Genome and evolution of the shade-requiring medicinal herb Panax ginseng.

Panax ginseng C. A. Meyer, reputed as the king of medicinal herbs, has slow growth, long generation time, low seed production and complicated genome structure that hamper its study. Here, we unveil the genomic architecture of tetraploid P. ginseng by de novo genome assembly, representing 2.98 Gbp with 59 352 annotated genes. Resequencing data indicated that diploid Panax species diverged in association with global warming in Southern Asia, and two North American species evolved via two intercontinental migrations. Two whole genome duplications (WGD) occurred in the family Araliaceae (including Panax) after divergence with the Apiaceae, the more recent one contributing to the ability of P. ginseng to overwinter, enabling it to spread broadly through the Northern Hemisphere. Functional and evolutionary analyses suggest that production of pharmacologically important dammarane-type ginsenosides originated in Panax and are produced largely in shoot tissues and transported to roots; that newly evolved P. ginseng fatty acid desaturases increase freezing tolerance; and that unprecedented retention of chlorophyll a/b binding protein genes enables efficient photosynthesis under low light. A genome-scale metabolic network provides a holistic view of Panax ginsenoside biosynthesis. This study provides valuable resources for improving medicinal values of ginseng either through genomics-assisted breeding or metabolic engineering.

Development and authentication of Panax ginseng cv. Sunhong with high yield and multiple tolerance to heat damage, rusty roots and lodging.

Ginseng (Panax ginseng) has been used as a valuable medicinal plant in Asia, and the demand for ginseng production for health functional food is increasing worldwide after the COVID-19 crisis. Although a number of cultivars have been developed to increase ginseng production, none of them were widely cultivated in Korea because they could not resist various environmental stresses while being grown in one place for at least 4 years. To address this, Sunhong was developed as a ginseng cultivar with high yield and multiple stress tolerance by pure line selection. Sunhong showed high yield and heat tolerance comparable to Yunpoong, a representative high-yielding cultivar, and exhibited 1.4 times lower prevalence of rusty roots than Yunpoong, suggesting that Sunhong can keep its high yield and quality during long-term cultivation. In addition, distinct color and lodging resistance were expected to increase the convenience of cultivation. To supply pure seeds to farmers, we also established a reliable high-throughput authentication system for Sunhong and seven ginseng cultivars through genotyping-by-sequencing (GBS) analysis. The GBS approach enabled to identify a sufficient number of informative SNPs in ginseng, a heterozygous and polyploid species. These results contribute to the improvement of yield, quality, and homogeneity, and therefore promote the ginseng industry. Supplementary Information: The online version contains supplementary material available at 10.1007/s13580-023-00526-x.

Lactobacillus koreensis sp. nov., isolated from the traditional Korean food kimchi.

A lactic acid bacterium, strain DCY50(T), isolated from the traditional Korean food kimchi, was studied to determine its taxonomic position. The strain was Gram-stain-positive, catalase-negative, facultatively anaerobic, rod-shaped and motile. The genomic DNA G+C content was 49 mol% and the peptidoglycan structure was of the A4α (l-Lys-d-Asp) type. Chemotaxonomic markers of the strain were consistent with its classification in the genus Lactobacillus. Comparisons of 16S rRNA and rpoA gene sequences showed that strain DCY50(T) was most closely related to the type strains of Lactobacillus parabrevis (98.4 and 91.6 % similarity, respectively, for the 16S rRNA and rpoA genes), L. hammesii (98.0 and 91.2 %), L. brevis (97.6 and 93.3 %) and L. senmaizukei (97.4 and 90.5 %). DNA-DNA relatedness of strain DCY50(T) to these type strains was below 36 %. According to the genotypic and phenotypic data, strain DCY50(T) could be differentiated from all known Lactobacillus species and should be classified in a novel species, for which the name Lactobacillus koreensis sp. nov. is proposed; the type strain is DCY50(T) ( = KCTC 13530(T)  = JCM 16448(T)).

In silico gene expression analysis in Codonopsis lanceolata root.

