Selection of GmSWEET39 for oil and protein improvement in soybean.

Soybean [Glycine max (L.) Merr.] was domesticated from wild soybean (G. soja Sieb. and Zucc.) and has been further improved as a dual-use seed crop to provide highly valuable oil and protein for food, feed, and industrial applications. However, the underlying genetic and molecular basis remains less understood. Having combined high-confidence bi-parental linkage mapping with high-resolution association analysis based on 631 whole sequenced genomes, we mapped major soybean protein and oil QTLs on chromosome15 to a sugar transporter gene (GmSWEET39). A two-nucleotide CC deletion truncating C-terminus of GmSWEET39 was strongly associated with high seed oil and low seed protein, suggesting its pleiotropic effect on protein and oil content. GmSWEET39 was predominantly expressed in parenchyma and integument of the seed coat, and likely regulates oil and protein accumulation by affecting sugar delivery from maternal seed coat to the filial embryo. We demonstrated that GmSWEET39 has a dual function for both oil and protein improvement and undergoes two different paths of artificial selection. A CC deletion (CC-) haplotype H1 has been intensively selected during domestication and extensively used in soybean improvement worldwide. H1 is fixed in North American soybean cultivars. The protein-favored (CC+) haplotype H3 still undergoes ongoing selection, reflecting its sustainable role for soybean protein improvement. The comprehensive knowledge on the molecular basis underlying the major QTL and GmSWEET39 haplotypes associated with soybean improvement would be valuable to design new strategies for soybean seed quality improvement using molecular breeding and biotechnological approaches.

Investigation of Monosodium Glutamate Alternatives for Content of Umami Substances and Their Enhancement Effects in Chicken Soup Compared to Monosodium Glutamate.

This research aimed to compare the effects of monosodium glutamate (MSG) and its alternatives on sensory characteristics of chicken soup. High-performance liquid chromatography analysis was carried out to quantify umami substances in potential MSG alternatives. Two mushroom extracts (CE and MC), one tomato extract (TC), and one yeast extract (YE) powders were selected due to their high equivalent umami concentration (EUC). These extracts together with MSG were then applied individually at four different levels (CE, MC, TC, MSG: 0.05%, 0.1%, 0.2%, 0.4%; YE: 0.0125%, 0.025%, 0.05%, 0.1%) in chicken soup in order to compare their impact on major sensory attributes using the degree of difference from control (DODC) test. Our results showed that all four extracts at all the usage levels exhibited an enhancement effect on the overall flavor, meaty flavor, saltiness, and umami taste. The extent of enhancement depended on the type of the alternative and its usage level. Higher levels of MSG alternatives (except YE) suppressed the chicken flavor. YE had similar enhancement effects as MSG on umami and salty tastes already at lower usage levels. At the lowest concentration, TC showed a stronger enhancement effect than MSG, but its effect on most attributes decreased as the usage dose increased. Compared to CE, the other mushroom extract MC resembled MSG at most levels. Overall, the closest synergistic effect in chicken soup was noted with 0.1% MSG, 0.1% MC, and 0.025% YE. PRACTICAL APPLICATION: This study compared the enhancement effects of MSG and selected alternatives in chicken soup. Results will help food manufacturers who would like to replace MSG with natural umami substances in soup products to enhance flavor and reduce sodium chloride.

The bracteatus pineapple genome and domestication of clonally propagated crops.

Domestication of clonally propagated crops such as pineapple from South America was hypothesized to be a 'one-step operation'. We sequenced the genome of Ananas comosus var. bracteatus CB5 and assembled 513 Mb into 25 chromosomes with 29,412 genes. Comparison of the genomes of CB5, F153 and MD2 elucidated the genomic basis of fiber production, color formation, sugar accumulation and fruit maturation. We also resequenced 89 Ananas genomes. Cultivars 'Smooth Cayenne' and 'Queen' exhibited ancient and recent admixture, while 'Singapore Spanish' supported a one-step operation of domestication. We identified 25 selective sweeps, including a strong sweep containing a pair of tandemly duplicated bromelain inhibitors. Four candidate genes for self-incompatibility were linked in F153, but were not functional in self-compatible CB5. Our findings support the coexistence of sexual recombination and a one-step operation in the domestication of clonally propagated crops. This work guides the exploration of sexual and asexual domestication trajectories in other clonally propagated crops.

Gene expression profiles that shape high and low oil content sesames.

