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Panax ginseng is a major medicinal crop with pharmaceutical efficacy derived from ginsenoside metabolites. Despite its genome information, the inefficiency of ginseng transformation hinders the study of the molecular mechanism of ginseng plant metabolism. Thus, this protocol aimed to develop an easy and efficient system for ginseng transformation. We established a transformation system using ginseng callus cultured in a liquid medium, which has a higher ratio compared with cotyledon explant. In addition, Agrobacterium tumefaciens has been used for plant transformation. Compared with the LBA4404 strain, C58C1 was inappropriate for ginseng transformation using ginseng callus. We induced and maintained calli in liquid medium and cut them into small pieces before infection. After infection, we selected calli that survived from the antibiotic medium until identification of newly growing cells. In β-glucuronidase (GUS) gene assay, the expression of the GUS gene was observed in cells that were newly generated from explants. We treated calli with 0.05 M of MgSO4 before infection. After MgSO4 pre-treatment, the transformation efficiency of growing cells around infected callus was increased. Moreover, we constructed and introduced a visible reporter RUBY system to easily identify transformed cells. Using this system, we could identify the cells by a red color with naked eyes. Based on our transformation protocol, the success rate has increased to 77.27% in surviving lines during selection culture. This stable ginseng transformation could facilitate the overexpression and knockout of ginseng lines for functional or synthetic biological studies.

The present study examined the volatile profiles of

High temperature is one of the most significant abiotic stresses reducing crop yield and quality by inhibiting plant growth and development. Global warming has recently increased the frequency of heat waves, which negatively impacts agricultural fields. Despite numerous studies on heat stress responses and signal transduction in model plant species, the molecular mechanism underlying thermomorphogenesis in

The whole-genome sequence ofThe complete chloroplast genome sequence of

Panax ginseng Meyer is a valuable medicinal crop. However, the species’ propagation is limited by its long reproductive cycle and low seed yield. The present study focused on P. ginseng plant regeneration via somatic embryogenesis and evaluated the genetic stability of regenerated plantlets. We assessed the effects of carbon source type and concentration on somatic embryo induction, maturation, and germination. Somatic embryogenesis was optimal in Murashige and Skoog (MS) medium supplemented with 5% sucrose; however, maturation peaked in 1/2 MS containing low concentrations of sucrose ranging from 1 to 2%. Germination and plant regeneration were optimal in germination medium supplemented with 2% sucrose based on high germination rates, efficient plantlet production, and balanced growth characteristics. Molecular marker analysis suggested that the genetic fidelity of the regenerated plants was comparable with that of the control. High-performance liquid chromatography (HPLC) analysis showed that in vitro-grown roots (IGRs) accumulated more ginsenoside than those of the control, but the ginsenoside content of 2 year old IGRs was similar to that of the controls after acclimatization. Our study provides valuable insights into the optimization of genetically stable micropropagation and could promote the distribution of superior P. ginseng cultivars with high product yields and quality.

Panax ginseng Meyer is an important medicinal crop; however, most ginseng farmers cultivate native species that are not genetically fixed. Although several cultivars have been developed in Korea, distribution to farmers remains insufficient given their low propagation characteristics. This study compared the efficiency of seed production and micropropagation via somatic embryogenesis. Seeds were collected from cultivars, and zygotic embryo-derived explants were inoculated and cultured in a series of media for micropropagation. Seed production and characteristics of commercial cultivars were evaluated. The number of seeds from a 4-year-old individual cultivar was 23.1–58.8, and seed characteristics varied with cultivars. The genotype had a notable effect on somatic embryogenesis and plant regeneration. The number of somatic embryos and shoots obtained from a single seed of cultivars was 71.3–100.2 and 50.7–61.3, respectively. The number of in vitro grown roots (IGRs) per a single seed was 37.1–41.1 in one year. IGRs were successfully acclimatized and sprouted normally in the field. Flow cytometry analysis suggested that micropropagated plants had no ploidy variations. The results demonstrated the utility of somatic embryogenesis in the in vitro micropropagation of P. ginseng cultivars. Our findings can enhance the distribution of cultivars among farmers in the future.

