Japonica Samples Research Articles

Molecular population genetic analysis of Eriocheir japonica inhabiting Andeok Valley

Eriocheir japonica inhabit rivers flowing into the East Sea, South Sea, and the coastal waters of Jeju-do in South Korea, Japan, and Sakhalin. The larvae migrate to freshwater, mature, and move to estuaries to spawn. Jeju-do has habitats that prevent the migration of aquatic organisms. Populations with low within-group genetic diversity experience increased extinction risk. Industrialization causes climate change and also creates species extinctions. The goal of this study was to investigate the genetic diversity of E. japonica inhabiting Andeok Valley to elucidate their genetic stability and encourage future conservation plans. E. japonica samples used for the study were from the Andeok Valley in South Korea. One walking leg was used for DNA extraction. DNA was amplified using polymerase chain reaction to sequence mitochondrial cytochrome c oxidase subunit 1 (CO1) and cytochrome b (CytB) genes. Haplotype and sequence diversity analyses were performed. Sequencing results for CO1 and CytB genes, included 658 and 651 base pairs, respectively. The median network analysis showed that the population inhabiting Andeok Valley had a genetically simple phylogenetic structure. This region provides simple habitat conditions, preventing gene transfer. The probability of extinction of E. japonica that inhabits Andeok Valley is markedly high.

Mitochondrial DNA and Microsatellite Analyses Showed Panmixia between Temporal Samples in Endangered Anguilla japonica in the Pearl River Basin (China).

The Japanese eel (Anguilla japonica) is a commercially important species in East Asia, the abundance of which has rapidly decreased in recent decades. The fishery resource in the Pearl River basin has mainly deteriorated due to overexploitation and habitat degradation. Knowledge on its genetic status is indispensable for resource management. In this study, we explored the temporal genetic structure of A. japonica on the basis of the concatenated sequences of two mitochondrial fragments (mtDNA) and eight microsatellite markers. A total of nine temporal samples (N = 127) were collected during 2019 and 2021 from Jiangmen City, China, which is located in the Pearl River estuary. mtDNA sequence analysis showed a high level of haplotype diversity, and yielded 124 haplotypes with ranging from 9 to 19 in temporal samples. All microsatellite loci were polymorphic among each of the nine temporal samples, with 150 alleles identified across all samples. Pairwise FST values were low and nonsignificant according to both mtDNA and microsatellite markers. STRUCTURE analysis showed that all temporal samples were not clearly differentiated from each other. The yielded outcomes supported a panmictic pattern in different temporal A. japonica samples. Therefore, our results call for the management of A. japonica as a single unit and joint conservation strategy of the species, since overexploitation in any region will decrease its global resource.

Activated Carbon-Based Immunochromatographic Strip Test for the Rapid Qualitative Analysis of Swertiamarin and Sweroside.

Swertia japonica (S. japonica) is a medicinal plant that belongs to the Gentianaceae family. Several reports confirm the biological effects of the S. japonica extract. This plant is used mainly as a digestive stimulant, appetite stimulant, and gastrointestinal disease remedy in Japan. Secoiridoid glycosides are a group of compounds related to the beneficial effects of this plant. We developed an immunochromatographic strip test for major secoiridoid glycosides, such as swertiamarin (SM) and sweroside (SS) detection. We fabricated an immunoprobe using activated carbon as a reporter molecule and a monoclonal antibody against SM and SS (MAb D2) as a detection molecule. The test and control zones of the strip test contained SM-cBSA and Goat pAb anti-mouse IgM HRP conjugate, respectively. The immunoprobe reacted competitively with free SM and/or SS and immobilized SM-cBSA. The results were read and interpreted by the black spot intensity in the test zone. We succeeded in developing a strip test system with a detection limit (LOD) of 12.5 µg/mL. The selectivity and reliability evaluation revealed that the strip test is suitable for detecting SM and SS in S. japonica. The result was ready to be read in 30 min. This method can be a useful tool for the screening of biologically active S. japonica samples for further preparation of traditional medicine. To the best of our knowledge, this is the first immunochromatographic strip test developed for the detection of SM and SS in S. japonica samples.