Expressed sequence tags (ESTs) provide valuable tools that can be used to predict the genes involved in primary and secondary metabolite synthesis. To the best of our knowledge, ESTs have not yet been developed for Codonopsis. lanceolata, and therefore, the EST referenced in this report is the first transcript for C. lanceolata. A cDNA library was constructed using the roots of C. lanceolata plants that were grown in a field. The selected 881 cDNA clones were sequenced and processed with an EST pipeline, resulting in 636 unique sequences, including 517 singletons and 119 contig sequences. Using bioinformatics tools, 81% of the EST sequence was putatively annotated. Data for unique transcripts were mined from biological databases and functionally classified using gene ontology (GO), the Kyoto Encyclopedia of Genes and Genomes Orthology, KEGG pathway maps, and protein family. The GO-based analyses were examined in terms of biotic and abiotic stress response, transport, cellular component organization, biogenesis, and secondary metabolic processes. The KEGG-based analyses of most transcripts were sorted by carbohydrate metabolism, energy metabolism, and biosynthesis of secondary metabolites. Five randomly-selected putative genes were used for an expression study using various stresses such as salt, H(2)O(2), salicylic acid, and methyl jasmonic acid. Mined data were organized in "The Codonopsis EST Database" (www.bioherbs.khu.ac.kr/Codonopsis).

Identification and characterization of class I chitinase in Panax ginseng C. A. Meyer.

The role of plant chitinases in protecting plants against a variety of fungal pathogens is well established. In the present study, a cDNA clone containing a class I chitinase (Chi-1) gene, designated as PgChi-1, has been isolated from the oriental medicinal plant Panax ginseng. PgChi-1 is predicted to encode a protein of 34.9 kDa consisting of 323 amino acid residues. PgChi-1 was found to be expressed constitutively in all of the studied organs of ginseng plant. Under various abiotic stress treatments including Cu, H2O2, mannitol, SA, JA, and NaCl, the expression of PgChi-1 in plantlets and hairy roots increased significantly compared to the control. When different parts of root were analyzed, maximum level was observed in taproot. In addition, levels of PgChi-1 expression were compared between healthy root and fungal, bacterial, and nematode infected root. Significant increase of PgChi-1 was noticed in pathogen infected roots than healthy roots. This study revealed that PgChi-1 may protect the P. ginseng under both biotic and abiotic stress conditions.

Effects of red ginseng extract on the epididymal sperm motility of mice exposed to ethanol.

The protective effects of red ginseng extract and ginseng wine against ethanol-induced male reproductive toxicity were evaluated in male mice using computer-assisted sperm analysis. Mice were divided into 4 groups of 10 and fed plain saline, 6 g/kg per d of ethanol in saline, red ginseng extract plus ethanol, or a fermented preparation of red ginseng extract daily for 5 weeks. We found that the average seminal vesicle weight was significantly lower in the ethanol-treated group compared to the control group, while those of the ginseng-treated groups tended to be higher than the ethanol-treated group. We found a significant decrease in sperm motility and progressiveness in mice treated with ethanol for 5 weeks, while administration of ethanol plus red ginseng extract appeared to minimize the negative effects of ethanol toxicity on male fertility. Serum testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were insignificantly lower in the ethanol-treated group than in the control group.

Phenological growth stages of Korean ginseng (Panax ginseng) according to the extended BBCH scale.

BACKGROUND: Phenological studies are a prerequisite for accomplishing higher productivity and better crop quality in cultivated plants. However, there are no phenological studies on Panax ginseng that improve its production yield. This study aims to redefine the phenological growth stages of P. ginseng based on the existing Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH) scale and proposes a disease control reference. METHODS: This study was conducted at the Korea Ginseng Corporation Experiment Station in Gyeonggi province, South Korea. Phenological observations were performed once weekly or twice monthly, based on the developmental stages. The existing BBCH scale with a three-digit code was used to redefine and update P. ginseng's phenological growth codes. RESULTS: The phenological description is divided into eight principal growth stages: three for vegetative growth (perennating bud, aerial shoot, and root development), four for reproductive growth (reproductive organ development, flowering, fruit development, and fruit maturation), and one for senescence according to the extended BBCH scale. A total of 58 secondary growth stages were described within the eight principal growth stages. Under each secondary growth stage, four mesostages are also taken into account, which contains the distinct patterns of the phenological characteristics in ginseng varieties and the process of transplanting seedlings. A practical management program for disease control was also proposed by using the BBCH code and the phenological data proposed in this work. CONCLUSION: The study introduces an extended BBCH scale for the phenological research of P. ginseng.

