Examining the Alterations in Metabolite Constituents and Antioxidant Properties in Mountain-Cultivated Ginseng (Panax ginseng C.A. Meyer) Organs during a Two-Month Maturation Period.

The current research was the first to prove the existence of fluctuations in the metabolite constituents and antioxidant properties in different organs (leaves, stems, and roots) of the mountain-cultivated ginseng (MCG) plant during a two-month maturation period. Four metabolites, including fatty acids, amino acids, ginsenosides, and phenolic phytochemicals, exhibited considerable differences in organs and maturation times with the following order: leaves > stems > roots. The predominant metabolite contents were found in leaves, with fatty acid (1057.9 mg/100 g) on 31 May, amino acid (1989.2 mg/100 g) on 13 July, ginsenosides (88.7 mg/g) on 31 May, and phenolic phytochemical (638.3 µg/g) on 31 May. Interestingly, ginsenoside content in leaves were highest, with 84.8 → 88.7 → 82.2 → 78.3 mg/g. Specifically, ginsenosides Re, Rd, and F2 showed abundant content ranging from 19.1 to 16.9 mg/g, 8.5 to 14.8 mg/g, and 9.5 to 13.1 mg/g, respectively. Phenolic phytochemicals exhibited remarkable differences in organs compared to maturation periods, with the highest total phenolic content and total flavonoid content recorded at 9.48 GAE and 1.30 RE mg/g in leaves on 31 May. The antioxidant capacities on radical, FRAP, and DNA protection differed significantly, with leaves on 31 May exhibiting the highest values: 88.4% (DPPH), 89.5% (ABTS), 0.84 OD593 nm (FRAP) at 500 µg/mL, and 100% DNA protection at 50 µg/mL. Furthermore, principal cluster analysis revealed metabolite variability as follows: ginsenoside (83.3%) > amino acid (71.8%) > phenolic phytochemical (61.1%) > fatty acid (58.8%). A clustering heatmap highlighted significant changes in metabolite components under the maturation times for each organ. Our findings suggest that MCG leaves on 31 May may be a potential source for developing nutraceuticals, offering highly beneficial components and strong antioxidants.

Enhancement of isoflavone aglycone, amino acid, and CLA contents in fermented soybean yogurts using different strains: Screening of antioxidant and digestive enzyme inhibition properties.

This study was the first to evaluate changes in isoflavone, amino acid, conjugated linoleic acid (CLA), antioxidant effect, and digestive enzyme inhibition during fermentation of soy-milk to soy-yogurt with L. brevis and L. plantarum. Total average isoflavones were reduced (1318.2 â†’ 971.1 µg/g) with an increase of aglycones (60.2 â†’ 804.9 µg/g, genistein > daidzein > glycitein) in soy powder yogurts (SPYs). Amino acids increased considerably, as did ornithine (average 4.1 â†’ 551.0 mg/g), and CLA showed high variations from not-detected (ND) to 0.5, 0.9 mg/g (cis-9, trans-11) and ND to 0.3, 0.2 mg/g (trans-10, cis-12). Digestive enzyme inhibitions (α-glucosidase, α-amylase, and pancreatic lipase) displayed high activities (average 50.6 â†’ 67.2, 5.2 â†’ 46.4, 10.6 â†’ 51.4%). Moreover, the antioxidant abilities against radicals were elevated as follows: ABTS > DPPH > hydroxyl (average 63.5 â†’ 86.5, 50.2 â†’ 70.3, 39.3 â†’ 55.2%). Specifically, SPY using mixed strains exhibited the greatest enzymatic inhibition and antioxidant capacities.

Policosanol profiles and adenosine 5'-monophosphate-activated protein kinase (AMPK) activation potential of Korean wheat seedling extracts according to cultivar and growth time.

