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Water stress and sulfur (S) deficiency are two constraints increasingly faced by crops due to climate change and low-input agricultural practices. To investigate their interaction in the grain legume pea (Pisum sativum), sulfate was depleted at the mid-vegetative stage and a moderate 9-d water stress period was imposed during the early reproductive phase. The combination of the stresses impeded reproductive processes in a synergistic manner, reducing seed weight and seed number, and inducing seed abortion, which highlighted the paramount importance of sulfur for maintaining seed yield components under water stress. On the other hand, the moderate water stress mitigated the negative effect of sulfur deficiency on the accumulation of S-rich globulins (11S) in seeds, probably due to a lower seed sink strength for nitrogen, enabling a readjustment of the ratio of S-poor (7S) to 11S globulins. Transcriptome analysis of developing seeds at the end of the combined stress period indicated that similar biological processes were regulated in response to sulfur deficiency and to the combined stress, but that the extent of the transcriptional regulation was greater under sulfur deficiency. Seeds from plants subjected to the combined stresses showed a specific up-regulation of a set of transcription factor and SUMO ligase genes, indicating the establishment of unique regulatory processes when sulfur deficiency is combined with water stress. [ABSTRACT FROM AUTHOR]

Increasing seed oil content is one of the most important breeding goals for soybean due to a high global demand for edible vegetable oil. However, genetic improvement of seed oil content has been difficult in soybean because of the complexity of oil metabolism. Determining the major variants and molecular mechanisms conferring oil accumulation is critical for substantial oil enhancement in soybean and other oilseed crops. In this study, we evaluated the seed oil contents of 219 diverse soybean accessions across six different environments and dissected the underlying mechanism using a high-resolution genome-wide association study (GWAS). An environmentally stable quantitative trait locus (QTL), GqOil20, significantly associated with oil content was identified, accounting for 23.70% of the total phenotypic variance of seed oil across multiple environments. Haplotype and expression analyses indicate that an oleosin protein-encoding gene (GmOLEO1), colocated with a leading single nucleotide polymorphism (SNP) from the GWAS, was significantly correlated with seed oil content. GmOLEO1 is predominantly expressed during seed maturation, and GmOLEO1 is localized to accumulated oil bodies (OBs) in maturing seeds. Overexpression of GmOLEO1 significantly enriched smaller OBs and increased seed oil content by 10.6% compared with those of control seeds. A time-course transcriptomics analysis between transgenic and control soybeans indicated that GmOLEO1 positively enhanced oil accumulation by affecting triacylglycerol metabolism. Our results also showed that strong artificial selection had occurred in the promoter region of GmOLEO1, which resulted in its high expression in cultivated soybean relative to wild soybean, leading to increased seed oil accumulation. The GmOLEO1 locus may serve as a direct target for both genetic engineering and selection for soybean oil improvement. [ABSTRACT FROM AUTHOR]

The R2R3-type MYB transcription factors have been shown to increase flavonoids accumulation by regulating the expression of key enzyme genes related to flavonoid biosynthesis pathway. However, the roles and underlying mechanisms of the soybean GmMYB12 gene in regulation of flavonoids accumulation and tolerance to abiotic stresses are rarely known. In the present study, the GmMYB12 gene was isolated and its function was characterized. Sequence and yeast one-hybrid analyses showed that GmMYB12 contained two MYB domains and belonged to R2R3-MYB protein with transactivation activity. Subcellular localization analysis in onion epidermal cells indicated that GmMYB12 was localized to the nucleus. Overexpression of GmMYB12 increased the production of downstream flavonoids and the expression of related genes in the flavonoid biosynthesis pathway. It also improved resistance to salt and drought stresses during seed germination, root development, and growing stage. Further component and enzymatic analyses showed significant increases of proline content, pyrroline-5-carboxylate synthase (P5CS), superoxide dismutase (SOD), and peroxidase (POD) activities, as well as significant reduction of H2O2 and malonaldehyde (MDA) content under salt and drought stresses in transgenic plants. Meanwhile, the expression level of AtP5CS, AtSOD and AtPOD genes was up-regulated against salt and drought stresses. Together, our finding indicated that changing the expression level of GmMYB12 in plants alters the accumulation of flavonoids and regulates plantlet tolerance to abiotic stress by regulating osmotic balance, protecting membrane integrity and maintaining ROS homeostasis. The GmMYB12 gene has the potential to be used to increase the content of valuable flavonoids and improve the tolerance to abiotic stresses in plants. [ABSTRACT FROM AUTHOR]

Glyphosate is the most extensively used herbicide in the world. However, concerns regarding its safety, side effects, and impact on other organisms have increased in recent years. This is the first study to analyze a large set of recent and historical Escherichia coli isolates varying in pathogenicity and beta-lactam resistance from different host species for their susceptibility to glyphosate isopropylamine salt (IPA), the active ingredient of the herbicide, and to a complete glyphosate-containing formulation (Roundup LB Plus). For this, minimum inhibitory concentrations (MIC) were determined for 238 E. coli isolates by broth microdilution in Mueller Hinton I media followed by the statistical analyses using Mann-Whitney-U test, multivariable analysis of variance (ANOVA) and a multivariable proportional-odds ordinal regression model. While the overall MIC distribution was narrow and lacked a highly resistant sub-population for both substances, statistical analyses revealed small but significant associations between glyphosate resistance levels and different factors tested. Mean MIC values for the entire dataset showed a higher level of resistance to the complete glyphosate-containing formulation (40 mg/ml IPA) than to pure glyphosate (10 mg/ml IPA) in E. coli. Isolates that originated from poultry had significantly higher MIC values for both pure glyphosate and the complete formulation. Pathogenic and non-extended-spectrum beta-lactamase (non-ESBL) E. coli isolates each showed significantly higher MIC values compared to commensals and ESBL-producing E. coli in pure glyphosate, but not in the complete formulation. Recently sampled isolates showed statistically higher MICs than the isolates of the historic standard E. coli collection of reference in pure glyphosate, when tested by nonparametric Mann-Whitney-U test, but not in the multivariable model. Further investigations are necessary to confirm whether these associations have a casual relationship with the glyphosate use or are the consequence of co-selection due to the increased application rates of antibiotics, heavy metals or other biocides. A possible accumulation of pathogenic bacteria in livestock animals fed with glyphosate-containing feed should also be considered. [ABSTRACT FROM AUTHOR]

