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Abstract Background Magnolia kobus belongs to the Magnoliaceae family and the genus Magnolia. The flowers are widely used in herbal tea and dietary supplements. It is effectively used in traditional medicines and its chemical constituents have various biological activities, such as anti-obesity, antioxidant, anticancer, anti-inflammatory, and anti-hyperlipidaemic activities. The flower developmental process of M. kobus involves a complex regulatory network and is intensively related to the quality and relative composition of primary and secondary metabolites of the flower. Hence, the identification of metabolic changes during different developmental stages of the flower was important for enhancing organoleptic and nutritional characteristics. To our knowledge, to date, none of the studies has investigated the relationship between metabolic changes (primary and secondary metabolites) during different developmental stages of the M. kobus flower. Results This study investigated metabolic changes in 47 hydrophilic and 13 lipophilic compounds and biological activities in Magnolia kobus flowers during four developmental stages. Metabolites were identified using HPLC, GC–qMS, and GC–TOFMS. Throughout flower development, metabolite accumulation showed significant differences based on metabolomics approaches. The dynamic changes of 21 differential metabolites occurred between the young and mature stages. Flower buds exhibited high levels of phenylpropanoids and phytosterols. The total phenolic and flavonoid contents were most abundant in the buds and decreased from the bud to the old stages. Flower bud extracts showed the most powerful effects in three antioxidant assays and antibacterial effects against 10 pathogenic bacterial strains. Conclusions These findings emphasise the value of M. kobus flower buds as an edible natural source and provide valuable insight into the metabolic changes that occur in M. kobus flowers during development. Graphical Abstract

IntroductionMicroalgae biotechnology utilizes species like Porphyridium cruentum for their valuable phycobiliproteins and exopolysaccharides, which have potential industrial applications and health benefits, particularly in skin condition improvement.MethodsA mutant of P. cruentum LIMS-PS-1061 was developed through ethyl methanesulfonate mutagenesis and subsequent colony screening to study changes in its biomass production and pigment composition under different lighting conditions.Results and discussionThe mutant exhibited a 33.9% increase in dry weight under white light compared to the wild type. Despite maintaining the total pigment content, specific components changed significantly: chlorophyll content decreased 2.20- and 3.61-fold under white and blue light respectively, while phycobiliproteins increased 1.59- and 1.23-fold under the same conditions. These alterations suggest a compensatory mechanism for maintaining photosynthetic capacity. Furthermore, the exopolysaccharides of P. cruentum upregulated genes related to skin moisturization, barrier enhancement, and elasticity, and promoted wound healing through fibroblast migration. This supports the proposed mechanism of action for P. cruentum’s exopolysaccharides in improving human skin conditions by integrating the effects of aquaporin 3, filaggrin, involucrin, loricrin, elastin, and fibrillin-1.

Abstract Background Soybean is a valuable source of edible protein and oil, as well as secondary metabolites that can be used in food products, cosmetics, and medicines. However, because soybean isoflavone content is a quantitative trait influenced by polygenes and environmental interactions, its genetic basis remains unclear. Results This study was conducted to identify causal quantitative trait loci (QTLs) associated with soybean isoflavone contents. A mutant-based F2 population (190 individuals) was created by crossing the Korean cultivar Hwanggeum with low isoflavone contents (1,558 µg g−1) and the soybean mutant DB-088 with high isoflavone contents (6,393 µg g−1). A linkage map (3,049 cM) with an average chromosome length of 152 cM was constructed using the 180K AXIOM® SoyaSNP array. Thirteen QTLs related to agronomic traits were mapped to chromosomes 2, 3, 11, 13, 19, and 20, whereas 29 QTLs associated with isoflavone contents were mapped to chromosomes 1, 3, 8, 11, 14, 15, and 17. Notably, the qMGLI11, qMGNI11, qADZI11, and qTI11, which located Gm11_9877690 to Gm11_9955924 interval on chromosome 11, contributed to the high isoflavone contents and explained 11.9% to 20.1% of the phenotypic variation. This QTL region included four candidate genes, encoding β-glucosidases 13, 14, 17–1, and 17–2. We observed significant differences in the expression levels of these genes at various seed developmental stages. Candidate genes within the causal QTLs were functionally characterized based on enriched GO terms and KEGG pathways, as well as the results of a co-expression network analysis. A correlation analysis indicated that certain agronomic traits (e.g., days to flowering, days to maturity, and plant height) are positively correlated with isoflavone content. Conclusions Herein, we reported that the major QTL associated with isoflavone contents was located in the interval from Gm11_9877690 to Gm11_9955924 (78 kb) on chromosome 11. Four β-glucosidase genes were identified that may be involved in high isoflavone contents of soybean DB-088. Thus, the mutant alleles from soybean DB-088 may be useful for marker-assisted selection in developing soybean lines with high isoflavone contents and superior agronomic traits.

