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The leaves of roselle (Hibiscus sabdariffa L.) have been used as a traditional folk medicine that has diuretic and mild laxative effects. Roselle is cultivated in many countries for medicines and food. However, studies on the variation of functional compounds in different accessions are relatively limited. In this study, we investigated the phenolic compound content of leaf extracts from 49 different roselle accessions from a worldwide collection by ultra-high performance liquid chromatography mass spectrometry. The aim was to chemically classify and assess the pharmacological properties of the phenolic compounds in the leaves. The phenolic compounds were identified as neochlorogenic acid, cryptochlorogenic acid, rutin, rutin isomer, isoquercitin, kaempferol-3-O-rutinoside, kaempferol-3-O-glucoside, quercetin, quercetin isomer, and kaempferol. These compounds were present at significantly different levels among the accessions. The most abundant phenolic compound was rutin, which was found in 45 accessions where the total phenolic compound content was 18.75-46.51 mg/g (average 31.52 mg/g). The phenolic compounds were classified into four groups. The two accessions with the highest total phenolic compound content were from Ghana (PI286316) and Senegal (PI275413). The hierarchical cluster analysis of the 49 roselle accessions showed that they formed five groups according to their phenolic compound content. Our results will be useful for the selection of roselle genotypes with improved functional compounds.

Kenaf is a valuable fiber and medicinal crop. Here, we performed RNA-sequencing to obtain comprehensive transcriptome information and identify putative genes involved in biosynthetic pathways of anthocyanins and kaempferitrin in kenaf. A total of 39.6 Gb reads were generated for six kenaf accessions. Individual de novo assembly of each accession and clustering of whole transcriptome contigs finally yielded 299,880 representative transcripts having an average length of 1217 bp, of which 231,825 (77.3%) were annotated against various databases. Expression profiling of the transcripts identified 6,592 differentially expressed genes (DEGs) among the three accessions, C-14 (a local landrace) and two of its mutant varieties, Jeokbong (purple leaves and stems) and Baekma (light green leaves and stems). In addition to their altered coloration, leaves of these two mutants had significantly different anthocyanin and kaempferitrin contents. We constructed anthocyanin and kaempferitrin biosynthetic pathways and identified 671 transcripts mapping to the entire pathway. In addition, 29 DEGs assigned to eight structural genes and 41 DEGs related to three transcription factor families were identified in high anthocyanin- and kaempferitrin-accumulating mutant Jeokbong. Our results provide a large transcriptome library pool for understanding gene functions and should also be useful in further studies of flavonoid biosynthesis in kenaf.

Protons may have contributed to the evolution of plants as a major component of cosmic-rays and also have been used for mutagenesis in plants. Although the mutagenic effect of protons has been well-characterized in animals, no comprehensive phenotypic and genomic analyses has been reported in plants. Here, we investigated the phenotypes and whole genome sequences of Arabidopsis M2 lines derived by irradiation with proton beams and gamma-rays, to determine unique characteristics of proton beams in mutagenesis. We found that mutation frequency was dependent on the irradiation doses of both proton beams and gamma-rays. On the basis of the relationship between survival and mutation rates, we hypothesized that there may be a mutation rate threshold for survived individuals after irradiation. There were no significant differences between the total mutation rates in groups derived using proton beam or gamma-ray irradiation at doses that had similar impacts on survival rate. However, proton beam irradiation resulted in a broader mutant phenotype spectrum than gamma-ray irradiation, and proton beams generated more DNA structural variations (SVs) than gamma-rays. The most frequent SV was inversion. Most of the inversion junctions contained sequences with microhomology and were associated with the deletion of only a few nucleotides, which implies that preferential use of microhomology in non-homologous end joining was likely to be responsible for the SVs. These results show that protons, as particles with low linear energy transfer (LET), have unique characteristics in mutagenesis that partially overlap with those of low-LET gamma-rays and high-LET heavy ions in different respects.

Terpenes constitute a large class of secondary metabolites in plants. The Oryza sativa terpene synthase is a vital gene in plant defense response. In this study, the molecular and physiological functions of Oryza sativa terpene synthase 30 (OsTPS30, LOC_Os08g07080) were investigated after exposure of the seeds and plants to gamma-rays. The OsTPS30 expression was slightly induced at 200 Gray (Gy), but was significantly induced at 400 Gy. The total terpenoid was synthesized more in OsTPS30- overexpressing (OX-OsTPS30) Arabidopsis thaliana plants than in wild-type (WT) plants. The OX-OsTPS30 plants exhibited resistance to gamma-rays, as compared to WT. The OX-OsTPS30 plants had significantly increased height and weight after gamma irradiation. Additionally, the activity of antioxidant enzymes was increased more in OX OsTPS30 plants than in WT plants after gamma irradiation. Furthermore, the OsTPS30-GFP fusion protein was mostly localized in the chloroplast, suggesting that OsTPS30 is putative MEP pathway-related terpene synthase.

Purpose: Gamma-rays and carbon ions are frequently used for mutation breeding in diverse plant species, whereas proton ions have been rarely used for this purpose. This study assessed the p...

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.

Roses are one of the most important floricultural crops, and their essential oils have long been used for cosmetics and aromatherapy. We investigated the volatile compound compositions of 12 flower-color mutant variants and their original cultivars. Twelve rose mutant genotypes were developed by treatment with 70 Gy of 60Co gamma irradiation of six commercial rose cultivars. Essential oils from the flowers of the 18 genotypes were analyzed by gas chromatography&ndash, mass spectrometry. Seventy-seven volatile compounds were detected, which were categorized into six classes: Aliphatic hydrocarbons, aliphatic alcohols, aliphatic ester, aromatic compounds, terpene alcohols, and others. Aliphatic (hydrocarbons, alcohols, and esters) compounds were abundant categories in all rose flowers. The CR-S2 mutant had the highest terpene alcohols and oil content. Three (CR-S1, CR-S3, and CR-S4) mutant genotypes showed higher ester contents than their original cultivar. Nonacosane, 2-methylhexacosane, and 2-methyltricosane were major volatile compounds among all genotypes. Hierarchical cluster analysis (HCA) of the rose genotypes gave four groups according to grouping among the 77 volatile compounds. In addition, the principal component analysis (PCA) model was successfully applied to distinguish most attractive rose lines. These findings will be useful for the selection of rose genotypes with improved volatile compounds.