Panax ginseng Meyer grows in east Russia and Asia. There is a high demand for this crop due to its medicinal properties. However, its low reproductive efficiency has been a hindrance to the crop’s widespread use. This study aims to establish an efficient regeneration and acclimatization system for the crop. The type of basal media and strength were evaluated for their effects on somatic embryogenesis, germination, and regeneration. The highest rate of somatic embryogenesis was achieved for the basal media MS, N6, and GD, with the optimal nitrogen content (≥35 mM) and NH4+/NO3− ratio (1:2 or 1:4). The full-strength MS medium was the best one for somatic embryo induction. However, the diluted MS medium had a more positive effect on embryo maturation. Additionally, the basal media affected shooting, rooting, and plantlet formation. The germination medium containing 1/2 MS facilitated good shoot development; however, the medium with 1/2 SH yielded outstanding root development. In vitro-grown roots were successfully transferred to soil, and they exhibited a high survival rate (86.3%). Finally, the ISSR marker analysis demonstrated that the regenerated plants were not different from the control. The obtained results provide valuable information for a more efficient micropropagation of various P. ginseng cultivars.

Ginseng, an important crop in East Asia, exhibits multiple medicinal and nutritional benefits because of the presence of ginsenosides. On the other hand, the ginseng yield is severely affected by abiotic stressors, particularly salinity, which reduces yield and quality. Therefore, efforts are needed to improve the ginseng yield during salinity stress, but salinity stress-induced changes in ginseng are poorly understood, particularly at the proteome-wide level. In this study, we report the comparative proteome profiles of ginseng leaves at four different time points (mock, 24, 72, and 96 h) using a label-free quantitative proteome approach. Of the 2484 proteins identified, 468 were salt-responsive. In particular, glycosyl hydrolase 17 (PgGH17), catalase-peroxidase 2, voltage-gated potassium channel subunit beta-2, fructose-1,6-bisphosphatase class 1, and chlorophyll a-b binding protein accumulated in ginseng leaves in response to salt stress. The heterologous expression of PgGH17 in Arabidopsis thaliana improved the salt tolerance of transgenic lines without compromising plant growth. Overall, this study uncovers the salt-induced changes in ginseng leaves at the proteome level and highlights the critical role of PgGH17 in salt stress tolerance in ginseng.

Ginseng (Panax ginseng) has a low genetic diversity and a narrow pool of genetic resources. Mutagenesis is one of the most powerful methods for inducing genetic variation in this species, but little research has been performed in ginseng. In this study, various tissues, including dehiscent seeds, 1-year-old roots, and somatic embryos were irradiated at different doses of gamma rays (20–400 Gy and zero dose as a control) to determine the most optimal concentration and tissues for the successful use of mutagenesis in ginseng breeding. The results revealed that high gamma doses (> 100 Gy) were detrimental to all irradiated tissues. A gradual and significant reduction in germination, emergence, and seedling growth were found as gamma irradiation dose increased. The reduction in survival rates and seedling growth by irradiation at serial doses showed that the LD50 of ginseng was 20–80 Gy, although the irradiation doses were tissue dependent. Based on our results, the optimal doses of gamma rays for inducing mutation in ginseng are

Ginseng (Panax ginseng C. A. Meyer), due to its relatively longer cultivation time, is often exposed to environmental stresses such as heat, salt, and drought. Particularly, salt-stress-derived oxidative damages greatly affect photosynthetic efficiency and consequently cause reduction of growth, development, and yield of ginseng. Thus, efforts have been made to understand the salt-stress-induced changes at proteome levels; however, the overall understanding of possible salt-responsive proteins in ginseng is still limited because of their low-abundance. A growing body of evidence suggests that plants secrete various low-abundant proteins localized in the intra- and extracellular spaces during stress conditions, and those proteins may have a key role for salt tolerance. Therefore, here, we report the ginseng fluids proteome to identify the potential salt-responsive proteins. This approach led to the identification of 261 secreted fluid proteins, and functional categorization revealed that identified proteins were majorly associated with photosynthesis, protein synthesis, cell binding, and various metabolisms. Further validation using qRT-PCR analysis showed similar expression profiles of heat-shock protein 70, glycosyl hydrolase 17, and fructose-bisphosphate aldolase class-I with proteome results. Overall, ginseng fluid proteomic analysis successfully identified the potential salt-responsive proteins, which might be helpful for understanding of salt-tolerance mechanisms in ginseng.

