Genome-Wide Identification of B-Box Gene Family and Candidate Light-Related Member Analysis of Tung Tree (Vernicia fordii).

Light is one of the most important environmental factors for plant growth. In the production process of tung oil tree cultivation, due to the inappropriate growth of shading conditions, the lower branches are often dry and dead, which seriously affects the yield of tung oil trees. However, little is known about the key factors of light-induced tree photomorphogenesis. In this study, a total of 22 VfBBX family members were identified to provide a reference for candidate genes in tung tree seedlings. All members of the VfBBX family have different numbers of highly conserved B-box domains or CCT domains. Phylogenetic evolution clustered the VfBBX genes into four categories, and the highest density of members was on chromosome 6. Interspecific collinearity analysis suggested that there were six pairs of duplicate genes in VfBBX members, but the expression levels of all family members in different growth and development stages of the tung tree were significantly divergent. After different degrees of shading treatment and physiological data determination of tung tree seedlings, the differential expression level and chlorophyll synthesis genes correlation analysis revealed that VfBBX9 was a typical candidate nuclear localization transcription factor that was significantly differentially expressed in light response. This study systematically identified the VfBBX gene family and provided a reference for studying its molecular function, enhanced the theoretical basis for tung tree breeding, and identified excellent varieties.

Longitudinal impact of distinct infection trajectories on all-cause mortality of older people in Taiwan: a retrospective, nationwide, population-based study.

BACKGROUND: Infectious diseases are the leading cause of deaths in adults aged 65 years or older. Studies of adverse infection outcomes have been limited to specific infections and acute episodes and have not investigated longitudinal trends of cumulative infections. We aimed to identify distinct trajectories of longitudinal infection episodes in older adults and to assess their corresponding risk of all-cause mortality. METHODS: In this retrospective cohort study, we included people aged 65 years or older who were admitted to hospital between Jan 1 and Dec 31, 2011, with one of the following infections: urinary tract, pneumonia, sepsis, cellulitis, cholecystitis, peritonitis, endocarditis, and meningitis. Participants were identified from Taiwan's National Health Insurance Research Database. We analysed infection episodes on a quarterly basis during a 5-year period (2011-15) and used group-based trajectory modelling to identify distinct trajectories. We examined the associations between infection trajectories and all-cause mortality using Kaplan-Meier curves and the Cox proportional hazard model. FINDINGS: Among 79 666 eligible older adults, we identified four distinct infection trajectories over the 5-year follow-up: infrequent (58 619 [73·6%]), increasing (9746 [12·2%]), decreasing (9069 [11·4%]), and frequent (2232 [2·8%]). Compared with people with infrequent infections, the adjusted hazard ratios for all-cause mortality were 2·96 (95% CI 2·82-3·11) in participants with frequent infections, 2·15 (2·09-2·22) in those with increasing infections, and 1·85 (1·80-1·91) in those with decreasing infections. INTERPRETATION: Older adults with multiple infection episodes, irrespective of type, pathogens, and distinct infection pattern, had greater risk of all-cause mortality compared with those with infrequent infections. Further research to define the overall infection burden in older adults is needed for risk stratification and to inform prevention strategies. FUNDING: The Interdisciplinary Research Center for Healthy Longevity of National Yang Ming Chiao Tung University from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education, the National Science and Technology Council, and the Ministry of Science and Technology in Taiwan.

Genome-wide identification and transcriptional profiling of the basic helix-loop-helix gene family in tung tree (Vernicia fordii).

The basic helix-loop-helix (bHLH) transcription factor gene family is one of the largest gene families and is extensively involved in plant growth, development, biotic and abiotic stress responses. Tung tree (Vernicia fordii) is an economically important woody oil plant that produces tung oil rich in eleostearic acid. However, the characteristics of the bHLH gene family in the tung tree genome are still unclear. Hence, VfbHLHs were first searched at a genome-wide level, and their expression levels in various tissues or under low temperature were investigated systematically. In this study, we identified 104 VfbHLHs in the tung tree genome, and these genes were classified into 18 subfamilies according to bHLH domains. Ninety-eight VfbHLHs were mapped to but not evenly distributed on 11 pseudochromosomes. The domain sequences among VfbHLHs were highly conserved, and their conserved residues were also identified. To explore their expression, we performed gene expression profiling using RNA-Seq and RT-qPCR. We identified five, 18 and 28 VfbHLH genes in female flowers, male flowers and seeds, respectively. Furthermore, we found that eight genes (VfbHLH29, VfbHLH31, VfbHLH47, VfbHLH51, VfbHLH57, VfbHLH59, VfbHLH70, VfbHLH72) were significant differential expressed in roots, leaves and petioles under low temperature stress. This study lays the foundation for future studies on bHLH gene cloning, transgenes, and biological mechanisms.

