Dong Yun Hyun

Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate

In this study, methyl jasmonate (MJ)-induced changes of triterpene saponins in ginseng (Panax ginseng C.A. Meyer) hairy roots and expression profiling of relevant responsive genes were analyzed. The transcription of PgSS (squalene synthase), PgSE (squalene epoxidase), and PNA (dammarenediol synthase-II) genes in hairy root cultures elicited by MJ treatment increased as compared with the controls, whereas that of PNX (cycloartenol synthase) decreased slightly. In order to select candidate genes encoding for cytochrome P450-dependent hydroxylase or glucosyltransferase associated with triterpene biosynthesis, RT-PCR analysis was conducted following MJ elicitation. No differences were observed in any expression among the five genes associated with the cytochrome P450 family, when compared to that of control. For candidates of the glucosyltransferase gene,expression of EST IDs PG07020C06, PG07025D04, and PG07029G02 was upregulated. In an effort to assess the effects of MJ elicitation on the biosynthesis of triterpene saponin, protopanaxadiol saponin (Rb group) and protopanaxatriol saponin (Rg group) contents in hairy roots were evaluated by HPLC analysis. With 7 days of MJ elicitation, levels of all ginseonsides of the two-groups increased much higher than that observed in the control. In particular, protopanaxadiol-type saponin contents increased by 5.5–9.7 times that of the control, whereas protopanaxatriol-type saponin contents were increased by 1.85–3.82-fold. In the case of Rg1 ginsenoside after MJ elicitation, the content was affected negatively in ginseng hairy root cultures.

Molecular authentication of ginseng cultivars by comparison of internal transcribed spacer and 5.8S rDNA sequences

Molecular authentication among three Panax species and within cultivars and accessions of P. ginseng was investigated using the DNA sequence in the ribosomal ITS1–5.8S–ITS2 region. Four single-nucleotide polymorphisms were identified between P. ginseng and other Panax species. In the electrophoresis profile, obtained after digestion with the enzyme TaqI, three fingerprinting patterns were obtained from cultivars and accessions of Panax species. Consequently, this authentication procedure based upon the restriction fragment length polymorphism in the ribosomal ITS1–5.8S–ITS2 region can now be utilized to differentiate these Panax species as well as major Korean cultivars such as Gopoong and Kumpoong from other cultivars and accessions in Panax species at the DNA level.

Comparison of Growth Characteristics and Ginsenoside Content of Ginseng (Panax ginseng C. A. Meyer) Cultivated with Greenhouse and Traditional Shade Facility

Growth characteristics, root yield and ginsenoside contents of 3-year-old ginseng in greenhouse shaded by sloped-curtain made of aluminum were compared to traditional shade facility in order to develop cultural practice for organic ginseng. Light transmittance ratio in greenhouse with sloped-curtain shade was distinctly lower than that of traditional shade from sunrise to 9 a.m., while its ratio in greenhouse was higher than traditional shade since 9 a.m. due to the reflection of light. Air temperature of greenhouse was higher than that of traditional shade on the first ten days of August due to more reflected light. Root yield of greenhouse was 44% higher than that of traditional cultivation because of the inflow of reflected light and the decrease of disease of Alternaria and Anthracnose by blocking rainfall. Dry matter partitioning ratio of rhizome and lateral root were increased in ginseng cultivated at greenhouse due to longer survival time in leaf than traditional cultivation. Total ginsenoside contents cultivated at greenhouse was decreased in the part of taproot, while it was increased in the part of lateral and fine root compare to traditional cultivation. Individual ginsenoside contents between greenhouse and traditional cultivation showed significant difference more frequent in fine root than taproot and lateral root. Total ginsenoside contents including , , Rc, Rd, Re, Rf, , and in whole root of 3-year-old ginseng did not showed significant difference by greenhouse and traditional cultivation.

Characterization of a dammarenediol synthase in Centella asiatica (L.) Urban.

To elucidate the exact function of CabAS in Centella asiatica, which was previously reported as a putative beta-amyrin synthase [Plant Cell Rep, 24:304-311, 2005], this gene was functionally expressed in the lanosterol synthase-deficient yeast mutant (erg7). After inducing the CabAS gene with galactose, a peak consistent with the dammarenediol standard was detected in LC/APCIMS analyses and the accumulated product was confirmed as dammarenediol. CabAS should therefore be renamed to C. asiatica dammarenediol synthase (CaDDS). The confirmation of this gene function may allow us to better understand the generation of numerous triterpene carbon skeletons.