BACKGROUND: Sesame (Sesamum indicum) can accumulate over 60% oil in its seed. However, low oil content genotypes with an oil content of less than 50% are also observed. To gain insights into how genes shape this variation, we examined 22 seed and carpel transcriptomes from 3 varieties of sesame with high and low oil content. RESULTS: A total of 34.6~52.2% of the sesame genes were expressed with a RPKM greater than 5 in the 22 tissue samples. The expressed gene numbers tended to decrease in the seed but fluctuated in the carpels from 10 to 30 days post-anthesis (DPA). Compared with that of the low oil content sesames, the high oil content sesame exhibited more positive gene expression during seed development. Typically, genes involved in lipid biosynthesis were enriched and could distinguish the high and low genotypes at 30 DPA, suggesting the pivotal role of seed oil biosynthesis in the later stages. Key homologous lipid genes that function in TAG biosynthesis, including those that encoded glycerol-3-phosphate acyltransferase (GPAT), acyl-CoA:diacylglycerol acyltransferase (DGAT), and phospholipid:diacylglycerol acyltransferase (PDAT), were strengthened asynchronously at different stages, but the lipid transfer protein (LTP)-encoding genes, including SIN_1019175, SIN_1019172 and SIN_1010009, usually were highlighted in the high oil content sesames. Furthermore, a list of 23 candidate genes was identified and predicted to be beneficial for higher oil content accumulation. Despite the different gene expression patterns between the seeds and carpels, the two tissues showed a cooperative relationship during seed development, and biological processes, such as transport, catabolic process and small molecule metabolic process, changed synchronously. CONCLUSIONS: The study elucidated the different expression profiles in high and low oil content sesames and revealed key stages and a list of candidate genes that shaped oil content variation. These findings will accelerate dissection of the genetic mechanism of sesame oil biosynthesis.

Population Pharmacokinetics and Pharmacodynamics of Piperacillin/Tazobactam in Patients with Nosocomial Infections.

OBJECTIVE: The study was to establish a population pharmacokinetic (PPK) model of piperacillin (PIP) and tazobactam (TAZ) that explain pharmacokinetic variability and to propose optimized dosage regimens in patients with nosocomial infections. METHODS: In total, 310 PIP and 280 TAZ concentration-time points were collected at steady state over multiple dosing intervals from 50 patients who received PIP/TAZ infused within 30 min or over 3 h. Drug analysis was performed by high-performance liquid chromatography (HPLC). Nonlinear mixed effects modeling was employed to develop PPK model and 1000 Monte Carlo simulation was used to predict the probability of target attainment (PTA) with a target time of non-protein-bound concentration above MIC > 50 % of the dosing interval. RESULTS: A model with one-compartment model had the best predictive performance for the PPK model. The population estimates of PIP were 13.8 L/h (31.1 %) for clearance (CL) and 21.7 L (38 %) for volume of distribution (V). The population estimates of TAZ were 9.3 L/h (29.1 %) for CL and 16 L (35.3 %) for V. Influence of creatinine clearance (CLcr) and body weight were identified as important covariates for PIP/TAZ CL and V, respectively. A 30-min infusion of 4 g every 6 h achieved robust (≥90 %) PTAs for MIC ≤ 16 mg/L. As an alternative mode of administration, a 3-h infusion of 4 g every 6 h achieved robust PTAs for Pseudomonas aeruginosa and Klebsiella pneumoniae. CONCLUSIONS: Prolonged infusions achieved better PTAs compared with shorter infusions at similar daily doses. This benefit was most pronounced for MICs between 16 and 40 mg/L.

Identification of QTLs associated with oil content and mapping FAD2 genes and their relative contribution to oil quality in peanut (Arachis hypogaea L.).