Evolution and function of ubiquitin-specific proteases (UBPs): Insight into seed development roles in plants.

The tung oil produced by the tung tree (Vernicia fordii) provides resources for the manufacture of biodiesel. Ubiquitin-specific proteases (UBPs) are the largest group of deubiquitinases and play key roles in regulating development and stress responses. Here, 21 UBPs were identified in V. fordii, roughly one-half the number found in Manihot esculenta and Hevea brasiliensis. Most UBP duplications are produced from whole-genome duplication (WGD), and significant differences in gene retention existed among Euphorbiaceae. The great majority of UBP-containing blocks in V. fordii, V. montana, Ricinus communis, and Jatropha curcas exhibited extensive conservation with the duplicated regions of M. esculenta and H. brasiliensis. These blocks formed 14 orthologous groups, indicating they shared WGD with UBPs in M. esculenta and H. brasiliensis, but most of these UBPs copies were lost. The UBP orthologs contained significant functional divergence which explained the susceptibility of V. fordii to Fusarium wilt and the resistance of V. montana to Fusarium wilt. The expression patterns and experiments suggested that Vf03G1417 could affect the seed-related traits and positively regulate the seed oil accumulation. This study provided important insights into the evolution of UBPs in Euphorbiaceae and identified important candidate VfUBPs for marker-assisted breeding in V. fordii.

Dissection of leucine-rich repeat receptor-like protein kinases: insight into resistance to Fusarium wilt in tung tree.

The tung tree is a woody oil plant native to China and widely distributed in the subtropics. The three main species commonly known as Vernicia are V. fordii, V. montana, and V. cordata. The growth and development of V. fordii are affected by a large number of plant pathogens, such as Fusarium wilt caused by Fusarium sp. In contrast, V. montana shows significant resistance to Fusarium wilt. The leucine-rich repeat receptor-like protein kinase (LRR-RLK) is the largest class of receptor-like kinases associated with plant resistance to Fusarium wilt. Here, we identified 239 VmLRR-RLKs in V. montana, and found that there were characteristic domains of resistance to Fusarium wilt in them. Phylogenetic analysis suggested that the VmLRR-RLKs are divided into 14 subfamilies, indicating that homologous genes in the same group may have similar functions. Chromosomal localization analysis showed that VmLRR-RLKs were unevenly distributed on chromosomes, and segment duplications were the main reason for the expansion of VmLRR-RLK family members. The transcriptome data showed that six orthologous pairs were up-regulated in V. montana in response to Fusarium wilt, while the corresponding orthologous genes showed low or no expression in V. fordii in resistance Fusarium wilt, further indicating the important role of LRR-RLKs in V. montana's resistance to infection by Fusarium spp. Our study provides important reference genes for the future use of molecular breeding to improve oil yield and control of Fusarium wilt in tung tree.

General and genetic toxicology studies of Aleurites moluccana (L.) Willd. seeds in vitro and in vivo assays.