Orchids with colorful leaves and flowers have significant ornamental value. Here, we used &gamma, irradiation-based mutagenesis to produce a Dendrobium bigibbum mutant that developed purple instead of the normal green leaves. RNA sequencing of the mutant plant identified 2513 differentially expressed genes, including 1870 up- and 706 downregulated genes. The purple leaf color of mutant leaves was associated with increased expression of genes that encoded key biosynthetic enzymes in the anthocyanin biosynthetic pathway. In addition, the mutant leaves also showed increased expression of several families of transcription factors including the MYB2 gene. Transient overexpression of D. biggibumMYB2 in Nicotiana benthamiana was associated with increased expression of endogenous anthocyanin biosynthesis genes. Interestingly, transient overexpression of orthologous MYB2 genes from other orchids did not upregulate expression of endogenous anthocyanin biosynthesis genes. Together, these results suggest that the purple coloration of D. biggibum leaves is at least associated with increased expression of the MYB2 gene, and the MYB2 orthologs from orchids likely function differently, regardless of their high level of similarity.

Roses are one of the most important floricultural crops, and their essential oils have long been used for cosmetics and aromatherapy. We investigated the volatile compound compositions of 12 flower-color mutant variants and their original cultivars. Twelve rose mutant genotypes were developed by treatment with 70 Gy of 60Co gamma irradiation of six commercial rose cultivars. Essential oils from the flowers of the 18 genotypes were analyzed by gas chromatography–mass spectrometry. Seventy-seven volatile compounds were detected, which were categorized into five classes: hydrocarbons, terpenoids, alcohols, esters, and others. Hydrocarbons, alcohols, and esters were major components in all rose flowers. The mutant genotypes CR-S8 and CR-S9 showed higher contents of hydrocarbons than the original cultivar. In addition, CR-S1, CR-S3, and CR-S4 mutant genotypes showed higher ester contents than their original cultivar. Nonacosane, 2-methylhexacosane, and 2-methyltricosane were major volatile compounds among all genotypes. Hierarchical cluster analysis of the rose genotypes gave four groups according to grouping among the 77 volatile compounds. These findings will be useful for the selection of rose genotypes with improved volatile compounds.

Calnexin (CNX) is an integral membrane protein that functions as a chaperone in the endoplasmic reticulum for the correct folding of proteins under stress conditions, rendering organisms tolerant under adverse conditions. Studies have investigated the cytogenetic effects of gamma irradiation (Ɣ-IR) on plants, but information on the molecular response under Ɣ-IR remains limited. Previously, we constructed a cDNA library of an irradiation-sensitive bioindicator plant, Tradescantia BNL4430 (T-4430) under Ɣ-IR, in which the Calnexin-1 gene was highly upregulated at 50 mGy treatment. TrCNX1 encodes a 61.4 kDa protein with conserved signature motifs similar to already reported CNX1s. TrCNX1 expression was evaluated by semiquantitative reverse transcriptase PCR and quantitative real-time PCR and was ubiquitously expressed in various tissues and highly upregulated in flower petals under 50 mGy Ɣ-IR stress. The protective function of TrCNX1 was investigated by overexpression of TrCNX1 in an Escherichia coli BL21(DE3) heterologous system. Using plate assay, we showed that TrCNX1 increased the viability of E. coli transformants under both UV-B and Ɣ-IR compared with the control, demonstrating that TrCNX1 functions under irradiation stress. TrCNX1 may enhance irradiation stress tolerance in crops and act as a radio marker gene to monitor the effects of radiation.

Kenaf is an economically important crop that contains various functional compounds. The objective of this study was to evaluate the nutritional properties of leaves, stem bark, flowers, and seeds collected from three mutant kenaf cultivars (Jangdae, Baekma, and Jeokbong) and two original cultivars (Jinju and C14). For all analyzed tissues, there were no significant differences in the proximate compositions (moisture, crude protein, crude fiber, crude fat, ash, and nitrogen-free extract) among the tested genotypes, except for the leaf crude protein content, which was highest in Jinju plants. Of the tested minerals, potassium and calcium were the most abundant in all tissues. Additionally, we detected significant differences in the mineral contents (e.g., calcium, phosphorus, potassium, magnesium, zinc, and iron) of various tissues in all cultivars. Proline and phenylalanine were the major amino acids detected in leaves, and the highest total amino acid (TAA) and essential amino acid (EAA) contents were observed in Jinju plants. The TAA and EAA contents were lower in stem bark than in the other tissues, except in Baekma plants, in which the lowest levels were observed in flowers. Moreover, the TAA and EAA contents were approximately 20 times lower in white flowers (Baekma) than in ivory flowers. Furthermore, the highest TAA and EAA concentrations were observed in Jangdae seeds. These results may be useful for identifying the optimal cultivar and tissues for use in food products.

We aimed to investigate the biological responses induced by acute and chronic gamma irradiation in colored wheat seeds rich in natural antioxidants. After acute and chronic irradiation, the phenotypic effects on plant growth, germination rate, seedling height, and root length were examined, and the biochemical changes were investigated by analyzing the expression of antioxidant enzyme-related genes, antioxidant enzyme activities, and total antioxidant capacity. High dosages of chronic radiation reduced plant growth compared with the controls. Electron spin resonance measurement and 2,2-diphenyl-1-picrylhydrazyl activity analysis showed lower amount of free radicals in colored wheat seeds on chronic irradiation with low dosage of gamma rays compared to seeds subjected to acute irradiation. Expression levels of anthocyanin biosynthesis genes, antioxidant-related genes, and antioxidant enzyme activity in seeds and young leaves of seedling showed diverse effects in response to different dosages and types of gamma irradiation. This suggests that phenotype is affected by the dosage and type of gamma radiation, and the phytochemicals in colored wheat seeds involved in antioxidant activity to scavenge free radicals respond differently to irradiation types. This provides evidence that acute and chronic exposure to radiation have different effects on seeds and young leaves after germination.