Policosanols is a health promoting aliphatic alcohol known as lipid-lowing agent. To enable maximising the functional properties of wheat, this research investigates the policosanol profiles and adenosine 5'-monophosphate-activated protein kinase (AMPK) activation potential of Korean wheat seedlings according to cultivars and growth times. GC-MS revealed six policosanols that differed markedly in content between 17 cultivars, especially, octacosanol (8) showed the most predominant component (49-83%), varying significantly in average concentrations with growth times as 361.4 (3 days) â†’ 613.0 (6 days) â†’ 203.1 (9 days) â†’ 196.5 (12 days) â†’ 50.9 mg/100 g (19 days). The highest average policosanol (738.7 mg/100 g) exhibited after 6 days, while the lowest was 104.4 mg/100 g on 19 days. Moreover, the wheat cultivars including Shinmichal 1, Anbaek, Namhae, and Joah at 6 days may be recommended as potential sources because of high policosanols (921.7-990.6 mg/100 g). Western blot analysis revealed markedly higher AMPK activation in cells treated with the hexane extracts (150-370% at 100 µg/ml) and octacosanol (8) possessed potent AMPK activator (control; 100 â†’ 280% at 200 µg/ml). It is confirmed that the AMPK activation by wheat seedlings are positively related to the highest policosanol content at the 6 days of growth time, independent of the cultivar. Our results may be contributed to enhance the wheat value regarding development of new cultivars and functional foods.

Comparative analysis of isoflavone aglycones using microwave-assisted acid hydrolysis from soybean organs at different growth times and screening for their digestive enzyme inhibition and antioxidant properties.

The objectives of this research were to demonstrate the changes in isoflavone-aglycones, total phenolics, and biological properties (digestive enzyme inhibition; antioxidant) from six organs including leaves, leafstalks, roots, stems, seeds, and pods at different growth times of soybean plant. Three isoflavone-aglycones in microwave-assisted acid hydrolysis extracts were elucidated using UHPLC-ESI-Q-TOF-MS/MS and their contents exhibited remarkable differences in leaves (245.93-2239.33 µg/g), roots (854.96-4425.34 µg/g), and seeds (ND-2339.62 µg/g). Specifically, the collected samples on 15-Oct (leaves: 2239.33; seeds: 2339.62 µg/g) and 31-Aug (roots: 4425.34 µg/g) showed the highest isoflavone-aglycones, and daidzein was observed the most abundant component, comprising approximately 70%. Moreover, the inhibitions against α-glucosidase and α-amylase displayed the predominant effects in roots (89;91%) and leaves (81;85%) of samples on 31-Aug and 15-Oct at 300 µg/ml. The antioxidant activities on ABTS, DPPH, and hydroxyl radicals increased considerably with the increases of growth times in leaves and seeds, especially, ABTS showed the highest scavenging abilities: leaves (15-Oct;83%) > roots (31-Aug;75%) > seeds (15-Oct;68%). Therefore, our results suggest that soybean leaves, roots and seeds may be considered as excellent natural sources for nutraceuticals.

Kochia scoparia seed extract suppresses VEGF-induced angiogenesis via modulating VEGF receptor 2 and PI3K/AKT/mTOR pathways.

Context: Kochia scoparia (L.) Schrad (Amaranthaceae), known as a traditional medicine in China, Japan and Korea, is reported to have various biological activities. However, K. scoparia seed extract (KSE) functional roles on angiogenesis and prostate cancer inhibition have not been elucidated. Objective: This study elucidates the effects of KSE on vascular endothelial growth factor (VEGF)-induced angiogenesis in human umbilical vein endothelial cells (HUVECs) and inhibition of proliferation in prostate cancer cells. Materials and methods: HUVECs were treated with 10-20 µg/mL of KSE and 20-50 ng/mL of VEGF for 12-72 h. Anti-angiogenesis properties of KSE were determined by wound healing, trans-well, tube formation, rat aortic ring assay and western blotting. Prostate cancer and normal cells were incubated with 10-250 µg/mL of KSE for 24 h, and cell viability was measured by SRB assay. Phenolic compounds in KSE were analyzed using a HPLC-PDA system. Results: IC50 for cell viability of HUVECs, LNCaP, PC-3, RC-58T and RWPE-1 by KSE were 30.64, 89.25, 123.41, 141.62 and >250 µg/mL, respectively. Treatment with KSE (20 µg/mL) significantly suppressed VEGF-induced migration, invasion and capillary-like structure formation of HUVECs and microvessel sprouting from rat aortic rings. In addition, KSE down-regulated PI3K/AKT/mTOR levels and phosphorylation of VEGF receptor 2 in HUVECs. 3-OH-tyrosol (1.63 mg/g) and morin hydrate (0.17 mg/g) were identified in KSE. Conclusions: KSE inhibits angiogenesis in HUVECs as well as proliferation in human prostate cancer cells, suggesting KSE may be useful herbal medicine for preventing progression of prostate cancer and angiogenesis.