Agaricus bisporus is one of the world’s most popular edible mushrooms, including in South Korea. We performed de novo genome assembly with a South Korean white-colored cultivar of A. bisporus, KMCC00540. After generating a scaffold-level genomic sequence, we inferred chromosome-level assembly by genomic synteny analysis with the representative A. bisporus strains H97 and H39. The KMCC00540 genome had 13 pseudochromosomes comprising 33,030,236 bp mostly covering both strains. A comparative genomic analysis with cultivar H97 indicated that most genomic regions and annotated proteins were shared (over 90%), ensuring that our cultivar could be used as a representative genome. However, A. bisporus suffers from browning even from only a slight mechanical stimulus during transportation, which significantly lowers its commercial value. To identify which genes respond to a mechanical stimulus that induces browning, we performed a time-course transcriptome analysis based on the de novo assembled genome. Mechanical stimulus induces up-regulation in long fatty acid ligase activity-related genes, as well as melanin biosynthesis genes, especially at early time points. In summary, we assembled the chromosome-level genomic information on a Korean strain of A. bisporus and identified which genes respond to a mechanical stimulus, which provided key hints for improving the post-harvest biological control of A. bisporus.

Ginsenosides, a group of tetracyclic saponins, accounts for the nutraceutical and pharmaceutical relevance of the ginseng (Panax sp.) herb. Owing to the associated therapeutic potential of ginsenosides, their demand has been increased significantly in the last two decades. However, a slow growth cycle, low seed production, and long generation time of ginseng have created a gap between the demand and supply of ginsenosides. The biosynthesis of ginsenosides involves an intricate network of pathways with multiple oxidation and glycosylation reactions. However, the exact functions of some of the associated genes/proteins are still not completely deciphered. Moreover, ginsenoside estimation and extraction using analytical techniques are not feasible with high efficiency. The present review is a step forward in recapitulating the comprehensive aspects of ginsenosides including their distribution, structural diversity, biotransformation, and functional attributes in both plants and animals including humans. Moreover, ginsenoside biosynthesis in the potential plant sources and their metabolism in the human body along with major regulators and stimulators affecting ginsenoside biosynthesis have also been discussed. Furthermore, this review consolidates biotechnological interventions to enhance the biosynthesis of ginsenosides in their potential sources and advancements in the development of synthetic biosystems for efficient ginsenoside biosynthesis to meet their rising industrial demands.

Agaricus bisporus is a globally cultivated mushroom with high economic value. Despite its widespread cultivation, commercial button mushroom strains have little genetic diversity and discrimination of strains for identification and breeding purposes is challenging. Molecular markers suitable for diversity analyses of germplasms with similar genotypes and discrimination between accessions are needed to support the development of new varieties. To develop cleaved amplified polymorphic sequences (CAPs) markers, single nucleotide polymorphism (SNP) mining was performed based on the A. bisporus genome and resequencing data. A total of 70 sets of CAPs markers were developed and applied to 41 A. bisporus accessions for diversity, multivariate, and population structure analyses. Of the 70 SNPs, 62.85% (44/70) were transitions (G/A or C/T) and 37.15% (26/70) were transversions (A/C, A/T, C/G, or G/T). The number of alleles per locus was 1 or 2 (average = 1.9), and expected heterozygosity and gene diversity were 0.0–0.499 (mean = 0.265) and 0.0–0.9367 (mean = 0.3599), respectively. Multivariate and cluster analyses of accessions produced similar groups, with F-statistic values of 0.134 and 0.153 for distance-based and model-based groups, respectively. A minimum set of 10 markers optimized for accession identification were selected based on high index of genetic diversity (GD, range 0.299–0.499) and major allele frequency (MAF, range 0.524–0.817). The CAPS markers can be used to evaluate genetic diversity and population structure and will facilitate the management of emerging genetic resources.

The scientific and systematic classification of cultivation age is important for preventing age falsification and ensuring the quality of ginseng. Therefore, we applied deep learning to classify the cultivation age of ginseng. Deep learning, which is based on an artificial neural network, is one of the new class of models for machine learning, and is state-of-the-art. It is a powerful tool and has been used to solve complex problems in many fields. In the present study, powdered samples of 4-, 5-, and 6-year-old ginseng were measured using high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy. NMR data were analyzed with deep learning and partial least-squares discriminant analysis (PLS-DA) to improve accuracy. The accuracy of the PLS-DA was 87.1% and the accuracy of the deep learning model was 93.9%. NMR spectroscopy with deep learning can be a useful tool for discrimination of ginseng cultivation age.

