherols, phytosterols, and phenolic compounds, among others. From an agricultural perspective, growing of this crop is of interest due to its short growth cycle and low fertilizer and water input requirements. Camelina is also tolerant to cold and drought and is consequently well adapted to grow in semiarid regions. Camelina is mainly cultivated for its oil in Europe and North America. In this review, the processes applied for camelina oil extraction, composition, and attributes, as well as the food and nonfood applications of camelina oil are reviewed. Applications include animal feed, functional foods, materials, biofuels, and agrochemicals. Valorization of the camelina protein found in the meal after the oil extraction is also discussed. Practical Applications: The need to develop an integrated process consisting of a degumming step to extract the mucilage from the whole camelina seeds, followed by an oil extraction step, and finally by a protein extraction step is highlighted. There is also a need to develop food applications of camelina oil. More research works should also focus on the utilization of camelina oil in food applications and in specialty applications such as functional foods, nutraceuticals, cosmetics, and pharmaceutical applications.

Processes for the production of protein isolates from camerlina Camelina sativa were developed by modifying the procedure used for other seeds from the Brassicaceae family, such as rapeseed and mustard. The procedure consisted of defatting the seed followed by alkaline extraction at pH 11, ultra- and diafiltration using a 5-kDa membrane, isolelectric precipitation of the proteins at pH 5 and recovery of the acid soluble protein isolates (SPI) after further filtration and drying. Protein yields as precipitated protein isolate and SPI were 10.7 and 11.4%, containing 67 and 42%, respectively. Due to the high concentration of mucilage in camelina, a water-to-seed ratio of 30 had to be used for protein extraction from the hexane-defatted seed and most of the mucilage had to be removed prior to the defatting process. A rapid mucilage extraction process using water at 55 °C was developed. Camelina mucilage absorbs water and oil. Viscosity measurements of dried and redissolved mucilage showed the highest values at natural pH, and the viscosity increased rapidly above 1% solids concentration. It may be a useful product in the food and pharmaceutical industries.

Every year, the production of coal-bed natural gas in the Powder River Basin results in the discharge of large amounts of coal-bed methane water (CBMW) in Wyoming; however, no sustainable disposal methods for CBMW are currently available. A greenhouse study was conducted to evaluate the potential to use CBMW as a source of irrigation water for camelina [ (L.) Crantz]. We assessed the effects of three CBMW concentrations (0% [1:0], 50% [1:1], and 100% [0:1] tap water to CBMW) on selected soil properties, growth, seed oil, and fatty acid composition of three camelina cultivars: Blaine Creek, Ligena, and Pronghorn. The 100% CBMW reduced seed and estimated biofuel yields by 24 and 23%, respectively, but increased the oil content by 3%, relative to the control. Additionally, the 100% CBMW visibly affected soil through formation of surface crust due to elevated levels of sodium (653 mg Na kg). The 50% CBMW had no significant effects on the seed yield, estimated biofuel yield, and oil content, but the soil Na levels were still high (464 mg kg), which could pose a long-term impact on soil structure. The CBMW tended to reduce the total saturated fatty acid, but it had no significant effects on the total monounsaturated or polyunsaturated fatty acids of camelina seeds. Overall, CBMW diluted with an equal amount of good-quality water could be used to irrigate camelina in the short term. Afterward, only good-quality water would have to be used until the accumulated dissolved solids are flushed out.

Camelina sativa seed is an underutilized oil source that attracts a growing interest, but it requires more research on its composition and processing. Its high omega-3 content and growing demand for clean food processing technologies make conventional oil extraction less attractive. In this study, the effect of extraction methods on the bioactive lipid composition of the camelina seed lipid was investigated, and its bioactive lipid composition was modified at the extraction stage using ethanol-modified supercritical carbon dioxide (SC-CO2). Ethanol-modified SC-CO2 extractions were carried out at varying temperatures (50 and 70 °C), pressures (35 and 45 MPa), and ethanol concentrations (0–10%, w/w), and were compared to SC-CO2, cold press, and hexane extraction. The highest total lipid yield (37.6%) was at 45 MPa/70 °C/10% (w/w) ethanol. Phospholipids and phenolic content increased significantly with ethanol-modified SC-CO2 (p