The present study was conducted to evaluate the biological responses of soybean irradiated by gamma-rays. Four elite Korean soybean cultivars, Kwangankong, Shinpaldal-2, Shinhwakong, and Ilmikong, were irradiated with 100-400 Gy of gamma-rays. All cultivars showed significant reductions in morphological parameters. Seedling emergence rates of Kwangankong, Shinpaldal-2, Shinhwakong, and Ilmikong decreased by 46%, 31%, 44%, and 43% at 400 Gy, respectively. Plant height and fresh weight decreased with increasing dose. The median reduction dose (RD50) for plant height ranged from 184 to 278 Gy with an average of 212 Gy.The optimal dose of gamma irradiation for inducing mutation in the four elite soybean cultivars was in the range 200-300 Gy. Contents of malonaldehyde (MDA) in the gamma-ray irradiated plants were higher than those in the controls. With the exception of Kwangankong, activities of ascorbate peroxidase (APX) in other cultivars decreased at 100 Gy and started to increase significantly at 200 Gy and Shinhwakong cultivar had highest APX value observed at 300 Gy. No significant changes in peroxidase (POD) activity were observed inthe Kwangankong, Shinpaldal-2, and Shinhwakong, while, POD activity increased with increasing dose in Ilmikong. In addition, gamma-ray treatments elicited a marked reduction in chlorophyll a contents compared to chlorophyll b contents in Kwangankong, Shinpaldal-2, and Ilmikong. Growth characteristics (plant height and fresh weight) were highly negatively (P ＜ 0.01) correlated with the dose of gamma-rays, while the MDA content and APX activities were positively (P ＜ 0.05) correlated with dose.

The kenaf plant is used widely as food and in traditional folk medicine. This study evaluated the morphological characteristics, functional compounds, and genetic diversity of 32 kenaf cultivars from a worldwide collection. We found significant differences in the functional compounds of leaves from all cultivars, including differences in levels of chlorogenic acid isomer (CAI), chlorogenic acid (CA), kaempferol glucosyl rhamnoside isomer (KGRI), kaempferol rhamnosyl xyloside (KRX), kaemperitrin (KAPT) and total phenols (TPC). The highest TPC, KAPT, CA, and KRX contents were observed in the C22 cultivars. A significant correlation was observed between flowering time and DM yield, seed yield, and four phenolic compounds (KGRI, KRX, CAI, and TPC) (P < 0.01). To assess genetic diversity, we used 80 simple sequence repeats (SSR) primer sets and identified 225 polymorphic loci in the kenaf cultivars. The polymorphism information content and genetic diversity values ranged from 0.11 to 0.79 and 12 to 0.83, with average values of 0.39 and 0.43, respectively. The cluster analysis of the SSR markers showed that the kenaf genotypes could be clearly divided into three clusters based on flowering time. Correlations analysis was conducted for the 80 SSR markers; morphological, chemical and growth traits were found for 15 marker traits (corolla, vein, petal, leaf, stem color, leaf shape, and KGRI content) with significant marker-trait correlations. These results could be used for the selection of kenaf cultivars with improved yield and functional compounds.

The faba bean, one of the few crops that produces L-3,4-dihydroxy-phenylalanine (L-dopa) naturally, is also a good source of fatty acids and isoflavones. This study analyzed fatty acids, L-dopa and isoflavones in the leaves, immature pods and seeds of selected high-yielding faba bean genotypes spring-sown in Korea. The most abundant fatty acids in the leaves, immature pods and seeds were linolenic acid (58.6-63.7%), linoleic acid (46.9-58.9%), and linoleic acid (45.7-63.6%), respectively. The L-dopa content was highest in the leaf for all the faba bean genotypes except PI430715 and PI614810 where it was highest in the immature pods. The α-linolenic acid and L-dopa contents of the seed and immature pod of the PI430715 and PI614810 genotypes were the highest of all the genotypes. Four isoflavones (daidzin, genistin, daidzein and genistein) were detected with levels of daidzin and genistin generally higher than those of daidzein and genistein in each part of the faba bean. The L-dopa content was correlated (P ≤ 0.05) with those of five fatty acids (oleic, gondoic, phytanic, α-linolenic, and behenic acid). Overall, the PI430715 and PI614810 faba bean genotypes with their higher L-dopa and α-linolenic acid contents were the most functional. This study may help identify optimal genotypes for breeding new cultivars which provide functional compounds to treat Parkinson’s disease or use in foods.

The flowers of Chrysanthemum morifolium Ramat. have been used as an herbal tea and in traditional medicine, and the plant has been developed to produce horticultural cultivars of various colors and shapes. In this study, a new chrysanthemum cultivar with dark purple petals (C. morifolium cv. ARTI-Dark Chocolate; ADC) was developed by radiation-induced mutation breeding of its original cultivar with purple striped white petals (C. morifolium cv. Noble Wine, NW). The phenolic profile and antioxidant property of ADC were investigated and compared with NW and the commercially available medicinal herb, C. morifolium with yellow petals (CM), in order to find a scientific support to produce a new source of natural antioxidant. Flavonoid and phenolic acid profiles of the ethanol extracts of the three flowers were analyzed by high-performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry (HPLC-DAD-ESIMS), while antioxidant properties were evaluated using the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging assay. Among the tested flowers, ADC possessed the strongest antioxidant capacity and the highest phenolic contents. Flavonoids (acacetin, apigenin, luteolin, acacetin-7-O-β-glucoside, apigenin-7-O-β-glucoside, luteolin-7-O-β-glucoside, and linarin) and phenolic acids (chlorogenic acid and mixture of 1,4-, 1,5-, and 3,5-dicaffeoylquinic acids) were identified and quantified.