Summary: Emerging genome editing technologies hold great promise for the improvement of agricultural crops. Several related genome editing methods currently in development utilize engineered, sequence‐specific endonucleases to generate DNA double strand breaks (DSBs) at user‐specified genomic loci. These DSBs subsequently result in small insertions/deletions (indels), base substitutions or incorporation of exogenous donor sequences at the target site, depending on the application. Targeted mutagenesis in soybean (Glycine max) via non‐homologous end joining (NHEJ)‐mediated repair of such DSBs has been previously demonstrated with multiple nucleases, as has homology‐directed repair (HDR)‐mediated integration of a single transgene into target endogenous soybean loci using CRISPR/Cas9. Here we report targeted integration of multiple transgenes into a single soybean locus using a zinc finger nuclease (ZFN). First, we demonstrate targeted integration of biolistically delivered DNA via either HDR or NHEJ to the FATTY ACID DESATURASE 2‐1a (FAD2‐1a) locus of embryogenic cells in tissue culture. We then describe ZFN‐ and NHEJ‐mediated, targeted integration of two different multigene donors to the FAD2‐1a locus of immature embryos. The largest donor delivered was 16.2 kb, carried four transgenes, and was successfully transmitted to T1 progeny of mature targeted plants obtained via somatic embryogenesis. The insertions in most plants with a targeted, 7.1 kb, NHEJ‐integrated donor were perfect or near‐perfect, demonstrating that NHEJ is a viable alternative to HDR for gene targeting in soybean. Taken together, these results show that ZFNs can be used to generate fertile transgenic soybean plants with NHEJ‐mediated targeted insertions of multigene donors at an endogenous genomic locus. [ABSTRACT FROM AUTHOR]

Vegetable soybean (VS), known as edamame, is appreciated for its robust flavor and exceptional nutrient content. Although the United States is one of the world's leading producers of soy, only a fraction is harvested as edamame. Research involving varietal differences is very limited, particularly among U.S. consumers. Sensory attributes of three VS varieties grown in Northern California (with organic methods) were assessed: Giant Midori (GM), ButterBean (BB), and Kuroshinja (KU). Participants (n = 74) rated four characteristics: flavor, texture, appearance, and overall liking on unlabeled, 11‐point hedonic scales and were asked if they would purchase the variety. A small portion of these participants would again evaluate the varieties, using free choice profiling (FCP) methodology. After these evaluations, texture analyzer, colorimeter, and high‐performance liquid chromatography (free amino acids, sugars, and isoflavones) testing were performed. Among tested varieties, distinct differences in quality were found, with GM preferred first, followed by KU, and BB. GM was significantly harder, a more intense green, had more free sugars and isoflavones and fewer free amino acids, whereas KU and BB trended to be the opposite. Analyzing FCP data with a General Procrustes Analysis, varieties were separated in different factors for flavor and texture, but not appearance. Although repeated, and larger trials are needed, our study signifies that consumers are able to detect differences among VS varieties. Further research may influence producers to grow the variety likely to be most profitable and marketable. [ABSTRACT FROM AUTHOR]

Soybean (Glycine max) is the most important dicot crop worldwide, and is increasingly used as a model legume due to the wide availability of genomic soybean resources; however, the slow generation times of soybean plants are currently a major hindrance to research. Here, we demonstrate a method for accelerating soybean breeding in compact growth chambers, which greatly shortens the generation time of the plants and accelerates breeding and research projects. Our breeding method utilizes commonly used fluorescent lamps (220 µmol m–2 s–1 at the canopy level), a 14 h light (30°C)/10 h dark (25°C) cycle and carbon dioxide (CO2) supplementation at >400 p.p.m. Using this approach, the generation time of the best-characterized elite Japanese soybean cultivar, Enrei, was shortened from 102–132 d reported in the field to just 70 d, thereby allowing up to 5 generations per year instead of the 1–2 generations currently possible in the field and/or greenhouse. The method also facilitates the highly efficient and controlled crossing of soybean plants. Our method uses CO2 supplementation to promote the growth and yield of plants, appropriate light and temperature conditions to reduce the days to flowering, and the reaping and sowing of immature seeds to shorten the reproductive period greatly. Thus, the appropriate parameters enable acceleration of soybean breeding in the compact growth chambers commonly used for laboratory research. The parameters used in our method could therefore be optimized for other species, cultivars, accessions and experimental designs to facilitate rapid breeding in a wide range of crops. [ABSTRACT FROM AUTHOR]

In the present study, Agrobacterium tumefaciens-mediated transformation of Glycine max (L.) Merr. (soybean) cv. DS-9712 using half-seed explants was optimized for eight different parameters, including seed imbibition, medium pH, infection mode (sonication and vacuum infiltration), co-cultivation conditions, concentrations of supplementary compounds, and selection. Using this improved protocol, maximum transformation of 14% and regeneration efficiencies of 45% were achieved by using explants prepared from mature seeds imbibed for 36 h, infected with A. tumefaciens strain EHA105 at an optical density (OD600) of 0.8, suspended in pH 5.4 medium containing 0.2 mM acetosyringone and 450 mg L−1 L-cysteine, followed by sonication for 10 s, vacuum infiltration for 2 min, and co-cultivated for 3 d on 35 mg L−1 kanamycin-containing medium. Independent transgenic lines were confirmed to be transgenic after ß-glucuronidase histochemical assays, polymerase chain reaction, and southern hybridization analysis. The protocol developed in the present study showed high regeneration efficiency within a relatively short time of 76 d. This rapid and efficient protocol might overcome some hurdles associated with the genetic manipulation of soybean. [ABSTRACT FROM AUTHOR]