Background: Ginseng is one of the well-known medicinal plants, exhibiting diverse medicinal effects. Its roots possess anticancer and antiaging properties and are being used in the medical systems of East Asian countries. It is grown in low-light and low-temperature conditions, and its growth is strongly inhibited at temperatures above 25°C. However, the molecular responses of ginseng to heat stress are currently poorly understood, especially at the protein level. Methods: We used a shotgun proteomics approach to investigate the effect of heat stress on ginseng leaves. We monitored their photosynthetic efficiency to confirm physiological responses to a high-temperature stress. Results: The results showed a reduction in photosynthetic efficiency on heat treatment (35°C) starting at 48 h. Label-free quantitative proteome analysis led to the identification of 3,332 proteins, of which 847 were differentially modulated in response to heat stress. The MapMan analysis showed that the proteins with increased abundance were mainly associated with antioxidant and translation-regulating activities, whereas the proteins related to the receptor and structural-binding activities exhibited decreased abundance. Several other proteins including chaperones, G-proteins, calcium-signaling proteins, transcription factors, and transfer/carrier proteins were specifically downregulated. Conclusion: These results increase our understanding of heat stress responses in the leaves of ginseng at the protein level, for the first time providing a resource for the scientific community. Keywords: High temperature, Panax ginseng, Photosynthetic efficiency, Shotgun proteomics

Ginseng (Panax ginseng C.A. Meyer) is a medicinal crop that requires a long culture time before it is ready to harvest, thus generating high economic and environmental costs. Symbiotic bacteria that live within the plant provide the host plant with many advantages in terms of metabolism and disease resistance. Here, we isolated endophytic bacteria from various tissues of P. ginseng seedlings using a culture-dependent method and we compared their tissue distribution. In addition, their antimicrobial activity against two fungal pathogens was investigated. Based on 16S rRNA sequencing, we identified 21 bacterial strains from ginseng seedlings. Leaves and rhizomes showed higher bacterial species diversity than root bodies and tails. While Bacillus strains were detected in all tissues, Xanthomonas and Micrococcaceae strains were specifically isolated from rhizome and leaf tissues, respectively. Fourteen bacterial strains showed antimicrobial activities against Cylindrocarpon destructans and/or Botrytis cinerea, with different activities. Among them, two strains (PgKB29 and PgKB35) showed strong antimicrobial activities against both fungi. Taken together, these results provide a better understanding of endophytic bacteria in P. ginseng seedlings and suggest the possibility of biological control of fungal pathogens using endophytic bacteria.

Korean ginseng (Panax ginseng C.A. Meyer) is a largely sought after resource due to its substantial health benefits. However, large-scale culture of ginseng seeds is limited because of its long maturation time and triple-dormancy. To develop new strategies to improve the dehiscence, germination, and breaking dormancy, we investigated the effects of stratification, gibberellin (GA3), and/or kinetin treatments on the dehiscence, development and germination of P. ginseng. Indehiscent seeds for the dehiscence test and dehiscent seeds for the germination test were immersed in a solution containing 288.7 μM GA3 and/or 232.3 μM kinetin for 24 h, respectively, at 25 °C. After dehiscence, a cold stratification experiment was conducted at 2 °C for 15, 30, 45, 60, and 90 days in the dark. Exogenous GA3 and kinetin treatments significantly improved the dehiscence rate. In addition, both plant growth regulators (PGRs) appeared to enhance the zygotic embryo development of ginseng seeds. GA3 on its own had a slight effect on breaking dormancy and germination, but the effect was not sufficient to serve as a complete substitute for the cold stratification requirement due to the deep physiological dormancy characteristic of P. ginseng. Kinetin treatment had a significant effect on germination and breaking dormancy, although a short period of cold stratification was also required. The combination of GA3 and kinetin treatments was more effective at enhancing both seed dehiscence and germination than either PGR treatment alone.

Gibberellins (GAs) are an important group of phytohormones associated with diverse growth and developmental processes, including cell elongation, seed germination, and secondary growth. Recent genomic and genetic analyses have advanced our knowledge of GA signaling pathways and related genes in model plant species. However, functional genomics analyses of GA signaling pathways in Panax ginseng, a perennial herb, have rarely been carried out, despite its well-known economical and medicinal importance. Here, we conducted functional characterization of GA receptors and investigated their physiological roles in the secondary growth of P. ginseng storage roots. We found that the physiological and genetic functions of P. ginseng gibberellin-insensitive dwarf1s (PgGID1s) have been evolutionarily conserved. Additionally, the essential domains and residues in the primary protein structure for interaction with active GAs and DELLA proteins are well-conserved. Overexpression of PgGID1s in Arabidopsis completely restored the GA deficient phenotype of the Arabidopsis gid1a gid1c (atgid1a/c) double mutant. Exogenous GA treatment greatly enhanced the secondary growth of tap roots, however, paclobutrazol (PCZ), a GA biosynthetic inhibitor, reduced root growth in P. ginseng. Transcriptome profiling of P. ginseng roots revealed that GA-induced root secondary growth is closely associated with cell wall biogenesis, the cell cycle, the jasmonic acid (JA) response, and nitrate assimilation, suggesting that a transcriptional network regulate root secondary growth in P. ginseng. These results provide novel insights into the mechanism controlling secondary root growth in P. ginseng.