Identification of <i>Indica-Japonica</i> Subspecies Rice by Duplex Fluorescence PCR Detection for Chloroplast DNA and Endogenous <i>gos</i> Gene

Asia cultivated rice are classified into two subspecies, indica and japonica rice (hsien and keng rice called in China, separately). It is necessary to establish a convenient and effective method to identify two subspecies because of the different quality characteristics between the rice products of them, which lead to different processing uses and commodity values. The identification method of PCR based on the difference of chloroplast DNA is convenient and effective, to be attempted to establish. Based on 69 bp fragment deletion of chloroplast DNA (cpDNA) found in ORF100 region nucleotides within the cpDNA Pst I-12 fragment in indica rice but not in japonica, the primer pair and a probe located on the specific cpDNA fragment in japonica rice were designed to identify indica and japonica rice. Another primer pair and probe used to detect endogenous gene gos in rice were combined with above primer pair and probe for detecting cpDNA to establish duplex fluorescence PCR to amplify cpDNA and gos gene for improving detection accuracy, to avoid the false negative results caused by DNA extraction error. The duplex fluorescence PCR detection method was established using typical japonica rice (pearl rice) and typical indica rice (Taixian 11). The accuracy of the method was validated with 547 samples including 177 samples of rice seed and eaten rice known as conventional japonica varieties and japonica type hybrid combinations and 370 samples of rice seed and eaten rice known as conventional indica rice varieties and indica type hybrid combinations. In 177 japonica samples, 170 samples with both positive results of cpDNA marker and gos gene, a coincidence rate of 96.05% agreement japonica rice, and 7 samples with positive results of gos gene and negative of cpDNA marker, not agreement japonica rice with non-coincidence rate of 3.95%, were detected. Of the 370 samples of indica rice seed and eaten rice, with positive results of gos gene and negative of cpDNA marker were detected in 340 samples, the rate of coincidence to indica rice was 91.89%. Other 30 samples with both positive results of cpDNA marker and gos gene, were detected, and the non-coincidence rate with indica rice was 8.11%. These identification results were in good agreement with known indica and japonica varieties, and the coincidence rate of japonica varieties was higher than indica. This method can be used to identify indica and japonica subspecies rice, especially suitable to identify conventional rice varieties.

Arsenic, cadmium, lead, and mercury contents of mushroom species in Korea and associated health risk

ABSTRACT Arsenic, cadmium, lead, and mercury contents of Ganoderma lucidum, Phellinus linteus, Umbilicaria esculenta, Trametes versicolor, Agaricus blazei, Hericium erinaciium, Paecilomyces japonica, Fuscoporia obliqua, Sarcodon imbricatu and Hypsizygus marmoreus mushroom samples from the South Korean market were analyzed using validated methods. Method validation was performed, and the limits of detection and quantification were determined to be 0.38–0.66 and 1.21–2.09 μg/kg, respectively. The recoveries of the target metals ranged from 91.3% to 103.5%. The contents of As, Cd, Hg, and Pb were significantly different among the ten mushroom species, and the ranged from 0.006 to 3.9, 0.025 to 5.8, 0.01 to 0.21, and 0.004 to 12.3 mg/kg dry weight, respectively. Additionally, the estimated daily exposures to these metals from consumption of mushroom used in this work were found to be safe in accordance with the provisional tolerable weekly intake. Considering the results in this study, it could be mentioned that the consumption of the ten mushrooms species from Korea presented no noticeable health risk to the general public.

Description of ten unrecorded bacterial species isolated from <i>Ostrea denselamellosa</i> and <i>Eriocheir japonica</i> from the Seomjin River

Ostrea denselamellosa and Eriocheir japonica samples were collected from the Seomjin River in 2019 as part of the “Research of Host-Associated Bacteria” research program. Almost 200 bacterial strains were isolated from the O. denselamellosa and E. japonica samples and subsequently identified by 16S rRNA gene sequencing. Among the bacterial isolates, ten strains possessed greater than 98.7% sequence similarity with published bacterial species that had not previously been recorded in Korea. These species were phylogenetically diverse, belonging to three phyla, four classes, seven orders, and eight genera. At the genus and class level, the previously unrecorded species belonged to Pseudoalteromonas, Aliivibrio, Rheinheimera, Leucothrix, and Shewanella of the class Gamma-proteobacteria, Olleya of the class Flavobacteriia, Algoriphagus of the class Cytophagia, and Lactococcus of the class Bacilli. The previously unrecorded species were further characterized by examining their Gram staining, colony and cell morphology, biochemical properties, and phylogenetic positions.