To understand translational capacity on a genome-wide scale across three developmental stages of immature soybean seed cotyledons, ribosome profiling was performed in combination with RNA sequencing and cluster analysis. Transcripts representing 216 unique genes demonstrated a higher level of translational activity in at least one stage by exhibiting higher translational efficiencies (TEs) in which there were relatively more ribosome footprint sequence reads mapping to the transcript than were present in the control total RNA sample. The majority of these transcripts were more translationally active at the early stage of seed development and included 12 unique serine or cysteine proteases and 16 2S albumin and low molecular weight cysteine-rich proteins that may serve as substrates for turnover and mobilization early in seed development. It would appear that the serine proteases and 2S albumins play a vital role in the early stages. In contrast, our investigation of profiles of 19 genes encoding high abundance seed storage proteins, such as glycinins, beta-conglycinins, lectin, and Kunitz trypsin inhibitors, showed that they all had similar patterns in which the TE values started at low levels and increased approximately 2 to 6-fold during development. The highest levels of these seed protein transcripts were found at the mid-developmental stage, whereas the highest ribosome footprint levels of only up to 1.6 TE were found at the late developmental stage. These experimental findings suggest that the major seed storage protein coding genes are primarily regulated at the transcriptional level during normal soybean cotyledon development. Finally, our analyses also identified a total of 370 unique gene models that showed very low TE values including over 48 genes encoding ribosomal family proteins and 95 gene models that are related to energy and photosynthetic functions, many of which have homology to the chloroplast genome. Additionally, we showed that genes of the chloroplast were relatively translationally inactive during seed development.

The effect of exogenous spermine on chilling injury (CI) and antioxidant defense system of immature vegetable soybean (Glycine max L.) during cold storage were investigated. Freshly harvested immature soybeans were treated with 0.8 mmol/L spermine at room temperature for 20 min and then stored at 5 ± 1 °C or 1 ± 1 °C and 85-95% relative humidity for up to 60 days. Results showed that exogenous spermine alleviated the CI, delayed the gradual decreasing activities of superoxide dismutase (SOD) and catalase, and maintained a favourable balance in reactive oxygen species levels at storage period. Although cold temperature (1 ± 1 °C) inhibited the synthesis of l-(malonylamino)-cyclopropane-l-carboxylic acid (MACC), raised ethylene production, and resulted in membrane damage, exogenous spermine obviously hindered the accumulation of 1-aminocyclopropane-1-carboxylic acid (ACC). It was concluded that exogenous spermine alleviated CI of cold-stored immature soybeans through regulating the antioxidant system and ACC metabolism. [ABSTRACT FROM AUTHOR]

An improved direct transformation method via particle bombardment of split-immature zygotic embryo explants with intact embryonic axis is reported. This method involves abiotic stress (cold treatment and plasmolysis) treatments of explants prepared from immature embryos of 8-10 mm size for improved somatic embryogeneis. Transgenic events were produced using optimized bombardment conditions and selection with hygromycin or glufosinate. Transgenic somatic embryos developed within as little as 4 weeks after bombardment of explants. Transgenic plants were regenerated 4-5 months after bombardment and the entire process from bombardment to T1 seed production took 7-9 months. Plants regenerated from the system were fertile and showed more than 90% heritability of the transgene to the next generation. Transformation frequencies of 5.4 and 2.7% (based on the number of bombarded split-immature embryo explants) were observed with hygromycin and glufosinate selection, respectively. [ABSTRACT FROM AUTHOR]

Triticale (× Triticosecale Wittmack) is a man-made species developed by crossing wheat (Triticum spp.) and rye (Secale cereale L.). It incorporates favorable alleles from both progenitor species (wheat and rye), enabling adaptation to environments that are less favorable for wheat yet providing better biomass yield and forage quality. Triticale has huge potential for both grain and forage production, though research to improve the crop for better adaptation and grain quality is lagging behind that of other small grains. It is also gaining popularity as a cover crop to improve soil health and reduce nutrient leaching. Because of its genetic and flower structure, triticale is suitable for both line and hybrid breeding methods. Advances in the areas of molecular biology and the wealth of genomic resources from both wheat and rye can be exploited for triticale improvement. Gene mapping and genomic selection will facilitate triticale breeding by increasing selection precision and reducing time and cost. The objectives of this review are to summarize current triticale production status, breeding, and genetics research achievements and to highlight gaps for future research. [ABSTRACT FROM AUTHOR]

Due to recalcitrant nature of chickpea (Cicer arietinum L.) to androgenesis, the production of double haploid plants has been only reported by Grewal et al. (Plant Cell Rep 28:1289-1299, 2009) using some physical stresses such as anther centrifugation and electrical shock. In the present study, we successfully obtained haploid plants from cultured anthers of two chickpea cultivars, Bivanij and Arman, using high 2,4-D and silver nitrate containing media without applying of these time and labor consuming stresses. For induction of androgenesis, different concentrations of 2, 4-D (0, 2, 5 and 10 mg/l) and silver nitrate (0, 5, 10, 15, 25 and 50 mg/l) were used in embryo development medium. In Bivanij cultivar, anther induction medium containing 10 mg/l 2,4-D and 15 mg/l silver nitrate produced the highest number of embryos (0.188) and regenerated plants (0.1) per each cultured anther, while the highest frequencies of embryos (0.1) and regenerated plants (0.075 and 0.063) were obtained from Arman cultivar when 10 mg/l 2,4-D was combined with 15 and 50 mg/l silver nitrate in anther culture medium, respectively. In second part of this study, different cold (4 °C for 4 and 7 days) and heat (30 °C for 10 days, 32 °C for 2 days and 35 °C for 8 h) pretreatments were applied on cultured anthers of Bivanij cultivar. Incubation of cultured anthers at 32 °C for 2 days significantly enhanced the rate of embryo formation up to 0.222 embryos per each anther, while the highest number of regenerated plants/anther (0.0332) was obtained when cold treated anthers at 4 °C for 7 days incubated at 30 °C for 10 days. Taken together, these results provide a good basis for large-scale generation of DH plants in this important legume species. [ABSTRACT FROM AUTHOR]