Analysis of Growth Characteristics and Physiological Disorder of Korean Ginseng Affected by Application of Manure in Paddy-Converted Field

Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.ABSTRACT: This study was carried out to investigate the difference of the content of soil chemical components and growthcharacteristics in five years old ginseng affected by application of manure in paddy-converted field. As all livestock manureregardless of kinds increased along with the whole soil chemical component, including the pH and EC in 2008. Change in theEC of control plot was slightly increased but not exceeded 1 ds/m over the years. However, the changes in the EC of livestockmanure regardless of kinds and amounts were highly increased and irregularly exceeded 1.5 ds/m in 2012. The 5 years oldginseng root fresh weight, treatment of fertilizing pig manure compost 4 ton per 10 areas (PMC 4t on/10a) and fowl manurecompost 4 ton per 10 areas (FMC 4 ton/10a), were superior to the others. But there were no difference between PMC 4 ton/10a, FMC 4 ton/10a and control. The standing crop rate 39.6%, treatment of fertilizing cattle manure compost 4 ton per10 areas (CMC 4 ton/10a), was best in all livestock manure. However that was relatively lower than control. Physiologicaldisorder occurrence rates of livestock manure related with leaf and root of ginseng were also higher than that of control. Ifexcessively using non-decomposed livestock manure, It would be caused physiological disorder in many ways. It is a bigproblem to be producing the quality ginseng. More research is needed to find out the economic and effective fertilizer.Key Words : Korean Ginseng, Paddy-Converted Field, Livestock Manure, Physiological Disorder

Analysis of Growth Characteristics and Physiological Disorder of Korean Ginseng Affected by Application of Decomposing Plant Residues in Paddy-Converted Field

*Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.**Planning and Coordination Division, NIHHS, RDA, Suwon 441-440, Korea.ABSTRACT : This study was carried out to investigate the difference of the content of soil chemical components and growthcharacteristics in six years old ginseng affected by application of decomposing plant residues in paddy-converted field. Theresults show that aerial parts of ginseng are no difference between press cake (PC) 200 ㎏/10a and control but subterraneanparts of ginseng PC 200㎏/10a, especially quantity related root fresh weight and tap root diameter, are statically about 1.6times heavier and about 1.2 times thicker than the ginseng control. Furthermore, the survival rate of PC 200㎏/10a is67.1% rise significantly compare with the control 50.7%. But compared with the PC 200㎏/10a and the PC 400㎏/10a,ginseng root growth and survival rate of PC 400㎏/10a get worse and that increase physiological disorder occurrence ratethan PC 200㎏/10a. Even though there are no significant differences between the ginseng of decomposing plant residuesexcept press cake treatment and the ginseng of control in growth characteristics, it does tend to increase the survival rateand decrease the physiological disorder occurrence rate in most fertilizer treatment except for RSC 2 ton/10a, RSC 4 ton/10a and RH 4 kL/10a. Noted that EC is highly increased and exceeded 1.7 ds/m in RSC 2 ton/10a, RSC 4 ton/10a and RH4kL/10a. It would be caused physiological disorder in many ways and affected ginseng growth characteristics, survival rate.Key Words : Korean Ginseng, Paddy-Converted Field, Decomposing Plant Residues, Ginseng Physiological Disorder, Press Cake

Analysis of Mitochondrial DNA Sequence and Molecular Marker Development for Identification of Panax Species

This study describes the identification of Panax species using mitochondrial consensus primers. Initially, a total of thirty primers were tested in ten Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. However, these primers could not generated polymorphisms among the Korean ginseng cultivars, and simply represented species-specific polymorphisms for P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region. Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B) were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as `AA`, `BA` and `AB`, respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species.

Genome and evolution of the shade‐requiring medicinal herb Panax ginseng

Summary 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.

Effects of Salt in Soil Condition on Chlorophyll Fluorescence and Physiological Disorder in Panax ginseng C. A. Meyer

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).

Emergence Rate and Growth Characteristics of Ginseng Affected by Different Types of Organic Matters in Greenhouse of Direct-Sowing Culture

Shading and soil environment are the main factors of growth and yield in ginseng (Panax ginseng C. A. Meyer). Ginseng yield is directly related to survival rate because of increased missing plant for their growing period. Under field conditions, diseases and pests significantly affect plant survival rate. We evaluated the seedling establishment, growth and ginsenoside of the ginseng plants, under controlled management conditions in a plastic greenhouse, when their treated with different types of organic matter. Ginseng seeds were sown at a rate of three seeds per hole, and the seeding space measured . Compared to the control, treatment of cattle manure vermicompost (CMV) was shown to increase seedling establishment and decrease ginsenoside content. Root weights of plants treated with CMV were higher than those of plants treated with other types of organic matter. In addition, seedling establishment of 2-year-old ginseng plants was decreased when it was compared to that of 1-year-old ginseng plants. Our results indicated that organic matter type and rate were associated with seedling establishment, growth characteristic and ginsenoside content in greenhouse of ginseng direct-sowing culture.