Iodus 40, salicylic acid, heptanoyl salicylic acid and trehalose exhibit different efficacies and defence targets during a wheat/powdery mildew interaction

Prophylactic efficacies of Iodus 40 and salicylic acid (SA) against wheat powdery mildew caused by Blumeria graminis f. sp. tritici have been shown and compared with those of heptanoyl salicylic acid (HSA) and trehalose. Plantlets treated once exhibited 55%, 50%, 95%, and 38% protection levels, respectively. Two sprayings increased these levels up to 60%, 65%, 100%, and 60%, respectively. Biological effects of these resistance inducers on reactive oxygen species (ROS) metabolism and lipid peroxidation were also investigated. We found clear differences in the extent and the type of induced responses, with HSA exhibiting both the most numerous and the highest effects. HSA and SA induced a 5.5-fold increase of whole cell DAB staining due to hydrogen peroxide accumulation, whereas Iodus 40 and trehalose increased staining intensity at the penetration sites only. However, these effects were not correlated with any modification of catalase (CAT), oxalate oxidase (OXO) or lipoxygenase (LOX) activities, except for HSA which decreased CAT in non-inoculated conditions and increased LOX in infectious conditions. HSA also induced an increase in the rate of lipid peroxidation, whereas Iodus 40 induced a decrease. The effects of the inducers on germinating conidia and wheat epidermal cells responding to fungal penetration were also investigated. Papilla-linked autofluorescence was affected by SA and Iodus 40 whereas germination was slightly altered by Iodus 40. The newly described protective efficacies and the partial, distinct and non-overlapping activities of these inducers on the wheat/powdery mildew interaction are discussed.     

Changes in phytohormones and fatty acids in wheat and rice seedlings in response to Hessian fly (Diptera: Cecidomyiidae) infestation

Phytohormones and fatty acids (FAs) play important roles in plant resistance to insects and pathogens. In this study, we investigated the similarities and differences in the accumulations of phytohormones and FAs in the resistant wheat (Triticum aestivum L.) 'Molly' and the nonhost rice (Oryza sativa L.) 'Niponbare' in responses to Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae), larval attacks. Using chemical ionization-gas-chromatography/mass spectrometry, we analyzed the concentrations of 13 phytohomones and FAs at the attack site of wheat and rice plants at 1, 6, 24, or 48 h after the initial attack. Hessian fly attack resulted in increases of salicylic acid (SA), 12-oxo-phytodienoic acid (OPDA), palmitic acid (FA16:0), but a decrease of abscisic acid in both wheat and rice plants. In addition, the accumulation of jasmonic acid (JA) increased, whereas the accumulation of cinnamic acid (CA) decreased in wheat plants, but no changes were observed in the accumulation of JA, and the accumulation of CA increased in rice plants after Hessian fly attack. However, the accumulations of benzoic acid, strearic acid (FA18:0), and oleic acid (FA18:1) increased in rice plants, but no changes were observed in wheat plants after Hessian fly attack. Hessian fly-induced changes were more rapid in wheat plants in comparison with those in rice plants. Our study suggests that SA and OPDA may be involved in resistance of wheat and rice plants to Hessian fly and that the R gene-mediated resistance responses are more rapid than nonhost resistance responses.     

Changes in the content and composition of lipid fatty acids in tobacco leaves and roots at low-temperature hardening

Changes in the fatty acid (FA) composition of leaf and root lipids of heat-loving tobacco (Nicotiana tabacum L., cv. Samsun) plants during low-temperature hardening (8°C for 6 days) were studied. Hardening could improve leaf but not root cold tolerance. As this took place, the relative content of polyunsaturated (18:2n-6 and 18:3n-3) FAs increased and the proportion of saturated and monounsaturated FAs decreased. In contrast, in the roots hardening slightly increased the concentration of saturated FAs (16:0 and 18:0) and reduced the level of unsaturated FAs (18:1n-9, 18:2n-6, and 18:3n-3). At the same time, root lipids contained much C_20–24 FAs, and their content increased during hardening. It was suggested that an increased FA saturation and elevated proportion of C_20–24 FAs in the root lipids resulting in the lower membrane fluidity could be a reason for incapability of heat-loving tobacco plant roots of hardening and plant death at the lowtemperature stress.     