White root rot disease, caused by the pathogen Rosellinia necatrix, is one of the world's most devastating plant fungal diseases and affects several commercially important species of fruit trees and crops. Recent global outbreaks of R. necatrix and advances in molecular techniques have both increased interest in this pathogen. However, the lack of information regarding the genomic structure and transcriptome of R. necatrix has been a barrier to the progress of functional genomic research and the control of this harmful pathogen. Here, we identified 10,616 novel full-length transcripts from the filamentous hyphal tissue of R. necatrix (KACC 40445 strain) using PacBio single-molecule sequencing technology. After annotation of the unigene sets, we selected 14 cell cycle-related genes, which are likely either positively or negatively involved in hyphal growth by cell cycle control. The expression of the selected genes was further compared between two strains that displayed different growth rates on nutritional media. Furthermore, we predicted pathogen-related effector genes and cell wall-degrading enzymes from the annotated gene sets. These results provide the most comprehensive transcriptomal resources for R. necatrix, and could facilitate functional genomics and further analyses of this important phytopathogen.

Panax ginseng, commonly known as ginseng, is a well-known medicinal plant that has been used as traditional medicine in China and Korea. Research in the past few decades supports the pharmacological effects of ginseng. For example, ginseng roots (extracts) exhibit multiple medicinal effects, such as anticancer, antiaging, and protection against circulatory shock, in humans. In this review, we summarize the progress made so far in the ginseng proteomics, starting from sample preparation to establishments of proteomes and databases. Both gel-based (1-DE and 2-DE in combination with LC–MS/MS) and gel-free proteomics technologies have been applied on wide range of samples, collected during different growth and developmental stages and under normal or adverse stress conditions. In particular, comparative proteome analysis has been carried out to investigate the protein profiles of Oriental, American and Indian ginsengs using majorly root and leaf tissues. Moreover, identification of stress-responsive proteins was a key focus that led to the detection of some of the common proteins such as heat shock protein (HSP), ATPase, enolase, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and ribonuclease/ginseng major protein (GMP). Acquired proteomics-based knowledge has been very fruitful in providing better insight into the ginseng biology, opening a door for comparative and translation research of other important medicinal plants. However, due to the fact that proteins undergo various post-transcriptional and post-translational modifications, obtained proteomics data do not always complement the transcriptomics data perfectly; therefore, future efforts would require the utilization of an integrated/holistic molecular-genetic (or omics) approach to explore the biology of this golden plant.

Background Licorice (Glycyrrhiza spp. L.) is used as a natural sweetener and medicinal herb in European and Asian countries. Molecular studies have been conducted to find differences between wild and cultivated species because most wild species are highly resistant to abiotic and biotic stresses compared with their cultivated species. However, few molecular markers have been developed for studying the genetic diversity and population structure of licorice species and to identify differences between cultivars. Thus, the present study aimed to develop a set of genomic simple sequence repeat (SSR) markers for molecular studies of these species. Methods In the present study, we developed polymorphic SSR markers based on whole-genomesequence data of Glycyrrhiza lepidota. Then, based on the sequence information, the polymorphic SSR markers were developed. The SSR markers were applied to 23 Glycyrrhiza individual plants. We also evaluated the phylogenetic relationships and interspecies transferability among samples. Results The genetic diversity analysis using these markers identified 2–23 alleles, and the major allele frequency, observed heterozygosity, genetic diversity, and polymorphism information content were 0.11–0.91, 0–0.90, 0.17–0.94, and 0.15–0.93, respectively. Interspecies transferability values were 93.5%, 91.6%, and 91.1% for G. echinata, G. glabra, and G. uralensis, respectively. Phylogenetic analysis clustered cultivated (group 1) and wild (group 2) species into three and two subgroups, respectively. The reported markers represent a valuable resource for the genetic characteri z ation of Glycyrrhiza spp. for theanalysis of its genetic variability, and as a tool for licorice transferability. This is the first intraspecific study in a collection of Glycyrrhiza spp. germplasm using SSR markers.

