• Korean Journal of Plant Resources
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• v.26 no.3
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• pp.362-369
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• 2013

Agastache rugosa, a member of the mint family (Labiatae), is a perennial herb widely distributed in East Asian countries. It is used in traditional medicine for the treatment of cholera, vomiting, and miasma. This study assessed the genetic diversity and population structures on 65 accessions of Korean mint A. rugosa germplasm based on inter simple sequence repeat (ISSR) markers. The selected nine ISSR primers produced reproducible polymorphic banding patterns. In total, 126 bands were scored; 119 (94.4%) were polymorphic. The number of bands generated per primer varied from 7 to 18. A minimum of seven bands was generated by primer 874, while a maximum of 18 bands was generated by the primer 844. Six primers (815, 826, 835, 844, 868, and 874) generated 100% polymorphic bands. This was supported by other parameters such as total gene diversity ($H_T$) values, which ranged from 0.112 to 0.330 with a mean of 0.218. The effective number of alleles ($N_E$) ranged from 1.174 to 1.486 with a mean value of 1.351. Nei's genetic diversity (H) mean value was 0.218, and Shannon's information index (I) mean value was 0.343. The high values for total gene diversity, effective number of alleles, Nei's genetic diversity, and Shannon's information index indicated substantial variations within the population. Cluster analysis showed characteristic grouping, which is not in accordance with their geographical affiliation. The implications of the results of this study in developing a strategy for the conservation and breeding of A. rugosa and other medicinal plant germplasm are discussed.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.05a
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• pp.12-12
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• 2010

We developed droplet-vitrification protocol, a combination of droplet-freezing and solution-based vitrification, and applied to germplasm collections of garlic, potato, lily as well as cell lines, including hairy roots, somatic embryos. To establish a garlic cryobank, four Korean garlic field collections at Danyang, Suwon, Mokpo and Namhae were cryopreserved last five years. The protocol applied consisted of preculture for 3-4 days at $10^{\circ}C$ on solid MS medium with 0.3M sucrose, loading for 40 min in liquid medium with 35% PVS3, dehydration with PVS3 for 150 min, cooling in $5{\mu}l$ droplets of PVS3 placed on aluminum foil strips by dipping these strips in liquid nitrogen, warming them by plunging the foil strips into pre-heated($40^{\circ}C$) 0.8M sucrose solution for 30s. A total of over 900 accessions of garlic were stored in liquid nitrogen for long-term conservation using unripe inflorescences, cloves or bulbils. Twelve alternative plant vitrification solutions were designed by modifying cryoprotectant concentrations from the original PVS2 and PVS3. The results suggest that PVS2-based vitrification solutions with increased glycerol and sucrose and/or decreased DMSO and EG concentrations can be applied for medium size explants which are tolerant to chemical toxicity and moderately sensitive to osmotic stress. PVS3 and variants can be used widely when samples are heterogeneous, of large size and/or very sensitive to chemical toxicity and tolerant to osmotic stress.

• Journal of Plant Biotechnology
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• v.40 no.3
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• pp.147-155
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• 2013

In this experiment, we report for the first time mass propagation by in vitro mircrotuberization of Codonopsis lanceolata. We first examined the effect of cytokinins on multiple shoot induction. 2.0 $mg{\cdot}L^{-1}$ of kinetin not only gave the highest rate of shoot induction (19.1%) but also the elongation of shoot (17.1 mm). Secondly, we investigated the effect of sugars on in vitro microtuberization from nodal segments. The diameter of tuberous roots was enlarged in the half-strength MS medium supplemented with 145.9 mM sucrose. Histological analysis revealed that the number of parenchymatous cell containing starch grains increased in the tuberous roots. In addition, unlike in non-tuberous root, vascular bundles were scattered inner cortex layer. Thirdly, in order to preserve and stimulate the germination, microtubers were stored at $4^{\circ}C$ refrigerator during 9 months and then transplanted to the artificial soils (vermiculrite : peatmoss = 1:1 v/v), resulting that the rates of survival and germination were 75% and 70%, respectively. These results indicated that mass propagation of C. lanceolata was achieved by in vitro microtuber formation, suggesting that this protocol might be applied for not only the propagation of elite clones but also conservation of C. lanceolata germplasm.

• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.135-141
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• 2017

Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. For conservation and germplasm utilization of the plant, it is imperative to obtaining information regarding the genetic diversity of the plant populations. Although C. tricuspidata is an important medicinal plant registered in South Korea, no molecular markers are currently available to distinguish Korean-specific ecotypes from other ecotypes of different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via the amplification refractory mutation system (ARMS)-PCR analyses. Molecular authentication of twelve C. tricuspidata ecotypes from different regions was performed, using DNA sequences in the trnL-F chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific C. tricuspidata ecotypes from different regions.

• Journal of Forest and Environmental Science
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• v.31 no.3
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• pp.214-224
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• 2015

The economy of India and so also of many Asian countries depends on bamboos and their uses are not only in domestic items but also in rural housing and raw materials to several industries and germplasm characterization is an important link between the conservation and utilization of plant genetic resources. Classical taxonomic studies of the bamboos are based on floral morphology and growth habit, which can cause problems in identification due to erratic flowering coupled with different biotic agencies and environmental factors. Identification and genetic relationships among accessions of Thamnocalamus falconeri were investigated using morphology and random amplified polymorphic DNAs (RAPD) technique. Analysis started by using 51 vegetative characters and forty two 10-mer primers that allowed us to distinguish different genotypes hailing from different eco- zones of Garhwal Himalayas (India). The selected primers (12) were used for identification and for establishing a profiling system to estimate genetic diversity. A total of 79.33% polymorphism was estimated by using 12 selected primers. The genetic similar analysis was conducted based on binary digits i.e. presence (1) or absence (0) of bands, which revealed a wide range of variability among the species whereas genetic relatedness was quite high based on vegetative characters. Cluster analysis clearly showed two major clusters for both of the markers viz. morphology and RAPD belonging to 10 accessions of T. falconeri. Two major clusters were further divided into minor clusters. Cluster based on RAPD marker showed grouping of accessions of closed locality whereas analogy was reported for vegetative traits. The RAPD technique has the potential for use in species identification and genetic relationships studies of bamboo for breeding program.

• Horticultural Science & Technology
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• v.16 no.4
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• pp.508-510
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• 1998

This study was carried out to establish the in vitro culture system of potato germplasms for minimizing the occurrence of variation and maximizing the culture period. We used osmoticum such as sorbitol or mannitol with sucrose in the absence of plant growth regulators. The growth of potato germplasms in the medium containing osmoticum was increased when the growth temperature was lowered. After six months storage in low temperature, plant heights of tetraploid was somewhat higher than those of diploid with the exception of stn-16 and the difference due to media was not observed. But after twelve months storage, survival rates of plants cultured in LSM 1(sucrose and sorbitol) was higher than those of plants cultured in LSM 2(sucrose and mannitol). The survival rate of stn-16, diploid wild species, was approximately 75% and it was considerably high. In Atlantic, tetraploid cultivated variety, every individual was survived.

• Korean Journal of Breeding Science
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• v.43 no.5
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• pp.399-404
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• 2011

Genetic diversity among 28 Cymbidium varieties was evaluated by using a sequence-related amplified polymorphism (SRAP) marker system. The SRAP marker which was based on the open reading frames (ORFs) regions was developed primarily for Brassica species, but has been applied to various crops. A total of 30 SRAP primer combinations were initially screened. Twenty-eight SRAP primer combinations showed high polymorphism among the 28 Cymbidium varieties, which were consisted of breeding varieties and their parents in National Institute of Horticultural & Herbal Science (NIHHS). The amplified DNA fragments were separated by denaturing acrylamide gels and detected silver staining method. One hundred ninety six polymorphic bands (7 per primer) were generated and ranged from 0.3 to 1.0 kb in size. Polymorphic fragments were scored for calculating simple matching coefficient of genetic similarity and cluster analysis with multi-variate statistical package (MVSP) 3.1. The mean genetic similarity coefficient value was 0.588. The results showed that the correlation between $F_1$ varieties and their parents was high. These studied SRAP markers will be useful tools for genotype identification, germplasm conservation, genetic relationships in Cymbidium.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.36-36
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• 2021