Apart from being utilized as a commercial fiber at maturity, kenaf shoots have potential as a food and feed source because of their diverse bioactivities. Previous studies have focused on mature stems because of their high biomass, whereas the antioxidant activities (AA) and the destination of AA contributors of kenaf stems and their high-yielding byproduct leaves during the growth stage have rarely been studied. Therefore, we investigated changes in AA and its relative components in kenaf leaves and stems during the four vital growth stages. Higher ABTS radical cation and DPPH radical scavenging abilities and ferric reducing antioxidant power, total phenolic content, total flavonoid content, and total polysaccharide content were observed at all leaf stages and in the late stem stages. Chlorogenic acid (CGA) and kaempferol glycosides, especially kaempferitrin (Kfr), were identified as representative phenolic acids and flavonoids in both kenaf leaves and stems. The content of CGA in both leaves and stems increased corresponding to the plant’s growth stage, whereas kaempferol glycosides were enhanced in leaves but declined in stems. The highest correlation was observed between TPC and AA in all organs. Further evaluation of CGA and Kfr verified that CGA was the predominant contributor to AA, surpassing Kfr. These findings suggest that kenaf leaves increase antioxidant levels as they grow and can be a useful source of stem harvesting byproducts.

Lentil is a leguminous crop with a high content of health-beneficial polyphenols. Lentil sprouts are popularly consumed in fresh vegetable markets, although their phytochemical qualities are not well understood. In this study, we investigated the accumulation of phenolics in lentil sprouts in response to photosynthetic and stress light qualities, including fluorescent light (FL), red LED (RL), blue LED (BL), ultraviolet A (UV-A), and ultraviolet B (UV-B). Three lentil cultivars, Lentil Green (LG), French Green (FG), and Lentil Red (LR), were used to evaluate sprouts grown under each light condition. The adequate light intensities for enhancing the antioxidant activity of lentil sprouts were found to be 11 W/m2 under photosynthetic lights (FL, RL, BL), and 1 W/m2 under stress lights (UV-A, UV-B). Subsequently, HPLC-ESI/Q-TOF MS analysis was conducted for the quantitative analysis of the individual phenolics that were accumulated in response to light quality. Four main phenolic compounds were identified: ferulic acid, tricetin, luteolin, and kaempferol. Notably, tricetin accumulation was significantly enhanced under BL across all three lentil cultivars examined. Furthermore, the study revealed that the other phenolic compounds were highly dependent on FL, BL, or UV-B exposure, exhibiting cultivar-specific variations. Additionally, the antioxidant activities of lentil extracts indicated that BL was most effective for LG and FG cultivars, whereas FL was most effective for enhancing antioxidant activity of LR cultivars as the sprouts grew.

Soybean [Glycine max (L.) Merr.] isoflavones, which are secondary metabolites with various functions, are included in food, cosmetics, and medicine. However, the molecular mechanisms regulating the glycosylation and malonylation of isoflavone glycoconjugates remain unclear. In this study, we conducted an RNA-seq analysis to compare soybean genotypes with different isoflavone contents, including Danbaek and Hwanggeum (low-isoflavone cultivars) as well as DB-088 (high-isoflavone mutant). The transcriptome analysis yielded over 278 million clean reads, representing 39,156 transcripts. The analysis of differentially expressed genes (DEGs) detected 2654 up-regulated and 1805 down-regulated genes between the low- and high-isoflavone genotypes. The putative functions of these 4459 DEGs were annotated on the basis of GO and KEGG pathway enrichment analyses. These DEGs were further analyzed to compare the expression patterns of the genes involved in the biosynthesis of secondary metabolites and the genes encoding transcription factors. The examination of the relative expression levels of 70 isoflavone biosynthetic genes revealed the HID, IFS, UGT, and MAT expression levels were significantly up/down-regulated depending on the genotype and seed developmental stage. These expression patterns were confirmed by quantitative real-time PCR. Moreover, a gene co-expression analysis detected potential protein–protein interactions, suggestive of common functions. The study findings provide valuable insights into the structural genes responsible for isoflavone biosynthesis and accumulation in soybean seeds.

The concentration of phenolic compounds in rapeseed is important because they are either anti-nutritional compounds or directly related to antioxidant activity. In this study, single nucleotide polymorphisms (SNPs) were identified using genotyping-by-sequencing (GBS), and an association study was conducted to investigate phenolic content in 95 rapeseed mutant lines derived from gamma rays and their original cultivar. A total of 3,196,318 SNPs were detected, resulting in the identification of a set of 70,208 union SNPs used to perform association studies. Seven compounds were identified, sinapine being the major phenolic compound (91.2–99.2%) in all genotypes. An association study was conducted for seven compounds and total phenolic content (TPC). It identified 241 SNPs that were significantly associated with these compounds, total sinapine content (TSC), and TPC. Based on the SNP markers detected, BnaC02g20420D was associated with dihexose, BnaC08g30570D with progoitrin, BnaA01g06890D with methyl sinapate, BnaA09g16810D with sinapine(4-O-8′)guaiacyl, BnaCnng39930D with trans-sinapine 1, BnaA07g31720D with trans-sinapine 2, BnaC03g31950D with sinapoyl malate, and BnaAnng27700D with TPC. These were selected as candidate genes that may play a key role in rapeseed. The SNP markers associated with these key phenolic compounds can be used as targets in breeding programs to reduce anti-nutritional components.