Background Licorice (Glycyrrhiza spp. L.) is used as a natural sweetener and medicinal herb in European and Asian countries. Molecular studies have been conducted to find differences between wild and cultivated species because most wild species are highly resistant to abiotic and biotic stresses compared with their cultivated species. However, few molecular markers have been developed for studying the genetic diversity and population structure of licorice species and to identify differences between cultivars. Thus, the present study aimed to develop a set of genomic simple sequence repeat (SSR) markers for molecular studies of these species. Methods In the present study, we developed polymorphic SSR markers based on whole-genomesequence data of Glycyrrhiza lepidota. Then, based on the sequence information, the polymorphic SSR markers were developed. The SSR markers were applied to 23 Glycyrrhiza individual plants. We also evaluated the phylogenetic relationships and interspecies transferability among samples. Results The genetic diversity analysis using these markers identified 2–23 alleles, and the major allele frequency, observed heterozygosity, genetic diversity, and polymorphism information content were 0.11–0.91, 0–0.90, 0.17–0.94, and 0.15–0.93, respectively. Interspecies transferability values were 93.5%, 91.6%, and 91.1% for G. echinata, G. glabra, and G. uralensis, respectively. Phylogenetic analysis clustered cultivated (group 1) and wild (group 2) species into three and two subgroups, respectively. The reported markers represent a valuable resource for the genetic characteri z ation of Glycyrrhiza spp. for theanalysis of its genetic variability, and as a tool for licorice transferability. This is the first intraspecific study in a collection of Glycyrrhiza spp. germplasm using SSR markers.

Bacillus cereus PgBE311, isolated from the root tissue of a 5-year-old Panax ginseng plant, showed activities against the fungal pathogens Cylindrocarpon destructans and Botrytis cinerea . Here, we report the genome sequence of B. cereus PgBE311.

Bacillus cereus PgBE311, isolated from the root tissue of a 5-year-old Panax ginseng plant, showed activities against the fungal pathogens Cylindrocarpon destructans and Botrytis cinerea. Here, we report the genome sequence of B. cereus PgBE311., Bacillus cereus PgBE311, isolated from the root tissue of a 5-year-old Panax ginseng plant, showed activities against the fungal pathogens Cylindrocarpon destructans and Botrytis cinerea. Here, we report the genome sequence of B. cereus PgBE311. The bacterium contains antibiotic-related gene clusters and has the potential to stimulate plant growth.

Chryseobacterium indologenes PgBE177, isolated from the root tissue of a 4-year-old Panax quinquefolius plant, showed antagonistic activity against Pseudomonas syringae pv. tomato DC3000, a bacterial pathogen., Chryseobacterium indologenes PgBE177, isolated from the root tissue of a 4-year-old Panax quinquefolius plant, showed antagonistic activity against Pseudomonas syringae pv. tomato DC3000, a bacterial pathogen. Here, we report the whole-genome sequence of C. indologenes PgBE177. The bacterium contains bacteriocin gene clusters and has the potential to stimulate plant growth.

Ginseng is one of the well-known medicinal plants, exhibiting diverse medicinal effects. Its roots possess anticancer and antiaging properties and are being used in the medical systems of East Asian countries. It is grown in low-light and low-temperature conditions, and its growth is strongly inhibited at temperatures above 25°C. However, the molecular responses of ginseng to heat stress are currently poorly understood, especially at the protein level.We used a shotgun proteomics approach to investigate the effect of heat stress on ginseng leaves. We monitored their photosynthetic efficiency to confirm physiological responses to a high-temperature stress.The results showed a reduction in photosynthetic efficiency on heat treatment (35°C) starting at 48 h. Label-free quantitative proteome analysis led to the identification of 3,332 proteins, of which 847 were differentially modulated in response to heat stress. The MapMan analysis showed that the proteins with increased abundance were mainly associated with antioxidant and translation-regulating activities, whereas the proteins related to the receptor and structural-binding activities exhibited decreased abundance. Several other proteins including chaperones, G-proteins, calcium-signaling proteins, transcription factors, and transfer/carrier proteins were specifically downregulated.These results increase our understanding of heat stress responses in the leaves of ginseng at the protein level, for the first time providing a resource for the scientific community.