ETHNOPHARMACOLOGY RELEVANCE: Aleurites moluccana is popularly used for the diseases like ulcers, fever, headache, asthma, conjunctivitis, gonorrhea, inflammation, hepatitis, and rheumatism. The seed, also known as "noz da Índia", has been popularly consumed for weight loss purposes but reports of toxicity have been associated with its ingestion. In the literature, there are not enough studies to elucidate its toxicology, so evaluating the general and genetic toxicological of A. moluccana seeds can provide data to ensure their intake. AIM OF THE STUDY: The objective of the present study was to elucidate the oral toxicity, mutagenicity, genotoxicity and cytotoxicity of A. moluccana seeds in vitro and in vivo assays. MATERIALS AND METHODS: The chemical composition of the aqueous extract of A. moluccana seeds (AEAMS) was analyzed in relation to phenolic compounds, tannins, flavonoids and fatty acid. For the in vitro assays, the cytotoxic potential was assessed by the MTS assay whereas the mutagenic potential was assessed by the Ames test. For in vivo assays, was conducted an acute oral toxicity study, with "Up-and-Down Procedure" and repeated dose toxicity with "Repeated Dose 28-Day Oral Toxicity". To assess genetic damage, mutagenic potential was assessed by the micronucleus test whereas the polychromatic erythrocyte/normochromatic erythrocyte ratio was obtained with bone marrow cells to determine the cytotoxic potential and genotoxic potential was assessed by the comet assay using peripheral blood cells. RESULTS: AEAMS did not show cytotoxic and mutagenic potential in vitro. No clinical signs of toxicity were observed in animals after the acute oral toxicity test, suggesting that the LD50 of aqueous extract of A. moluccana seeds > 2000 mg/kg in a single dose by intragastric route. However, in toxicity at repeated doses for 28 days, the doses initially established (250; 500 and 750 mg/kg/day by intragastric route) caused mortality in the animals and the reestablished doses (25, 50 and 100 mg/kg/day by intragastric route) showed no changes in parameters or clinical signs attributed to toxicity. Furthermore, AEAMS also did not show mutagenic, genotoxic and cytotoxic potential in vivo. CONCLUSIONS: AEAMS did not show cytotoxic, genotoxic and mutagenic potential in vitro and in vivo. And although the AEAMS has an LD50 > 2000 mg/kg, and does not have physiological, biochemical, hematological, histopathological changes or clinical signs related to toxicity when administered in low concentrations and for a short period, in high concentrations and continued use caused toxicity and mortality in Wistar rats. In order to obtain complementary results, is recommended highly that further mid and long-term toxicological studies are investigated, and in no-rodent specie.

Vernicia fordii (Hemsl.) Airy Shaw extract stimulates insulin secretion in pancreatic ß-cells and improves insulin sensitivity in diabetic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Vernicia fordii (Hemsl.) Airy Shaw (V. fordii) is also known as the tung tree and its leaves and fruit are used as an oriental treatment for dyspepsia, edema, and skin diseases, which are known as diabetic complications. AIM OF THE STUDY: In this study, we aimed to investigate the methanolic extract (VF5) of the leaves of V. fordii as an insulin secretagogue and its probable mechanism and verify the effect in HFD-fed mice. MATERIALS AND METHODS: The insulin secretagogue activity of different doses of VF5 (0.1, 0.3 and 1.0 µg/ml) was assessed using in vitro insulin secretion assay and confirmed the anti-diabetic effect in mice fed HFD for 4 weeks with different doses of VF5 (10, 20 and 50 mg/kg oral) for another 6 weeks. Glbenclamide (30 mg/kg, oral) was used as positive control drug. The possible mechanisms were evaluated by using Gö6983 (10 µM), U73122 (10 µM) and nifedipine (10 µM). The major constituents of VF5 were analyzed by UPLC-QToF-MS and 1H and 13C NMR spectroscopy. RESULTS: UPLC-QToF-MS and NMR spectroscopy analysis indicated that one of the main active components of VF5 was tigliane-diterpene esters. VF5 functioned as an insulin secretagogue and enhanced mitochondria respiration and insulin homeostasis. We confirmed that VF5 preserved the ß-cell and reduced the ß-cell expansion which caused by metabolic stress under HFD. The antidiabetic role of VF5 in HFD fed mice was assessed by glucose tolerance test (GTT) and insulin tolerance test (ITT), fasting plasma insulin level, fasting blood glucose level, AKT signal in peripheral tissue in the absence of toxic effects. Mechanistically, insulinotropic effect of VF5 was mediated by activation of PKCα via intracellular Ca2+ influx and enhanced mitochondria function. CONCLUSION: VF5 exhibits potent insulin secretagogue function and improves insulin sensitivity and protection of pancreatic ß-cells from metabolic stress without toxicity. Taken together, our study suggests that VF5 could be potentially used for treating diabetes and metabolic diseases through improving ß-cell function.

