• Korean Journal of Plant Resources
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• v.21 no.5
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• pp.347-351
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• 2008

Rhizomes of Acorus gramineus Soland have been used as sedatives, analgesics, and stomachics in Korean medicine. Even though A. gramineus produced in Korea is known as having better efficacy of a remedy than that in China, its cultivation area has not increased because of the lack of seedlings. To solve this problem, seed propagation method was tested. Seeds were harvested and sowed three times in June, 2005. The best results showed at greenish yellow seed stage harvested in mid June, and its germination ratio was 90.7%. Seeds were well stored at $5^{\circ}C$ for 30 days. As for in vitro culture, multiple shoots were induced first in MS basal medium supplemented with $2.0mg{\cdot}L^{-1}$ NAA plus $0.1mg{\cdot}L^{-1}$ BA, and then roots were induced in MS basal medium containing $0.1mg{\cdot}L^{-1}$ BA or $0.5mg{\cdot}L^{-1}$ NAA plus $1.0mg{\cdot}L^{-1}$ BA. Growth of A. gramineus seedlings in Wonyesangto, potting material sold in markets for horticultural plants, was superior to vermiculite.

• Weed & Turfgrass Science
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• v.5 no.1
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• pp.17-22
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• 2016

Yellow tuft symptoms of a dense cluster on zoysiagrass (Zoysia japonica Steud.) occurred extensively at cultivated fields of zoysiagrass sods in Jangsung. The dense cluster of zoysiagrass showed significant morphological changes such as the tufts of shortening of internodes. The disease symptom was spread on a large scale throughout stolon nodes with multiple short leaves and it thrives in broom-like shaped clusters, exhibiting light green or yellow color on their leaves. The dense cluster of zoysiagrass had approximately 5.8 times more leaves on each node of its stolon then healthy zoysiagrass. Also, these zoysiagrass had poorly developed root and stolon caused by the tufts of a dense cluster of shoots. The dense cluster of zoysiagrass were collected for the putative causal agent incubation and upon close observation, it was found that the sporangia took the shape of a lemon, each sporangium was pointed at the end of its axis and was measured to be $60{\sim}96{\times}42{\sim}51{\mu}m$. These findings were analogous to the mycological characteristics of sporangia formed by the pathogen Sclerophthora macrospora. The symptoms of yellow tuft were prevalent in spring and autumn. Therefore, this study aims to present fundamental data in relation to yellow tuft on zoysiagrass in Korea.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.175-182
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• 2009

Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

• Horticultural Science & Technology
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• v.16 no.2
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• pp.239-241
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• 1998

The experiments were carried out to improve culture efficiency of rhizome and mericlone propagation through settlement of problems occurring during culture period of temperate Cymbidium species. Shooting efficiency from rhizome of C. forrestii 'Nokwoon' was improved, when cultured in $H_3P_4$ medium (Hyponex 3+peptone 4g/L) supplemented with 170mg/L $NaH_2PO_4{\cdot}H_2O$ and 0.4mg/L Thiamin e HCl, but the other varieties were not influenced to shooting efficiency by additives. Medium in which rhizome of C. nishiuchianum 'Hodukjiwha' was cultured became less browned in $H_3P_4$ medium added with 150mg/L PVP, but the other treatments of antioxidants was failed to prevent the medium browning. Re-formation of rhizome from young shoots of C. forrestii 'Sojub', 5.5cm in length occured in $H_3P_4$ enriched with 2.0 mg/L NAA and 1.0 mg/L BA under darkness, but axillary buds were elongated in the medium with 1.0 mg/L NAA and 3.0 mg/L BA under light condition. On the other hand, rhizomes from young shoot of C. forrestii 'Seosinmae' and 'Songmae', 5.5cm and 2.5cm in length respectively were reformed in 2.0 mg/L NAA and 5.0mg/L kinetin under darkness, but multishoot from young shoot were emerged in 2.0mg/L NAA and 3.0mg/L BA.