In addition to attracting pollinators and protecting plants from photooxidative damages, carotenoids play important roles in signaling related to plant development and physiological changes through their cleaved derivatives, apocarotenoids. Yellow chrysanthemum flower petals [ Chrysanthemum morifolium (Ramat.)] are the result of a deleted or suppressed carotenoid cleavage dioxygenase gene ( CmCCD4a ). Therefore, investigating the global gene expression patterns in chrysanthemum mutants producing yellow petals represents an appropriate strategy to analyze the effects of signaling mediated by carotenoid derivatives. We compared the transcriptomes of chrysanthemum cultivars producing white petals with purple stripes (‘Noble Wine’) and its mutant producing yellow petals with purple stripes (‘ARTI-Yellow Star’). Read mapping and polymerase chain reaction analysis revealed that CmCCD4a is absent in the ARTI-Yellow Star genome, while at least three CmCCD4a gene family members are actively transcribed in ‘Noble Wine’ plants. These results indicate that the deletion of CmCCD4a may be responsible for the development of yellow petals in ARTI-Yellow Star flowers. During comparisons of gene expression patterns between Noble Wine and ARTI-Yellow Star, we detected numerous significantly up-regulated plastid protein-coding genes as well as significantly or mildly up-regulated carotenoid biosynthesis genes in ARTI-Yellow Star plants. The up-regulated plastid protein-coding genes belong to a broad range of functional categories related to translation, photosynthesis, reactive oxygen species scavenging, oxidative phosphorylation, transcription, processing, terpenoid metabolism, and chlorophyll metabolism. Our results present a possible relationship between CmCCD4a and regulation of plastid protein-coding genes.

Stearic acid is the most important saturated fatty acid compound in soybean oil. A typical soybean cultivar has 2–3% stearic acid content of total oil. Recently, a new high stearic acid mutant Hfa180 was developed from gamma-ray irradiation with “Ilmikong.” The objectives of this study were to identify the mutant gene associated with high stearic acid content in Hfa180 and to develop a Kompetitive Allele-Specific PCR (KASP) assay for selection of soybeans with elevated levels of stearic acid in seeds. Target sequencing of the whole GmSACPD-C gene encoding D9-stearoyl-ACP-desaturase revealed that Hfa180 contained a three-base-pair deletion (GAG) at genomic positions 190 to 192 of the 1st exon of GmSACPD-C. The three-base deletion caused a variant with a single amino acid (glutamine) deletion at position 64 in the predicted GmSACPD-C amino acid in Hfa180, as well as an average 60% reduction in GmSACPD-C gene expression in Hfa180 compared to that of the wild type. Genetic linkage analysis using the F2 population derived from a crossing between Hfa180 and the wild-type “Deapung” confirmed that the mutation allele of GmSACPD-C was perfectly correlated with elevation of seed stearic acid content. In addition, a KASP marker was developed to identify the mutation allele on the GmSACPD-C gene, which was successfully applied to select high stearic acid soybean lines. The identification of the mutant gene of the high stearic acid mutant Hfa180 and development of the KASP marker could be useful for improving stearic acid content in soybean oil.

Blackberries (Rubus fruticosus L.) are a good source of antioxidants and contain appreciable levels of functional compounds. This study was carried out to evaluate fruit qualities (pH, soluble solids content, titratable acidity, and minerals), functional compounds (total phenolic content, total flavonoid content, anthocyanins, and ellagic acid), and antioxidant activity among five mutated blackberry lines over three ripening stages to examine the effect of fruit maturation on functional compounds and antioxidant activity. The content of sugars, total phenolic content, total flavonoid content, and anthocyanins (cyanidin-3-O-glucoside, cyanidin-3-O-xyloside, cyanidin-3-O-malonylglucoside, and cyanidin-3-O-dioxalylglucoside) increased as fruit development advanced, whereas the ellagic acid content decreased as the fruit became more mature. Among the minerals tested, potassium showed the highest concentrations at all stages of development. The antioxidant activity was correlated with total phenolic content (P≤0.01), total flavonoid content (P≤0.01), and total anthocyanins (P≤0.05). The results indicate that Maple and B201 lines contained the highest anthocyanin content and exhibited greater antioxidant activity.

Kenaf (Hibiscus cannabinus L.) is an annual herbaceous crop and the third largest fiber crop of economic importance, after cotton and jute. In view of recent global environmental challenges, multipurpose uses of kenaf have been developed, such as animal feed, bio-plastics, bio-fuels, and phytoremediation. We cloned genes related to monolignol biosynthesis using a reverse transcriptase PCR (RT-PCR) strategy. ‘Auxu’ cultivars in the seedling phase were used as the experimental source material. The 4-coumarate:CoAligase (4CL), cinnamoyl-CoA reductase (CCR), ferulate 5-hydroxylase (F5H), caffeic acid O-methyltransferase (COMT), cinnamyl alcohol dehydrogenase (CAD), encoding hydroxycinnamoyl transferase (HCT), ρ-coumarate 3-hydroxylase (C3H) and caffeoyl-CoA-O-methyltransferase (CCoAOMT), cDNAs had open reading frames of 1623, 1017, 1179, 921, 1107, 1323, 1527 and 744 bp, encoding polypeptides of 540, 338, 392, 306, 368, 441, 508 and 247 amino acid residues, respectively. We submitted the five gene sequences (4CL, CCR, F5H, COMT, and CAD) to the GenBank database. A phylogenetic tree showed that lignin biosynthesis genes had the closest relationships with Gossypium and Theobroma cacao. Expression levels of the genes 4CL, HCT, C3H, CCoAOMT, CCR, F5H, COMT, and CAD in stems from plants harvested at four different growth stages were evaluated. Expression levels of 4CL, HCT, C3H, CCoAOMT, CCR, F5H, COMT, and CAD were gradually increased during the vegetative growth of kenaf stem. Similarly with the gene expression, lignin amounts were also increased during the stem growth. Thus, these six genes may be directly involved in lignin biosynthesis. These results will be useful basic data for molecular breeding to enhance bio-mass production.

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&ndash, 84.02%) and a relatively low activity group with green leaf varieties (57.47&ndash, 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 &lt, 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.