Phytochemical Analysis and Antinociceptive Properties of Hydroalcoholic Extracts of Aleurites moluccanus Bark.

Aleurites moluccanus is used in folk medicine to treat many diseases including pain and inflammatory processes in general. Considering the potential of the leaf extract, evidenced in a previous study, the present study investigates the antinociceptive and anti-inflammatory properties of the hydroethanolic extract of A. moluccanus bark and isolated compounds in animal models of pain. The antinociceptive and anti-inflammatory activities of A. moluccanus bark were evaluated through hyperalgesia induced by carrageenan, PGE2, cytokines, bradykinin, epinephrine, Freund's complete adjuvant, and lipopolysaccharide. Five compounds were isolated from the dichloromethane bark extract: acetyl aleuritolic acid, atraric acid, spruceanol, (5ß,10α)-12-hydroxy-13-methoxy-8,11,13-podocarpatrien-3-one and sonderianol. To optimize the extraction conditions, ethanol 50, 70, and 90°GL were used as extracting solvent, in a 1 : 20 (w/v) drug : solvent ratio, under stirring at room temperature for 4 h. The extracts were named AMC50, AMC70, and AMC90, respectively. These extracts were administered to mice (250 mg/kg, p. o.) with reduced mechanical hyperalgesia activity in the carrageenan test. Of these, AMC90 showed the best results. Pure (5ß,10α)-12-hydroxy-13-methoxy-8,11,13-podocarpatrien-3-one showed a beneficial effect for up to 48 hours after the administration of carrageenan, while acetyl aleuritolic acid was effective only in the first hour. AMC90 was able to reverse the analgesia induced only by prostaglandin E2 and tumor necrosis factor. We also induced hyperalgesia using the lipopolysaccharide and Freund's complete adjuvant models, with positive results. These results support the antinociceptive and anti-inflammatory activity of A. moluccanus bark extract. The observed effects are partly due to the presence of acetyl aleuritolic acid, atraric acid, and (5ß,10α)-12-hydroxy-13-methoxy-8,11,13-podocarpatrien-3-one.

Deciphering the roles of leucine-rich repeat receptor-like protein kinases (LRR-RLKs) in response to Fusarium wilt in the Vernicia fordii (Tung tree).

Leucine-rich repeat receptor-like protein kinases (LRR-RLKs) are vital for plant growth and development, signal transduction, immunity, and play diverse roles in plant defense responses. However, the LRR-RLK genes have not been systematically studied in Vernicia fordii (tung tree), especially its response to Fusarium wilt. Here, we carried out an integrative analysis of LRR-RLKs among five Euphorbiaceae species: Hevea brasiliensis (rubber tree), Manihot esculenta (cassava), Jatropha curcas (physic nut), Ricinus communis (castor bean), and V. fordii, which contained 223, 311, 186, 138, and 167 LRR-RLKs, respectively. Maximum-likelihood tree was estimated using LRR-RLKs of Arabidopsis thaliana as a template, and they allowed us to divide Euphorbiaceae LRR-RLKs into 22 groups. There are 126 segmental and 30 tandem duplications in these Euphorbiaceae genomes by synteny analysis. The tissue-specific expression patterns revealed that V. fordii LRR-RLKs (VfLRR-RLKs) were differentially expressed in various tissues, and some of them exhibited specific expression in meristems tissues, which suggested their potential functions during organ formation and cell fate specification. Two VfLRR-RLK pairs (Vf01G2125 and Vf03G1740, Vf06G2687 and Vf10G1659), which generated by tandem duplication events, were associated with possible resistance to Fusarium wilt infection. The qRT-PCR confirmed these four VfLRR-RLKs contained opposite expression profiles during pathogen infection in V. fordii and V. montana. Taken together, our data systematically analyzed the LRR-RLK family in Euphorbiaceae genomes for the first time. We highlight the putative roles of VfLRR-RLKs in response to Fusarium wilt infection, and VfLRR-RLKs may be further applied in marker-assisted breeding to control Fusarium wilt in V. fordii.

Physicochemical Characterization of Candlenut (Aleurites moluccana)-derived Biodiesel Purified with Deed Eutectic Solvents.