Red and blue artificial light sources are commonly used as photosynthetic lighting in smart farm facilities, and they can affect the metabolisms of various primary and secondary metabolites. Although the soybean plant contains major flavonoids such as isoflavone and flavonol, using light factors to produce specific flavonoids from this plant remains difficult because the regulation of light-responded flavonoids is poorly understood. In this study, metabolic profiling of soybean seedlings in response to red and blue lights was evaluated, and the isoflavone–flavonol regulatory mechanism under different light irradiation periods was elucidated. Profiling of metabolites, including flavonoids, phenolic acids, amino acids, organic acids, free sugars, alcohol sugars, and sugar acids, revealed that specific flavonol, isoflavone, and phenolic acid showed irradiation time-dependent accumulation. Therefore, the metabolic gene expression level and accumulation of isoflavone and flavonol were further investigated. The light irradiation period regulated kaempferol glycoside, the predominant flavonol in soybeans, with longer light irradiation resulting in higher kaempferol glycoside content, regardless of photosynthetic lights. Notably, blue light stimulated kaempferol-3-O-(2,6-dirhamnosyl)-galactoside accumulation more than red light. Meanwhile, isoflavones were controlled differently based on isoflavone types. Malonyl daidzin and malonyl genistin, the predominant isoflavones in soybeans, were significantly increased by short-term red light irradiation (12 and 36 h) with higher expressions of flavonoid biosynthetic genes, which contributed to the increased total isoflavone level. Although most isoflavones increased in response to red and blue lights, daidzein increased in response only to red light. In addition, prolonged red light irradiation downregulated the accumulation of glycitin types, suggesting that isoflavone’s structural specificity results in different accumulation in response to light. Overall, these findings suggest that the application of specific wavelength and irradiation periods of light factors enables the regulation and acquisition of specialized metabolites from soybean seedlings.

Abstract Faba bean (Vicia faba L.), a globally important grain legume providing a stable source of dietary protein, was one of the earliest plant cytogenetic models. However, the lack of draft genome annotations and unclear structural information on mRNA transcripts have impeded its genetic improvement. To address this, we sequenced faba bean leaf transcriptome using the PacBio single-molecule long-read isoform sequencing platform. We identified 28,569 nonredundant unigenes, ranging from 108 to 9669 bp, with a total length of 94.5 Mb. Many unigenes (3597, 12.5%) had 2–20 isoforms, indicating a highly complex transcriptome. Approximately 96.5% of the unigenes matched sequences in public databases. The predicted proteins and transcription factors included NB-ARC, Myb_domain, C3H, bHLH, and heat shock proteins, implying that this genome has an abundance of stress resistance genes. To validate our results, we selected WCOR413-15785, DHN2-12403, DHN2-14197, DHN2-14797, COR15-14478, and HVA22-15 unigenes from the ICE-CBF-COR pathway to analyze their expression patterns in cold-treated samples via qRT-PCR. The expression of dehydrin-related genes was induced by cold stress. The assembled data provide the first insights into the deep sequencing of full-length RNA from faba bean at the single-molecule level. This study provides an important foundation to improve gene modeling and protein prediction.

Isoflavones are major secondary metabolites that are exclusively produced by legumes, including soybean. Soy isoflavones play important roles in human health as well as in the plant defense system. The isoflavone content is influenced by minor-effect quantitative trait loci, which interact with polygenetic and environmental factors. It has been difficult to clarify the regulation of isoflavone biosynthesis because of its complex heritability and the influence of external factors. Here, using a genotype-by-sequencing-based genome-wide association mapping study, 189 mutant soybean genotypes (the mutant diversity pool, MDP) were genotyped on the basis of 25,646 high-quality single nucleotide polymorphisms (SNPs) with minor allele frequency of >0.01 except for missing data. All the accessions were phenotyped by determining the contents of 12 isoflavones in the soybean seeds in two consecutive years (2020 and 2021). Then, quantitative trait nucleotides (QTNs) related to isoflavone contents were identified and validated using multi-locus GWAS models. A total of 112 and 46 QTNs related to isoflavone contents were detected by multiple MLM-based models in 2020 and 2021, respectively. Of these, 12 and 5 QTNs were related to more than two types of isoflavones in 2020 and 2021, respectively. Forty-four QTNs were detected within the 441-Kb physical interval surrounding Gm05:38940662. Of them, four QTNs (Gm05:38936166, Gm05:38936167, Gm05:38940662, and Gm05:38940717) were located at Glyma.05g206900 and Glyma.05g207000, which encode glutathione S-transferase THETA 1 (GmGSTT1), as determined from previous quantitative trait loci annotations and the literature. We detected substantial differences in the transcript levels of GmGSTT1 and two other core genes (IFS1 and IFS2) in the isoflavone biosynthetic pathway between the original cultivar and its mutant. The results of this study provide new information about the factors affecting isoflavone contents in soybean seeds and will be useful for breeding soybean lines with high and stable concentrations of isoflavones.