Radiation randomly induces chromosomal mutations in plants. However, it was recently found that the frequency of flower-color mutants could be specifically increased by upregulating anthocyanin pathway gene expression before radiation treatments. The mechanisms of chlorophyll biosynthesis and degradation are active areas of plant study because chlorophyll metabolism is closely connected to photosynthesis. In this study, we determined the dark/light treatment conditions that resulted in upregulation of the expression levels of six chlorophyll pathway genes, uroporphyrinogen III synthase (HEMD), uroporphyrinogen III decarboxylase (HEME2), NADPH-protochlorophyllide oxidoreductase (POR) A (PORA), chlorophyll synthase (CHLG), chlorophyllase (CLH2), and red chlorophyll catabolite reductase (RCCR), and measured their effects on the γ-irradiation-induced frequencies of leaf-color mutants in two Cymbidium cultivars. To degrade chlorophyll in rhizomes, 60–75 days of dark treatment were required. To upregulate the expressions of chlorophyll pathway genes, 10 days of light treatment appeared to be optimal. Dark/light treatments followed by γ-irradiation increased chlorophyll-related leaf mutants by 1.4- to 2.0-fold compared with γ-ray treatment alone. Dark/light treatments combined with γ-irradiation increased the frequency of leaf-color mutants in Cymbidium, which supports the wider implementation of a plant breeding methodology that increases the mutation frequency of a target trait by controlling the expression of target trait-related genes.

γ-rays have been used to develop many mutations in diverse ornamental plants, including chrysanthemum, of which most changes have been flower color. Flower color is determined by plant pigments, including anthocyanins, which are affected by mutation of genes in the anthocyanin biosynthesis pathway. Sucrose and plant hormones also crosstalk in complex signaling pathways and modulate anthocyanin biosynthesis. In this study, we determined the optimal sucrose and plant hormone treatments for expression of anthocyanin biosynthesis genes and compared the mutation frequency among sucrose, methyl jasmonate (MeJA), and/or γ-ray treatments in chrysanthemum. Anthocyanin biosynthesis genes were highly expressed with 50 mM sucrose treatment for 18 h. The highest expression levels of anthocyanin biosynthesis genes were observed under sucrose with MeJA treatment followed by MeJA treatment alone. Sucrose with abscisic acid (ABA) treatment, and ABA or GA3 treatment alone, had no effect on expression of anthocyanin biosynthesis genes. Sucrose with MeJA pre-treatment followed by γ-irradiation resulted in the highest mutation frequency (14.5%) among all treatments and a 1.5-fold increase in mutation frequency compared with γ-ray treatment alone. The ratio of anthocyanin-related flower-color mutants with sucrose and/or MeJA pre-treatments followed by γ-irradiation was higher than that with γ-ray treatment alone. Sucrose and/or MeJA modulated the expression of anthocyanin biosynthesis genes and highly expressed genes may be affected more readily by γ-rays, thus resulting in an increased frequency of anthocyanin-related flower-color mutants in chrysanthemum.

Apios americana, a leguminous plant, is used as food in some countries. Although the biological activities of Apios extract have been reported, there have been no reports about the anti-inflammatory mechanism of lupinalbin A on the RAW264.7 cells. In this study, we investigated the anti-inflammatory effect of A. americana lupinalbin A on lipopolysaccharide (LPS)-treated RAW264.7 cells. Lupinalbin A significantly inhibited nitric oxide production and inducible nitric oxide synthase expression in LPS-treated RAW264.7 cells. The expression of cytokines, including interleukin-6, tumor necrosis factor-α, and chemokine of monocyte chemoattractant protein, was reduced under lupinalbin A exposure in LPS-treated RAW264.7 cells. In addition, lupinalbin A significantly decreased LPS-induced interferon (IFN)-β production and STAT1 protein levels in RAW264.7 cells. Taken together, these results suggest that A. americana lupinalbin A exerts anti-inflammatory effects via the inhibition of pro-inflammatory cytokines and blocking of IFN-β/STAT1 pathway activation.

Anthocyanin derivatives are well-known secondary constituents contained in fruits. The inhibitory activity of anthocyanin derivatives toward soluble epoxide hydrolase (sEH) was tested for potential applications in the treatment of cardiovascular diseases. Anthocyanin derivatives 1-5 showed dose-dependent inhibitory activity toward sEH, with IC50 values ranging from 4.3±0.2 to 25.3±2.6μM. Lineweaver-Burk plots showed that all anthocyanin derivatives preferentially interacted with allosteric sites instead of active sites as noncompetitive (1-3) and mixed (4 and 5) inhibitors. Furthermore, the cavity located next to the active site may interact with anthocyanin derivatives (1-5) by molecular docking. Among the tested derivatives, (4) bonded with key amino acids at two loops around the binding site for 10ns. Finally, anthocyanin derivatives (1-5) are potential inhibitors of sEH, and anthocyanin-rich fruits may be useful for the targeted treatment of cardiovascular diseases via sEH inhibition.

The goal of this study was to identify phytochemicals with inhibitory activity against tyrosinase. Nine compounds 1-9 were isolated from the tubers of Apios americana. This is the first report of aromadendrin 5-methyl ether (1) being isolated from the Apios species. Among them, compounds 2 and 8 showed inhibitory activity toward tyrosinase. Based on a Dixon plot, the potential Ki values of competitive inhibitors 2 and 8 were calculated as 10.3 ± 0.8 µM and 44.2 ± 1.7 µM, respectively. An IC50 value of 13.2 ± 1.0 µM was calculated for the slow-binding inhibitor 2 after preincubation with tyrosinase. Additionally, the predicted binding sites between the receptor and ligand, as well as secondary structure changes, in the presence of 2 were examined by molecular simulation.

In this study, the flavonoid and isoflavone contents in four checks with seed coat colour altered from yellow to black were determined, and their antioxidant activity was measured. Three anthocyanins (delphinidin-3-O-β-D-glucoside, D3G; cyanidin-3-O-β-D-glucoside, C3G; and petunidin-3-O-β-D-glucoside, Pt3G) were detected in the four checks, while their controls (cvs. ‘Danbaek’ and ‘Daepung’) were not found. Among them, D-16 derived from cv. ‘Danbaek’ showed the highest anthocyanin contents. The isoflavone contents of all checks showed lower levels compared to their controls. Among six isoflavones, glycitein was only detected in DP-39. To analyse the radical scavenging activities among the four checks, we conducted DPPH, FRAP and UWLA assays. Significant differences in antioxidant activities were obtainable in the four checks, which contained significant differences in levels of anthocyanins and isoflavones. Based on these results, gamma irradiation may change the isoflavone and anthocyanin contents of soybean. Our four checks selected in this study may be useful for breeding soybean varieties to alter their nutritional values.