Changes in the content of gamma-linolenic C18:3 (n-6) and stearidonic C18:4 (n-3) acids in developing seeds of viper's bugloss Echium vulgare L

Changes in the composition of fatty acids (FA) were determined in lipid extracts isolated from developing ovaries containing ovules and developing seeds of Echium vulgare L. The samples were collected successively over 20 days beginning with the first day after flowering. The contents of the n-6 FA family members, i.e., gamma-linolenic (GLA) (C(18:3)) and linoleic (LA) (C(18:2)) acids changed in a parallel manner and reached the maximum of 13.9% and 24%, respectively, on the 12th day, after which they fell systematically down to 8.6% and 18.2%, respectively, on the 20th day after flowering. Starting with day 13, the content of alpha-linolenic acid (ALA) (C(18:3) n-3) begins to grow intensively, from 24.2% to 39.3% on the 20th day after flowering. The increase in the content of stearidonic acid (SDA) (C(18:4) n-3), up to 10.5% on the 20th day after flowering, occurred steadily as the seeds developed, and was independent of the changes in the content of GLA and LA. The pattern of changes in the content of SDA, GLA, LA and ALA during the development of seeds, and the occurrence of SDA in the seed oil of other plants, demonstrate that the biosynthesis of SDA in the seeds is critically dependent on the presence of ALA. The above condition indicates that SDA biosynthesis in the seeds of Echium vulgare follows the scheme LA --> simultaneous, competitive, action of Delta(6) and Delta(15) desaturases, leading to the formation of GLA and ALA, respectively, and then ALA (Delta(6) des) --> SDA. The biosynthesis according to the scheme: GLA (Delta(15) des) --> SDA is highly unlikely.     

Fatty Acids and Squalene Carried by Alpha Fetoprotein, and Fetal and Adult Serum Albumin from Chicken. Comparison with These from Mammals

Alpha fetoprotein (C-AFP), serum albumin (C-fSA) from chicken embryos, and SA from hens were purified using gentle chromatographic and electrophoretic methods, and their fatty acids (FAs) and squalene contents were analyzed using gas chromatography and mass spectrometry. In 7-day-old chick embryo, AFP carries 16% and fSA 6% free FAs, the rest being carried as phospholipids, which differs from rat and pig AFP, where all fatty acids are carried like free fatty acids. At this stage C-AFP contains 3.6% arachidonic acid, which falls to 1.7% in 14-day-old embryos. Both of these figures are significantly lower than in humans, rats, calves, and pigs. C-AFP does not transport docosahexaenoic acid, in notable contrast to the mammals mentioned above. The finding of squalene in the two fetal proteins is reported for the first time. During the interval between 7 and 14 days, the proportion of C16:1 n-7 and of C18:2 n-6 increases 10-fold, that of C18:0 quadruples, and that of C18:1 n-9 decreases 3-fold. Squalene increases in this period from 2.2% to 10.0%. The C-fSA of a 7-day-old embryo transports only one FA with more than two unsaturated carbons, arachidonic acid (2.4%). It also contains squalene (1.2%). Similarly, only arachidonic acid (2.5%), but not squalene is found in hen SA. The percentage of saturated and monounsaturated FAs divided by the percentage of polyunsaturated FAs, and the ratios of the percentage of FAs with C14–C18 with respect to those with C20–C22 transported by C-AFP are very different from those of studied mammals.     