Sorghum (Sorghum bicolor L.) is a promising biomass crop with high yields of cellulose, hemicellulose, and lignin. Sorghum biomass has emerged as an eco-friendly industrial material useful for producing biofuels and bioplastics. This study conducted genotyping-by-sequencing (GBS)-based genome-wide association studies (GWAS) to establish the genetic basis of traits associated with biomass. Specifically, the researchers evaluated agronomic traits and phenolic compounds using 96 sorghum genotypes. Six phenolic compounds, luteolinidin diglucoside, luteolin glucoside, apigeninidin glucoside, luteolinidin, apigeninidin, and 5-O-Me luteolinidin, were found to be the major phenolic compounds in all genotypes. Out of our six detected phenolic compounds (luteolinidin diglucoside, luteolin glucoside, apigeninidin glucoside, luteolinidin, apigeninidin, and 5-O-Me luteolinidin), luteolinidin was the major phenolic compound in all genotypes. Next, a GWAS analysis was performed to confirm significant associations between 192,040 filtered single-nucleotide polymorphisms (SNPs) and biomass-related traits. The study identified 40 SNPs on 10 chromosomes that were significantly associated with heading date (4 SNPs), plant height (3 SNPs), dry yield (2 SNPs), and phenolic compounds (31 SNPs). The GWAS analysis showed that SbRio.10G099600 (FUT1) was associated with heading date, SbRio.09G149200 with plant height, SbRio.06G211400 (MAFB) with dry yield, SbRio.04G259800 (PDHA1) with total phenolic content and luteolinidin diglucoside, and SbRio.02G343600 (LeETR4) with total phenolic content and luteolinidin, suggesting that these genes could play key roles in sorghum. These findings demonstrate the potential value of sorghum as a biomass resource and the potential for selecting sorghum genotypes with reduced phenolic contents for use in the bioindustry.

The biochemical compounds in kenaf leaves and flowers mainly consist of flavonoids, including flavonoid glycosides and floral anthocyanins. In the present study, we performed comparative transcriptome analysis using RNA-sequencing and identified putative genes involved in flower coloration in different flower developmental stages of three kenaf mutants including Baekma (white flower), Jangdae (ivory flower), and Bora (purple flower). A total of 36.1 Gb reads were generated for two kenaf accessions and 38,601 representative transcripts with an average length of 1350 bp were yielded, of which 33,057 (85.64%) were annotated against two databases. Expression profiling of the transcripts identified 1044 and 472 differentially expressed genes (DEGs) among three mutants in the young bud and full bloom stages, respectively. KEGG enrichment analysis of these DEGs revealed that the representative pathway was “biosynthesis of secondary metabolites”, including phenylpropanoid biosynthesis and flavonoid biosynthesis. Consequently, we investigated genes related to the phenylpropanoid pathway, which included 45 DEGs from a total of 1358. Our results provide useful information for understanding gene functions of flower coloration in kenaf, which will be useful in further studies.

The enzyme phosphoribosyl pyrophosphate synthase (PRPS) catalyzes the conversion of ribose 5-phosphate into phosphoribosyl diphosphate; the latter is a precursor of purine and pyrimidine nucleotides. Here, we investigated the function of PRPS from the single-celled green alga Chlamydomonas reinhardtii in its response to DNA damage from gamma radiation or the alkylating agent LiCl. CrPRPS transcripts were upregulated in cells treated with these agents. We generated CrPRPS-overexpressing transgenic lines to study the function of CrPRPS. When grown in culture with LiCl or exposed to gamma radiation, the transgenic cells grew faster and had a greater survival rate than wild-type cells. CrPRPS overexpression enhanced expression of genes associated with DNA damage response, namely RAD51, RAD1, and LIG1. We observed, from transcriptome analysis, upregulation of genes that code for key enzymes in purine metabolism, namely ribonucleoside-diphosphate pyrophosphokinase subunit M1, adenylate kinase, and nucleoside-diphosphate kinase. We conclude that CrPRPS may affect DNA repair process via regulation of de novo nucleotide synthesis.

Faba leaves are an unusual vegetable which contain not only a range of functional phytochemicals, but also certain undesirable flavors, which limit their consumption. In this study, several cooking methods (microwaving, roasting, steaming, and boiling), which are expected to reduce the odd flavors, were evaluated in terms of both health benefit effects and odd flavor factors, including antioxidant activities and the content of non-volatile and volatile organic compounds (VOCs). A cooking time of 5 min was selected because of the high content of l-dopa (l-3,4-dihydroxyphenylalanine) and aim of reducing the undesirable flavors of the cooked faba leaves. Microwaving and steaming significantly increased the l-dopa content by 24% and 19%, respectively. Roasting specifically increased the content of flavonols, exhibiting a 28% increase of kaempferol-3-O-arabinoside-7-O-rhamnoside, representatively, whereas boiling decreased about 50% of most phytochemicals evaluated. Microwaving and steaming treatments significantly increased the antioxidant activities. The l-dopa content and antioxidant activities of the processed faba leaves were strongly positively correlated with either an R2 = 0.863 of DPPH radical scavenging activity or an R2 = 0.856 value of ABTS radical scavenging activity, showing that l-dopa was a key antioxidant. All cooking methods potentially improved the flavor of the cooked faba leaves compared with that of the fresh leaves, because they significantly reduced the contents of VOCs such as alcohols, aldehydes, and ketones. These VOCs were the main components (>90%) in the fresh leaves. Adverse aromatic hydrocarbons were newly formed by the microwaving treatment, typically producing p-xylene, which is known to be a harmful dose-dependent compound, but they were not detected in leaves processed by the other cooking methods; therefore, although microwaving efficiently increased antioxidant activity, the chemical safety of the aromatic hydrocarbons produced need further study.