The relative availabilities of nitrogen and sulphur can modulate seed storage protein composition in grain legumes and cereals. Soybean contains two major seed storage proteins, glycinins and β-conglycinins that account for approximately 70% of total protein and their composition is affected by nitrogen and sulphur supplies. The present study demonstrated the effect of sulphur (Gypsum @ 20 kg S ha-1) and recommended a dose of nitrogen (Urea @ 31.25 kg N ha-1) alone or in combination on accumulation patterns of various protein fractions and protein quality in soybean seeds under agro-climatic conditions of Punjab. Application of nitrogen or sulphur alone decreased the relative proportion of β-subunit of purified β-conglycinin fraction than control. The acidic polypeptides of glycinin fraction of globulin and 11S/7S ratio were increased in all the treatments in comparison to control, and the maximum increase was reported in the combined application of gypsum with the recommended dose of urea. The lower proportion of total 7S, increased acidic polypeptides and improved 11S:7S ratio by combined treatment of nitrogen and sulphur suggests that gypsum @ 20 kg S ha-1 can be beneficial when applied along the recommended nitrogen dose to improve soybean protein quality. [ABSTRACT FROM AUTHOR]

Key message : GmMYB58 and GmMYB205 are key positive regulators that are involved in isoflavonoid biosynthesis in seeds of Glycine max, and they activate the expression of several structural genes in the isoflavonoid pathway. Abstract: MYB transcription factors (TFs) are major regulators involved in flavonoid/isoflavonoid biosynthesis in many plant species. However, functions of most MYB TFs remain unknown in flavonoid/isoflavonoid pathway in Glycine max. In this study, we identified 321 MYB TFs by genome-wide searching, and further isolated and functionally characterized two MYB TFs, GmMYB58 and GmMYB205. The deduced GmMYB58 and GmMYB205 proteins contain highly conserved R2R3 repeat domain at the N-terminal region that is the signature motif of R2R3-type MYB TFs. GmMYB58 and GmMYB205 were highly expressed in early seed development stages than in the other tested organs. GmMYB58 and GmMYB205 GFP fusion proteins were found to be localized in the nucleus when they were transiently expressed in Arabidopsis thaliana mesophyll protoplast. Both GmMYB58 and GmMYB205 can activate the promoter activities of GmCHS, GmIFS2, and GmHID in the transient trans-activation assays, and the activation of GmHID by both GmMYB58 and GmMYB205 was further confirmed by yeast one-hybrid assay. In addition, over-expression of GmMYB58 and GmMYB205 resulted in significant increases in expression levels of several pathway genes in soybean hairy roots, in particular, IFS2 by more than fivefolds in GmMYB205-over-expressing lines. Moreover, isoflavonoid contents were remarkably enhanced in the GmMYB58 and GmMYB205 over-expressing hairy roots than in the control. Our results suggest that GmMYB58 and GmMYB205 are seed-specific TFs, and they can enhance isoflavonoid biosynthesis mainly through the regulation of GmIFS2 and GmHID in G. max. [ABSTRACT FROM AUTHOR]

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Lutein benefits human health significantly, including that of the eyes, skin and heart. Therefore, increasing lutein content in soybean seeds is an important objective for breeding programmes. However, no information about soybean lutein-related quantitative trait loci (QTL) has been reported, as of 2016. The aim of the present study was to identify QTLs underlying the lutein content in soybean seeds. A population including 129 recombinant inbred lines was developed from the cross between ‘Dongnong46’ (lutein 13·10 µg/g) and ‘L-100’ (lutein 23·96 µg/g), which significantly differed in seed lutein contents. This population was grown in ten environments including Harbin in 2012, 2013, 2014 and 2015; Hulan in 2013, 2014 and 2015; and Acheng in 2013, 2014 and 2015. A total of 213 simple sequence repeat markers were used to construct the genetic linkage map, which covered approximately 3623·39 cM, with an average distance of 17·01 cM between markers. In the present study, eight QTLs associated with lutein content were found initially, which could explain 1·01–19·66% of the observed phenotypic variation in ten different tested environments. The phenotypic contribution of qLU-1 (located near BARC-Satt588 on chromosome 9 (Chr 9; linkage group (LG) K)) was >10% across seven tested environments, while qLU-2 (located near Satt192 of Chr 12 (LG H)) and qLU-3 (located near Satt353 of Chr12 (LGH)) could explain 5–10% of the observed phenotypic variation in more than seven environments, respectively. qLU-5, qLU-6, qLU-7 and qLU-8 could be detected in more than four environments. These eight QTLs were novel, and have considerable potential value for marker-assistant selection of higher lutein content in soybean lines. [ABSTRACT FROM PUBLISHER]

Medicinal plants are a significant source of biological active and health protective compounds. Post-harvest treatments can affect, in different ways, the content and biological activity of such compounds. One of the most common post-harvesting methods is drying. In this study, we investigated the effect of drying method on the content of natural pigments (chlorophylls, carotenoids and anthocyanins) and on the antioxidant capacity of two traditionally used herbs, the Melissa officinalis (lemon balm) and the Urtica dioica (stinging nettle) both of them landraces collected from plants grown in Nitra region, West of Slovakia. The freeze-dried samples of both herbs exhibited the highest content of chlorophyll a (7.55 ± 0.13 mg/g dry mass for lemon balm and 9.41 ± 0.17 mg/g dry mass for stinging nettle), chlorophyll b (3.12 ± 0.28 mg/g dry mass for lemon balm and 3.34 ± 0.24 mg/g dry mass for stinging nettle) and carotenoids (2.11 ± 0.24 mg/g dry mass for lemon balm and 2.62 ± 0.06 mg/g dry mass for stinging nettle). The content of chlorophylls and carotenoids correlated with the DPPH antioxidant (radical scavenging) capacity. Higher antioxidant capacity of the lemon balm extracts compared to nettle samples was attributed to the higher content of polyphenol compounds anthocyanins. Despite the higher cost, the freeze drying (lyophilisation) was recommended as the most suitable drying method, mainly for reasons of preserving maximum pigment content and antioxidant capacity. [ABSTRACT FROM AUTHOR]