Characterization of Aspergillus species based on fatty acid profiles

Cellular fatty acid (FA) composition was utilized as a taxonomic tool to discriminate between different Aspergillus species. Several of the tested species had the same FA composition and different relative FA concentrations. The most important FAs were palmitic acid (C16:0), estearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2), which represented 95% of Aspergillus FAs. Multivariate data analysis demonstrated that FA analysis is a useful tool for differentiating species belonging to genus Aspergillus. All the species analyzed showed significantly FA acid profiles (p < 0.001). Furthermore, it will be possible to distinguish among Aspergillus spp. in the Flavi Section. FA composition can serve as a useful tool for the identification of filamentous fungi.     

Effect of chestnut and quebracho tannins on fatty acid profile in rumen liquid- and solid-associated bacteria: an in vitro study

Tannins are phenolic compounds that interfere with biohydrogenation (BH) of polyunsaturated fatty acids (FAs). The aim of the present in vitro study was to investigate the effects of two different sources of tannins on FA profiles of rumen bacteria, with particular reference to rumenic and vaccenic acid. A control diet (C; composed of 300 g/kg of wheat straw, 132 g/kg of soyabean meal, 96 g/kg of barley meal, 152 g/kg of maize meal, 300 g/kg of maize gluten and 20 g/kg of mineral vitamin premix, all expressed on dry matter (DM)) and four diets, obtained by adding to C two different types of tannins from chestnut (TC) and from quebracho (TQ) at two concentration levels (49 and 82 g/kg DM), were compared. The content of the main unsaturated FAs (C18:1 cis9, C18:1 trans11, C18:2 cis9, cis12 and C18:3 cis9, cis12, cis15) from solid-associated bacteria (SAB) and liquid-associated bacteria (LAB) was affected by the presence of tannins in the diets. In particular, C18:1 trans11 content was significantly increased, especially with TC1, whereas the decreasing of C18:1 cis9 was unaffected, regardless of the presence or the kind of tannins added to feeds. SAB contained higher amounts of intermediates of polyunsaturated FA BH (as C18:1 trans11 and C18:2 cis9, trans11) than LAB that were characterized by a higher amount of C18:0. In the concentration range adopted in this study, the effect of TC and TQ on changes of bacterial FA profile was comparable. Tannins seem to be a good means to modulate the FA profile of rumen bacteria, favouring the accumulation of C18:1 trans11 during in vitro rumen fermentation.     

A novel unsaturated fatty acid hydratase toward C16 to C22 fatty acids from Lactobacillus acidophilus

Hydroxy FAs, one of the gut microbial metabolites of PUFAs, have attracted much attention because of their various bioactivities. The purpose of this study was to identify lactic acid bacteria with the ability to convert linoleic acid (LA) to hydroxy FAs. A screening process revealed that a gut bacterium, Lactobacillus acidophilus NTV001, converts LA mainly into 13-hydroxy-cis-9-octadecenoic acid and resulted in the identification of the hydratase responsible, fatty acid hydratase 1 (FA-HY1). Recombinant FA-HY1 was purified, and its enzymatic characteristics were investigated. FA-HY1 could convert not only C18 PUFAs but also C20 and C22 PUFAs. C18 PUFAs with a cis carbon-carbon double bond at the Δ12 position were converted into the corresponding 13-hydroxy FAs. Arachidonic acid and DHA were converted into the corresponding 15-hydroxy FA and 14-hydroxy FA, respectively. To the best of our knowledge, this is the first report of a bacterial FA hydratase that can convert C20 and C22 PUFAs into the corresponding hydroxy FAs. These novel hydroxy FAs produced by using FA-HY1 should contribute to elucidating the bioactivities of hydroxy FAs.     