BACKGROUND: Dietary supplementation with oilseeds can reduce methane emission in ruminants, but only a few common seeds have been tested so far. This study tested safflower (Carthamus tinctorius), poppy (Papaver somniferum), hemp (Cannabis sativa), and camelina (Camelina sativa) seeds in vitro using coconut (Cocos nucifera) oil and linseed (Linum usitatissimum) as positive controls. RESULTS: All the tested oilseeds suppressed methane yield (mL g-1 dry matter, up to 21%) compared to the non-supplemented control when provided at 70 g oil kg-1 dry matter, and they were as effective as coconut oil. Safflower and hemp were more effective than linseed (21% and 18% vs. 10%), whereas the effects of poppy and camelina were similar to linseed. When methane was related to digestible organic matter, only hemp and safflower seeds and coconut oil were effective compared to the non-supplemented control (up to 11%). The level of methanogenesis and the ratios of either the n-6:n-3 fatty acids or C18:2 :C18:3 in the seed lipids were not related. CONCLUSION: Unconventional oilseeds widen the spectrum of oilseeds that can be used in dietary methane mitigation. In vivo confirmation of their methane mitigating effect is still needed, and their effects on animal performance still must be determined. © 2017 Society of Chemical Industry.

Camelina seed is a new alternative omega-3 source attracting growing interest. However, it is susceptible to oxidation due to its high omega-3 content. The objective of this study was to improve the oxidative stability of the camelina seed oil at the extraction stage in order to eliminate or minimize the use of additive antioxidants. Camelina seed oil extracts were enriched in terms of natural antioxidants using ethanol-modified supercritical carbon dioxide (SC-CO2 ) extraction. Oxidative stability of the camelina seed oils extracted by ethanol modified SC-CO2 was studied by differential scanning calorimeter (DSC), and compared with cold press, hexane, and SC-CO2 methods. Nonisothermal oxidation kinetics of the oils obtained by different extraction methods were studied by DSC at varying heating rates (2.5, 5, 10, and 15 °C/min). Increasing ethanol level in the ethanol-modified SC-CO2 increased the oxidative stability. Based on oxidation onset temperatures (Ton ), SC-CO2 containing 10% ethanol yielded the most stable oil. Oxidative stability depended on the type and content of the polar fractions, namely, phenolic compounds and phospholipids. Phenolic compounds acted as natural antioxidants, whereas increased phospholipid contents decreased the stability. Study has shown that the oxidative stability of the oils can be improved at the extraction stage and this may eliminate the need for additive antioxidants.

Summary The CRISPR/Cas9 nuclease system is a powerful and flexible tool for genome editing, and novel applications of this system are being developed rapidly. Here, we used CRISPR/Cas9 to target the FAD2 gene in Arabidopsis thaliana and in the closely related emerging oil seed plant, Camelina sativa, with the goal of improving seed oil composition. We successfully obtained Camelina seeds in which oleic acid content was increased from 16% to over 50% of the fatty acid composition. These increases were associated with significant decreases in the less desirable polyunsaturated fatty acids, linoleic acid (i.e. a decrease from ~16% to

Camelina (Camelina sativa L. Crantz) shows potential to provide an alternative renewable energy source and enhance crop diversification in temperate semiarid regions. Information on the effect of N, P, K, and S on yield and quality of camelina for biodiesel feedstock in the northern Great Plains (NGP) of the United States is limited. The objective of this experiment was to determine the effects of the above nutrients on seed and oil yields, test weight, oil concentration and agronomic nitrogen use efficiency (ANUE) of camelina on a clay loam soil in central Montana. Results showed that fertilizer treatments significantly affected seed yield, oil concentration and oil yield of camelina. The seed and oil yields ranged from 677 to 1306 kg ha–¹ and from 234 to 445 kg ha–¹, respectively. Although the highest seed and oil yields were obtained from the application of 134–22–22–28 kg ha–¹ N–P₂O₅–K₂O–S, they were statistically in the same group with yields achieved from the application of only 45 kg ha–¹ N. Application of P and S fertilizers increased camelina seed yield compared with the control treatment. There was no response to K fertilization. Simultaneous application of N and S did not show synergistic effects in enhancing ANUE. The ANUE reduced with increasing N application rates. From trend analysis, application of 60 kg ha–¹ N produced agronomic maximum seed and oil yields. Therefore, optimizing camelina seed and oil yields production with regard to nutrient management using current variety should focus on N fertilization.