Quantitative Interrelation between Atractylenolide I, II, and III in Atractylodes japonica Koidzumi Rhizomes, and Evaluation of Their Oxidative Transformation Using a Biomimetic Kinetic Model.

Analytical methods based on ultraperformance liquid chromatography/ion-trap mass spectrometry (UPLC/ion-trap MS) were developed for quantification of atractylenolide I, II, and III in the methanol extract of Atractylodes japonica rhizomes with a C18 column in an acidified water/acetonitrile gradient eluent in an LC system, and ion-trap MS coupled with electrospray ionization was employed under positive-ion mode. The three atractylenolides were quantified in all A. japonica samples, and the content of atractylenolide I, II, and III showed a significant correlation to each other. Such high correlation was explained by the mechanistic insights into the biosynthetic pathway of atractylenoide III and I from atractylenoide II by using the biomimetic cytochrome P450 model, [Fe(tmp)](CF3SO3) (tmp = meso-tetramesitylporphyrin). Atractylenolides could be transformed by oxidation via the oxidative enzyme in the A. japonica plant. The present study first reports the first oxidative transformation of atractylenolides using the heme iron model complex.

Out of Asia: Biogeography of fungal populations reveals Asian origin of diversification of the Laccaria amethystina complex, and two new species of violet Laccaria

Purple Laccaria are ectomycorrhizal basidiomycetes associated with temperate forests all over the Northern Hemisphere in at least two taxa: Laccaria amethysteo-occidentalis in North America, and L. amethystina complex in Eurasia, as shown by Vincenot et al. (2012). Here, we combine a further study of the genetic structure of L. amethystina populations from Europe to southwestern China and Japan, using neutral Single Sequence Repeat (SSR; microsatellite) markers; and a systematic description of two novel Asian species, namely Laccaria moshuijun and Laccaria japonica, based on ecological, morphological, and molecular criteria (rDNA sequences). Population genetics provides evidence of the ancient isolation of three regional groups, with strong signal for speciation, and suggests a centre of origin of modern populations closest to present-day Chinese populations. Phylogenetic analyses confirm speciation at the molecular level, reflected in morphological features: L. moshuijun samples (from Yunnan, China) display strongly variable cheilocystidia, while L. japonica samples (from Japan) present distinctive globose to subglobose spores and clavate cheilocystidia. This study of a species complex primarily described with an extremely wide ecological and geographical range sheds new light on the biodiversity and biogeography of ectomycorrhizal fungi.

Effects of water-washing Saccharina japonica on fast pyrolysis in a bubbling fluidized-bed reactor

The pyrolysis characteristics of non-edible Saccharina japonica (S. japonica) obtained from an offshore high-density aquaculture facility were systematically investigated using a thermogravimetric analyzer and a bubbling fluidized-bed reactor. As high mineral contents in S. japonica cause ash fouling or agglomeration of char and bed material in fluidized-bed reactor, S. japonica was water-washed as pre-treatment to remove minerals and sticky materials. The pretreated S. japonica samples were mostly decomposed between 200 and 350 °C at heating rates of 5–20 °C/min. The calculated activation energy increased from 36.31 to 393.01 kJ/mol when pyrolysis conversion increased from 5 to 60%. The yields of bio-oil were 25.57–31.27 wt% under the investigated pyrolysis conditions (temperature: 350–450 °C; fluidization velocity: 2.0 × Umf − 4.5 × Umf). The highest bio-oil yield (31.27 wt%) from water-washed S. japonica was obtained at 375 °C and 4.0 × Umf, and the HHVs of the organic and aqueous phases in the bio-oil were 31.47 and 5.41 MJ/kg, respectively. The major compounds in the bio-oil were analyzed by GC-MS and the carbon number distribution of the bio-oils was determined through simulated distillation by TGA.