Expression of the Isochrysis C18-delta9 polyunsaturated fatty acid specific elongase component alters Arabidopsis glycerolipid profiles

A cDNA isolated from the prymnesiophyte micro-alga Isochrysis galbana, designated IgASE1, encodes a fatty acid elongating component that is specific for linoleic acid (C18:2n-6) and alpha-linolenic acid (C18:3n-3). Constitutive expression of IgASE1 in Arabidopsis resulted in the accumulation of eicosadienoic acid (EDA; C20:2n-6) and eicosatrienoic acid (ETrA; C20:3n-3) in all tissues examined, with no visible effects on plant morphology. Positional analysis of the various lipid classes indicated that these novel fatty acids were largely excluded from the sn-2 position of chloroplast galactolipids and seed triacylglycerol, whereas they were enriched in the same position in phosphatidylcholine. EDA and ETrA are precursors of arachidonic acid (C20:4n-6), eicosapentaenoic acid (C20:5n-3), and docosahexaenoic acid (C22:6n-3) synthesized via the so-called omega6 Delta8 desaturase and omega3 Delta8 desaturase biosynthetic pathways, respectively. The synthesis of significant quantities of EDA and ETrA in a higher plant is therefore a key step in the production of very long chain polyunsaturated fatty acid in oil-seed species. The results are further discussed in terms of prokaryotic and eukaryotic pathways of lipid synthesis in plants.     

Relationships between hepatic stearoyl-CoA desaturase-1 activity and mRNA expression with liver fat content in humans

Stearoyl-CoA desaturase-1 (SCD1) has gained much interest as a future drug target to treat fatty liver and its consequences. However, there are few and inconsistent human data about expression and activity of this important enzyme. We investigated activity and expression of SCD1 and their relationships with liver fat (LF) content in human liver samples. Fifty subjects undergoing liver surgery were studied. SCD1 activity was estimated from the ratio of oleate (C18:1) to stearate (C18:0) within lipid subfractions. Furthermore, SCD1 mRNA expression and LF content were measured. Similarly to previous studies, we observed a strong positive correlation between LF content and the C18:1/C18:0 ratio in the combined fatty acid (FA) fractions (r = 0.96, P < 0.0001), which could be interpreted as higher SCD1 activity with increasing LF. However, hepatic SCD1 mRNA expression did not correlate with LF (r = 0.16, P = 0.13). To solve these conflicting data, we analyzed the FA composition of hepatic lipid subfractions. With increasing LF content the amount of FAs from the triglyceride (TG) fraction increased (r = 0.96, P < 0.0001), whereas the FAs from the phospholipid (PL) fraction remained unchanged (r = -0.17, P = 0.19). Of these two major lipid fractions, the C18:1/C18:0 ratio in TG was 16-fold higher than in PL. Supporting the SCD1 mRNA expression data, the C18:1/C18:0 ratio of the TG or PL fraction did not correlate with LF (r = 0.26, P = 0.12 and r = 0.08, P = 0.29). We provide novel information that SCD1 activity and mRNA expression appear not to be elevated in subjects with high LF content. We suggest that the FA composition of lipid subclasses, rather than of mixed lipids, should be analyzed to estimate SCD1 activity.     

Ala54Thr Fatty Acid-Binding Protein 2 (FABP2) Polymorphism in Recurrent Depression: Associations with Fatty Acid Concentrations and Waist Circumference

Background

Fatty acid (FA)-alterations may mediate the mutual association between Major Depressive Disorder (MDD) and cardiovascular disease (CVD). However, etiology of observed FA-alterations in MDD and CVD remains largely unclear. An interesting candidate may be a mutation in the fatty acid–binding protein 2 (FABP2)-gene, because it regulates dietary FA-uptake. Therefore, we aimed to test the hypotheses that in MDD-patients the FABP2 Ala54Thr-polymorphism would be (I) more prevalent than in sex- and age-matched controls, (II) associated with observed alterations in FA-metabolism, and (III) associated with CVD-risk factor waist circumference.

Methods

We measured concentrations of 29 different erythrocyte FAs, FABP2-genotype, and waist circumference in recurrent MDD-patients and matched never-depressed controls.