The aim of this study was to evaluate the effect of nitrogen fertilization (0, 40, 80, 120, 160 kg N ha⁻¹) on changes in selected physical, thermal, and geometric parameters of camelina seeds. Samples 0 kg N ha⁻¹ and 40 kg N ha⁻¹ formed a homogenous group with lower values of 1,000 seed weight and differed significantly from the second homogeneous group of samples 80, 120, and 160 kg N ha⁻¹ with higher 1,000 seed weight. Nitrogen fertilization did not exert a significant effect on bulk density, true density, or porosity. Fertilization had no significant effect on the thermal properties of camelina seeds. Based on all linear dimensions and shape factors, samples 0 and 40 kg N ha⁻¹ formed one homogenous group and were significantly different from the remaining samples. Samples 0 and 40 kg N ha⁻¹ were characterized by the lowest values of linear dimensions, including length, width, surface area, profile specific perimeter, and convex perimeter, the lowest values of selected shape factors, including diameter range, standard deviation of all radii, Blair‐Bliss coefficient, and Danielsson coefficient, as well as the highest values of the radius ratio and roundness. PRACTICAL APPLICATIONS: Nitrogen fertilization strongly influences the properties of camelina seeds. Camelina seeds subjected to 120 kg N ha⁻¹ were characterized by the highest dimensions, such as: length, width, surface area, and high values of 1,000 seed weight and they may be most desirable to be used in practice.

Sowing date and sowing method can have a profound influence on the productivity of alternative crops like camelina in semiarid agroecosystems. The objective of this study was to determine the effects of sowing date, sowing method, and cultivar on morphology, phenology, grain yield, oil concentration, oil, and biodiesel production of camelina. A 2‐year study was carried out at the University of Nevada, Reno Main Station Field Laboratory, during the spring to early summer of 2016 and 2017. Treatments were two sowing dates (SD) of 18 March 2016 (early SD) and 17 April 2016 (late SD) in Year 1 and 11 April 2017 (early SD) and 11 May 2017 (late SD) in Year 2. The change in SD in the second year was due to the excessively wet field condition preventing land preparation and sowing. There were two sowing methods (SM) imposed (broadcast and drill) and three cultivars of camelina (Blaine Creek, Columbia, and Pronghorn) arranged in a 3 × 2 × 2 factorial in a randomized complete block design experiment with four replications. Responses were considered different if p

As nonfood oilseed varieties are being rapidly developed, new varieties may affect agricultural production efficiency and life-cycle assessment results. Current, detailed feedstock production information is necessary to accurately assess impacts of the biofuel life-cycle. The life-cycle impacts of four nonfood oilseeds (carinata [ L. Braun], camelina [ L. Crantz], canola or rapeseed [ L.], and sunflower [ L.]) were modeled using Argonne National Laboratory's GREET model to compare feedstocks for renewable biojet production using cold-press oil extraction. Only feedstock-related inputs were varied, allowing isolation of feedstock influence. Carinata and camelina performed slightly better than other oilseed crops at most product stages and impact categories as a result of current, low-input agricultural information and new feedstock varieties. Between 40 to 50% of SO and NO emissions, ∼25% of greenhouse gas (GHG) emissions, and ∼40% of total energy consumption for the biojet production impact occurred during feedstock production. Within the first standard deviation, total well-to-tank emissions varied between ∼13% (GHG) and ∼35% (SO) for all feedstocks emphasizing the importance of accurate agricultural production information. Nonfood oilseed feedstock properties (e.g., oil content, density) and agricultural management (e.g., fertilization, yield) affect life-cycle assessment results. Using biofuels in feedstock production and focusing on low-impact management would assist producers in improving overall product sustainability.

