Background : A series of studies were conducted to optimize adventitious root induction in vitro from explants of Astragalus membranaceus using various nutrient media supplemented with plant hormones.
Methods and Results : Levels of active components were analyzed from adventitious roots induced under different media conditions. Among the different media conditions, Murashige and Skoog medium supplemented with 1.0㎎• ℓ −1 indole-3-butyric acid resulted in the greatest adventitious root induction rate. The amount of the major active component of the adventitious roots of Ama1, calycosin-7-O-β-D-glucopyranoside was higher than that of other adventitious root samples.
Conclusions : These results suggest that the adventitious roots of A. membranaceus could be used for the commercial production of medicines.
In this study, genetic diversity of wild Codonopsis lanceolata collected in Korea were analysed using SSR makers. Wild C. lanceolata roots were collected in Jeollanam-do Jangheung-gun Choentae Mountain as in roots. The wild C. lanceolata plants were cultivated in Chungbuk National University greenhouse and the leaves were sampled from 36 plants. The genomic DNA of C. lanceolata was extracted using CTAB. PCR was performed using a program of 35 cycles at 94℃ for 30 sec, 60℃ for 30 sec, and 72℃ for 30 sec with an pre-denaturation of 94℃ for 5 min and a final extension of 72℃ for 30 min. The PCR reaction mixture contains 5 pmole of primers and 20 ng of DNA template in a 20 μL reaction volume. The genotype of the analyzed samples were very different. Therefore, the wild C. lanceolata collected in Korea look genetically diverse.
Angelica gigas, also called Dang Gui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. In Korea, we are using the roots of A. gigas., but, they are using Angelica sinensis in China and using Angelica acutiloba. in Japan to obtain many active constituents such as dercursin, decursinol angelate, nodakenetin, nodakenin, umbelliferone, β-sisterol, or α-pinene. The plants of the Angelica family are used to improve gynecological health. The biggest problem in the cultivation of A. gigas is bolting. If the bolting occurs, A. gigas can not be used as a medicinal component because the roots are lignified. In this study, 11 A. gigas genetic resources in Korea; 1. Hwangje variety, 2. Sungwoo Jongmyo company, 3. Bonghwa No. 1, 4. Bonghwa No. 2, 5. Bonghwa No. 3, 6. Bonghwa No. 4, 7. Jechun local variety, 8. Jirisan local variety, 9. Manchu variety in Eumseong, 10. Manchu variety in Bonghwa, 11. Jinbu local variety, were collected and performed phylogenetic analysis using RAPD molecular markers.
Platycodon grandiflorum A. is a perennial plant belongs to Campanulaceae family. This plant has been used herbal medicine ingredient in East Asia. Because of the high saponin content, it is an economically important medicinal plant in Korea. It has been reported that saponins of P. grandiflorum were mainly synthesized in root tissues. The studies about root growth of the plant were few. Expansin is an important protein playing a role in root growth of plants, and is known as a nonenzymatic protein. Expansins are novel plant cell wall loosening proteins leading to turgor-driven cell extension. Expansin encoding genes exist in multigene family, and there are more than 30 genes in Arabidopsis thaliana. and more than 50 genes in Oryaza sativa. Therefore, identification of the genes was difficult in P. grandiflorum because of the lack of genome sequence. Recently, the development of next generation sequencing (NGS) technologies make it possible to obtain the target genes sequences rapidly and precisely. In this study, to identify the expansin encoding genes in P. grandiflorum, we used RNA-seq analysis with Illumina HiSeq platform. We analyzed whole transcriptome of P. grandiflorum through the RNA-seq analysis based on next generation seuqencing. CLC Genomics Workbench software (Clc Bio inc.) was used for assembly. We assembled 122,663 contigs and search 123 contigs were identified from the search using 61 expansin gene
Codonopsis lanceolata is a perennial climber. The roots are used as medicinal materials or vegetables. Recently, demand for C. lanceolata is increasing as a healthy food. C. lanceolata is distributed in India and East Asia such as China, Japan as well as Korea. In South Korea, this plant is widely cultivated in Gangwon-do province. No C. lanceolata varieties were developed in Korea. The objective of this study is to analyze genetic diversity of C. lanceolata cultivated in Korea using SSR makers. C. lanceolata roots were collected in each region were cultivated in Chungbuk National University greenhouse. Samples were obtained from fresh leaves of 5 plants from each collection region. The genomic DNA was extracted using CTAB. Genetic diversity was analysed using 4 sets of C. lanceolata SSR makers. PCR was performed in total 20 μL reaction volume containing 20 ng of DNA template, 5 pmole of primers. The genotypes of the analyzed samples were very similar. That means that the genetic diversity of C. lanceolata cultivated in Korea is very low.