Gamma-ray irradiation is an important tool in ornamental plant breeding, particularly to induce flower-color variation. Standard-type chrysanthemum ‘Migok’ and spray-type ‘Argus’ have seven and four gamma-ray mutants, respectively, with novel flower colors. Genetic relationships among 26 chrysanthemum varieties or mutants were analyzed using amplified fragment length polymorphisms (AFLPs) with 20 fluorescently-labeled E+3/M+3 primer combinations (PCs). Clustering analysis was carried out using similarity coefficients and unweighted pair group method with arithmetic averages (UPGMA). The 20 PCs produced 2,837 bands, 95.6% of which were polymorphic. E-ACC/M-CAG had the most polymorphic bands (199) and high polymorphic information content, marker index, and resolving power values. Similarity coefficients ranged from 0.63 to 0.97 overall but were 0.73–0.83 in the 11 radio-mutants. The 26 varieties and radio-mutants were divided into four major UPGMA groups; the 11 radio-mutants formed two subgroups and 10 standard-type varieties or radio-mutants were closely clustered into another. The most promising mutant-specific marker candidates were PC E-ACG/M-CAG (47.5%) and E-ACA/M-CAT (44%) for the ‘Migok’ and ‘Argus’ families, respectively.

Protective effects of extracts of the new cultivar MNU-32 developed from blackberry (Rubus fruticosus) cultivar V-9 against H2O2-induced oxidative stress in HepG2 cells were investigated. DPPH radical scavenging activities of MNU-32 and V-9 extracts were 570.00 μg/mL (IC50) and 644.55 μg/mL (IC50), respectively. Inhibition effects of MNU-32 extracts against intracellular ROS generation, cell cytotoxicity, and lactate dehydrogenase release were higher than for the V-9 extract in H2O2-treated HepG2 cells. Compared with the V-9 extract, MNU-32 extract treatments produced higher levels of lipid peroxidation attenuation and greater protection against DNA damage, restored the activity of superoxide dismutase in H2O2-treated HepG2 cells, and increased protein expression levels of superoxide dismutase in HepG2 cells in a dose and time-dependent manner. MNU-32 blackberry extracts exerted protective effects against H2O2-induced cell damage due to higher anthocyanin contents than the V-9 blackberry extract.

A new compound 1, 5-methoxy-2,5,7,4′-tetrahydroxy-coumaronochromone, along with seven known compounds (2–8), were isolated from Apios americana using open column chromatography. Their structures were established based on an analysis of 1D and 2D NMR, and MS spectra. Among these, two compounds 1 and 2 showed inhibitory activity on soluble epoxide hydrolase (sEH) at a concentration below 50 μM. The respective competitive (1) and mixed (2) inhibitors were revealed to have Ki values of 21.0 ± 0.8 and 14.5 ± 1.5 μM, based on the Dixon plot. The potential inhibitor (2) was visually presented in a predicted binding pose in the receptor by molecular docking. Additionally, molecular dynamics were performed for a detailed understanding of their complex by Gromacs 4.6.5 package.

The focus of this study was the mechanism of starch accumulation in Chlamydomonas reinhardtii high-starch mutants. Three C. reinhardtii mutants showing high-starch content were generated using gamma irradiation. When grown under nitrogen-deficient conditions, these mutants had more than twice as much starch than a wild-type control. The mechanism of starch over-accumulation in these mutants was studied with comparative transcriptome analysis. In all mutants, induction of phosphoglucomutase 1 (PGM1) expression was detected; PGM1 catalyzes the inter-conversion of glucose 1-phosphate and glucose 6-phosphate in both starch biosynthetic and glycolytic pathway. Interestingly, transcript levels of phosphoglucose isomerase 1 (PGI1), fructose 1,6-bisphosphate aldolase 1 and 2 (FBA1 and FBA2) were down-regulated in all mutants; PGI1, FBA1, and FBA2 act on downstream of glucose 6-phosphate conversion in glycolytic pathway. Therefore, down-regulations of PGI1, FBA1, and FBA2 may lead to accumulation of upstream metabolites, notably glucose 6-phosphate, resulting in induction of PGM1 expression through feed-forward regulation and that PGM1 overexpression caused starch over-accumulation in mutants. These results suggest that PGI1, FBA1, FBA2, and PGM1 correlate with each other in terms of coordinated transcriptional regulation and play central roles for starch over-accumulation in C. reinhardtii.

The protective effect of cyanidin-3-glucoside (C3G) against carbon tetrachloride (CCl4)-induced liver injury in mice was investigated. Administration of C3G attenuated the levels of serum aspartate aminotransferase, alanine aminotransferase, and liver lipid peroxidation in CCl4-treated mice. Histopathological examination of mouse livers showed that C3G reduced the incidence of liver lesions induced by CCl4. Moreover, C3G prevented DNA damage and decreased the protein levels of γ-H2AX in CCl4-treated mouse livers. C3G also increased the activity of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, which were decreased due to CCl4 administration in CCl4-treated mouse livers. C3G inhibited the expression levels of IL-6 and iNOS in CCl4-treated mice. C3G apparently protects the liver from CCl4-induced hepatic damage through antioxidant and anti-inflammatory mechanisms.

Induction of mutations using chemical mutagens has proved to be a useful tool in crop improvement and has advantages over transgenic approaches in view of legislative restrictions and intellectual property. Among the chemical mutagens, ethyl methanesulfonate (EMS) has been widely used to generate novel traits. In this study, we constructed an EMS mutant population consisting of 3945 M2 mutant lines using a Korean landrace of Capsicum annuum ‘Yuwol-cho’. In total, 1480 M2 mutant lines were evaluated for novel traits. The mutant lines showed phenotypic variations such as plant growth (small size and dwarfism), development of leaves (variegation, colour and morphological changes), flowers (inflorescence, morphological and organ colour changes), and fruits (morphological and colour changes). Most of these mutant phenotypes were inherited recessively. Many of the mutant phenotypes were unique in pepper, while some were similar to those of known mutants in other plant species. These mutant lines will be useful for the study of gene function in C. annuum.