Rg3-enriched Korean Red Ginseng extract inhibits blood-brain barrier disruption in an animal model of multiple sclerosis by modulating expression of NADPH oxidase 2 and 4.

BACKGROUND: Multiple sclerosis (MS) and its animal model, the experimental autoimmune encephalomyelitis (EAE), are primarily characterized as dysfunction of the blood-brain barrier (BBB). Ginsenoside-Rg3-enriched Korean Red Ginseng extract (Rg3-KRGE) is known to exert neuroprotective, anti-inflammatory, and anti-oxidative effects on neurological disorders. However, effects of Rg3-KRGE in EAE remain unclear. METHODS: Here, we investigated whether Rg3-KRGE may improve the symptoms and pathological features of myelin oligodendroglial glycoprotein (MOG)35-55 peptide - induced chronic EAE mice through improving the integrity of the BBB. RESULTS: Rg3-KRGE decreased EAE score and spinal demyelination. Rg3-KRGE inhibited Evan's blue dye leakage in spinal cord, suppressed increases of adhesion molecule platelet endothelial cell adhesion molecule-1, extracellular matrix proteins fibronection, and matrix metallopeptidase-9, and prevented decreases of tight junction proteins zonula occludens-1, claudin-3, and claudin-5 in spinal cord following EAE induction. Rg3-KRGE repressed increases of proinflammatory transcripts cyclooxygenase-2, inducible nitric oxide synthase, interleukin (IL)-1 beta, IL-6, and tumor necrosis factor-alpha, but enhanced expression levels of anti-inflammatory transcripts arginase-1 and IL-10 in the spinal cord following EAE induction. Rg3-KRGE inhibited the expression of oxidative stress markers (MitoSOX and 4-hydroxynonenal), the enhancement of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2) and NOX4, and NADPH activity in the spinal cord of chronic EAE mice. Furthermore, apocynin, a NOX inhibitor, mimicked beneficial effects of Rg3-KRGE in chronic EAE mice. CONCLUSION: Our findings suggest that Rg3-KRGE might alleviate behavioral symptoms and pathological features of MS by improving BBB integrity through modulation of NOX2/4 expression.

Generation and gene ontology based analysis of expressed sequence tags (EST) from a Panax ginseng C. A. Meyer roots.

Panax ginseng C. A. Meyer is a perennial herb from the Araliaceae family. Traditionally used as a medicinal plant in Oriental medicine for more than thousand years. Ginsenosides are the major therapeutic components in ginseng roots. Roots of the ginseng plant have more commercial value and based on the age. No genomic data available till now. In this study, transcriptome analysis for hairy root, 14 year root, 4 year root get insight in to ginsenoside pathway and genes responsible for long survival and stress. Totally 6,757 Expressed Sequence Tags (EST) was obtained from cDNA libraries. Clustering of those ESTs returned 1,037 contigs and 3,445 singlets for a total of 4,482 putative unigenes. Use of bioinformatics methods 85% of EST sequence was well annotated towards reeds one dimensional concept. The unique transcripts were functionally classified by using Gene Ontology (GO) hierarchy, Kyoto Encyclopedia of Genes and Genomes (KEGG), KEGG orthology and structural domain data from biological database. Isoprenoid and putative ginsenoside pathway genes were discussed. EST dataset provides a wide outlook of the genes expressed in hairy roots, 14 years root and 4 years root. The dataset contains more than 1,365 EST sequences related to plant secondary metabolism and 745 sequences related to stresses. This study will improve the genetic engineering of ginseng plant and ginsenosides rich plant development. One dimensional data will lead to the two and three dimensional data.