Extended Low Temperature and Cryostorage Longevity of Salix Seeds with Desiccation Control.

Salix xerophila, S. maximowiczii, and S. koreensis are species of willow native to Korea that are important for bioenergy production. However, the native range of these species has narrowed in recent years due to the impact of climate change. Seeds of these Salix species lose viability within 4 weeks at ambient temperature, and within 4 months at -4°C. Preservation techniques are urgently needed to protect these valuable resources. The effects of seed water content (SWC; 3%, 6%, 9%, 12%, 18%, and 24%) and temperature (ambient, 4°C, -18°C, -80°C, and -196°C) on storage stability were investigated for up to 48, 52, or 60 months, depending on species. Optimal storage temperature and SWC varied between species. S. xerophila seed could be stored without deterioration for 60 months with 9% SWC at -80°C, but rapidly lost viability when stored at -18°C. In S. maximowiczii and S. koreensis, 100% and 90% of normal germination, respectively, was maintained with 18% SWC at -18°C or -80°C. Thus, for some Salix species, storage at -18 and -80°C may provide an economical alternative to cryopreservation or medium-term storage for the maintenance of seedbanks or breeding stocks.

Extending Populus seed longevity by controlling seed moisture content and temperature.

The poplar species Populus davidiana and P. koreana are widely grown in plantations and as biofuel resources, but little is known about ex-situ seed conservation in poplar. To identify the optimal long-term seed storage conditions for these species, we evaluated the viability of seeds with different seed water contents (SWCs) at various storage temperatures and time periods. P. davidiana seeds with <6% SWC could be stored at room temperature (RT) for 4 weeks, while P. koreana seeds showed no storability at RT. P. davidiana seeds with 3% SWC showed 74% viability after 36 months of storage at 4°C, while those with 9-18% SWC showed >89% viability after 48 months of storage at -18°C. Long-term storage at -80°C was best for P. davidiana seeds with a wide range of SWCs (3-24%), with 91-98% of normal germination after 48 months of storage. However, the normal germination of P. koreana seeds with 3-24% SWC declined to <20% after 36 months of storage, even at -18°C and -80°C. No significant difference was observed between seeds immersed vs. not immersed in liquid nitrogen for both species. Our findings increase the possibility for long-term seed conservation for both Populus species.

Rice OsMYB5P improves plant phosphate acquisition by regulation of phosphate transporter.

Myeloblastosis (MYB) transcription factors play central roles in plant developmental processes and in responses to nutrient deficiency. In this study, OsMYB5P, an R2R3-MYB transcription factor, was isolated and identified from rice (Oryza sativa L. 'Dongjin') under inorganic phosphate (Pi)-deficient conditions. OsMYB5P protein is localized to the nucleus and functions as a transcription activator in plant development. Overexpression of OsMYB5P in rice and Arabidopsis (Arabidopsis thaliana Col-0) increases tolerance to phosphate starvation, whereas OsMYB5P knock-out through RNA interference increases sensitivity to Pi depletion in rice. Furthermore, shoots and roots of transgenic rice plants overexpressing OsMYB5P were longer than those of wild plants under both normal and Pi-deficient conditions. These results indicate that OsMYB5P is associated with the regulation of shoot development and root- system architecture. Overexpression of OsMYB5P led to increased Pi accumulation in shoots and roots. Interestingly, OsMYB5P directly bound to MBS (MYB binding site) motifs on the OsPT5 promoter and induced transcription of OsPT5 in rice. In addition, overexpression of OsMYB5P in Arabidopsis triggered increased expression of AtPht1;3, an Arabidopsis Pi transporter, in shoots and roots under normal and Pi-deficient conditions. Together, these results demonstrate that overexpression of OsMYB5P increases tolerance to Pi deficiency in plants by modulating Pi transporters at the transcriptional level in monocots and dicots.

Direct Effects on Seed Germination of 17 Tree Species Under Elevated Temperature and CO2 Conditions.

