SummaryCamelina (Camelina sativa), a Brassicaceae oilseed, has received recent 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, a transcriptome reference was built from 2047 Sanger ESTs and more than 2 million 454-derived sequence reads, representing genes expressed in developing camelina seeds. The transcriptome of approximately 60K transcripts from 22 597 putative genes includes camelina homologues of nearly all known seedexpressed 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 thaliana and camelina genes was also exploited to engineer high oleic lines by RNAi with Arabidopsis FAD2 and FAE1 sequences. It is expected that these transcriptomic data will be useful for breeding and engineering of additional camelina seed traits and for translating findings from the model Arabidopsis to an oilseed crop.

Nucleotide sequence analysis of the flanking regions of the carBC genes of Pseudomonas sp. strain CA10 revealed that there were two open reading frames (ORFs) ORF4 and ORF5, in the upstream region of carBC. Similarly, three ORFs, ORF6 to ORF8, were found in the downstream region of carBC. The deduced amino acid sequences of ORF6 and ORF8 showed homologies with ferredoxin and ferredoxin reductase components of bacterial multicomponent dioxygenase systems, respectively. ORF4 and ORF5 had the same sequence and were tandemly linked. Their deduced amino acid sequences showed about 30% homology with large (␣) subunits of other terminal oxygenase components. Functional analysis using resting cells harboring the deleted plasmids revealed that the products of ORF4 and -5, ORF6, and ORF8 were terminal dioxygenase, ferredoxin, and ferredoxin reductase, respectively, of carbazole 1,9a-dioxygenase (CARDO), which attacks the angular position adjacent to the nitrogen atom of carbazole, and that the product of ORF7 is not indispensable for CARDO activity. Based on the results, ORF4, ORF5, ORF6, and ORF8 were designated carAa, carAa, carAc, and carAd, respectively. The products of carAa, carAd, and ORF7 were shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be polypeptides with molecular masses of 43, 36, and 11 kDa, respectively. However, the product of carAc was not detected in Escherichia coli. CARDO has the ability to oxidize a wide variety of polyaromatic compounds, including dibenzo-p-dioxin, dibenzofuran, biphenyl, and polycyclic aromatic hydrocarbons such as naphthalene and phenanthrene. Since 2,2,3-trihydroxydiphenyl ether and 2,2,3-trihydroxybiphenyl were identified as metabolites of dibenzo-p-dioxin and dibenzofuran, respectively, it was considered that CARDO attacked at the angular position adjacent to the oxygen atom of dibenzo-p-dioxin and dibenzofuran as in the case with carbazole.Microbial degradation of xenobiotics such as polycyclic aromatic hydrocarbons, heterocyclic polyaromatics, and halogenated aromatics and aliphatics is quite useful to detoxify and mineralize these pollutants and thus is an important goal of research related to bioremediation. Among these pollutants, polychlorinated dibenzo-p-dioxin (DD), dibenzofuran (DBF), and carbazole (CAR) are well known to possess strong mutagenic and toxic activities (2) and also to be recalcitrant molecules (7). A critical point in the degradative pathways of these compounds is the initial oxidation of the chemically stable aromatic ring. In the aerobic degradation of many aromatic compounds, the multicomponent aromatic ring dioxygenases are very important enzyme complexes, consisting of two or three protein components (17, 28). Multicomponent dioxygenases catalyze a redox reaction in which molecular oxygen atoms are incorporated into the aromatic ring at the expense of the oxidation of NAD(P)H (17, 28). The recalcitrance of these xenobiotic compounds is considered to be due to requirement of rare and unique aromatic ring oxygenases for degrada...

Significance Many plants produce valuable fatty acids in seed oils that provide renewable alternatives to petrochemicals for production of lubricants, coatings, or polymers. However, most plants producing these unusual fatty acids are unsuitable as crops. Metabolic engineering of oilseed crops, or model species, to produce the high-value unusual fatty acids has produced only low yields of the desired products, and previous research has indicated fatty acid degradation as a potential major factor hindering oilseed engineering. By contrast, we here present evidence that inefficient utilization of unusual fatty acids within the endoplasmic reticulum can induce posttranslational inhibition of acetyl–CoA carboxylase activity in the plastid, thus inhibiting fatty acid synthesis and total oil accumulation.

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HighlightTranscriptomic data mining and combinatorial gene stacking was used for metabolic engineering of Camelina to produce oils containing medium-chain length fatty acids that mimic jet fuel hydrocarbon components.