Isolation and characterization of a Glutamate decarboxylase (GAD) gene and their differential expression in response to abiotic stresses from Panax ginseng C. A. Meyer.

Glutamate decarboxylase (GAD) catalyzes the conversion of L-glutamate to γ-aminobutyric acid (GABA). A full-length cDNA encoding GAD (designated as PgGAD) was isolated and characterized from the root of Panax ginseng C. A. Meyer. The length cDNA of PgGAD was 1881 bp and contained a 1491 bp open reading frame (ORF) encoding a glutamate decarboxylase protein of 496 amino acids, possessing a Ser-X-X-Lys active site, which belongs to the GAD group. The deduced amino acid sequence of the PgGAD was classified in the plant GAD family and has 76-85% high similarity with other plants as like petunia, Arabidopsis, tomato. Secondary structure of PgGAD was predicted by using SOPMA software program. Southern blot analysis of genomic DNA suggests that, there is more than one copy of the PgGAD gene. The organ specific gene expression pattern also studied in P. ginseng seedlings, in which the stem showed elevated expression than root, leaf, bud and rhizomes. Along with this, we also confirmed the gene expression of PgGAD under various abiotic stresses like temperature stress, osmotic stress, anoxia, oxidative stress, and mechanical damage. Temporal analysis of gene expression except exposure of oxidative stress revealed an enhanced expression after each stresses. The enzyme activity of PgGAD was stimulated to 2-fold under cold stress.

Development of molecular markers for the determination of the new cultivar 'Chunpoong' in Panax ginseng C. A. Meyer associated with a major latex-like protein gene.

Chunpoong is one of the most valuable cultivars of Panax ginseng C. A. MEYER, and is widely grown in Korea and China. Insertion/deletion (InDel) markers and single nucleotide polymorphism (SNP) markers are useful tools for marker-assisted selections. The SNP marker for determinate Chunpoong was previously developed from the nad7 gene of mtDNA by Wang et al. (2009) but was effective only on a limited range of cultivars. In this study, we studied the reasons for this limited application and developed new useful markers for application in Chunpoong-breeding programs. The new markers of InDel and SNP were designed in the major latex-like protein (MLP-like) gene which was highly expressed in 4-year-old Chunpoong expressed sequence tags (ESTs). To validate the marker in polymerase chain reaction (PCR), we used an InDel marker for identification of Chunpoong in the 70 Panax samples based on a double-blind test, and the success rate was 100%. For rapid and reliable assay of Chunpoong in numerous samples, we utilized an EvaGreen dye and melting curve method on real-time PCR. Furthermore, we designed an SNP marker that depended on the InDel region for more efficient detection of Chunpoong in real-time PCR. Compared with PCR-based assays, our Chunpoong SNP marker and real-time PCR assay offers a significant savings of time and labor in the analysis of large numbers of Chunpoong samples.

Korean Red Ginseng alleviates dehydroepiandrosterone-induced polycystic ovarian syndrome in rats via its antiinflammatory and antioxidant activities.

BACKGROUND: Beneficial effects of Korean Red Ginseng (KRG) on polycystic ovarian syndrome (PCOS) remains unclear. METHODS: We examined whether pretreatment (daily from 2 hours before PCOS induction) with KRG extract in water (KRGE; 75 and 150 mg/kg/day, p.o.) could exert a favorable effect in a dehydroepiandrosterone (DHEA)-induced PCOS rat model. RESULTS: Pretreatment with KRGE significantly inhibited the elevation of body and ovary weights, the increase in number and size of ovarian cysts, and the elevation of serum testosterone and estradiol levels induced by DHEA. Pretreatment with KRGE also inhibited macrophage infiltration and enhanced mRNA expression levels of chemokines [interleukin (IL)-8, monocyte chemoattractant protein-1), proinflammatory cytokines (IL-1ß, IL-6), and inducible nitric oxide synthase in ovaries induced by DHEA. It also prevented the reduction in mRNA expression of growth factors (epidermal growth factor, transforming growth factor-beta (EGF, TGF-ß)) related to inhibition of the nuclear factor kappa-light-chain-enhancer of activated B cell pathway and stimulation of the nuclear factor erythroid-derived 2-related factor 2 pathway. Interestingly, KRGE or representative ginsenosides (Rb1, Rg1, and Rg3(s)) inhibited the activity of inflammatory enzymes cyclooxygenase-2 and iNOS, cytosolic p-IkB, and nuclear p-nuclear factor kappa-light-chain-enhancer of activated B in lipopolysaccharide-induced RAW264.7 cells, whereas they increased nuclear factor erythroid-derived 2-related factor 2 nuclear translocation. CONCLUSION: These results provide that KRGE could prevent DHEA-induced PCOS via antiinflammatory and antioxidant activities. Thus, KRGE may be used in preventive and therapeutic strategies for PCOS-like symptoms.