Cryopreservation has been broadly used as an efficient method for a long-term conservation for many types of plants especially vegetatively propagated plants. Among several cryopreservation methods, a droplet-vitrification was the most widely applicable and efficient method. Studies have developed protocols for strawberry using droplet-vitrification method and suggested the practical use of the protocol for large number of germplasm with a little modification. In this study, the droplet vitrification method of shoot tip has been tested on 31 accessions provided around the world. Shoot tips were precultured on Murashige and Skoog (MS) liquid medium supplemented with 0.3~0.5M sucrose. Precultured explants were osmoprotected with loading solution, 35% of PVS3 (C4, 17.5% glycerol and 17.5% sucrose) for 40 min and exposed to dehydration solution, PVS3 (B1, 50% glycerol and 50% sucrose) for 60 min. Then, the explants were transferred onto droplets containing 2.5 uL PVS3 on sterilized aluminum foils prior to direct immersion in liquid nitrogen (LN) for 1hr. The cryopreserved shoot tips were rapidly warmed in a water bath at 40C and then unloaded in MS with 0.8M sucrose for 40 min. The shoot tips were cultured in NH₄NO₃-free MS post culture medium for 2 weeks. Subsequently, the explants were moved to the MS medium for 6 weeks and evaluated the regrowth rate. By this droplet-vitrification protocol, twenty-four accessions showed at least 40% regrowth rate. Out of 24 accessions, 'Nonsan1ho' had the highest regeneration rate of 85.8% and 'Jumbo pureberry' had the lowest with 42.1%.

• Horticultural Science & Technology
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• v.31 no.3
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• pp.337-343
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• 2013

The aims of this study were to compare genetic distances among 14 Phalaenopsis varieties using simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) marker systems and to determine the discrimination using SSR. A total of 111 SSR primers and 30 SRAP combinations were initially screened. Twelve SSR primers and thirty SRAP combinations showed high polymorphism among the 14 Phalaenopsis varieties including domestic breeding varieties, conserved in National Institute of Horticultural & Herbal Science (NIHHS). The amplified DNA fragments were separated by denaturing acrylamide gels and detected by silver staining method. A total of 474 polymorphic bands, including 55 by SSRs and 419 by SRAPs, were identified and used for genetic diversity analysis. Polymorphic bands were scored for calculating a simple matching coefficient of genetic similarity and cluster analysis with multi-variate statistical package (MVSP) 3.1. Fourteen Phalaenopsis varieties were classified into three major groups at similarity coefficient value of 0.683 and 0.66 using SRAP and SSR, respectively. Also we could discriminate these domestic breeding Palaenopsis varieties using only SSR 20 and SSR 22. The results indicate that SSR analysis is effective for discrimination among Phalaenopsis varieties and SRAP is useful for genetic diversity when there is no sequence information. These studied SSR and SRAP markers will be useful tools for genotype identification, germplasm conservation and genetic relationship study in Phalaenopsis.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.388-395
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• 2015

Somatic embryogenesis is as an excellent technology for potential use in plant mass production, germplasm conservation, or genetic engineering. We examined the effect of cold storage using 3 embryogenic callus lines with different levels of embryogenesis competence derived from immature zygotic embryo cultures of Kalopanax setemlobus. Somatic embryo induction, germination and plant conversion were evaluated after 1, 3 and 6 months storage at $4^{\circ}C$ in the dark. Most cold-stored embryogenic calli formed somatic embryos normally even after 6 months; however, the induction rate was gradually decreased by increasing the storage period. The most competent line tended to show a slight decline in somatic embryo induction rate, as compared with other lines after cold storage. In general, cold storage resulted in reduced somatic embryo germination and plant regeneration, although 93% somatic embryo germination and 91% plant conversion were achieved regardless of the storage period. Cold storage led to cell browning and degradation. Additionally, the cell structures were confirmed by the aceto-carmine and evans blue dye evaluation. Collectively, our results showed that embryogenic callus of K. septemlobus could be preserved at $4^{\circ}C$ without subculture for 6 months, and suggested the need for storage of relatively more competent embryogenic calli lines to support somatic embryo induction.