Licorice (Glycyrrhiza glabra) is an important medicinal crop often used as health foods or medicine worldwide. The molecular genetics of licorice is under scarce owing to lack of molecular markers. Here, we have developed cleaved amplified polymorphic sequence (CAPS) and high-resolution melting (HRM) markers based on single nucleotide polymorphisms (SNP) by comparing the chloroplast genomes of two Glycyrrhiza species (G. glabra and G. lepidota). The CAPS and HRM markers were tested for diversity analysis with 24 Glycyrrhiza accessions. The restriction profiles generated with CAPS markers classified the accessions (2-4 genotypes) and melting curves (2-3) were obtained from the HRM markers. The number of alleles and major allele frequency were 2-6 and 0.31-0.92, respectively. The genetic distance and polymorphism information content values were 0.16-0.76 and 0.15-0.72, respectively. The phylogenetic relationships among the 24 accessions were estimated using a dendrogram, which classified them into four clades. Except clade III, the remaining three clades included the same species, confirming interspecies genetic correlation. These 18 CAPS and HRM markers might be helpful for genetic diversity assessment and rapid identification of licorice species.

This study was conducted to investigate growth characteristics to develop the technique to select resistant cultivar by wet injury at an early stage through the automatic irrigation maintaining 30, 20, 10 kPa respectively using native variety, Chunpoong, Yunpoong, Gumpoong and Sunun. The aerial growth was decreased at 10 kPa compared to 30 kPa. In addition, the survival rate was decreased by 66.6%, 62.3%, 33.8% at 30, 20, 10 kPa, respectively. The survival rate of Chunpoong and Gumpoong were higher than others at 10 kPa. While root growth characteristics such as root length, root weight, number of lateral root and side root were tended to decrease, root diam was no significant or increased. And the more humid condition is, the more the incidence rate of rusty root and rough skin were tended to increase. The epidermal thickness of Chunpoong and Gumpoong was increased but the figures of native variety, Yunpoong and Sunun were decreased at 10 kPa compared to 30 kPa. But, the tissue stiffness of root was decreased at 10 kPa compared to 30 kPa.

Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

Background : In order to determine the effects of planting methods on root growth of ginseng varieties, two different methods, direct seed sowing and transplanting were compared in terms of their effects on different root growth characteristics. Methods and Results : Higher fresh root weight was observed in ginseng grown by direct seed sowing. Direct seed sowing of three cultivars (Sunhyang, Chungsun and K-1) resulted in higher yield, whereas no difference was observed in the yield of one cultivar (Chungsun). Gumpoong was highly tolerant to physiological stress, as it showed fewer symptoms of rusty and rough skin root diseases in both direct seed sowing and transplanting. The average main root length per total root length of ginseng grown by direct seed sowing was 33.6%, whereas that of ginseng grown by the average of those by transplanting was 22.4%. Other root growth characteristics, including root length, main root diameter, and number of side roots, improved when the direct seed sowing method was used. Conclusions : To our knowledge, this is the first study reporting the differences in root growth parameters of ginseng varieties grown by direct seed sowing or transplanting at the same planting density. Because of the advantages of direct sowing during ginseng planting, developing new varieties and improving cultivation methods are imperative.

Background : Excessively high concentration of sodium ion causednutrient deficiency and significantly decrease growth. This study was carried out to determine the limiting concentration range of sodium ion in the soil of ginseng field. Methods and Results : The growth of the ginseng cultivar Chunpoong reduced with increase in salinity, and the rate of growth reduction was higher in shoots than that of roots. Particularly, ginseng plants cultivated at high level of nitrate nitrogen or sodium may suffer delayed development and stunted growth. Chlorophyll damage occurred on the leaves of ginseng planted in relatively high levels (> ) of sodium ion, as determined by the fluorescence reaction. The incidence of physiological disorder in ginseng cultivated at 249 sites was correlated with the concentration of sodium ion in the soils. About 74% of ginseng fields in which physiological disorders occurred had concentrations of sodium ion in soil greater than . In contrast, the concentration of sodium ions at 51 of 85 sites where no damage occurred was relatively (). Conclusions : The concentration of sodium ion in soil of ginseng fields can be classified into three levels optimum (), permissible allowance (0.15 - 0.2) and excessive (> 0.2).

This study was carried out to investigate the effects of the Cirsium japonicum var. ussuriense (C. japonicum) extract on serum level of hormones from induced osteoporosis by ovariectomized rats. Two month-old rats were ovariecto- mized (OVX), remained untreated for 8 weeks, and were subsequently administered C. japonicum (200 ㎎/㎏) every day for 8 weeks. We examined the effects of treated C. japonicum on ovariectomy-related changes in Insulin-like Growth Factors (IGF), Insulin-like Growth Factor binding protein-3 (IGBF-3), Estrogen, Calcium, and Phosporus. After 8 weeks, the serum levels of IGF-I, -II, and IGFBP-3 were higher presented as compared to the other two groups (P < 0.05), in the C. japonicum extract treatment on OVX rats. There were differences between OVX and C. japonicum extract treated OVX rats in serum levels of Ca 2+ , but Ca 2+ levels for the normal group was higher than for the other two groups. The C. japonicum extract increased both serum IGFs and IGFBP-3 levels on induced osteoporotic rat by ovariectomized. Thus, these results revealed that the C. japonicum extract is a possible role for improvement of osteoporosis induced-ovariectomized rats and has a great potential as an alternative tool for the treatment of osteoporosis.