Isolation of S-adenosyl-L-methionine synthetase gene from Panax ginseng C.A. meyer and analysis of its response to abiotic stresses.

A cDNA clone containing a S-adenosyl-L-methionine synthetase (SAMS) gene, named as PgSAM, was isolated from a commercial medicinal plant Panax ginseng. PgSAM is predicted to encode a precursor protein of 307 amino acid residues, and its sequence shares high homology with a number of other plant SAMS. PgSAM is expressed at different levels in various organs of ginseng. The expression of PgSAM in adventitious roots and hairy roots of P. ginseng were analyzed using reverse transcriptase (RT)-PCR and real-time PCR under various abiotic stresses. Salt, salicylic acid, abscisic acid and chilling stresses induced PgSAM significantly at different time points within 2-72 h post-treatment. This study revealed that PgSAM may help to protect the plants against various abiotic stresses.

Comparative analysis of the transcriptomes and primary metabolite profiles of adventitious roots of five Panax ginseng cultivars.

BACKGROUND: Various Panax ginseng cultivars exhibit a range of diversity for morphological and physiological traits. However, there are few studies on diversity of metabolic profiles and genetic background to understand the complex metabolic pathway in ginseng. METHODS: To understand the complex metabolic pathway and related genes in ginseng, we tried to conduct integrated analysis of primary metabolite profiles and related gene expression using five ginseng cultivars showing different morphology. We investigated primary metabolite profiles via gas chromatography-mass spectrometry (GC-MS) and analyzed transcriptomes by Illumina sequencing using adventitious roots grown under the same conditions to elucidate the differences in metabolism underlying such genetic diversity. RESULTS: GC-MS analysis revealed that primary metabolite profiling allowed us to classify the five cultivars into three independent groups and the grouping was also explained by eight major primary metabolites as biomarkers. We selected three cultivars (Chunpoong, Cheongsun, and Sunhyang) to represent each group and analyzed their transcriptomes. We inspected 100 unigenes involved in seven primary metabolite biosynthesis pathways and found that 21 unigenes encoding 15 enzymes were differentially expressed among the three cultivars. Integrated analysis of transcriptomes and metabolomes revealed that the ginseng cultivars differ in primary metabolites as well as in the putative genes involved in the complex process of primary metabolic pathways. CONCLUSION: Our data derived from this integrated analysis provide insights into the underlying complexity of genes and metabolites that co-regulate flux through these pathways in ginseng.

Characteristics of Korean ginseng varieties of Gumpoong, Sunun, Sunpoong, Sunone, Cheongsun, and Sunhyang.