BACKGROUND: Peanut is one of the major source for human consumption worldwide and its seed contain approximately 50% oil. Improvement of oil content and quality traits (high oleic and low linoleic acid) in peanut could be accelerated by exploiting linked markers through molecular breeding. The objective of this study was to identify QTLs associated with oil content, and estimate relative contribution of FAD2 genes (ahFAD2A and ahFAD2B) to oil quality traits in two recombinant inbred line (RIL) populations. RESULTS: Improved genetic linkage maps were developed for S-population (SunOleic 97R × NC94022) with 206 (1780.6 cM) and T-population (Tifrunner × GT-C20) with 378 (2487.4 cM) marker loci. A total of 6 and 9 QTLs controlling oil content were identified in the S- and T-population, respectively. The contribution of each QTL towards oil content variation ranged from 3.07 to 10.23% in the S-population and from 3.93 to 14.07% in the T-population. The mapping positions for ahFAD2A (A sub-genome) and ahFAD2B (B sub-genome) genes were assigned on a09 and b09 linkage groups. The ahFAD2B gene (26.54%, 25.59% and 41.02% PVE) had higher phenotypic effect on oleic acid (C18:1), linoleic acid (C18:2), and oleic/linoleic acid ratio (O/L ratio) than ahFAD2A gene (8.08%, 6.86% and 3.78% PVE). The FAD2 genes had no effect on oil content. This study identified a total of 78 main-effect QTLs (M-QTLs) with up to 42.33% phenotypic variation (PVE) and 10 epistatic QTLs (E-QTLs) up to 3.31% PVE for oil content and quality traits. CONCLUSIONS: A total of 78 main-effect QTLs (M-QTLs) and 10 E-QTLs have been detected for oil content and oil quality traits. One major QTL (more than 10% PVE) was identified in both the populations for oil content with source alleles from NC94022 and GT-C20 parental genotypes. FAD2 genes showed high effect for oleic acid (C18:1), linoleic acid (C18:2), and O/L ratio while no effect on total oil content. The information on phenotypic effect of FAD2 genes for oleic acid, linoleic acid and O/L ratio, and oil content will be applied in breeding selection.

Flavonol content, oil%, and fatty acid composition variability in seeds of Teramnus labialis and T. uncinatus accessions with nutraceutical potential.

Teramnus labialis and T. uncinatus are both underutilized legume species. Teramnus labialis is used as food in India while T. uncinatus has potential use in pasture mixes. Photoperiod-sensitive Teramnus accessions were grown in the greenhouse from 2010 to 2011 and evaluated for flavonol content, oil%, and fatty acid compositions. Significant variations for seed numbers produced, flavonol content, oil%, and fatty acid compositions were detected. Seed numbers ranged from 16 to 3,792 in both species. Teramnus accessions produced more quercetin (ranging from 0.615 to 2.228 mg/g) in their seeds than the other flavonols. However kaempferol and isorhamnetin content ranged from 0 to 0.066 and 0 to 0.086 mg/g (dry seed weight basis), respectively among all accessions. Oil% ranged from 2.65 to 5.64% and more oleic, linoleic, and linolenic acids ranging from 6.69 to 25.97, 31.82 to 41.44, and 17.7 to 32.66%, respectively, were produced among all Teramnus accessions. The seeds from all Teramnus accessions also produced the least saturated fatty acid compositions (ranging from 0.08 to 15.36%). Several significant correlations were also detected for these traits among the accessions. Quercetin showed highly significant positive correlations with kaempferol (r = 0.59, p < .0001), oil% (r = 0.58, p < .0001), and oleic acid (r = 0.31, p < .001). Quercetin also showed a significant negative correlation with linoleic acid (r = -0.49, p < .0001). These correlations are important because useful breeding procedures could be conducted on improving flavonol, oil%, and fatty acid compositions in Teramnus labialis and T. uncinatus accessions.

Identification and characterization of transcript polymorphisms in soybean lines varying in oil composition and content.

BACKGROUND: Variation in seed oil composition and content among soybean varieties is largely attributed to differences in transcript sequences and/or transcript accumulation of oil production related genes in seeds. Discovery and analysis of sequence and expression variations in these genes will accelerate soybean oil quality improvement. RESULTS: In an effort to identify these variations, we sequenced the transcriptomes of soybean seeds from nine lines varying in oil composition and/or total oil content. Our results showed that 69,338 distinct transcripts from 32,885 annotated genes were expressed in seeds. A total of 8,037 transcript expression polymorphisms and 50,485 transcript sequence polymorphisms (48,792 SNPs and 1,693 small Indels) were identified among the lines. Effects of the transcript polymorphisms on their encoded protein sequences and functions were predicted. The studies also provided independent evidence that the lack of FAD2-1A gene activity and a non-synonymous SNP in the coding sequence of FAB2C caused elevated oleic acid and stearic acid levels in soybean lines M23 and FAM94-41, respectively. CONCLUSIONS: As a proof-of-concept, we developed an integrated RNA-seq and bioinformatics approach to identify and functionally annotate transcript polymorphisms, and demonstrated its high effectiveness for discovery of genetic and transcript variations that result in altered oil quality traits. The collection of transcript polymorphisms coupled with their predicted functional effects will be a valuable asset for further discovery of genes, gene variants, and functional markers to improve soybean oil quality.