A high-oil candlenut (Aleurites moluccana)-derived biodiesel was purified using new generation solvents called Deep eutectic solvents (DESs) and physicochemical properties of the purified biodiesel were assessed according to the accepted international standards for biodiesel (ASTM D6751; EN 14214). Binary and ternary choline chloride-based DESs containing glycerol and ethylene glycol as hydrogen bond donors were synthesized. The results of the investigation showed that the quality of DESs-purified biodiesel generally improved compared with the unpurified biodiesel and fulfilled the accepted standard limits for biodiesel except carbon residue and oxidation stability properties. Choline chloride/ethylene glycol (ChCl/Eg, 1:2 molar ratio) having a DES/biodiesel ratio of 1:1 (DES2-11) has enhanced the Fatty Acid Methyl Ester (FAME) content of unpurified biodiesel from 81.89% to 98.13%, and the oxidation stability from 2.36 h to 3.06 h. 1H NMR examination showed the successful synthesis of DESs; GC-MS analysis of candlenut oil and its biodiesel revealed oil and fatty acid methyl ester compositions, respectively. The similarity in FAME compositions between DESs-purified and unpurified biodiesel as revealed through GC-MS examination suggests the selective removal of impurities and the efficiency of DESs in improving the quality of candlenut-derived biodiesel.

Hidden in plain sight: Systematic investigation of Leucine-rich repeat containing genes unveil the their regulatory network in response to Fusarium wilt in tung tree.

Leucine-rich repeat-containing genes (LRRs) have been reported to play important roles in responses to diseases. However, we poorly understood the response of LRRs to Fusarium wilt infection in tung tree (Vernicia fordii), which is an important economic tree. Here, 437 LRR-containing proteins containing nine types of LRR domains were identified in V. fordii genome. Phylogenetic analysis suggested that nine types of LRR domains could not be divided into separate classes, implying that these LRR domains had a common origin. Totally, 27 LRRs were related to possible resistance to Fusarium wilt after 2, 8, and 13 days post-inoculation. We further found that Vf06G1605 was up-regulate under Fusarium wilt infection after these three time points, Vf10G1602 and Vf02G1413 were up-regulated at 8, and 13 dpi, while Vf07G2320 was down-regulated at these three time points. The WGCNA and promoter elements suggested that WRKY possibly regulate the responses of LRRs to Fusarium wilt infection. This study highlighted the phylogeny and function of LRRs in V. fordii and provided a systematic analysis of these genes in the V. fordii genome. Our results presented here might clearly illustrate physiological mechanisms of resistance to Fusarium wilt infection and the target of marker-assisted breeding in V. fordii.

Weight loss supplement causing acute heart block in a child.

A 16-year-old male was admitted to the paediatric ICU with acute onset of vomiting, somnolence, and chest pain, and electrocardiogram showing 2nd degree heart block after ingesting an Aleurites moluccana (Candlenut) seed as a herbal weight loss supplement. Electrocardiogram showed progressively worsening heart block with down-sloping of the ST segments, resembling digoxin toxicity. After 2 days of ICU observation, his symptoms began to improve and eventually resolved. The side effects of herbal supplements are often unknown but by analysing cases such as these, physicians can develop a better understanding of these substances to help guide management.

Aleurites moluccanus and its main active constituent, the flavonoid 2″-O-rhamnosylswertisin, in experimental model of rheumatoid arthritis.