Legume crops are grown throughout the world and provide an excellent food source of digestible protein and starch, as well as dietary fibre, vitamins, minerals, and flavonoids. Fruit and seeds from legumes are also an important source of vegetables for a well-balanced diet. A trend in elevated temperature as a result of climate change increases the risk of a heat stress-induced reduction in legume crop yield. High temperatures during the crop reproductive development phase are particularly detrimental to fruit/seed production because the growth and development of the reproductive tissues are sensitive to small changes in temperature. Hormones are signalling molecules that play important roles in a plant's ability to integrate different environmental inputs and modify their developmental processes to optimize growth, survival, and reproduction. This review focuses on the hormonal regulation of reproductive development and heat stress-induced alteration of this regulation during (i) pollination, (ii) early fruit set, and (iii) seed development that affects fruit/seed yield in legume and other model crops. Further understanding of hormone-regulated reproductive growth under non-stress and heat-stress conditions can aid in trait selection and the development of gene modification strategies and cultural practices to improve heat tolerance in legume crops contributing to improved food security. [ABSTRACT FROM AUTHOR]

This study aimed to investigate potential pathways associated with arbutin synthesis and potential synthetic routes to 6′- O-caffeoylarbutin (CA) in Vaccinium dunalianum Wight (Ericaceae) via de novo transcriptome sequencing. De novo transcriptome sequencing of leaf buds of V. dunalianum was performed. After quality control and transcriptome assembly, unigenes were predicted and their functions were annotated using Blast2GO. Furthermore, the coding sequences of unigenes were predicted based on NR, SWISS-PROT, KEGG and COG databases. Subsequently, pathways related to arbutin synthesis and potential synthetic routes to CA were analyzed based on KEGG and ExPASy databases. In total, 55537512 clean reads (11.7 G) were generated, and there were 4998376080 nt clean nucleotides. The majority of unigenes were related to the pathways of metabolism, such as carbohydrate and lipid metabolism. Besides, a set of enzymes were related to uridine diphosphate (UDP)-glucose synthesis (e.g. K01784) via some pathways, such as amino sugar and nucleotide sugar metabolism, starch and sucrose metabolism, etc. Some other enzymes were associated with hydroquinone synthesis (e.g. K15849) via tyrosine metabolism. Additionally, chlorogenic acid, caffeoyl-CoA and 1- O-hydroxycinnamoyl-β-D-glucose (HCA-Glc) were predicted to participate in CA synthesis as acyl donors. Some enzymes in pathways related to glycometabolism (e.g. K01784) and tyrosine metabolism (e.g. K15849) may participate in arbutin synthesis. Additionally, chlorogenic acid, caffeoyl-CoA and HCA-Glc may take part in CA synthesis. [ABSTRACT FROM AUTHOR]

Although the Early Soybean Production System (ESPS) in the Midsouthern USA increased seed yield under irrigated and non-irrigated conditions, heat stress and drought still lead to poor seed quality in heat sensitive soybean cultivars. Our breeding goal was to identify breeding lines that possess high germination, nutritional quality, and yield potential under high heat and dryland production conditions. Our hypothesis was that breeding lines derived from exotic germplasm might possess physiological and genetic traits allowing for higher seed germinability under high heat conditions. In a 2-year field experiment, breeding lines derived from exotic soybean accessions, previously selected for adaptability to the ESPS in maturity groups (MG) III and IV, were grown under non-irrigated conditions. Results showed that three exotic breeding lines had consistently superior germination across 2 years. These lines had a mean germination percentage of >80%. Two (25-1-1-4-1-1 and 34-3-1-2-4-1) out of the three lines with ⩾80% germination in both years maintained high seed protein, oleic acid, N, P, K, B, Cu, and Mo in both years. Significant (P < 0.05) positive correlations were found between germination and oleic acid and with K and Cu in both years. Significant negative correlations were found between germination and linoleic acid, Ca, and hard seed in both years. There were positive correlations between germination and N, P, B,Mo, and palmitic acid only in 2013. A negative correlation was found between germination and green seed damage and linolenic acid in 2013 only. Seed wrinkling was significantly negatively correlated with germination in 2012 only. A lower content of Ca in the seed of high germinability genotypes may explain the lower rates of hard seed in those lines, which could lead to higher germination. Many of the differences in yield, germination, diseases, and seed composition between years are likely due to heat and rainfall differences between years. The results also showed the potential roles of seed minerals, especially K, Ca, B, Cu, and Mo, in maintaining high seed quality. The knowledge gained from this research will help breeders to select for soybean with high seed nutritional qualities and high germinability. [ABSTRACT FROM AUTHOR]

For plant regeneration through somatic embryogenesis, immature cotyledonary nodes and embryonic axes explants of soybean cultivars JS 335 were cultured on six different medium fortified with MS salts, B5 vitamins and combinations of growth regulators. Between explants, immature embryonic axes demonstrated better regeneration potential as compared to cotyledonary node. For immature embryonic axes culture, better morphogenic callus induction, embryogenesis, organogenesis and plantlet regeneration was observed on MS with 4 B1.86N, MS 10N, MS .1B1N and MS.2B1N as compared to other media formulations. Cotyledonary nodes exhibited higher callus induction, embryogenesis, organogenesis on media with MS4 B1.86N, MS .1B1N, MS 3B6N and higher plantlet regeneration on media with MS.5B1N. [ABSTRACT FROM AUTHOR]