Thermogravimetric characteristics and pyrolysis kinetics of high-density-aquacultured Saccharina Japonica: Effects of water-washing

The thermogravimetric characteristics and pyrolysis kinetics of high-density-aquacultured Saccharina japonica were investigated using a thermogravimetric analyzer and micro-tubing reactor, respectively. The effect of water washing S. japonica samples on pyrolysis product distribution was studied at 360, 370 and 380°C with various residence times from 1.0 to 3.0min. The kinetic rate constants of pyrolysis for untreated and water-washed S. japonica samples were determined using nonlinear least-squares regression of experimental data assuming first-order kinetics. The predominant reaction pathway of water-washed S. japonica was liquid to gas product (k3) at each temperature, while that of untreated S. japonica was biomass to liquid product (k2) at 360 and 370°C, then liquid to gas product (k3) at 380°C. However, the k2/k1 ratio of water-washed S. japonica (1.2166–1.3247) was more than two times that of untreated S. japonica (0.5763–0.6346).

Isothermal kinetics of torrefied Cryptomeria japonica in CO2 gasification

Cryptomeria japonica (CJ) samples were heated in a thermogravimetric analyzer (TGA) with CO2 as agent to study the gasification kinetics in the isothermal process. The samples were torrefied at 250 and 350 °C for 1 h. before being tested in TGA. Three isothermal conditions of 750, 800, and 850 °C were tested in this study to estimate the kinetic parameters of the Arrhenius form with the proposed models. The homogeneous model (HM), shrinking core model (SCM), and linear model (LM) were used, and the predicted results obtained from these models were compared with experimental data. The reaction rate of gasification was enhanced as temperature was raised, and a correlation of kinetic parameters with temperature was obtained. The simulated results of the linear model (LM) fit well with experimental data. This work is useful for design gasification in downdraft gasifier.

Rapid and quantitative determination of 10 major active components in Lonicera japonica Thunb. by ultrahigh pressure extraction-HPLC/DAD

An ultrahigh pressure extraction (UPE)-high performance liquid chromatography (HPLC)/diode array detector (DAD) method was established to evaluate the quality of Lonicera japonica Thunb. Ten active components, including neochlorogenic acid, chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, rutin, luteoloside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and quercetin, were qualitatively evaluated and quantitatively determined. Scanning electron microscope images elucidated the bud surface microstructure and extraction mechanism. The optimal extraction conditions of the UPE were 60% methanol solution, 400 MPa of extraction pressure, 3 min of extraction time, and 1:30 (g/mL) solid:liquid ratio. Under the optimized conditions, the total extraction yield of 10 active components was 57.62 mg/g. All the components showed good linearity (r2 ≥ 0.9994) and recoveries. This method was successfully applied to quantify 10 components in 22 batches of L. japonica samples from different areas. Compared with heat reflux extraction and ultrasonic-assisted extraction, UPE can be considered as an alternative extraction technique for fast extraction of active ingredient from L. japonica.

Cellulose extraction from Zoysia japonica pretreated by alumina-doped MgO in AMIMCl

In this study, alumina-doped MgO was produced as a solid alkali for lignocellulose pretreatment. Pretreatment with alumina-doped MgO disrupted the lignocellulose structure and significantly reduced the lignin content of the Z. japonica. After pretreatment, Z. japonica showed significant solubility in 1-allyl-3-methylimidazolium chloride (AMIMCl). The similar high solubility of pretreated Z. japonica samples by original alumina-doped MgO and used alumina-doped MgO also proved that alumina-doped MgO had strong stability, which can be recycled and used repeatedly. The regenerated cellulose was similar to microcrystalline cellulose according to FTIR and NMR analyses. Compared to microcrystalline cellulose, only the crystallinity of the regenerated cellulose decreased.