Effects on seed germination characteristics of 17 tree species were investigated under elevated temperature and CO2. Seeds of 5 needle-leaf and 12 broad-leaf species were germinated under four conditions: 24°C + 400 µmol CO2 mol air-1, 24°C + 750 µmol CO2 mol air-1, 27°C + 400 µmol CO2 mol air-1, and 27°C + 750 µmol CO2 mol air-1. The elevated temperature and CO2 affected germination percent (GP) of 7 tree species seeds.GPs of Pinus densiflora, P. thunbergii, Betula ermanii, and Maackia amurensisseeds were affected by the elevated temperature, while only that of P.jezoensis seed was influenced by the elevated CO2. GPs of Malus baccata and Zelkova serrataseeds were influenced by both the elevated temperature and CO2. In addition, the elevated temperature and CO2also affected mean germination time (MGT) of 12 tree species seeds. Particularly, MGTs of P. thunbergii and Rhododendron tschonoskii seeds were influenced by both factors. In conclusion, elevated temperature and CO2 affected seed germination characteristics, which were reflected by significant differences among tree species. Specifically, these two factors exerted stronger influence on germination pattern such as MGT rather than seed germination percent.

Three-dimensional surface topography of the needle stomatal complexes of Pinus rigida and its hybrid species by complementary microscopy.

Three-dimensional surface topography of needle stomatal complexes was investigated in Pinus rigida, Pinus taeda, and their interspecific hybrid Pinus rigitaeda. The stomatal complexes of P. rigida appeared to be sunken and ca. 15 microm deep by white light scanning interferometry. Stomatal grooves were evident in P. taeda along the stomata and amounted to ca. 5 microm deep. The centers of stomata maintained the similar height to the stomatal apertures. Meanwhile, the stomatal complexes of P. rigitaeda (ca. 15 microm deep) were characterized by distinct stomatal grooves and sunken stomatal chambers. In addition, field emission scanning electron microscopy revealed the stomatal complexes of P. rigida partially filled with epicuticular waxes. It was common to observe distinct stomatal grooves and chamber-filled stomata on P. taeda needles. The stomatal complexes of P. rigitaeda had the distinct stomatal grooves and were partially filled with wax tubules and rodlets. Surface roughness measurements of stomatal complexes showed higher levels of roughness from P. rigida and P. rigitaeda than that from P. taeda. These results indicate that the hybrid species P. rigitaeda showed intermediacy in surface characteristics between the parent species, suggesting the genetic control of needle stomatal complexes in the hybrid species.

Bis(di-2-pyridyl-methane-diol-κN,O,N')nickel(II) dibenzoate.

The title compound, [Ni(C(11)H(10)N(2)O(2))(2)](C(7)H(5)O(2))(2), consists of an Ni(II) ion coordinated by two tridentate chelating (2-py)(2)C(OH)(2) ligands (py is pyrid-yl) and two benzoate anions. The Ni(II) ion is located on a twofold rotation axis, and its geometry is distorted octa-hedral. The gem-diol ligand (2-py)(2)C(OH)(2) adopts an η(1):η(1):η(1) coordination mode. There are O-H⋯O hydrogen bonds between the gem-diol ligands and benzoate anions.

Gel-based proteomics approach for detecting low nitrogen-responsive proteins in cultivated rice species.

Nitrogen fertilization is essential for increasing rice production to meet the food demands of increasing world's population. We established an in vivo hydroponic rice seedling culture system to investigate physio-biochemical/molecular responses of various rice japonica and indica cultivars to low nitrogen (N). Three-week-old seedlings grown in Yoshida's nutrient solution manifested stable and reproducible symptoms, such as reduced shoot growth and length under low N. Out of 12 genetically selected cultivars, 11 cultivars showed varied degrees of growth reduction response to applied N (4 and 40 ppm N for treatment and control, respectively), whereas one cultivar (no. 12) showed similar growth as the control though its leaf width was smaller than control. Leaves of a representative low N-responsive cultivar (BG90-2) were sampled for revealing protein profiles between low and normal (control) N application by two-dimensional gel electrophoresis (2-DGE). Forty-one proteins were identified with MALDI-TOF-MS and nESI-LC-MS/MS. Assignment of proteins into major (energy metabolism, photosynthesis and oxidative stress) and minor functional categories, revealed many novel low N-responsive proteins, including those having energy/photosynthesis- and defense/stress- and iron homeostasis-related functions. Results suggest the usefulness of proteomics in identifying novel N-responsive proteins and may provide potential markers for rice response to low N.