Phomopsis seed decay of soybean [Glycine max(L.) Merr.] is a seedborne fungal disease caused byDiaporthelongicollathat causes yield losses and reduced seed quality. Biocontrol of this pathogen by a strain ofSarocladium kilienseisolated from a culture ofD. longicollawas investigatedin vitroandin vivo. A zone of inhibition formed between the two fungiin vitro, but was poorly sustained, and inhibition of conidial germination ofD. longicollaby a culture filtrate ofS. kiliensewas not significant, thoughD. longicollamycelial growth was inhibited by a 25% culture filtrate. Co-inoculation of both fungi failed to reduce seed rot or increase seed germination in greenhouse and growth chamber experiments, respectively. Co-inoculation of both fungi also failed to reduce pycnidial development in colonies ofD. longicollagrowing on leaf pieces, but soaking of the leaf pieces withS. kilienseconidia for one or three days prior to inoculation withD. longicollaeliminated pycnidial development completely. Although it was not possible to reproduce the parasitism ofS. kilienseonD. longicollaunder the experimental and environmental conditions used, the potential to useS. kilienseas a protectant biocontrol for soybean fungal pathogens should be further investigated. [ABSTRACT FROM PUBLISHER]

To facilitate direct harvest, preharvest herbicide applications are used to desiccate weeds and to accelerate and promote uniform dry bean (Phaseolus vulgaris L.) maturation. Field trials were conducted near Richville, MI, in 2013 and 2014 to evaluate the effects of preharvest herbicide treatments on desiccation, yield, and canned black bean quality and color. Three black bean cultivars, Zorro, Eclipse, and Zenith, were planted on two different dates in each of 2 yr. Three preharvest herbicide treatments, paraquat, glyphosate, and saflufenacil, were applied at a standard (pods = 80% yellow) and an early (pods = 50% yellow) application timing. A nontreated control was established for each cultivar. Differences in black bean desiccation were greatest 3 d after treatment (DAT), with paraquat and saflufenacil showing the quickest desiccation at the early application timing. By 7 DAT, desiccation for most preharvest treatments was >95%. Saflufenacil applied at the early timing reduced yield by as much as 55% compared with the nontreated control. Yield was generally not affected when preharvest herbicide treatments were applied at the standard application timing. Black bean cultivar influenced canned bean color retention (Zenith > Zorro > Eclipse). Preharvest herbicide treatments reduced color retention when applied at the early application timing; glyphosate reduced color retention by as much as 24%. Because of variability in bean maturation rate within a field, preharvest herbicide treatments need to be applied as close to physiological maturity as possible to minimize yield losses and maintain color retention of canned black bean. [ABSTRACT FROM AUTHOR]

Most proteins produced in the endoplasmic reticulum (ER) of eukaryotic cells fold via disulfide formation (oxidative folding). Oxidative folding is catalyzed by protein disulfide isomerase (PDI) and PDI-related ER protein thiol disulfide oxidoreductases (ER oxidoreductases). In yeast and mammals, ER oxidoreductin-1s (Ero1s) supply oxidizing equivalent to the active centers of PDI. In this study, we expressed recombinant soybean Ero1 (GmERO1a) and found that GmERO1a oxidized multiple soybean ER oxidoreductases, in contrast to mammalian Ero1s having a high specificity for PDI. One of these ER oxidoreductases, GmPDIM, associated in vivo and in vitro with GmPDIL-2, was unable to be oxidized by GmERO1a. We therefore pursued the possible cooperative oxidative folding by GmPDIM, GmERO1a, and GmPDIL-2 in vitro and found that GmPDIL-2 synergistically accelerated oxidative refolding. In this process, GmERO1a preferentially oxidized the active center in the a9 domain among the a, a9, and b domains of GmPDIM. A disulfide bond introduced into the active center of the a9 domain of GmPDIM was shown to be transferred to the active center of the a domain of GmPDIM and the a domain of GmPDIM directly oxidized the active centers of both the a or a9 domain of GmPDIL-2. Therefore, we propose that the relay of an oxidizing equivalent from one ER oxidoreductase to another may play an essential role in cooperative oxidative folding by multiple ER oxidoreductases in plants. [ABSTRACT FROM AUTHOR]

The evolutionary forces that maintain genetic variation in quantitative traits within populations remain poorly understood. One hypothesis suggests that variation is under purifying selection, resulting in an excess of low-frequency variants and a negative correlation between minor allele frequency and selection coefficients. Here, we test these predictions using the genetic loci associated with total expression variation (eQTLs) and allele-specific expression variation (aseQTLs) mapped within a single population of the plant Capsella grandiflora. In addition to finding eQTLs and aseQTLs for a large fraction of genes, we show that alleles at these loci are rarer than expected and exhibit a negative correlation between phenotypic effect size and frequency. Overall, our results show that the distribution of frequencies and effect sizes of the loci responsible for local expression variationwithin a single outcrossing population are consistent with the effects of purifying selection. [ABSTRACT FROM AUTHOR]

The EFSA GMO Panel previously assessed the two single events that are combined to produce soybean MON 87769 × MON 89788 and did not identify safety concerns. No new data on these single events, leading to a modification of the original conclusions on safety, were identified. The molecular, agronomic, phenotypic and compositional data on soybean MON 87769 × MON 89788 did not give rise to safety concerns. The Panel considers that there is no reason to expect interactions between the single events to impact on food and feed safety. There were no concerns regarding the potential toxicity or allergenicity of soybean MON 87769 × MON 89788, and no evidence that the genetic modification significantly changes the overall allergenicity. Because of the lack of data on dietary exposure to refined bleached deodorised oil from soybean MON 87769 × MON 89788, the EFSA GMO Panel could not complete the human health and nutrition assessment. There are no concerns regarding the use of feedingstuffs derived from defatted toasted MON 87769 × MON 89788 soybean meal. There are no indications of an increased likelihood of establishment and spread of feral soybean plants. Potential interactions of soybean MON 87769 × MON 89788 with biotic and abiotic environments were not considered relevant to this application. The unlikely, but theoretically possible, transfer of recombinant genes from soybean MON 87769 × MON 89788 to environmental bacteria is not of safety concern. The post-market environmental monitoring plan and reporting intervals conform with the scope of this application. In conclusion, the Panel could not complete the food and feed safety assessment of soybean MON 87769 × MON 89788 because of the lack of an appropriate nutritional assessment. The Panel concludes that soybean MON 87769 × MON 89788 is unlikely to have adverse effects on the environment in the context of application EFSA-GMO-NL-2010-85. [ABSTRACT FROM AUTHOR]