Codonopsis lanceolata is used as a natural medicine or vegetables. It originates in East Asia such as Korea, Japan and China. Similar to Panax ginseng, C. lanceolata contains saponins as effective components. C. lanceolata is cultivated in many regions of South Korea. But, no variety was developed yet and the origin discrimination in the distribution market of C. lanceolata became a problem. In this study, we collected 20 C. lanceolata regional groups; Hoengseong, Wonju, Samcheok, Chuncheon, Pyeongchang, Hongcheon, Yongin, Yangpyeong, Danyang, Chungju, Bonghwa, Ulleung, Yeongju, Sancheong, Muju, Gwangyang, Sinan, Hwasun, Jeju-si and Seogwipo-si, and tested the genetic relationship using RAPD molecular markers. The genomic DNA was extracted using CTAB and the RAPD analysis was performed using 32 primers of Operon Technologies. NTsys-PC program was used for the phylogenetic analysis of the data.
Platycodon grandiflorum is a herbal flowering perennial plant belongs to Campanulaceae family. The saponins derived from P. grandiflorum were termed platycosides and platycodin D, which is the most abundant saponin in the plant and pharmacologically active component, was intensively studied. Platycodin D is synthesized from triterpenoids by several enzymes including cytochrome P450. Cytochrome P450 is known to exist in superfamily in plant kingdom and essential roles in saponin biosynthetic pathway by hydroxylation or oxidation of triterpene skeletons. However, the key genes of P450 involved in biosynthesis of saponin was not identified because of its low conservation rate in amino acid sequence level among plant species and gene superfamilies. Recently, next generation sequencing (NGS) technology is rapidly developed as a method to discover target genes. In this study, we tried to identify P450 genes involved in saponin biosynthetic pathway from the various tissues of P. grandiflorum using RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obteined 122,663 contigs and found out 191 putative P450 genes. The phylogenetic relationship was analyzed and putative genes related to platicoside biosysthesis were selected and cloned for further analysis.
Platycodon grandiflorum is a perennial herbal plant belongs to Campanulaceae family. It has very important genetic value as a major plant in Asterids order. The major ingredients are platycosides, terpenoid saponins. In Korean industrial plants market, it was produced 5,633 tons in 2013, and the total amount of production was less than only five species, omija, ginger, raspberry, yam and deodeok. P. grandiflorum is called ‘Gilgyung’ and is used as a fresh vegetable and an ornamental plant. Nowadays, various components of P. grandiflorum were already published. But, genetic research is in the starting stage. In this study, 11 cultivars; 1. MariesⅡ, 2. Hakone double white, 3. Hakone double blue, 4. Fuji white, 5. Fuji pink, 6. Fuji blue, 7. Astra white, 8. Astra pink, 9. Astra blue, 10. Astrasemi double blue, 11. Jangback, were analyzed using 60 Operon Universal RAPD primers. The results were phylogenetically analyzed and related to the morphological characteristics of the cultivars.
Platycodon grandiflorum is a species of herbaceous flowering perennial plant of the family Campanulaceae. The major ingredients are platycosides, terpenoid saponins. It contains 1-4 % of the dry weight and there are about 20 types of platycosides. Among them, platycodin D have various pharmacological effects on cough and cold. Platycosides are synthesized from oleanane by mevalonic acid pathway and cytochrome P450s and UGTs are important enzymes in the saponin biosynthesis. UGT is glucose transfer enzyme and act on the final step of the secondary metabolite biosynthesis. In this study, we tried to identify UGT genes involved in saponin biosynthetic pathway from the various tissues of P. grandiflorum and non germinated seeds using RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. The phylogenetic relationship was analyzed and putative genes related to platicoside biosysthesis were selected and cloned for further analysis.