Camelina (Camelina sativa L. Crantz) is an oilseed crop with the potential for dryland crop production in the Great Plains. However, pod shattering may cause significant yield losses. We determined the impact of different harvest times on camelina seed yield (SY), water use efficiency, protein and oil content, and estimated biodiesel yield. Spring (Blaine Creek) and winter (BX WG1) camelina cultivars were harvested at three different stages, corresponding to the three-digit Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH) scales of 805 (early harvest; 50% ripe pods), 807 to 808 (mid-harvest; 70–80% ripe pods), and 809 (late harvest; >90% ripe pods). In addition, different harvest methods were assessed to identify and quantify other sources of yield loss. Seed moisture contents at early, mid, and late harvests were 14.2, 9.8, and 6.8%, respectively. On average, the SY of early harvest was 9.5 and 23.6% greater than the mid- and late harvests, respectively. Total seed loss incurred during direct-combine harvest was 11.7%, which was attributable to the mechanical disturbance imposed on the pods and the combine settings. Camelina seed oil content was greatest at mid-harvest, but the estimated biodiesel yield was not significantly different between the early and mid-harvests. In general, direct combining when 75% of camelina pods are ripe will provide a balance between SY loss and the seed protein and oil contents.

Interest from the US commercial aviation industry and commitments established by the US Navy and Air Force to use renewable fuels has spurred interest in identifying and developing crops for renewable aviation fuel. Concern regarding greenhouse gas emissions associated with land-use change and shifting land grown for food to feedstock production for fuel has encouraged the concept of intensifying current prominent cropping systems through various double cropping strategies. Camelina (Camelina sativa L.) and field pennycress (Thlaspi arvense L.) are two winter oilseed crops that could potentially be integrated into the corn (Zea mays L.)–soybean [(Glycine max (L.) Merr.] cropping system, which is the prominent cropping system in the US Corn Belt. In addition to providing a feedstock for renewable aviation fuel production, integrating these crops into corn–soybean cropping systems could also potentially provide a range of ecosystem services. Some of these include soil protection from wind and water erosion, soil organic C (SOC) sequestration, water quality improvement through nitrate reduction, and a food source for pollinators. However, integration of these crops into corn–soybean cropping systems also carries possible limitations, such as potential yield reductions of the subsequent soybean crop. This review identifies and discusses some of the key benefits and constraints of integrating camelina or field pennycress into corn–soybean cropping systems and identifies generalized areas for potential adoption in the US Corn Belt.

349 Camelina is an ancient crop believed to have evolved as a weed in fi elds planted with fl ax, hence the name “false fl ax” (Budin et al., 1995; Gugel and Falk, 2006). According to Matthaus and Zubr (2000), camelina was cultivated for oil in Europe during the Bronze and Iron Ages; however, its production dwindled during the Middle Ages. Th ere has been recent interest in camelina production because of increased demand for biofuel and other industrial applications from non-edible oilseeds. Several attractive features of camelina make it a potential oilseed crop. It is a low-cost bioenergy crop and the oil has been used successfully as fuel for diesel transport engines (Bernardo et al., 2003). According to Shonnard et al. (2010), when camelina jet fuel was fl ight tested, it met all the requirements for engine performance. In addition, greenhouse gases emitted during combustion of camelina-based fuels were lower than that of petroleum based fuel. Pinzi et al. (2009) indicated that cold weather aff ects the performance of most biofuels; however, fuel derived from camelina is able to withstand lower temperatures because of its high polyunsaturated fatty acid content. Besides biodiesel potential, camelina seeds have an average oil content of 350 to 450 g kg–1, and the proportion of unsaturated fatty acid in the oil is approximately 900 g kg–1 (Gugel and Falk, 2006). Th e high content of unsaturated fatty acid makes camelina oil fastdrying which is useful for making environmentally friendly polymers, varnishes, paints, cosmetics, and dermatological products (Zaleckas et al., 2012). Agronomically, camelina has wide environmental adaptation because it can grow under diff erent climatic and soil conditions (Zubr, 2003). According to Moser and Vaughn (2010), camelina is able to grow well in semiarid regions and in low-fertile and saline soils. Camelina requires low agricultural inputs and its production cost is relatively low (Budin et al., 1995; Moser and Vaughn, 2010). Th ough camelina fi ts well in crop production systems in the semiarid regions in the Great Plains, there Crop Economics, Production & Management