BACKGROUND: Ginseng (Panax ginseng Meyer) is an important medicinal herbs in Asia. However, ginseng varieties are less developed. METHOD: To developed ginseng varieties, a pure line selection method was applied in this study. RESULTS: Gumpoong was testing of 4-yr-old specimens in 2002, the proportions of the below-ground roots that were rusty colored for Gumpoong was 1.29 in Daejeon and 1.45 in Eumseong, whereas the proportions for its yellow berry variant were 2.60 and 2.45 in the two regions, respectively. Thus the Gumpoong was resistant to root rust. Sunpoong has a high yielding property. Its average root weight is 70.6 g for 6-yr-old roots. Its yield is 2.9 kg/1.62m(2) and the rate of heaven- and earth-grade product is 20.9%, which is very high compared to 9.4% for Yunpoong. Sunone is resistance to root rot and the survival rate of 4-yr-old roots was 44.4% in 1997, whereas that of the violet-stem variant landrace was 21.7%. Sunhyang has content of arginyl-fructosyl-glucose (AFG), which produces the unique scent of red ginseng, is 95.1 µmol/g and greater than the 30.8 µmol/g of Chunpoong in 6-yr-old plants. Sunun and Cheongsun are being nurtured to protect genetic resources. CONCLUSION: Developed ginsneg varieties will be used as the basis for the protection of genetic resources and breeding.

Molecular cloning and characterization of nucleoside diphosphate (NDP) kinases from Chinese cabbage (Brassica campestris).

Nucleoside diphosphate kinases (NDPKs) are key metabolic enzymes that catalyze the synthesis of non-adenosine nucleoside triphosphates (NTP) by transfer of the terminal phosphate between NDP and NTP. Recently we isolated three NDPK cDNAs from Chinese cabbage cDNA library. BcNDK1 has 636 bp and encodes a putative 17.4 kDa protein, BcNDK2 has 854 bp and encodes a putative 25.5 kDa protein, and BcNDK3 is 986 bp long and encodes a putative 25.7 kDa protein. The precursor proteins of BcNDK2 and BcNDK3 have long N-terminal extensions containing putative chloroplast or mitochondrial targeting sequences. A phylogenic tree showed that the 3 BcNDKs are highly homologous to other plant NDPK genes, especially those of Arabidopsis. Expression of the BcNDK genes depended on the developmental stage and the conditions of seed germination. Most notably, expression of BcNDK2 increased dramatically in seedlings transferred to the light after germinating in the dark. In addition, BcNDK3 differed from BcNDK1 in being highly expressed in the hooks and cotyledons of seedlings. Although all BcNDKs were highly expressed in petals, BcNDK1 was also expressed in pistils. Expression of each of the BcNDKs increased as the flower bud matured. These results indicate that NDPKs are involved in physiological pathways activated by a variety of environmental conditions and at different developmental stages.

A simple and rapid technique for the authentication of the ginseng cultivar, Yunpoong, using an SNP marker in a large sample of ginseng leaves.

Yunpoong is an important Korean ginseng (Panax ginseng C. A. Meyer) cultivar, but no molecular marker has been available to identify Yunpoong from other cultivars. In this study, we developed a single nucleotide polymorphism (SNP) marker for Yunpoong based on analysis of expressed sequence tags (ESTs) in an exon region of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. This SNP marker had high specificity to authenticate Yunpoong in twelve different main ginseng cultivars. For application of the molecular marker, a rapid identification method was established based on the NaOH-Tris method and real-time polymerase chain reaction (PCR) in order to ensure more efficiency in the cultivar selection. The biggest feature of the NaOH-Tris method was that it made the extraction of DNA very simple and rapid in young leaf tissues. We only spent 1 min to extract DNA and directly used it to do PCR. In this report, the conventional DNA extraction method was used to develop molecular marker process, and the NaOH-Tris method was applied in screening large numbers of cultivars. Moreover, the greatest advantage of the real-time PCR compared with traditional PCR, is time saving and high efficiency. Thus, this strategy provides a rapid and reliable method for the specific identification of Yunpoong in a large number of samples.

Morphological Characteristics of Ginseng Leaves in High-Temperature Injury Resistant and Susceptible Lines of Panax ginseng Meyer.