Results

FABP2-genotype distribution did not significantly differ between the 137 MDD-patients and 73 matched controls. However, patients with the Ala54Thr-polymorphism had (I) higher concentrations of especially eicosadienoic acid (C20:2ω6; P=.009) and other 20-carbon FAs, and associated (II) lower waist circumference (P=.019). In addition, FABP2-genotype effects on waist circumference in patients seemed (I) mediated by its effect on C20:2ω6, and (II) different from controls.

Conclusions

Although Ala54Thr-polymorphism distribution was not associated with recurrent MDD, our results indicate that FABP2 may play a role in the explanation of observed FA-alterations in MDD. For Ala54Thr-polymorphism patients, potentially adaptive conversion of increased bioavailable dietary precursors into eicosadienoic acid instead of arachidonic acid might be related to a low waist circumference. Because this is the first investigation of these associations, replication is warranted, preferably by nutrigenetic studies applying lipidomics and detailed dietary assessment.     

Effects of Nitrogen fertilisation and regrowth period on fatty acid concentrations in perennial ryegrass (Lolium perenne L.)

Information on lipids in forages is scarce, and effects of nitrogen (N) application level and regrowth period on the fatty acid (FA) concentration and composition of perennial ryegrass (Lolium perenne L.) were studied. N was applied at 0, 45 and 100 kg ha^?1, and swards were cut after various regrowth periods, resulting in six treatments designed as randomised blocks with three replicates. The stages of development ranged from vegetative to elongating swards, with herbage yield levels from 1.9 to 4.2 t dry matter (DM) ha^?1. Concentrations of individual FA were determined by gas chromatography, and canopy characteristics and herbage quality were assessed. The treatments resulted in canopies with contrasting DM yields and protein concentrations. Five FAs, representing 98% of total FAs, were studied in detail. On an average, the concentration of these major FAs in fresh grass was 15.1 g kg^?1 DM, and 69% of the major FAs consisted of C18:3. Regrowth period affected (P < 0.05) the total FA concentration, and significantly (P < 0.01) lower concentrations of C18:3 and C16:1 were found after a longer period of regrowth. N application resulted in higher (P < 0.001) concentrations of all FAs. The FA composition was not affected by N application, but a longer regrowth period significantly (P < 0.001) decreased the proportion of C18:3 and increased those of C18:2 and C16:0. A strong, positive overall linear relation was found between the concentrations of total FAs and C18:3 with the crude protein concentration in the herbage. These studies demonstrate opportunities to affect the FA concentration and composition of FA in herbage through management strategies, which could affect milk FA composition.     

Fatty acid composition of the parasitic mite Varroa destructor and its host the worker prepupae of Apis mellifera

The present study analyzes the fatty acid (FA) profile of lipids isolated from Varroa destructor Anderson & Trueman, a parasitic mite of the honey bee (Apis mellifera L.), uninfected and infected worker prepupae of the Carnolian subspecies Apis mellifera carnica Pollmann, and bee bread fed to the worker brood. Significant differences are observed in the FA profiles of lipids isolated from parasites, hosts and bee bread. Parasitism by V. destructor (henceforth, varroosis) induces visible changes in the lipid profile of worker prepupae. In infected prepupae, the percentage of total saturated FAs is lower and the percentage of unsaturated FAs is higher than in uninfected insects. These differences result from significant changes in the percentages of FAs that are most abundant in the evaluated groups (i.e. C16:0, C18:1 9c, C18:2n‐6 and C18:3n‐3 FAs). In mites and in uninfected and infected prepupae, the predominant FAs are oleic acid (41.07 ± 2.26%, 42.79 ± 1.21% and 45 ± 0.20%, respectively) and palmitic acid (22.62 ± 0.87%, 39.48 ± 0.43% and 36.84 ± 0.22%, respectively). Highly significant differences in FA composition are noted between bee bread and worker brood. The results suggest specific mechanisms of FA uptake, accumulation and metabolism in the food chain of this parasitic association, beginning from the food processed by nurse bees for larval feeding, through host organisms (worker brood) to V. destructor mites.     