Gamma rays as a type of ionizing radiation constitute a physical mutagen that induces mutations and could be effectively used in plant breeding. To compare the effects of gamma and ionizing irradiation according to exposure time in common wheat (Keumgang, IT 213100), seeds were exposed to 60Co gamma rays at different dose rates. To evaluate the amount of free radical content, we used electron spin resonance spectroscopy. Significantly more free radicals were generated in the case of long-term compared with short-term gamma-ray exposure at the same dose of radiation. Under short-term exposure, shoot and root lengths were slightly reduced compared with those of the controls, whereas long-term exposure caused severe growth inhibition. The expression of antioxidant-related and DNA-repair-related genes was significantly decreased under long-term gamma-ray exposure. Long-term exposure caused higher radiosensitivity than short-term exposure. The results of this study could help plant breeders select an effective mutagenic induction dose rate in wheat.

Lentil (Lens culinaris; Fabaceae), one of the major pulse crops in the world, is an important source of proteins, prebiotics, lipids, and essential minerals as well as functional components such as flavonoids, polyphenols, and phenolic acids. To improve crop nutritional and medicinal traits, hybridization and mutation are widely used in plant breeding research. In this study, mutant lentil populations were generated by γ-irradiation for the development of new cultivars by inducing genetic diversity. Molecular networking via Global Natural Product Social Molecular Networking web platform and dipeptidyl peptide-IV inhibitor screening assay were utilized as tools for structure-based discovery of active components in active mutant lines selected among the lentil population. The bioactivity-based molecular networking analysis resulted in the annotation of the molecular class of phosphatidylcholine (PC) from the most active mutant line. Among PCs, 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine (18:0 Lyso PC) was selected for further in vivo study of anti-obesity effect in a high-fat diet (HFD)-induced obese mouse model. The administration of 18:0 Lyso PC not only prevented body weight gain and decreased relative gonadal adipose tissue weight, but also attenuated the levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and leptin in the sera of HFD-induced obese mice. Additionally, 18:0 Lyso PC treatment inhibited the increase of adipocyte area and crown-like structures in adipose tissue. Therefore, these results suggest that 18:0 Lyso PC is a potential compound to have protective effects against obesity, improving obese phenotype induced by HFD.

Irregular rooting of rosemary stem cuttings, causing differences in either stem maturation or responses to growth conditions, restricts uniform production. Here, rooting efficiency of apical, middle, and basal cuttings from rosemary stems was evaluated by controlling light conditions to prevent irregular rooting. The types of light applied to the cuttings were natural sunlight (NSL), fluorescent, red, and blue (BL) light. Among these light sources, BL significantly induced root growth of not only basal cuttings, but also apical and middle cuttings, whereas NSL induced poor root formation in apical and middle cuttings. In particular, the roots of apical cuttings exposed to BL grew twice as fast as those exposed to other types of light. The overexpression of BL-induced IAA synthetic genes confirmed the rooting patterns. IAA synthetic genes were significantly upregulated by BL in the apical and middle cuttings. Irradiating with 50 μmol photons m−2 s−1 BL resulted in similar root production levels among the cutting positions with high biomass, guaranteeing the successful production of uniform cuttings. Thus, the application of proper high-intensity BL promoted healthy, similar-quality rosemary cuttings among stem cutting positions.

The leaves and seeds of the faba bean are good sources of L-3,4-dihydroxyphenylalanin (L-dopa), and are usually eaten with thermal cooking methods. However, little information is available on the effect of thermal treatments on their nutritional value. We compared the changes in color, contents of L-dopa, vitamin C (Vc), total phenolics (TP), total flavonoids (TF) and antioxidant activity after dry heating or steaming faba bean leaves and seeds. The young leaves provided higher values of all the estimate factors, regardless of the thermal treatment. Steaming significantly degraded nutritional values of the leaves, but less changed in seeds, whereas dry heat maintained these attributes. The contents of L-dopa, Vc, TP and TF were shown to have strongly positive correlations with antioxidant activity in the leaves, whereas only L-dopa content was positively correlated with antioxidant activity of the seeds. Faba leaves contained relatively high L-dopa which possessed strong antioxidant activity compared to the Vc. As L-dopa is an important contributor to the antioxidant activity of faba leaves and seeds, consuming L-dopa from leaves may provide beneficial effects not only regarding Parkinson’s Disease.