Analysis of Dioscorea japonica by Hydrophilic Interaction Liquid Chromatography

Dioscorea japonica Thunb., traditionally used in folk medicine in Korea for the treatment of hyperglycemia, is known to have biological activity. However, few analytical methods suitable for rapid assessment of constituents of D. japonica rhizomes have been reported. A high-performance liquid chromatography method coupled with variable wavelength ultraviolet detection was developed for the determination of eight constituents of D. japonica rhizome. The separation was performed efficiently on a hydrophilic interaction liquid chromatography column by gradient elution with acetonitrile and water. Calibration curves showed excellent linear regression correlation levels (R 2 > 0.9999) within the range of tested concentrations. Intra- and inter-day variations, evaluated by determining relative standard deviations, were below 1.71%. Constituent recovery rates were 93.37–104.24% with relative standard deviations of 0.55–1.98% for spiked D. japonica samples. The developed method was suitable for the qualitative and quantitative determination of the major components in D. japonica.

Quality control of Lonicera japonica stored for different months by electronic nose

The objective of this study was to investigate the potential of the electronic nose (E-nose) technique for monitoring the storage time and quality of L. japonica. An E-nose was used to detect odors of L. japonica samples during a storage period of 16 months. Linear discriminant analysis (LDA) and radial basis function (RBF) neural network were performed to differentiate L. japonica samples stored for different months. The content of chlorogenic acid of L. japonica was determined to confirm the quality changes and investigate its correlation with the odor response values. Results showed that L. japonica samples of different storage months could be classified correctly by LDA and RBF neural network. The change trends of the odor response and the content of chlorogenic acid had both declined along with the storage time. Also, there was a significant correlation (p=0.000) between the odor index and the content of chlorogenic acid. In conclusion, the odor intensity could reflect the quality of L. japonica to a certain degree. The E-nose technique could be used as a rapid, simple, sensitive and effective method for the quality control of L. japonica.

Dissolution of cellulose from AFEX-pretreated Zoysia japonica in AMIMCl with ultrasonic vibration

In this study, 1-allyl-3-methylimidazolium chloride (AMIMCl), an ionic liquid, was synthesized and characterized by a series of test methods. Pretreatment of Zoysia japonica by ammonia fiber expansion (AFEX) was shown to reduce significantly the mass of hemicellulose and lignin in biomass, thereby breaking the lignocellulosic structure. Z. japonica samples pretreated with AFEX showed reasonable solubility in AMIMCl upon ultrasonic treatment. The rate of cellulose regeneration from Z. japonica samples pretreated with AFEX increased with increase in applied power of ultrasonication within a certain power range from 0 to 110W. The regeneration rate of cellulose from AFEX-pretreated Z. japonica reached a maximum of 97% when the ultrasonic power was 110W. Fourier transform infrared spectroscopy and nuclear magnetic resonance analyses indicated that the regenerated cellulose was similar to microcrystalline cellulose.

Determination of perchlorate and iodide concentrations in edible seaweeds

Perchlorate and iodide concentrations were determined in brown (Undaria pinnatifida and Laminaria japonica) and red (Porphyra sp.) edible seaweeds, which are commonly consumed by Korean people, with the use of ion chromatography, coupled with a tandem mass spectrometer. Seaweeds (i.e., good sources of iodine) are among the most important plant life in the ocean and commonly consumed as food and nutritional supplement in South Korea. All seaweed samples were purchased from different regions in South Korea. The detected concentrations of perchlorate were as follows: 19.7–620.7 μg kg−1 dry weight (n = 11, mean concentration = 149.2 μg kg−1 dry weight) for L. japonica and 7.3–21.7 μg kg−1 dry weight (mean concentration = 10.6 μg kg−1 dry weight) for U. pinnatifida. Of the 11 samples of Porphyra sp., only 1 sample showed 6.7 μg kg−1 dry weight perchlorate. The concentrations of iodide in all seaweed samples varied from 0.44 to 6,800 mg kg−1 dry weight. L. japonica samples (n = 11) had significantly higher iodide concentrations, with a mean of 5,261 mg kg−1 dry weight. The bioconcentration factor values for perchlorate and iodide in the three different seaweeds varied widely and showed similar variation trends. The trend for perchlorate and iodide was Porphyra sp. < U. pinnatifida < L. japonica. The results have provided growing evidence that perchlorate frequently occurs in food products.