The nature of the vegetative to reproductive transition in the shoot apical meristem of Camelina sativa summer annual cultivar CO46 and winter annual cultivar Joelle was confirmed by treating seedlings with or without 8 weeks of vernalization. True to their life cycle classification, Joelle required a vernalization treatment to induce bolting and flowering, whereas CO46 did not. In this study, whole genome sequence, RNAseq, and resequencing of PCR‐amplified transcripts for a key floral repressor were used to better understand factors involved in the flowering habit of summer and winter biotypes at the molecular level. Analysis of transcriptome data indicated that abundance for one of the three genes encoding the floral repressor FLOWERING LOCUS C (FLC; Csa20 g015400) was 16‐fold greater in Joelle compared to CO46 prior to vernalization. Abundance of this transcript decreased only slightly in CO46 postvernalization, compared to a substantial decrease in Joelle. The results observed in the winter annual biotype Joelle are consistent with repression of FLC by vernalization. Further characterization of FLC at both the genome and transcriptome levels identified a one base deletion in the 5th exon coding for a keratin‐binding domain in chromosome 20 of CO46 and Joelle. The one base deletion detected in chromosome 20 FLC is predicted to result in a frameshift that would produce a nonfunctional protein. Analysis of whole genome sequence indicated that the one base deletion in chromosome 20 FLC occurred at a greater ratio in the summer biotype CO46 (2:1) compared to the winter biotype Joelle (1:4); similar trends were also observed for RNAseq and cDNA transcripts mapping to chromosome 20 FLC of CO46 and Joelle.

Cold-pressed linseed and camelina oils have been available on the global market for many years. They are known for their high contents of polyunsaturated fatty acids (PUFAs), mainly α-linolenic (ALA). This study aimed to evaluate the quality and oxidative stability of linseed and camelina cold-pressed oils purchased in retail outlets in Poland. The fatty acid composition, peroxide value, acid value, anisidine value, chlorophyll pigments, carotenoid pigments, and oxidative stability of these oils were analyzed. It was found that all analyzed oils meet the standards of requirements of the Codex Alimentarius (2009) for cold-pressed oils. The content of chlorophyll and carotenoid pigments of linseed and camelina oils was significantly different. The percentage composition of PUFAs was high in both types of oils, but it was significantly higher in linseed than in camelina oils. The results of Rancimat and PDSC tests show low oxidative stability of all tested oils, although camelina oils were more stable than linseed oils. 1 Practical applications Based on the study, it can be concluded that linseed and camelina oils should be included in the human diet to provide an optimal ratio of omega-6 to omega-3 fatty acids. The differences between tested oils of the same seed species were significant. This study shows that quick methods of testing an oils oxidative stability (Rancimat and PDSC) may be successfully applied in oil studies. The study evaluates the quality of cold-pressed linseed and camelina oils purchased in retail outlets in Poland. The research shows a correlation (r = −0.673) between content of polyunsaturated fatty acids and oxidative stability of tested oils. Oxidative stability was determined by Rancimat test and fatty acid composition by GC-FID.