ETHNOPHARMMACOLOGICAL RELEVANCE: Aleurites moluccana is used in folk medicine to treat pain, fever, asthma, hepatitis, gastric ulcer and inflammatory process in general, and the nut oil had been topically applied to treat arthritis and other joint pain, however the seeds are classified as toxic for oral use. AIM: Faced with the need for new alternative to treat the symptoms and modify rheumatoid arthritis (RA) the aim of this work was to evaluate the effects of A. moluccanus' leaves dried extract in rats and mice submitted to complete Freund adjuvant (CFA)-induced RA. MATERIAL AND METHODS: Wistar Rats and Swiss mice were submitted to CFA-induced RA in the right hindpaw. They received A. moluccanus extract (orally; p.o.), dexamethasone (subcutaneously), 2″-O-rhamnosylswertisin (p.o.) or vehicle (p.o.), from the 14th day after the CFA injection for up to 8 days. The mechanical hypersensitivity was evaluated using the von Frey filaments and the paw-oedema was measured using a plethysmometer. The rats' injected hindpaw was used to perform the histological analysis. RESULTS: A. moluccanus was able to significantly reduce the mechanical hypersensitivity in both ipsi- and contralateral hindpaws of mice injected with CFA, in a dose dependent manner. Furthermore, the paw-oedema was progressively reduced by A. moluccanus. Similar results were obtained for the positive-control drug dexamethasone and the isolated compound 2″-O-rhamnosylswertisin. Besides the effects mentioned above, the extract was also effective to repair the joint damage in CFA-induced RA rats, including reduction of fibrosis, cartilage degradation and bone erosion scores. CONCLUSION: These results together with the literature data reinforce the anti-hypersensitivity and anti-inflammatory activity of A. moluccanus extract. Part of the observed effects is due to the presence of the compound 2″-O-rhamnosylswertisin. The fact that the extract acted as a disease modifier point this herbal product as a promisor and safe tool to treat RA and other associated chronic diseases.

Tung Tree (Vernicia fordii) Genome Provides A Resource for Understanding Genome Evolution and Improved Oil Production.

Tung tree (Vernicia fordii) is an economically important woody oil plant that produces tung oil rich in eleostearic acid. Here, we report a high-quality chromosome-scale genome sequence of tung tree. The genome sequence was assembled by combining Illumina short reads, Pacific Biosciences single-molecule real-time long reads, and Hi-C sequencing data. The size of tung tree genome is 1.12 Gb, with 28,422 predicted genes and over 73% repeat sequences. The V. fordii underwent an ancient genome triplication event shared by core eudicots but no further whole-genome duplication in the subsequent ca. 34.55 million years of evolutionary history of the tung tree lineage. Insertion time analysis revealed that repeat-driven genome expansion might have arisen as a result of long-standing long terminal repeat retrotransposon bursts and lack of efficient DNA deletion mechanisms. The genome harbors 88 resistance genes encoding nucleotide-binding sites; 17 of these genes may be involved in early-infection stage of Fusarium wilt resistance. Further, 651 oil-related genes were identified, 88 of which are predicted to be directly involved in tung oil biosynthesis. Relatively few phosphoenolpyruvate carboxykinase genes, and synergistic effects between transcription factors and oil biosynthesis-related genes might contribute to the high oil content of tung seed. The tung tree genome constitutes a valuable resource for understanding genome evolution, as well as for molecular breeding and genetic improvements for oil production.

Characteristics and correlations of leaf stomata in different Aleurites montana provenances.

Stomata are important indexes in studies of plant origin, evolution, and classification and are important organs in plant phylogenetic relationship studies. Nine Aleurites montana provenances were used in this study to examine stomatal density, stomatal area, stomatal perimeter, long axis length, and short axis length. The correlation and cluster analyses were conducted among the morphological parameters of the pores. The results showed that there were significant differences in different A. Montana provenances in terms of stomatal morphology parameters. The average stomatal density, stomatal area, stomatal perimeter, stomatal long axis length, and stomatal short axis length of each provenance was between 224.16-307.10 stomata/mm2, 80.42-99.76 µm, 36.39-41.32 µm, 15.89-18.44 µm, and 6.53-7.46 µm, respectively, and the coefficient of variation was between 9.18%-20.15%, 17.57%-33.77%, 9.31%-18.79%, 9.71%-18.48%, and 10.26%-21.57%, respectively. Correlation analysis shows that there was a significant negative correlation between stomatal density and stomatal shape parameters (stomatal area, stomatal perimeter, stomatal long axis, stomatal short axis) and there was a significant positive correlation between stomatal parameters. There was no significant correlation between geographical environment factors and stomatal characteristics. There is a close relationship between stomatal morphology and stomatal conductance whereby dense small stomata can quickly adapt to changes in the environment; considering the characteristics A. montana stomata in terms of stomatal stability and ability to adapt to the environment, the Jianyang, Zhenghe, Fuding, Shaxian provenances were identified as being more suitable for planting at different sites. This study provides a theoretical basis for the genetic improvement and breeding of high quality A. montana provenances.

Identification of the major diacylglycerol acyltransferase mRNA in mouse adipocytes and macrophages.