Gene transcription in the leaves of rice undergoing salt-induced morphological changes (Oryza sativa L.).

We describe the gene expression profile of third leaves of rice (cv. Nipponbare) seedlings subjected to salt stress (130 mM NaCl). Transcripts of Mn-SOD, Cu/Zn-SOD,cytosolic and stromal APX, GR and CatB were regulated, whereas expression of thylakoid-bound APX and CatA were down-regulated. The levels of the compatible solute proline and of transcripts of its biosynthetic gene, Delta1-pyrroline-5-carboxylate synthetase (P5CS), were strongly increased by salt stress. Interestingly, a potential compatible solute, gamma-aminobutyric acid (GABA), was also found to be strongly induced by salt stress along with marked up-regulation of transcripts of GABA-transaminase. A dye-swap rice DNA microarray analysis identified a large number of genes whose expression in third leaves was altered by salt stress. Among 149 genes whose expression was altered at all the times assayed (3, 4 and 6 days) during salt stress, there were 47 annotated novel genes and 76 unknown genes. These results provide new insight into the effect of salt stress on the expression of genes related to antioxidant enzymes, proline and GABA as well as of genes in several functional categories.

Effect of textile wastewaters on Saccharomyces cerevisiae using DNA microarray as a tool for genome-wide transcriptomics analysis.

Textile mill effluents (TMEs) discharged from the textile industry can be considered as one class of hypothetical toxicants in the environment. To investigate the potential toxicity of TMEs, we applied cDNA microarray technology to examine the genome-wide expression profiles in model eukaryote, Saccharomyces cerevisiae. The results revealed a rich source of genetic information for the yeast cells that were exposed to the untreated and treated TMEs. Among the 5956 valid genes, 275 genes were up-regulated and 40 genes were down-regulated for the untreated TMEs. On the other hand, only 90 genes were up-regulated, and 29 genes were down-regulated upon exposure to the treated TMEs. The changes in gene expression were also confirmed by RT-PCR. The potent up- and down-regulation of genes suggest that yeast cells undergo genome-wide changes in mRNA expression, indicative of a stress response. Additionally, a classification into specific functional gene categories indicated that untreated and even treated TMEs still had toxicity. Especially, the genes related to oxidative stress, such as AHP1, ATX1, GRX1, TRX1 and TRX2, were up-regulated in treated TMEs that can directly reach to surface and ground waters, and sediments.

A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf.

By using an in vivo hydroponic rice seedling culture system, we investigated the physiological and biochemical responses of a model rice japonica cultivar Nipponbare to salt stress using proteomics and classical biochemical methods. Yoshida's nutrient solution (YS) was used to grow rice seedlings. YS-grown 18-day-old seedlings manifested highly stable and reproducible symptoms, prominently the wilting and browning of the 3rd leaf, reduced photosynthetic activity, inhibition in overall seedling growth, and failure to develop new (5th) leaf, when subjected to salt stress by transferring them to YS containing 130 mM NaCl for 4 days. As leaf response to salt stress is least investigated in rice by proteomics, we used the 3rd leaf as source material. A comparison of 2-DE protein profiles between the untreated control and salt-stressed 3rd leaves revealed 55 differentially expressed CBB-stained spots, where 47 spots were increased over the control. Of these changed spots, the identity of 33 protein spots (27 increased and 5 decreased) was determined by nESI-LC-MS/MS. Most of these identified proteins belonged to major metabolic processes like photosynthetic carbon dioxide assimilation and photorespiration, suggesting a good correlation between salt stress-responsive proteins and leaf morphology. Moreover, 2-DE immunoblot and enzymatic activity analyses of 3rd leaves revealed remarkable changes in the key marker enzymes associated with oxidative damage to salt stress: ascorbate peroxidase and lipid peroxidation were induced, and catalase was suppressed. These results demonstrate that hydroponic culture system is best suited for proteomics of salt stress in rice seedling.