Abstract Background Mesenchymal stem cells (MSCs) have broad-spectrum therapeutic effects in various diseases, and thus have many clinical applications. However, it is difficult to produce sufficient numbers of MSCs for clinical use, and improved culture systems are required. Here, we report the effects of calcium (Ca2+) and hypoxia on the proliferation of human umbilical cord blood-derived MSCs (hUCB-MSCs). In addition, we determined the optimal conditions of these two factors for the large-scale culture of hUCB-MSCs. Methods hUCB-MSCs were maintained under hypoxic conditions (3% O2) with 1.8 mM Ca2+ during long-term culture, and their proliferation was evaluated. To characterize the underlying mechanisms, the effects on hypoxia-inducible factor (HIF)-1α and the extracellular signal-regulated kinase (ERK) signaling pathways were investigated. The therapeutic effects in a mouse emphysema model were analyzed and compared with those of naive MSCs. Results The proliferation of Ca2+/hypoxia-treated hUCB-MSCs was increased compared with that observed using either calcium or hypoxia culture alone, without loss of stem cell marker expression or differentiation ability. The enhancement of the proliferation capacity of hUCB-MSCs by the synergistic effects of Ca2+ and hypoxia was dependent on the expression of HIF-1α and the ERK signaling pathway. The proliferation of Ca2+/hypoxia-treated hUCB-MSCs resulted in a delayed senescence phenotype and increased the expression levels of stemness genes such as Oct4 and Nanog compared to those observed in conventional culture conditions. In addition, Ca2+/hypoxia-treated MSCs transplantation in the mouse emphysema model showed the same therapeutic effects as observed with naive MSCs. Conclusions These findings suggest that a Ca2+/hypoxia-based expansion system has applications for the large-scale production of MSCs for therapeutic purposes.

Soybean seeds are consumed worldwide owing to their nutritional value and health benefits. In this study we investigated the metabolic properties of 208 soybean mutant diversity pool (MDP) lines by measuring the isoflavone and fatty acid contents of the seed. The total isoflavone content (TIC) ranged from 0.88 mg/g to 7.12 mg/g and averaged 3.08 mg/g. The proportion of oleic acid among total fatty acids (TFA) ranged from 0.38% to 24.66% and averaged 11.02%. Based on the TIC and TFA among the 208 MDP lines, we selected six lines with altered isoflavone content and six lines with altered oleic acid content compared with those of the corresponding wild-types for measuring gene expression. Each of twelve genes from the isoflavone and fatty acid biosynthesis pathways were analyzed at three different seed developmental stages. Isoflavone biosynthetic genes, including CHI1A, IFS1, and IFS2, showed differences in stages and expression patterns among individuals and wild-types, whereas MaT7 showed consistently higher expression levels in three mutants with increased isoflavone content at stage 1. Expression patterns of the 12 fatty acid biosynthetic genes were classifiable into two groups that reflected the developmental stages of the seeds. The results will be useful for functional analysis of the regulatory genes involved in the isoflavone and fatty acid biosynthetic pathways in soybean.

Rapeseed is the most important oil crop used in the food and biodiesel industries. In this study, based on single nucleotide polymorphism (SNP) identified from genotyping-by-sequencing (GBS), and an association study of flowering time, crude fat and fatty acid contents were investigated in 46 rapeseed mutant lines derived from gamma rays. A total of 623,026,394 clean data reads were generated with 6.6 million reads on average. A set of 37,721 filtered SNPs was used to perform gene ontology and phylogenetic analysis. Hierarchical cluster analysis of the rapeseed mutant lines gave eight groups based on flowering time and fatty acid compositions. Gene ontological analysis of the mutant lines showed that many genes displaying SNPs are involved in cellular processes, cellular anatomy, and binding. A total of 40 SNPs were significantly associated with flowering time (1 SNP), crude fat content (2 SNPs), and fatty acid content (37 SNPs). A total of 21 genes were annotated from fatty acid content SNPs; among them, nine genes were significantly enriched in reproductive processes, such as embryonic development, fruit development, and seed development. This study demonstrated that SNPs are efficient tools for mutant screening and it provides a basis that the improving the oil qualities of rapeseed.

The development of soybean with high antioxidant activities for use in the food and cosmetics industries is a target of breeding programs. In soybean, antioxidants are associated with seed color, although the metabolic basis for seed coloration remains incompletely understood. We selected six γ-ray-induced mutant lines that exhibited black, partially black, brown, partially brown, or yellowish-white pigmentation in the seed coat. Antioxidant activity and contents of anthocyanins, flavan-3-ols, and isoflavones were evaluated in the seed coat and cotyledons. The lines with black or brown seeds showed the highest antioxidant activities. The cotyledons showed no significant differences in seed coat components or antioxidant activities among lines. Black and brown seed coat components showed the highest antioxidant activities. The black seed coat contained five anthocyanins, whereas seed coats of brown- and yellow-seeded lines entirely lacked anthocyanins. Both black and brown seeds were rich in flavan-3-ols, including catechin and epicatechin, which were the predominant antioxidant contributors in brown seeds. Isoflavone contents showed weaker correlations with antioxidant activity than anthocyanins and flavan-3-ols. These results demonstrated that antioxidant activities were determined by anthocyanins in black seeds and flavan-3-ols in brown and black seeds, whereas relatively low antioxidant activities in yellow seeds reflected their high isoflavone contents.

Transposable elements (TEs)—major components of eukaryotic genomes—have the ability to change location within a genome. Because of their mobility, TEs are important for genome diversification and evolution. Here, a simple rapid method, using the consensus terminal inverted repeat sequences of PONG, miniature inverted-repeat transposable element (MITE)-Tourist (M-t) and MITE-Stowaway (M-s) as target region amplification polymorphism (TE-TRAP) markers, was employed to investigate the mobility of TEs in a gamma-irradiated soybean mutant pool. Among the different TE-TRAP primer combinations, the average polymorphism level and polymorphism information content value were 57.98% and 0.14, respectively. Only the PONG sequence separated the mutant population into three major groups. The inter-mutant population variance, determined using the PONG marker (3.151 and 29%) was greater than that of the M-t (2.209 and 20%) and M-s (2.766 and 18%) markers, whereas the reverse was true for the intra-mutant population variations, with M-t and M-s values, being 15.151 (82%) and 8.895 (80%), respectively, compared with the PONG marker (7.646 and 71%). Thus, the MITE markers revealed more dynamic and active mobility levels than the PONG marker in gamma-ray irradiated soybean mutant lines. The TE-TRAP technique associated with sensitive MITEs is useful for investigating genetic diversity and TE mobilization, providing tools for mutant selection in soybean mutation breeding.