This study was conducted to evaluate the growth characteristics and fatty acid composition among 15 kenaf mutants derived from the kenaf germplasm C14 and 15 kenaf accessions originating from Russia, India, China, Iran, and Italy. The overall growth performance (plant height, stem diameter, flowering date, leaf, and flower size) of the stem color mutant lines derived from C14 are similar to those of the original variety. However, the flower color mutant lines derived from C14 showed flowering to occur 10 days later when compared with the original variety and showed smaller leaf sizes than the original variety. Late-ripened kenaf accessions (Jinju, Auxu, and Jnagdae) can yield more bio-mass compared with early or medium-maturing germplasm. The late maturity kenaf (Auxu, Jinju, and Jangdae) has a higher oil percentage than the early maturity germplasm. Linoleic, oleic, and palmitic acids were the predominant fatty acids in all kenaf seeds. The stem color mutant lines significantly surpassed the parental means of all saturated fatty acids. In addition, the flower color mutant lines showed broad ranges of variation in oleic acid. The 15 accessions showed a wide range of fatty acid compositions, spanning from 29.75 to 38.30% saturated fatty acids and 61.70 to 70.24% total unsaturated fatty acids, and the late maturity kenaf has a higher linoleic acid percentage than the early maturity germplasm. The flowering period was highly positively (P ≤ 0.01) correlated with the plant height, stem diameter, oil percent, and linolenic acid (C18:3), and it was significantly negatively (P ≤ 0.01) correlated with stearic acid (C18:0). These results will provide valuable information to assist the parental selection of kenaf breeding.

The agronomic performance and storage protein patterns of 722 soybean landraces collected from eight geographically different Korean locations were investigated. The days to 50% flowering, days to maturity, and 100-seed weight ranged from 68.9 to 71.9 (d), 140.1 to 146.6 (d), and 22.4 to 26.8 (g), respectively. High-throughput protein profiling electrophoresis was performed, and the banding patterns were analyzed. Among the 722 soybean landraces, lipoxygenase bands were found to be absent in 21 lines. Nei’s gene diversity (h) ranged from 0 to 0.2642, with an average value of 0.1565. The mean coefficient of gene differentiation (Gst) was 0.0944, and the estimated gene flow (Nm) in the population was 4.7971. In a correlation matrix between the agronomic traits and protein banding patterns, the acidic banding pattern was significantly associated with all the other factors. The phenotypic and genotypic differences of the collection areas were evaluated, and the excellent soybean lines with high-value proteins, including 11S globulins, or without antinutritional factors such as lipoxygenase and trypsin inhibitor were selected.

In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an NH4NO3-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

Brachypodium distachyon has been highlighted as a model monocot plant with small genome and short life cycle. Biofuels are being developed as renewable energy sources to replace fossil fuels. Bioethanol production is negatively correlated with lignin content. Here, Brachypodium was acutely or chronically irradiated at doses of 50, 100, 150, 200, and 250 Gy. The effect of radiation on plant growth and generation of mutant populations was explored. The lethal effect of radiation was higher in acutely irradiated M0 populations. A dose-dependent negative effect in plant height, tiller number, floral spikelet, and total seed number was observed, with a positive effect in days to heading. The phenotype of 1,773 M1 plants was evaluated, with 417 plants being selected to construct the M2 population. The 31 M2 plants that showed the least staining with phloroglucinol were selected. These mutants could be useful materials for studies such as identification of nucleotide substitutions in genes involved in lignin biosynthesis pathway, monitoring of mutant physiological traits, and evaluation of fitness for bioethanol production. As biological resources, the M2 populations generated in this work will contribute to studies of functional genomics of Brachypodium and to the breeding of grass crops.

Dandelion has been widely utilized for medicinal and edible purposes. This research was conducted to evaluate the effect of supplemental LED (light-emitting diode) light on germination, growth characteristics and anthocyanin content of dandelion (Taraxacum officinale) seedling using LED blue (460 nm), red (660 nm, R), blue + red (B:R=6:4) and fluorescent lamp light treatment. By LED illumination to T. officinale seed germination speed was delayed, and germination rate was the highest in the fluorescent light. The growth characteristics (plant height, number of leaves, root length and fresh weight) were greatly influenced by supplemental LED light compared with control treatment, and the growth promotion was the most effective in the red LED illumination. After 60 days of red and mixed LED light treatments, anthocyanin content of dandelion plants was significantly changed. The anthocyanin content was increased by 12~19 mg/100 g under the red LED and the mixed light conditions compared with the control and the blue LED. Results indicate that illumination with red and mix LEDs, compared with other light treatments, is beneficial for promotion of growth and anthocyanin content in dandelion.

Salinity is a major environmental stress to plants. In this study, the ability of plants to tolerate salt was investigated by studying growth, physiological characteristics, and expression levels of genes related to the salt-stress response in the salt-tolerant rice mutant (Till-II-877), which was derived from γ-ray irradiation. Compared to plants grown under normal conditions, the height and root length of wild type (WT) were reduced by approximately 40 and 29% following exposure to salt stress for 3 weeks, whereas Till-II-877 line showed 29 and 23% reductions in plant height and root length, respectively. No significant changes were observed in total chlorophyll content, and the malondialdehyde content of the mutant increased less than that of the WT under salt treatment. Gene expression was compared between the WT and mutant lines using microarray analysis. An unbiased analysis of the gene expression datasets allowed us to identify the pathways involved in salt-stress responses. Among the most significantly affected pathways, changes in gene expression were observed in α-linolenic acid and linoleic acid metabolism (in lipid metabolism), fructose and mannose metabolism and glycolysis-gluconeogenesis (in carbohydrate metabolism), cysteine and methionine metabolism (in amino acid metabolism), and carbon fixation (in the energy metabolism of photosynthetic organisms) under salt stress. These results show that the differential response of plants subjected to salt stress was due to changes in multiple metabolic pathways. These findings increase our understanding of the effects of salt stress in rice and may aid in the development of salt-tolerant rice cultivars.