Utilization of Dispersive Liquid-Liquid Microextraction Coupled with HPLC-UV as a Sensitive and Efficient Method for the Extraction and Determination of Oleanolic Acid and Ursolic Acid in Chinese Medicinal Herbs

Isomeric triterpenic acids of oleanolic acid (OA) and ursolic acid (UA) both have very low ultraviolet absorption and always exist in the same plant, so the separation and simultaneous determination of them have been a difficult task. In this study, a sensitive method combining dispersive liquid-liquid microextraction (DLLME) with HPLC-UV was developed for the extraction and determination of OA and UA in traditional Chinese medicinal herbs (CMHs). Variables influencing DLLME such as type and volume of extraction solvent, volume of dispersive solvent, ionic strength, aqueous phase pH, extraction time, centrifugation speed and time, and sample volume were investigated and optimized. Under the optimum conditions, both OA and UA attained favorable extraction efficiencies with enrichment factors 1378 and 933, respectively. The linear dynamic ranges of 0.07 - 30.4 μg?mL–1 for OA and 0.08 - 33.6 μg?mL–1 for UA were obtained with square correlation coefficients of 0.9963. The detection limits of OA and UA were both 0.02 μg?mL–1. The method recoveries ranged between 88.2% - 116.2% for OA and 85.7% - 108.2% for UA with the RSDs (n = 5) lower than 8.6%. The proposed method was successfully applied to concentrate and simultaneously determine these two triterpenic acids in Hedyotis diffusa and Eriobotrya japonica samples.

Effects of elevated CO2on organic matter decomposition capacities and community structure of sulfate-reducing bacteria in salt marsh sediment

Increasing atmospheric affects the soil carbon cycle by influencing microbial activity and the carbon pool. In this study, the effects of elevated on extracellular enzyme activities (EEA; -glucosidase, N-acetylglucosaminidase, aminopeptidase) in salt marsh sediment vegetated with Suaeda japonica were assessed under ambient atmospheric concentration (380 ppm) or elevated concentration (760 ppm) conditions. Additionally, the community structure of sulfate-reducing bacteria (SRB) was analyzed via terminal restriction fragments length polymorphism (T-RFLP). Sediment with S. japonica samples were collected from the Hwangsando intertidal flat in May 2005, and placed in small pots (diameter 6 cm, height 10 cm). The pots were incubated for 60 days in a growth chamber under two different concentration conditions. Sediment samples for all measurements were subdivided into two parts: surface (0-2 cm) and rhizome (4-6 cm) soils. No significant differences were detected in EEA with different treatments in the surface and rhizome soils. However, the ratio of -glucosidase activity to N-acetylglucosaminidase activity in rhizome soil was significantly lower (P than at 380 ppm , thereby suggesting that the contribution of fungi to the decomposition of soil organic matter might in some cases prove larger than that of bacteria. Community structures of SRB were separated according to different treatments, suggesting that elevated may affect the carbon and sulfur cycle in salt marshes.

16S rRNA gene phylogenesis of culturable predominant bacteria from diseased Apostichopus japonicus (Holothuroidea, Echinodermata)

Cultured Apostichopus japonicus in China suffers from a kind of skin ulceration disease that has caused severe economic loss in recent years. The disease, pathogens of which are supposed to be bacteria by most researchers, is highly infectious and can often cause all individuals in the same culture pool to die in a very short time. The 16S rRNA gene phylogenesis of the culturable bacteria from the lesions of diseased individuals was conducted to study the biodiversity of the bacterial communities in the lesions and to identify probable pathogen(s) associated with this kind of disease. S. japonica samples were selected from a hatchery located in the eastern part of Qingdao, China. Bacterial universal primers GM5F and DS907R were used to amplify the 16S rRNA gene of bacteria colonies, and touchdown PCR was performed to amplify the target sequences. The results suggest that γ- proteobacteria (Alteromonadales and Vibrionales) of CFB group, many strains of which have been also determined as pathogens in other marine species, are the predominant bacterial genera of the diseased Apostichopus japonicus individuals.