Oil, fatty acid, flavonoid, and resveratrol content variability and FAD2A functional SNP genotypes in the U.S. peanut mini-core collection.

Peanut seeds contain high amounts of oil and protein as well as some useful bioactive phytochemicals which can contribute to human health. The U.S. peanut mini-core collection is an important genetic resource for improving seed quality and developing new cultivars. Variability of seed chemical composition within the mini-core was evaluated from freshly harvested seeds for two years. Oil, fatty acid composition, and flavonoid/resveratrol content were quantified by NMR, GC, and HPLC, respectively. Significant variability was detected in seed chemical composition among accessions and botanical varieties. Accessions were further genotyped with a functional SNP marker from the FAD2A gene using real-time PCR and classified into three genotypes with significantly different O/L ratios: wild type (G/G with a low O/L ratio <1.7), heterozygote (G/A with O/L ratio >1.4 but <1.7), and mutant (A/A with a high O/L ratio >1.7). The results from real-time PCR genotyping and GC fatty acid analysis were consistent. Accessions with high amounts of oil, quercetin, high seed weight, and O/L ratio were identified. The results from this study may be useful not only for peanut breeders, food processors, and product consumers to select suitable accessions or cultivars but also for curators to potentially expand the mini-core collection.

Assessment of oil content and fatty acid composition variability in two economically important Hibiscus species.

The Hibiscus genus encompasses more than 300 species, but kenaf (Hibiscus cannabinus L.) and roselle (Hibiscus sabdariffa L.) are the two most economically important species within the genus. Seeds from these two Hibiscus species contain a relatively high amount of oil with two unusual fatty acids: dihydrosterculic and vernolic acids. The fatty acid composition in the oil can directly affect oil quality and its utilization. However, the variability in oil content and fatty acid composition for these two species is unclear. For these two species, 329 available accessions were acquired from the USDA germplasm collection. Their oil content and fatty acid composition were determined by nuclear magnetic resonance (NMR) and gas chromatography (GC), respectively. Using NMR and GC analyses, we found that Hibiscus seeds on average contained 18% oil and seed oil was composed of six major fatty acids (each >1%) and seven minor fatty acids (each <1%). Hibiscus cannabinus seeds contained significantly higher amounts of oil (18.14%), palmitic (20.75%), oleic (28.91%), vernolic acids (VA, 4.16%), and significantly lower amounts of stearic (3.96%), linoleic (39.49%), and dihydrosterculic acids (DHSA, 1.08%) than H. sabdariffa seeds (17.35%, 18.52%, 25.16%, 3.52%, 4.31%, 44.72%, and 1.57%, respectively). For edible oils, a higher oleic/linoleic (O/L) ratio and lower level of DHSA are preferred, and for industrial oils a high level of VA is preferred. Our results indicate that seeds from H. cannabinus may be of higher quality than H. sabdariffa seeds for these reasons. Significant variability in oil content and major fatty acids was also detected within both species. The variability in oil content and fatty acid composition revealed from this study will be useful for exploring seed utilization and developing new cultivars in these Hibiscus species.

Screening of the entire USDA castor germplasm collection for oil content and fatty acid composition for optimum biodiesel production.

Castor has tremendous potential as a feedstock for biodiesel production. The oil content and fatty acid composition in castor seed are important factors determining the price for production and affecting the key fuel properties of biodiesel. There are 1033 available castor accessions collected or donated from 48 countries worldwide in the USDA germplasm collection. The entire castor collection was screened for oil content and fatty acid composition by nuclear magnetic resonance (NMR) and gas chromatography (GC), respectively. Castor seeds on the average contain 48.2% oil with significant variability ranging from 37.2 to 60.6%. Methyl esters were prepared from castor seed by alkaline transmethylation. GC analysis of methyl esters confirmed that castor oil was composed primarily of eight fatty acids: 1.48% palmitic (C16:0), 1.58% stearic (C18:0), 4.41% oleic (C18:1), 6.42% linoleic (C18:2), 0.68% linolenic (C18:3), 0.45% gadoleic (C20:1), 84.51% ricinoleic (C18:1-1OH), and 0.47% dihydroxystearic (C18:0-2OH) acids. Significant variability in fatty acid composition was detected among castor accessions. Ricinoleic acid (RA) was positively correlated with dihydroxystearic acid (DHSA) but highly negatively correlated with the five other fatty acids except linolenic acid. The results for oil content and fatty acid composition obtained from this study will be useful for end-users to explore castor germplasm for biodiesel production.