This study aims to produce human milk fat substitutes (HMFS) rich in polyunsaturated fatty acids (PUFA), mainly the essential omega-3 linolenic acid, by acidolysis reaction between tripalmitin (PPP) and free fatty acids (FFA) from camelina oil, in stirred batch reactor and solvent-free media. The non-commercial heterologous Rhizopus oryzae lipase (rROL), immobilized on Lewatit VP OC 1600 or on Relizyme OD403/S, and the immobilized commercial lipase from Rhizomucor miehei (Lipozyme RM IM) were tested as biocatalysts. Both lipases are sn-1,3 selective. After 24 h reaction at 60°C, using biocatalyst loads of 5 wt% (in relation to tripalmitin), 48.9, 43.6, and 18.3 mol% of fatty acid incorporation in triacylglycerols (TAG) were obtained with Lipozyme RM IM and rROL immobilized on Lewatit or on Relizyme, respectively. rROL immobilized on Lewatit was selected as biocatalyst for the acidolysis, as alternative to the commercial immobilized lipases. With this biocatalyst, the molar incorporation increased with its initial water activity (0.55–0.95). Modeling acidolysis catalyzed by rROL immobilized on Lewatit was performed by response surface methodology, as a function of temperature (58–72°C) and molar ratio FFA:PPP (1.2:1–6.8:1). The highest PPP consumption was achieved at 60°C with a molar ratio of 2:1. The yield of HMFS (new TAG) increased from 42.6 wt% (5% biocatalyst load) to 52% with 8% load, after 24 h acidolysis. Practical applications: Camelina oil showed to be a good source of PUFA, mainly essential fatty acids, to incorporate in HMFS. After 24 h acidolysis under optimized conditions, catalyzed by the non-commercial sn-1,3 regioselective rROL immobilized on Lewatit VP OC 1600, the TAG fraction contains 67.7 mol% of palmitic acid at position 2. These structured lipids rich in PUFA can be used in blends with 1,3-dioleoyl-2-palmitoyl-glycerol (OPO) in order to mimic the human milk fat. The performance of this biocatalyst was comparable to that observed with Lipozyme RM IM. The replacement of high-cost commercial immobilized lipases by rROL immobilized on Lewatit may reduce the biocatalyst cost. In addition, since the best molar ratio FFA:PPP for rROL is 2:1, i.e., the stoichiometric value for the acidolysis catalyzed by sn-1,3 regioselective lipases, the use of this biocatalyst will reduce downstream costs related with unconverted FFA recovery. The performance of rROL immobilized on Lewatit VPOC 1600 is comparable to that of Lipozyme RM IM. After 24 h acidolysis, under optimized conditions, triacylglycerols contain 67.8 mol% palmitic acid at position-2.

Seed oil composed of wax esters with long-chain monoenoic acyl moieties represents a high-value commodity for industry. Such plant-derived sperm oil-like liquid wax esters are biodegradable and can have excellent properties for lubrication. In addition, wax ester oil may represent a superior substrate for biodiesel production. In this study, we demonstrate that the low-input oil seed crop Camelina sativa can serve as a biotechnological platform for environmentally benign wax ester production. Two biosynthetic steps catalysed by a fatty alcohol-forming acyl-CoA reductase (FAR) and a wax ester synthase (WS) are sufficient to achieve wax ester accumulation from acyl-CoA substrates. To produce plant-derived sperm oil-like liquid wax esters, the WS from Mus musculus (MmWS) or Simmondsia chinensis (ScWS) were expressed in combination with the FAR from Mus musculus (MmFAR1) or Marinobacter aquaeolei (MaFAR) in seeds of Arabidopsis thaliana and Camelina sativa. The three analysed enzyme combinations Oleo3:mCherry:MmFAR1∆c/Oleo3:EYFP:MmWS, Oleo3:mCherry:MmFAR1∆c/ScWS and MaFAR/ScWS showed differences in the wax ester molecular species profiles and overall biosynthetic performance. By expressing MaFAR/ScWS in Arabidopsis or Camelina up to 59% or 21% of the seed oil TAGs were replaced by wax esters, respectively. This combination also yielded wax ester molecular species with highest content of monounsaturated acyl moieties. Expression of the enzyme combinations in the Arabidopsis fae1 fad2 mutant background high in oleic acid resulted in wax ester accumulation enriched in oleyl oleate (18:1/18:1 > 60%), suggesting that similar values may be obtained with a Camelina high oleic acid line. peerReviewed