Hibiscus species are rich in phenolic compounds and have been traditionally used for improving human health through their bioactive activities. The present study investigated the phenolic compounds of leaf extracts from 18 different H. acetosella accessions and evaluated their biofunctional properties, focusing on antioxidant and antibacterial activity. The most abundant phenolic compound in H. acetosella was caffeic acid, with levels ranging from 14.95 to 42.93 mg/100 g. The antioxidant activity measured by the ABTS assay allowed the accessions to be classified into two groups: a high activity group with red leaf varieties (74.71–84.02%) and a relatively low activity group with green leaf varieties (57.47–65.94%). The antioxidant activity was significantly correlated with TAC (0.933), Dp3-Sam (0.932), Dp3-Glu (0.924), and Cy3-Sam (0.913) contents (p < 0.001). The H. acetosella phenolic extracts exhibited antibacterial activity against two bacteria, with zones of inhibition between 12.00 and 13.67 mm (Staphylococcus aureus), and 10.67 and 13.33 mm (Pseudomonas aeruginosa). All accessions exhibited a basal antibacterial activity level (12 mm) against the Gram-positive S. aureus, with PI500758 and PI500764 exhibiting increased antibacterial activity (13.67 mm), but they exhibited a more dynamic antibacterial activity level against the Gram-negative P. aeruginosa.

DNA methylation plays important roles in the regulation of gene expression and maintenance of genome stability in many organisms, including plants. In this study, we treated rice with gamma rays (GRs) and DNA methyltransferase inhibitors (DNMTis) to induce variations in DNA methylation and evaluated epigenetic diversity using methylation-sensitive amplified polymorphism (MSAP) and transposon methylation display (TMD) marker systems. Comparative and integrated analyses of the data revealed that both GRs and DNMTis alone have epimutagenic effects and that combined treatment enhanced these effects. Calculation of methylation rates based on band scoring suggested that both GRs and DNMTis induce epigenetic diversity by demethylation in a dose-dependent manner, and combined treatment can induce variations more synergistically. The difference in the changes in full and hemi-methylation rates between MSAP and TMD is presumed to be caused by the different genomic contexts of the loci amplified in the two marker systems. Principal coordinate, phylogenic, and population structure analyses commonly yielded two clusters of individuals divided by DNMTi treatment. The clustering pattern was more apparent in TMD, indicating that DNMTis have a stronger effect on hypermethylated repetitive regions. These findings provide a foundation for understanding epigenetic variations induced by GRs and DNMTis and for epigenetic mutation breeding.

Kenaf is a source of fiber and a bioenergy crop that is considered to be a third world crop. Recently, a new kenaf cultivar, "Jangdae," was developed by gamma irradiation. It exhibited distinguishable characteristics such as higher biomass, higher seed yield, and earlier flowering than the wild type. We sequenced and analyzed the transcriptome of apical leaf and stem using Pacific Biosciences single-molecule long-read isoform sequencing platform. De novo assembly yielded 26,822 full-length transcripts with a total length of 59 Mbp. Sequence similarity against protein sequence allowed the functional annotation of 11,370 unigenes. Among them, 10,100 unigenes were assigned gene ontology terms, the majority of which were associated with the metabolic and cellular process. The Kyoto encyclopedia of genes and genomes (KEGG) analysis mapped 8875 of the annotated unigenes to 149 metabolic pathways. We also identified the majority of putative genes involved in cellulose and lignin-biosynthesis. We further evaluated the expression pattern in eight gene families involved in lignin-biosynthesis at different growth stages. In this study, appropriate biotechnological approaches using the information obtained for these putative genes will help to modify the desirable content traits in mutants. The transcriptome data can be used as a reference dataset and provide a resource for molecular genetic studies in kenaf.

Ionizing radiation combined with in vitro tissue culture has been used for development of new cultivars in diverse crops. The effects of ionizing radiation on mutation induction have been analyzed on several orchid species, including Cymbidium. Limited information is available on the comparison of mutation frequency and spectrum based on phenotypes in Cymbidium species. In addition, the stability of induced chimera mutants in Cymbidium is unknown. In this study, we analyzed the radiation sensitivity, mutation frequency, and spectrum of mutants induced by diverse γ-ray treatments, and analyzed the stability of induced chimera mutants in the Cymbidium hybrid cultivars RB003 and RB012. The optimal γ-irradiation conditions of each cultivar differed as follows: RB003, mutation frequency of 4.06% (under 35 Gy/4 h); RB012, 1.51% (20 Gy/1 h). Re-irradiation of γ-rays broadened the mutation spectrum observed in RB012. The stability of leaf-color chimera mutants was higher than that of leaf-shape chimeras, and stability was dependent on the chimera type and location of a mutation in the cell layers of the shoot apical meristem. These results indicated that short-term γ-irradiation was more effective to induce mutations in Cymbidium. Information on the stability of chimera mutants will be useful for mutation breeding of diverse ornamental plants.