Plant leaf cuticle is related to the prevention of moisture loss, transpiration, and diffusion of light reflection. The purpose of this study was to examine the morphological characteristics of ginseng leaves in ginseng plants resistant and susceptible to high temperature injury (HTI) to be related with the leaf-burning. For the HTI resistant lines Yunpoong, high-temperature injury resistance (HTIR) 1, HTIR 2, and HTIR 3, and the HTI-susceptible line Chunpoong, the cuticle densities were 53.0%, 46.2%, 44.9%, 48.0%, and 17.0%; the adaxial leaf cuticle layers were 141.3, 119.7, 119.7, 159.4, and 85.0 nm in thickness; the abaxial leaf cuticle layers were 153.6, 165.8, 157.9, 199.6, and 119.4 nm in thickness; and the stomtal lengths were 21.7, 32.4, 29.4, 30.9, and 21.8 µm, respectively. All of these aspects suggest that HTI resistant lines have higher cuticle density, thicker adaxial and abaxial leaf cuticle layers, and longer of stomta length than the HTI-susceptible line, protecting leaves from moisture loss and excessive transpiration under high temperatures to be resistant against the leaf-burning.

Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxidase gene in Panax ginseng.

Squalene epoxidase catalyzes the first oxygenation step in phytosterol and triterpenoid saponin biosynthesis and is suggested to represent one of the rate-limiting enzymes in this pathway. Here, we investigated the roles of two squalene epoxidase genes (PgSQE1 and PgSQE2) in triterpene and phytosterol biosynthesis in Panax ginseng. PgSQE1 and PgSQE2 encoded deduced proteins of 537 and 545 amino acids, respectively. Amino acid sequences deduced from PgSQE1 and PgSQE2 share 83% homology, but the N-terminal regions (first 60 amino acids) are highly different. PgSQE1 mRNA abundantly accumulated in all organs. PgSQE2 was only weakly expressed and preferentially in petioles and flower buds. Methyl jasmonate (MeJA) treatment enhanced the accumulation of PgSQE1 mRNA in roots, but rather suppressed expression of PgSQE2. Precursor (squalene) treatment coordinately upregulated the expression of both PgSQE1 and PgSQE2. In situ hybridization analysis established that both PgSQE1 and PgSQE2 mRNAs accumulated preferentially in vascular bundle tissue and resin ducts of petioles. RNA interference of PgSQE1 in transgenic P. ginseng completely suppressed PgSQE1 transcription. Concomitantly, the interference of PgSQE1 resulted in reduction of ginsenoside production. Interestingly, silencing of PgSQE1 in RNAi roots strongly upregulated PgSQE2 and PNX (cycloartenol synthase) and resulted in enhanced phytosterol accumulation. These results indicate that expression of PgSQE1 and PgSQE2 were regulated in a different manner, and that PgSQE1 will regulate ginsenoside biosynthesis, but not that of phytosterols in P. ginseng.

Microbacterium soli sp. nov., an alpha-glucosidase-producing bacterium isolated from soil of a ginseng field.

Five Gram-type-positive, aerobic, rod-shaped, non-motile strains of Microbacterium (DCY 17(T), Ms1, Ms2, Ms3 and Ms4) were isolated from soil from a ginseng field in Daejeon, South Korea. On the basis of 16S rRNA gene sequence similarity, these strains were shown to be related to Microbacterium esteraromaticum DSM 8609(T) (96.1 %), M. xylanilyticum DSM 16914(T) (96.0 %), M. aquimaris JS54-2(T) (95.6 %), M. insulae DS-66(T) (95.5 %), M. ketosireducens IFO 14548(T) (95.5 %) and M. arabinogalactanolyticum DSM 8611(T) (95.4 %). Chemotaxonomic data revealed that the type strain, DCY 17(T), possesses menaquinones MK-12, MK-11 and MK-13 and the predominant fatty acids C(15 : 0) anteiso (32.5 %), C(15 : 0) iso (27.5 %), C(16 : 0 ) iso (17.0 %), C(17 : 0) anteiso (13.2 %), C(17 : 0) iso (6.1 %) and C(14 : 0) iso (2.1 %). The DNA G+C content of strain DCY 17(T) is 70.2 mol% and those of strains Ms1 to Ms4 are in the range 68.9-73.5 mol%. The physiological and biochemical tests suggested that these strains represent a novel species. Based on these data, DCY 17(T) (=KCTC 19237(T) =LMG 24010(T)) is classified as the type strain of a novel Microbacterium species, for which the name Microbacterium soli sp. nov. is proposed.