Licorice (Glycyrrhiza glabra) is an important medicinal crop often used as health foods or medicine worldwide. The molecular genetics of licorice is under scarce owing to lack of molecular markers. Here, we have developed cleaved amplified polymorphic sequence (CAPS) and high-resolution melting (HRM) markers based on single nucleotide polymorphisms (SNP) by comparing the chloroplast genomes of two Glycyrrhiza species (G. glabra and G. lepidota). The CAPS and HRM markers were tested for diversity analysis with 24 Glycyrrhiza accessions. The restriction profiles generated with CAPS markers classified the accessions (2–4 genotypes) and melting curves (2–3) were obtained from the HRM markers. The number of alleles and major allele frequency were 2−6 and 0.31–0.92, respectively. The genetic distance and polymorphism information content values were 0.16–0.76 and 0.15–0.72, respectively. The phylogenetic relationships among the 24 accessions were estimated using a dendrogram, which classified them into four clades. Except clade III, the remaining three clades included the same species, confirming interspecies genetic correlation. These 18 CAPS and HRM markers might be helpful for genetic diversity assessment and rapid identification of licorice species.

Variovorax paradoxus KB5, isolated from the inside of Arabidopsis thaliana leaves, showed antibacterial activity against the phytopathogen Pseudomonas syringae pv. tomato DC3000. Here, we report a draft genome sequence of V. paradoxus KB5, which contains a delftibactin-like nonribosomal peptide biosynthetic gene cluster.

White root rot disease, caused by the pathogen Rosellinia necatrix, is one of the world’s most devastating plant fungal diseases and affects several commercially important species of fruit trees and crops. Recent global outbreaks of R. necatrix and advances in molecular techniques have both increased interest in this pathogen. However, the lack of information regarding the genomic structure and transcriptome of R. necatrix has been a barrier to the progress of functional genomic research and the control of this harmful pathogen. Here, we identified 10,616 novel full-length transcripts from the filamentous hyphal tissue of R. necatrix (KACC 40445 strain) using PacBio single-molecule sequencing technology. After annotation of the unigene sets, we selected 14 cell cycle-related genes, which are likely either positively or negatively involved in hyphal growth by cell cycle control. The expression of the selected genes was further compared between two strains that displayed different growth rates on nutritional media. Furthermore, we predicted pathogen-related effector genes and cell wall-degrading enzymes from the annotated gene sets. These results provide the most comprehensive transcriptomal resources for R. necatrix, and could facilitate functional genomics and further analyses of this important phytopathogen.

This study was performed to identify optimal harvesting time of ginseng seeds and to examine the effect of GA3 treatment for improvement of seed stratification rate. Ginseng seeds harvested from Land race, Chunpoong and Yun- poong cultivar in July 20 were tested for stratification rate. It was shown that stratification rates of land race, Yunpoong and Chunpoong cultivar were 94.1%, 93.1%, and 82.6%, respectively. Seeds of Chunpoong cultivar harvested 10-15 days later showed a comparable stratification rate to that of Land race, indicating that late harvest of Chunpoong seeds is beneficial for the increase of stratification rate. The higher stratification rate was found in mature seeds (92.3%) than immature seeds (37.8%), both of which were harvested in July 20. Stratification rate of mature seeds harvested in July 15 was 87.5%, dem- onstrating optimal harvesting time of ginseng seeds with higher stratification rate is after mid-July. An exponential growth of endosperms of ginseng seeds was observed from early June to mid-June and then slow growth was observed. There was no obvious growth of embryos from fertilization to mid-August. After the this time, embryos quickly grew until late October. Thus, appropriate stratification control is essential during the period (from early September to late October) in order to optimize embryo growth and development. While no increase of stratification rate was observed in seeds treated with 50 ppm of GA3, significant increases were observed in seeds treated with 100 ppm of GA3. At this concentration of GA3, the stratification rate of Land race, Chunpoong and Yunpoong cultivar was 95.0%, 95.3%, and 96.5%, respectively.