The pink or red ketocarotenoids, canthaxanthin and astaxanthin, are used as feed additives in the poultry and aquaculture industries as a source of egg yolk and flesh pigmentation, as farmed animals do not have access to the carotenoid sources of their wild counterparts. Because soybean is already an important component in animal feed, production of these carotenoids in soybean could be a cost-effective means of delivery. In order to characterize the ability of soybean seed to produce carotenoids, soybean cv. Jack was transformed with the crtB gene from Pantoea ananatis, which codes for phytoene synthase, an enzyme which catalyzes the first committed step in the carotenoid pathway. The crtB gene was engineered together in combinations with ketolase genes (crtW from Brevundimonas sp. strain SD212 and bkt1 from Haematococcus pluvialis) to produce ketocarotenoids; all genes were placed under the control of seed-specific promoters. HPLC results showed that canthaxanthin is present in the transgenic seeds at levels up to 52 μg/g dry weight. Transgenic seeds also accumulated other compounds in the carotenoid pathway, such as astaxanthin, lutein, β-carotene, phytoene, α-carotene, lycopene, and β-cryptoxanthin, whereas lutein was the only one of these detected in non-transgenic seeds. The accumulation of astaxanthin, which requires a β-carotene hydroxylase in addition to a β-carotene ketolase, in the transgenic seeds suggests that an endogenous soybean enzyme is able to work in combination with the ketolase transgene. Soybean seeds that accumulate ketocarotenoids could potentially be used in animal feed to reduce or eliminate the need for the costly addition of these compounds. [ABSTRACT FROM AUTHOR]

Physicochemical, functional and microstructural characteristics of the seeds from three vegetable soybean varieties ('Asmara', 'Randolph', and 'Owens') grown in Virginia and their responses to cooking process were investigated. In the three varieties, carbohydrate is the predominant component ranging from 42.4 to 48.1 %, followed by protein (34.2-35.4 %), oil (13.1-17.5 %) and ash (4.21-4.88 %). The major sugars in the beans are sucrose (5.94-12.2 %) and fructose (1.61-2.31 %). Cooking process significantly (P < 0.05) decreased the levels of carbohydrate and sucrose, but increased protein and fiber contents. Among the three varieties, raw 'Asmara' was the heaviest (40.7 g/100 seeds), while 'Owens' beans had the greatest hardness (3615 g). 'Owens' had the highest water holding capacity (246.7 mL/100 g sample), the lowest oil holding capacity (148.1 mL/100 g sample), protein solubility (11.9 %), emulsifying activity (40.8 %) and foaming capacity (8.97 %). Cooking significantly (P < 0.05) decreased the lightness, hardness, protein solubility, emulsifying activity and foaming capacity of the beans but increased their water absorption and holding capacities. 'Owens' showed the least total solid loss (3.09 %) during the cooking. Bean morphology was also affected by cooking with larger granules and rough surface. Of the three varieties investigated, 'Owens' exhibited significant differences in the physicochemical and functional properties as well as the response to cooking process. [ABSTRACT FROM AUTHOR]

Soybean flavonoids, a group of important signaling molecules in plant-environment interaction, ubiquitously exist in soybean and are tightly regulated by many genes. Here we reported that GmMYB100, a gene encoding a R2R3 MYB transcription factor, is involved in soybean flavonoid biosynthesis. GmMYB100 is mainly expressed in flowers, leaves and immature embryo, and its level is decreased after pod ripening. Subcellular localization assay indicates that GmMYB100 is a nuclear protein. GmMYB100 has transactivation ability revealed by a yeast functional assay; whereas bioinformatic analysis suggests that GmMYB100 has a negative function in flavonoid biosynthesis. GmMYB100-overexpression represses the transcript levels of flavonoid-related genes in transgenic soybean hairy roots and Arabidopsis, and inhibits isoflavonoid (soybean) and flavonol ( Arabidopsis) production in transgenic plants. Furthermore, the transcript levels of six flavonoid-related genes and flavonoid (isoflavonoid and flavone aglycones) accumulation are elevated in the GmMYB100-RNAi transgenic hairy roots. We also demonstrate that GmMYB100 protein depresses the promoter activities of soybean chalcone synthase and chalcone isomerase. These findings indicate that GmMYB100 is a negative regulator in soybean flavonoid biosynthesis pathway. [ABSTRACT FROM AUTHOR]

Transgenic soya bean ( Glycine max) plants overexpressing a seed-specific bacterial phytoene synthase gene from Pantoea ananatis modified to target to plastids accumulated 845 μg β carotene g−1 dry seed weight with a desirable 12:1 ratio of β to α. The β carotene accumulating seeds exhibited a shift in oil composition increasing oleic acid with a concomitant decrease in linoleic acid and an increase in seed protein content by at least 4% (w/w). Elevated β-carotene accumulating soya bean cotyledons contain 40% the amount of abscisic acid compared to nontransgenic cotyledons. Proteomic and nontargeted metabolomic analysis of the mid-maturation β-carotene cotyledons compared to the nontransgenic did not reveal any significant differences that would account for the altered phenotypes of both elevated oleate and protein content. Transcriptomic analysis, confirmed by RT- PCR, revealed a number of significant differences in ABA-responsive transcripton factor gene expression in the crtB transgenics compared to nontransgenic cotyledons of the same maturation stage. The altered seed composition traits seem to be attributed to altered ABA hormone levels varying transcription factor expression. The elevated β-carotene, oleic acid and protein traits in the β-carotene soya beans confer a substantial additive nutritional quality to soya beans. [ABSTRACT FROM AUTHOR]