Effect of age on the fatty acid content of Blumeria graminis conidia

Blumeria (=Erysiphe) graminis f.sp. tritici (Bgt), the causal agent of wheat powdery mildew, is responsible for an important disease leading to considerable yield reductions in wheat worldwide. Conidia of the obligate plant pathogen Bgt were analysed for their total fatty acid (FA) composition as a function of their ontogeny. A total of 17 FAs were detected (C(12)-C(24) saturated and unsaturated ones), including the presence of unusual long-chain monoenoic FAs. In young conidia, the major FAs were C(18:2) (23%), C(16:0) (16%), C(18:0) (15.2%) and C(18:1) (14.3%). In old conidia, the main FAs were C(24:1) (20.7%), C(22:0) (15%), C(22:1) (13.5%) and C(24:0) (9.7%). The amount of total FA was about 39 microg.mg of dry weight(-1) in young conidia and decreased clearly to 18 microg.mg of dry weight(-1) in older conidia. For the first time, we have demonstrated that the FA composition of conidia changes greatly with age. Medium-chain FAs (C(12)-C(18)) are predominant in very young conidia (75%), whereas long-chain FAs (C(22)-C(24)) are the major compounds in old conidia (74%). This study showed a significant elongation of FAs and a drastic decrease in the total FA amount during the ontogeny of conidia.     

Restoration of defective cross talk in ssi2 mutants: role of salicylic acid, jasmonic acid, and fatty acids in SSI2-mediated signaling

The Arabidopsis mutants ssi2 and fab2 are defective in stearoyl ACP desaturase, which causes altered salicylic acid (SA)- and jasmonic acid (JA)-mediated defense signaling. Both ssi2 and fab2 plants show spontaneous cell death, express PR genes constitutively, accumulate high levels of SA, and exhibit enhanced resistance to bacterial and oomycete pathogens. In contrast to constitutive activation of the SA pathway, ssi2 and fab2 plants are repressed in JA-mediated induction of the PDF1.2 gene, which suggests that the SSI2-mediated signaling pathway modulates cross talk between the SA and JA pathways. In this study, we have characterized two recessive nonallelic mutants in the ssi2 background, designated as rdc (restorer of defective cross talk) 2 and rdc8. Both ssi2 rdc mutants are suppressed in constitutive SA signaling, show basal level expression of PR-1 gene, and induce high levels of PDF1.2 in response to exogenous application of JA. Interestingly, while the rdc8 mutation completely abolishes spontaneous cell death in ssi2 rdc8 plants, the ssi2 rdc2 plants continue to show some albeit reduced cell death. Fatty acid (FA) analysis showed a reduction in 16:3 levels in ssi2 rdc8 plants, which suggests that this mutation may limit the flux of FAs into the prokaryotic pathway of glycerolipid biosynthesis. Both rdc2 and rdc8 continue to accumulate high levels of 18:0, which suggests that 18:0 levels were responsible for neither constitutive SA signaling nor repression of JA-induced expression of the PDF1.2 gene in ssi2 plants. We also analyzed SA and JA responses of the fab2-derived shs1 mutant, which accumulates levels of 18:0 over 50% lower than those in the fab2 plants. Even though fab2 shs1 plants were morphologically bigger than fab2 plants, they expressed PR genes constitutively, showed HR-like cell death, and accumulated elevated levels of SA. However, unlike the ssi2 rdc plants, fab2 shs1 plants were unable to induce high levels of PDF1.2 expression in response to exogenous application of JA. Together, these results show that defective cross talk in ssi2 can be restored by second site mutations and is independent of morphological size of the plants, cell death, and elevated levels of 18:0.     