BACKGROUND: Triacylglycerols (TAGs) are the major form of energy storage in eukaryotes. Diacylglycerol acyltransferases (DGATs) catalyze the final and rate-limiting step of TAG biosynthesis. Mammalian DGATs are classified into DGAT1 and DGAT2 subfamilies. It was unclear which DGAT was the major isoform expressed in animal cells. The objective was to identify the major DGAT mRNA expressed in cultured mouse adipocytes and macrophages and compared it to that expressed in tung tree seeds. METHODS: qPCR evaluated DGAT mRNA levels in mouse 3 T3-L1 adipocytes and RAW264.7 macrophages and tung tree seeds. RESULTS: TaqMan qPCR showed that DGAT2 mRNA levels were 10-30 fold higher than DGAT1 in adipocytes and macrophages, and DGAT mRNA levels in adipocytes were 50-100-fold higher than those in macrophages. In contrast, the anti-inflammatory tristetraprolin/zinc finger protein 36 (TTP/ZFP36) mRNA levels were 2-4-fold higher in macrophages than those in adipocytes and similar to DGAT1 in adipocytes but 100-fold higher than DGAT1 in macrophages. SYBR Green qPCR analyses confirmed TaqMan qPCR results. DGAT2 mRNA as the major DGAT mRNA in the mouse cells was similar to that in tung tree seeds where DGAT2 mRNA levels were 10-20-fold higher than DGAT1 or DGAT3. CONCLUSION: The results demonstrated that DGAT2 mRNA was the major form of DGAT mRNA expressed in mouse adipocytes and macrophages and tung tree seeds.

Molecular mechanism of the extended oil accumulation phase contributing to the high seed oil content for the genotype of tung tree (Vernicia fordii).

BACKGROUND: Oil from seeds of the tung tree (Vernicia fordii) has unique drying properties that are industrially important. We found that the extended oil accumulation period was related to the high seed oil content at maturity among tung tree population. In order to understand the molecular mechanism underlying the high oil content in tung tree seed, Tree H and L were adopted for the further investigation, with seed oil content of about 70 and 45%, respectively. We compared the transcriptomic changes of seed at various times during oil accumulation between the two trees. RESULTS: Transcriptomes analysis revealed that many genes involved in glycolysis, fatty acid synthesis, and tri-acyl glyceride assembly still kept high expression in the late period of seed oil accumulation for Tree H only. Many genes in fatty acid degradation pathway were largely up regulated in the late period of seed oil accumulation for Tree L only. Four transcription factors related to fatty acid biosynthesis had different expression pattern in the seed oil accumulation period for the two trees. WRI1 was down regulated and kept the low expression in the late period of seed oil accumulation for the two trees. PII, LEC1 and LEC1-LIKE extended the high expression in the late period of seed oil accumulation in Tree H only. CONCLUSIONS: The continued accumulation of oil in the late period of seed oil accumulation for Tree H was associated with relatively high expression of the relevant genes in glycolysis, fatty acid synthesis and tri-acyl glyceride assembly. PII, LEC1, and LEC1-LIKE rather than WRI1 should play an important role in the oil continual accumulation in the late period of seed oil accumulation in Tree H. This study provides novel insight into the variation in seed oil content and informs plant breeding strategies to maximize oil yield.

Stable carbon and nitrogen isotopes in kukui (Aleurites moluccanus) endocarp along rainfall and elevation gradients: Archaeological implications.

Stable carbon and nitrogen isotopes are often used to make inferences of past environments and social patterns. We analyze δ 13C and δ 15N values in contemporary kukui (Aleurites moluccanus) endocarp to examine the effects of site environment. Results from across environmental transects on Hawai'i Island show strong patterns for both stable isotopes. For δ 13C a robust linear relationship with elevation is exhibited, strengthened by the inclusion of rainfall. This relationship breaks down at a minimum threshold of annual rainfall, possible relating to physiological responses to drought. For δ 15N, the only significant relationship observed pertains to substrate age. The endocarp from kukui is one of the most readily identified plant remains in the Pacific archaeological records and is often targeted for radiocarbon dating. We discuss the potential implications of our results regarding ancient climate, inferred diets, and habitat composition.