Soybean (Glycine max (L.) Merr.) isoflavone is important for human health and plant defense system. To identify novel quantitative trait loci (QTL) and epistatic QTL underlying isoflavone content in soybean, F5:6, F5:7 and F5:8 populations of 130 recombinant inbred (RI) lines, derived from the cross of soybean cultivar ‘Zhong Dou 27′ (high isoflavone) and ‘Jiu Nong 20′ (low isoflavone), were analyzed with 95 new SSR markers. A new linkage map including 194 SSR markers and covering 2,312 cM with mean distance of about 12 cM between markers was constructed. Thirty four QTL for both individual and total seed isoflavone contents of soybean were identified. Six, seven, ten and eleven QTL were associated with daidzein (DZ), glycitein (GC), genistein (GT) and total isoflavone (TI), respectively. Of them 23 QTL were newly identified. The qTIF_1 between Satt423 and Satt569 shared the same marker Satt569 with qDZF_2, qGTF_1 and qTIF_2. The qGTD2_1 between Satt186 and Satt226 was detected in four environments and explained 3.41%-10.98% of the phenotypic variation. The qGTA2_1, overlapped with qGCA2_1 and detected in four environments, was close to the previously identified major QTL for GT, which were responsible for large a effects. QTL (qDZF_2, qGTF_1 and qTIF_2) between Satt144-Satt569 were either clustered or pleiotropic. The qGCM_1, qGTM_1 and qTIM_1 between Satt540-Sat_244 explained 2.02%–9.12% of the phenotypic variation over six environments. Moreover, the qGCE_1 overlapped with qGTE_1 and qTIE_1, the qTIH_2 overlapped with qGTH_1, qGCI_1 overlapped with qDZI_1, qTIL_1 overlapped with qGTL_1, and qTIO_1 overlapped with qGTO_1. In this study, some of unstable QTL were detected in different environments, which were due to weak expression of QTL, QTL by environment interaction in the opposite direction to a effects, and/or epistasis. The markers identified in multi-environments in this study could be applied in the selection of soybean cultivars for higher isoflavone content and in the map-based gene cloning. [ABSTRACT FROM AUTHOR]

Two artificial diets developed for rearing Lygus spp., a fresh yolk chicken egg based-diet (FYD) and a dry yolk chicken egg based-diet (DYD), were evaluated as an alternative food source for rearing the southern green stink bug, Nezara viridula (L.) (Heteroptera: Pentatomidae). Survival to adult was 97.3 and 74.67%, respectively, on the fresh and dry yolk diets. Insects fed FYD had 100% survival of nymphs from first through fourth instars. Adult development was significantly shorter on FYD (30.37±SE 0.30 d) as compared with DYD (32.77±SE 0.16 d). Increased male and female longevity, higher fecundity, and larger egg mass sizes were also observed with N. viridula-fed FYD. However, fertility and hatchability was higher on DYD. A complete cohort life table was constructed to describe the development of N. viridula on both diets. [ABSTRACT FROM AUTHOR]

β-Conglycinin, a major component of seed storage protein in soybean, comprises three subunits: α, α′ and β. The expression of genes for these subunits is strictly controlled during embryogenesis. The proximal promoter region up to 245 bp upstream of the transcription start site of the α subunit gene sufficiently confers spatial and temporal control of transcription in embryos. Here, the binding profile of nuclear proteins in the proximal promoter region of the α subunit gene was analysed. DNase I footprinting analysis indicated binding of proteins to the RY element and DNA regions including box I, a region conserved in cognate gene promoters. An electrophoretic mobility shift assay ( EMSA) using different portions of box I as a probe revealed that multiple portions of box I bind to nuclear proteins. In addition, an EMSA using nuclear proteins extracted from embryos at different developmental stages indicated that the levels of major DNA-protein complexes on box I increased during embryo maturation. These results are consistent with the notion that box I is important for the transcriptional control of seed storage protein genes. Furthermore, the present data suggest that nuclear proteins bind to novel motifs in box I including 5′- TCAATT-3′ rather than to predicted cis-regulatory elements. [ABSTRACT FROM AUTHOR]

Background: Orange-spotted grouper (Epinephelus coioides) with protogynous hermaphroditic features are one of the most economically important aquaculture species in Taiwan. However, larvae stage grouper are susceptible to infection by the bacterial pathogen Vibrio alginolyticus. To better understand the molecular mechanisms of the immune response to V. alginolyticus in Epinephelus coioides larvae, we used high-throughput deep sequencing technology to study the effect of infection on gene expression. Results: A total of 114,851,002 reads were assembled, consisting of 9,687,355,560 nucleotides; these were further assembled into 209,082 contigs with a mean length of 372 bp. Gene ontology (GO) analysis of the transcriptome revealed 12 cellular component subcategories, 16 molecular function subcategories, and 42 biological process subcategories (P value <0.05). A total of 32664 Epinephelus coioides genes were mapped to the Kyoto Encyclopedia of Genes and Genomes (KEGG); 1504 differentially expressed genes (DEGs) were subsequently identified, in 12 categories (P value <0.05). Vibrio infection affected the expression of genes involved in complementation, coagulation cascades, pathogen (Staphylococcus aureus) infection, phagosome activity, antigen processing, and the antigen presentation pathway. Conclusion; We conclude that the complement pathway of innate immunity and the hepicidin antimicrobial peptide may play important roles in the defense of Epinephelus coioides larvae against V. alginolyticus, and the immune response may activate at 4 h after bacterial infection. These results implicate the complement pathway signal pathway in immunity during V. alginolyticus infection at early developmental stages, enhancing our understanding of the mechanisms underlying the immune response to Vibrio infection in Epinephelus coioides. [ABSTRACT FROM AUTHOR]