Temporal dynamics of amino and fatty acid composition in the razor clam Ensis siliqua (Mollusca: Bivalvia)

Few studies have been conducted on the temporal dynamics of both amino acid (AA) and fatty acid (FA) profiles in marine bivalves. We investigated the seasonal variation of these compounds in the pod razor clam Ensis siliqua in relation to food availability, salinity, water temperature and reproductive cycle. AA content varied between 46.94 and 54.67 % dry weight (DW), and the AAs found in greater quantity were glutamic acid, glycine and aspartic acid. FA content varied between 34.02 and 87.94 mg g^?1 DW and the FAs found in greater quantity were 16:0 and 22:6n-3. Seasonal trends were observed for AAs and FAs. FAs increased with gametogenesis and decreased with spawning while AA content increased throughout spawning. The effect of increasing temperature and high food availability during the spawning season masked the loss of AAs resulting from gamete release. Still, a comparatively greater increase in the contents of glutamic acid and leucine with spawning indicate their possible involvement in a post-spawning gonad recovery mechanism. A post-spawning decrease in 14:0, 16:0, 16:1n-7, 18:1n-7 and 18:1n-9 is indicative of the importance of these FAs in bivalve eggs. An increase in 18:3n-3, 18:4n-3, 20:1n-9 and 20:2n-6 during gametogenesis suggests their involvement in oocyte maturation. The FA 22:4n-6, while increasing with spawning, appears to play a role in post-spawning gonad recovery. Salinity did not have an effect on the AA composition. None of the environmental parameters measured had an effect on FA composition.     

Fatty acid composition of membrane lipids and energy metabolism in mitochondria of pea seedlings under water deficit

The impacts of water deficit and melamine salt of bis(oximethyl)phosphonic acid (melaphen) on the fatty acid (FA) composition of membrane lipids and energy metabolism in mitochondria of 5-day-old pea (Pisum sativum L., cv. Flora-2) seedlings were studied. Insufficient watering resulted in the accumulation of saturated and a decrease in the content of unsaturated FAs with 18 and 20 carbon atoms. Seed treatment with 3 × 10^?10 M melaphen prevented these changes in the FA composition in the mitochondrial membrane lipids. Changes in the FA compositions of membrane lipids were correlated with changes in energy metabolism in mitochondria: the efficiency of oxidative phosphorylation and the rate of NAD-dependent substrate oxidation in the presence of ADP and FCCP (carbonyl cyanide-p-trifluoromethoxyphenylhydrazone) were reduced. A close correlation was observed between changes in the highest rates of NAD-dependent substrate oxidation and the relative content of FAs with 18 (r = 0.76489) and 20 (r = 0.9637) carbon atoms. The regulatory role of C₁₈ and C₂₀ unsaturated FAs in the mitochondrial energy metabolism of pea seedlings is discussed.     

Castor phospholipid:diacylglycerol acyltransferase facilitates efficient metabolism of hydroxy fatty acids in transgenic Arabidopsis

Producing unusual fatty acids (FAs) in crop plants has been a long-standing goal of green chemistry. However, expression of the enzymes that catalyze the primary synthesis of these unusual FAs in transgenic plants typically results in low levels of the desired FA. For example, seed-specific expression of castor (Ricinus communis) fatty acid hydroxylase (RcFAH) in Arabidopsis (Arabidopsis thaliana) resulted in only 17% hydroxy fatty acids (HFAs) in the seed oil. In order to increase HFA levels, we investigated castor phospholipid:diacylglycerol acyltransferase (PDAT). We cloned cDNAs encoding three putative PDAT enzymes from a castor seed cDNA library and coexpressed them with RcFAH12. One isoform, RcPDAT1A, increased HFA levels to 27%. Analysis of HFA-triacylglycerol molecular species and regiochemistry, along with analysis of the HFA content of phosphatidylcholine, indicates that RcPDAT1A functions as a PDAT in vivo. Expression of RcFAH12 alone leads to a significant decrease in FA content of seeds. Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12 restored FA levels to nearly wild-type levels, and this was accompanied by a major increase in the mass of HFAs accumulating in the seeds. We show the usefulness of RcPDAT1A for engineering plants with high levels of HFAs and alleviating bottlenecks due to the production of unusual FAs in transgenic oilseeds.