• FLOWER RESEARCH JOURNAL
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• v.17 no.1
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• pp.23-28
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• 2009

This study was conducted to examine the effect of plant growth regulators on the formation of adventitious bud callus(ABC) and plant regeneration in shoot tip culture of Zantedeschia spp. 'Florex Gold'. Treatment of $0.1mgL^{-1}\;N$-phenyl-N'-1,2,3-thiadiazol-5-ylurea(thidiazuron, TDZ) was more promotive for formation of ABC in shoot tip culture than 6-benzylaminopurine(BA) treatment, and short shoots were developed. Comparing to treatment of BA, mixing treatment of BA and 1-naphthaleneacetic acid(NAA) inhibited the formation of ABC and multiple shoot. The proliferation of ABC derived from sections(0.3 cm) of ABC produced by shoot tip culture in medium supplemented with $0.1mgL^{-1}\;TDZ$ was more effective in medium with $0.1mgL^{-1}\;TDZ$ or $2.0mgL^{-1}\;BA$ than the other treatments. The shoot regeneration and growth from sections of ABC was more promotive in treatment of $0.001mgL^{-1}\;TDZ$. Also, the root growth from sections of ABC was better in medium with $0.001mgL^{-1}\;TDZ$ or $0.2mgL^{-1}\;BA$. Consequently, in vitro mass production of Zantedeschia spp. 'Florex Gold' can be obtained via indirect organogenesis through plant regeneration and proliferation of ABC which was derived from shoot tip culture at $0.1mgL^{-1}\;TDZ$.

• Korean Journal of Plant Resources
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• v.31 no.4
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• pp.355-362
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• 2018

This study was performed to develop the mass propagation system using tissue culture technique to supply the seeds of Elephant garlic (Allium ampeloprasum L.) which has difficulty in propagation. Immature spathe of Elephant garlic was cultured on Murashige & Skoog (MS) medium supplemented with two plant growth regulators, naphthaleneacetic acid (NAA) and kinetin. After 6 weeks of culture, the highest number of shoot (14.9/explant) was obtained when the immature spathe with 10 cm length was cultured right after harvesting. In MS medium supplemented with 2 mg/L kinetin and 0.5 mg/L NAA, the most vigorous growth characteristics was observed, the shoot number was 14.9/explant, its length was 11.3 cm, and its fresh weight was 2.5 g. When the bulblets were cultured in MS medium with 2 mg/L kinetin and 0.5 mg/L NAA, the addition of 30 mg/L adenine improved their proliferation and growth significantly, the highest bulblet formation rate (48%) was obtained. The addition of 7% sucrose also increased the bulblet formation rate at the highest frequency of 98.2%. The shoots were shown be more vigorously proliferated at the secondary subculture stage rather than primary culture stage, their propagation rate was 80% after subculture.

• The Journal of the Convergence on Culture Technology
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• v.4 no.4
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• pp.325-329
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• 2018

There are two kinds of calla belonging to the Zantedeschia spp. depending on the growing condition. Between them, various colored calla with flower shape and flower color are very popular as cut flowers. However, colored calla is very low in natural reproduction rate, and it takes a long time to obtain a flowering bulb. So it is urgent to establish a large breeding system of good varieties. In this study, various plant growth hormone treatments were carried out to improve the growth of the plants in calla. Root formation was the highest at 81.5 % in basal MS medium, and the formation of shoot and multiple shoots were the highest at 100.0 % and 36.4 % in the combined treatment of BA 2.0 mg/L and IBA 1.0 mg/L, respectively. In fresh weight, the highest growth rate was observed with 11.2 times increase in BA 2.0 mg/L single treatment. Auxin and cytokinin mixed treatment were widely used in previous studies and positive effects on the growth of calla plants will help to establish the micro-propagation system.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.04a
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• pp.61-62
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• 2003

Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.53-62
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• 2019

The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.