Seed oil and fatty acid content in okra (Abelmoschus esculentus) and related species.

Approximately 1100 genebank accessions of okra (Abelmoschus esculentus) and 540 additional accessions that included six of its related species-A. caillei, A. crinitis, A. esculentus, A. ficulneus, A. manihot, A. moschatus and A. tuberculatus-were evaluated for seed oil content using time domain NMR (TD-NMR). Oil content in seed of A. caillei, A. esculentus, A. ficulneus, A. manihot, A. moschatus and A. tuberculatus was in the ranges 2.51-13.61%, 12.36-21.56%, 6.62-16.7%, 16.1-22.0%, 10.3-19.8% and 10.8-23.2%, respectively. Accession PI639680 (A. tuberculatus) had the highest seed oil content (∼23%). Accessions of A. esculentus with high seed oil content included PI nos. PI274350 (21.5%), PI538082 (20.9%) and PI538097 (20.9%). Values for the three accessions of A. manihot with the highest seed oil content were PI nos. PI639673 (20.4%), PI639674 (20.9%) and PI639675 (21.9%), all representing var. tetraphyllus. Average percent seed oil in materials of A. esculentus from Turkey and Sudan (17.35% and 17.36%, respectively) exceeded the averages of materials from other locations. Ninety-eight accessions (total of six species) were also examined for fatty acid composition. Values of linoleic acid ranged from 23.6-50.65% in A. esculentus. However, mean linoleic acid concentrations were highest in A. tuberculatus and A. ficulneus. Concentrations of palmitic acid were significantly higher in A. esculentus (range of 10.3-36.35%) when compared to that of other species, and reached a maximum in PI489800 Concentrations of palmitic acid were also high in A. caillei (mean = ∼30%). Levels of oleic acid were highest in A. manihot, A. manihot var. tetraphyllus and A. moschatus.

[Effect of intense pulsed light on transforming growth factor-beta1 mRNA expression in rat skin].

OBJECTIVE: To observe the effect of intense pulsed light (IPL) on transforming growth factor-beta1 mRNA (TGF-beta1 mRNA) expression in rat skin and explore the molecular mechanisms of photorejuvenation. METHODS: Fifteen SD rats were exposed to IPL in 3 dermal regions with triple pulses (duration of 4, 5, and 6 ms) at the energy density of 34 J/cm2 and pulse delay of 20 or 25 ms. On days 1, 3, 5, 7, 15, and 30 after the treatment, skin specimens from the treated and non-treated areas were obtained to detect TGF-beta1 mRNA expression with in situ hybridization. RESULTS: In the UPL-exposed skin areas, TGF-beta1 mRNA expression was detected in the epidermal keratinocytes and dermal cells 1 day after the exposure, reaching the highest expression level on day 7 followed by gradual decrement since day 15, and till day 30, only weak expression was found in the dermal cells. In the non-exposed regions, the cells remained negative for TGF-beta1 mRNA. CONCLUSION: IPL can enhance TGF-beta1 mRNA expression in the skin, suggesting that TGF-beta1 plays an important role in dermal remodeling in photorejuvenation.

[Effect of intense pulsed light on heat shock protein 70 expression in skin].

OBJECTIVE: To observe the effect of intense pulsed light (IPL) on heat shock protein 70 (HSP70) expression in skin and elucidate the biological mechanisms of photorejuvenation. METHODS: The skin of 15 SD rats was exposed to IPL in 3 regions at the energy density of 34 J/cm(2) in triple pulses with the duration of 4, 5 and 6 ms, respectively, and pulse delays of 20 and 25 ms. On days 1, 3, 5, 7, 15 and 30 after treatment, specimens of the treated and untreated skin were taken, respectively, for determination of HSP70 expression by immunohistochemistry. RESULTS: In treated regions, positive immunohistochemical staining was observed on day 1 in the epidermal keratinocytes, sebaceous gland cells and endothelial cells. The staining reached the highest intensity on day 7, gradually weakened on day 15, and disappeared on day 30. In the untreated areas, the cells were negative for immunohistochemical staining. CONCLUSION: Skin HSP70 expression can be enhanced by IPL, suggesting the role of HSP70 in photorejuvenation.