The effect of salinity (NaCl treatment) on the nutritive value of wheat sprouts was investigated by analyzing the expression of phenylpropanoid biosynthetic pathway genes and the levels of phenylpropanoid compounds. Treatment with various concentrations of NaCl (50, 100, and 200 mM) resulted in increased epicatechin levels but decreased accumulation of catechin hydrate, benzoic acid, and quercetin compounds in the sprouts compared with the control (0 mM). The trans-cinnamic acid, 4-hydroxybenzoic acid, ferulic acid, epicatechin, and total phenylpropanoid level in wheat sprout was the highest at 50 mM of NaCl treatment. Six-day-old wheat plantlets exposed to 50 mM NaCl for 6, 12, 24, 48, and 72 h, showed that the total phenylpropanoids accumulation was the highest at 48 h after the treatment and most of the treatments showed higher phenylpropanoid content than the control at the same time points. Although the shoot and root length and the fresh weight of wheat sprouts decreased with NaCl treatment, these results suggest that treatment of 50 mM NaCl improves the nutritional quality of wheat sprouts, due to increased phenylpropanoid concentrations.

Mutation breeding is useful for improving agronomic characteristics of various crops. In this study, we conducted a genetic diversity and association analysis of soybean mutants to assess elite mutant lines. On the basis of phenotypic traits, we chose 208 soybean mutants as a mutant diversity pool (MDP). We then investigated the genetic diversity and inter-relationships of these MDP lines using target region amplification polymorphism (TRAP) markers. Among the different TRAP primer combinations, polymorphism levels and polymorphism information content (PIC) values averaged 59.71% and 0.15, respectively. Dendrogram and population structure analyses divided the MDP lines into four major groups. According to an analysis of molecular variance (AMOVA), the percentage of inter-population variation among mutants was 11.320 (20.6%), whereas mutant intra-population variation ranged from 0.231 (0.4%) to 14.324 (26.1%). Overall, intra-population genetic similarity was higher than that of inter-populations. In an analysis of the association between TRAP markers and agronomic traits using three different statistical approaches based on the single factor analysis (SFA), the Q general linear model (GLM), and the mixed linear model (Q+K MLM), we detected six significant marker−trait associations involving five phenotypic traits. Our results suggest that the MDP has great potential for soybean genetic resources and that TRAP markers are useful for the selection of soybean mutants for soybean mutation breeding.

Salinity stress affects plants by reducing the water potential and causing ion imbalance or disturbances in ion homeostasis and toxicity. Salinity stress frequently causes both osmotic and ionic stress in plants, resulting in the increase or decrease of certain secondary metabolites in plants. In this study, the effect of NaCl treatment on the nutritional quality of tartary buckwheat plants was studied by conducting an HPLC analysis of phenylpropanoid and anthocyanin content. It was observed that there was no significant change of color in tartary buckwheat during salt treatment. The accumulation of most phenylpropanoid compounds increased slightly in response to the NaCl concentration. The total phenylpropanoid content in tartary buckwheat was the highest at 100 mM NaCl treatment. Seven-day-old wheat plantlets treated with 100 mM NaCl for 2, 4, 6, and 8 days showed the highest accumulation of total phenylpropanoids at day 8 after treatment, while the content of most phenylpropanoids was higher than that in the control during this period. Although the development of tartary buckwheat slightly decreased with NaCl treatment and the accumulation of anthocyanin compounds did not change in plants with a diffident NaCl concentration and time treatment, the results suggest that the salinity treatment of tartary buckwheat causes antioxidant activity improvement by inducing an accumulation of flavonoid and phenolic compounds. However, since the anthocyanin content did not increase, the antioxidant effect of the treatment is not expected to be significant.

Polygalacturonase (PG) gene is a typical gene family present in eukaryotes. Forty-nine PGs were mined from the genomes of Neurospora crassa and five Aspergillus species. The PGs were classified into 3 clades such as clade 1 for rhamno-PGs, clade 2 for exo-PGs and clade 3 for exo- and endo-PGs, which were further grouped into 13 sub-clades based on the polypeptide sequence similarity. In gene structure analysis, a total of 124 introns were present in 44 genes and five genes lacked introns to give an average of 2.5 introns per gene. Intron phase distribution was 64.5% for phase 0, 21.8% for phase 1, and 13.7% for phase 2, respectively. The introns varied in their sequences and their lengths ranged from 20 bp to 424 bp with an average of 65.9 bp, which is approximately half the size of introns in other fungal genes. There were 29 homologous intron blocks and 26 of those were sub-clade specific. Intron losses were counted in 18 introns in which no obvious phase preference for intron loss was observed. Eighteen introns were placed at novel positions, which is considerably higher than those of plant PGs. In an evolutionary sense both intron loss and gain must have taken place for shaping the current PGs in these fungi. Together with the small intron size, low conservation of homologous intron blocks and higher number of novel introns, PGs of fungal species seem to have recently undergone highly dynamic evolution.