Arabidopsis presumably has few sensors for gamma-rays and few signal transduction systems that respond to them. In an effort to assess their radiation sensitivity, wild-type ( Ler ) Arabidopsis plants were irradiated with various doses of gamma-rays at the vegetative (VE) and reproductive (RE) stages. 100 Gy treatment induced the higher production of siliques during both the VE and RE stages compared with non-irradiation. Treatments at doses over 200 Gy repressed shoot growth, and the plants perished under 800 Gy treatment. The results of physiological analysis using electron spin resonance (ESR) and transmission electron microscopy (TEM) revealed that increased doses of gamma-rays induce greater ROS generation. To establish the gene expression profiles after gamma irradiation and for an analysis of the antioxidant response, we employed an oligonucleotide microarray system. Different responses of genes related with ROS scavenging and signal transduction pathways by a gamma irradiation were observed. At least 33 and 42 out of all genes with significantly altered expression were associated with ROS scavenging and signal transduction pathways having an induction or repression ratio cutoff of at least 2-fold, respectively. CAT3 (At1g20620), Ferritin1 (At5g01600), Blue copper binding protein (At5g20230), and AOX putative (At1g32350) were up-regulated regardless of dosage at the VE stage. Reactive oxygen species signaling genes encoding phospholipase, zinc finger protein, WRKY DNA-binding protein, and calcium binding protein were highly expressed, evidencing changes greater than 2-fold. Our transcriptomic profile of the responses of Arabidopsis genes to gamma irradiation showed that plants evidenced altered expressions of many signal transduction and antioxidant genes, as have been seen with other environmental stresses.

The aim of this study was to investigate the protective ability of blackberry extract (BE) against oxidative stress in carbon tetrachloride (CCl(4))-treated rats. The results showed that treatment with BE attenuated lipid peroxidation that was increased by CCl(4) and also markedly recovered the activity of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR), that were decreased by CCl(4). BE also elevated the protein expression levels of NF-E2-related factor-2 (Nrf2), CuZnSOD, MnSOD, GPx-1/2, and heme oxygenase-1 (HO-1), but not that of catalase. Furthermore, the administration of BE significantly attenuated the levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) that were increased by CCl(4). Therefore, the present study suggests that BE possesses significant protective effects against in vivo oxidative stress.

In earlier investigations, Perilla frutescens (L.) Britt. cv. Chookyoupjaso (CJC) mutants were obtained following mutagenesis induced by 200 Gy of γ-rays. The aim of this study was to compare the CJC and 6 P. frutescens (L.) Britt. cv. Chookyoupjaso mutant lines (CJMs), with respect to their phytochemical profiles and to evaluate anti-inflammatory properties by selecting the most bioactive CJM. The methanol extracts of CJMs were tested for inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Among them, CJM-45 showed significant inhibition of NO production. This extract was further partitioned using ethyl acetate (EtOAc), butanol (BuOH), and water. The EtOAc fraction (EF-cjm45) was evaluated for antiinflammatory activities. These results indicated that the EF-cjm45 reduced NO production by inhibiting inducible nitric oxide synthase (iNOS) expression. The EF-cjm45 treatment also significantly diminished expression of MCP-1 and IL-6. In the EF-cjm45, perillaketone, isoegomaketone, ursolic acid, kaempferol, and rosmarinic acid were also found. This study reveals the potential therapeutic use of bioactive CJM-45 and justifies the wide application for this plant in traditional medicine.

Isoegomaketone (IK) is an essential oil component of Perilla frutescens (L.) Britt., and there have been no studies investigating its biological activities. We found that IK inhibits lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophages, and moreover, when IK was injected into animals prior to LPS administration, NO serum levels decreased in a dose-dependent manner. These results indicate that IK possesses anti-inflammatory activity both in vitro and in vivo. IK suppressed the phosphorylation of STAT-1 and the production of IFN-beta. Treatment with IK also inhibited the activation of NF-kappaB and activator protein-1, but more IK was required for inhibition than for STAT-1 inhibition, indicating that downregulation of inducible nitric oxide synthase gene expression by IK is mainly attributed to the blockade of STAT-1 activation. Furthermore, IK also induced the expression of heme oxygenase-1 (HO-1) through the activation of nuclear factor E2-related factor 2. Treatment with SnPP, a selective inhibitor of HO-1, reversed the IK-induced suppression of STAT-1 phosphorylation and NO production. Taken together, IK isolated from P. frutescens inhibits NO production in LPS-treated RAW 264.7 macrophages through simultaneous induction of HO-1 and inhibition of the IFN-beta-STAT-1 pathway.

In addition to its role as an energy source for plants, animals and humans, starch is also an environmentally friendly alternative to fossil fuels. In rice, the eating and cooking quality of the grain is determined by its starch properties. The floury endosperm of rice has been explored as an agronomical trait in breeding and genetics studies. In the present study, we characterized a floury endosperm mutant, flo(a), derived from treatment of Oryza sativa ssp. japonica cultivar Hwacheong with MNU. The innermost endosperm of the flo(a) mutant exhibited floury characteristics while the outer layer of the endosperm appeared normal. Starch granules in the flo(a) mutant formed a loosely-packed crystalline structure and X-ray diffraction revealed that the overall crystallinity of the starch was decreased compared to wild-type. The FLO(a) gene was isolated via a map-based cloning approach and predicted to encode the tetratricopeptide repeat domain-containing protein, OsTPR. Three mutant alleles contain a nucleotide substitution that generated one stop codon or one splice site, respectively, which presumably disrupts the interaction of the functionally conserved TPR motifs. Taken together, our map-based cloning approach pinpointed an OsTPR as a strong candidate of FLO(a), and the proteins that contain TPR motifs might play a significant role in rice starch biosynthetic pathways.