• Korean Journal of Plant Tissue Culture
• /
• v.21 no.2
• /
• pp.85-90
• /
• 1994

Attempts were made to regenerate plants from petiole explane of Forest Pimpinella barchycarpa via repetitive somatic embryogenesis. Effective induction of somatic emb교ogenesis was achieved on both MS and modified $B_{5}\;(mB_{5})$ media containing BA + 2,4-D or BA + 2,4-D + NAA under light condition (16-h photoperiod/day) cultures. The explants exposed to the ligt produced numerous somatic embryos while those kept under the dark did not form any on the same medium. Somatic embryos at different developmental stages were observed to arise within a individual explants. Plantlets could be regenerated on $mB_{5}$ basal medium or $mB_{5}$ containing 0.1 mg/L NAA Secondary adventive embryos were formed on the surface of the somatic embryos. Therefore, repetitive somatic embryogenesis could be achieved by secondary embryogenesis. Although the treatment of 2,4-D or NAA alone was effective in callus formation and growth, but not in induction of somatic embryogenesis. Some explants, cultured on NAA-containing media in darkness, produced only adventive roots. The embryogenic potential was maintained for two years when subcultured to BA and 2,4-D containing media with 5 weeks inteval. Regenerated plantlets were maintained on $mB_{5}$ or MS basal media for 4 to 6 more weeks and transferred to soil of an artificial mixture for acclimation. Most plantlets (more than 97%) survived, and grew without any deformity.

• Journal of Korean Society of Forest Science
• /
• v.13 no.1
• /
• pp.25-39
• /
• 1971

Recently successful induction of haploid plant by means of anther culture method has become a big topic among geneticists and plant breeders. The haploid plant can be used as a precious material for such basic researches as mutation or genetics. Once the haploid is obtained, production of homozygous plant is not a difficult problem. The method of producing homozygous plant can, also, be applied to the practical breeding works. When applied to the hybridization of self-fertilizing breeding period would be greatly shortened and in cross-fertilizing vegetables production of uniform hybrid seed would be very easily obtained. Last few years many scientists attempted anther cultures using various plant species, but it was successful only in several species. Unlike the other tissue cultures which use somatic organs or tissues as explants, anther culture seems to be very difficult because the plants or calli have to be induced from the haploid microspores or pollen grains. In the present experiment anther culture of fruit trees and ornamental shrubs of four genera and seven species was attemped. Anthers of Various stages ranging from tetrad and late microspore were cultured on the modified Murashige and Skoog's medium supplemented with various concentrations of auxins and kinetin as growth regulators. Handling of materials, sterilization, and other operations of culture were done by routine methods. The results were summarized as follows: 1. Calli were induced in the anthers of Forsythia Koreana Nak., Rhododendron mucronuratum Turcz., R. yedoense Max. var. Poukhanense Nak., and Prunus armeniaca L. var. ansu Max. No signs of callus were observed in Prunus persica Sieb. et Zucc. var. vurgaris Max., Pyrus ussuriensis var. macrostipes (Nak.), and Prunus salcina Lindley. 2. Calli were easily formed in any of the media with differing concentrations of auxins and kinetin. 3. In F. Koreana calli developed from anther surface and connective. Callus emerging out of anther locule was not observed. 4. Somatic calli arose from filament, connective, and inside of anther wall in R. mucronulatum. Many of the microspores accumulated starch grains. 5. The anther lobes located opposite the filament of R. yedoense turned easily to calli. This phenomenon was not observed in R. mucronulatum. Microspore embedded for a period in the medium became starch pollen. No callus was observed arising from microspore. 6. In P. armeniaca calli were not induced from somatic anther tissues. Instead, callus emerged out of anther locule rupturing the anther slit. Starch was not formed in the microspore. 7. In P. persica, Pyrus ussuriensis, and P. salcina, calli were not observed in the anthers examined more than 60 days after culture. Microspores of these species, however, were free of starch grains even after long period of subculture. 8. It was learned that somatic calli of the species examined arose usually from endothelium of anther wall, septum of two neighboring anther locules, parenchyma tissues of connectives, or anther lobes. 9. In the anther locule of P. armeniaca cultured long in medium, swollen microspores, polynucleate microspores, multicellular pollen grains, or callus mass were frequently observed, this indicating that the callus of this species was microspore-origin. 10. It was clarified that in P. armeniaca production of haploid plant by anther culture might be possible.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.28 no.3
• /
• pp.171-176
• /
• 2008

Efficient plant regeneration system of birdsfoot trefoil (Lotus corniculatus L.) was development. The factors affecting the somatic embryo formation, its proliferation and regeneration capacity of leaf and stem explants of Empire cultivar was investigated. The highest number of somatic embryos was obtained on B5 medium supplemented with 1 mg/L NAA and 1 mg/L BA. Depending on different explants, highest frequency of embryogenic callus and regeneration were observed in Empire with leaf explants. The response from stem explants was slower and callus induction was less than that from leaf explants. Regenerated shoots formed complete plantlets in on 1/2 MS medium supplemented with 1 mg/L IBA. Regenerated plants were morphologically uniform with normal shape and growth pattern.

• Journal of Plant Biotechnology
• /
• v.30 no.1
• /
• pp.35-40
• /
• 2003

In this study we investigated the effect of gametocides on the number of larger-sized pollen in anther, and also induced callus from the anther culture of Zoysia japonica Steud. Before culturing, we have observed pollens in anther through fluorescence and electron microscopes to know pollen dimorphism. There were two types of pollens observed. One type (30-36 ${\mu}{\textrm}{m}$ in diameter) consisted of vacuolated, larger-sized pollens and the other (15-20 ${\mu}{\textrm}{m}$ in diameter) smaller-sized ones with dense cytoplasm and plenty of amyloplasts. Within few hours, all the smaller-sized pollens were dead, while larger-sized ones were viable for one or two days. To induct larger-sized pollens, various gametocides were leaf-sprayed on three booting stages cultured under 4$0^{\circ}C$ /15$^{\circ}C$ (day/night) before anther culturing. Number of these larger pollens were few (less than 1%) in anther without spraying gametocides. GA$_3$increased the number of larger-sized pollens when applied at mid-booting stage. GA$_3$ with 50 mg/L treatment caused the highest percentage (25.4%) of the larger-sized pollen. Anthers with GA$_3$ treatment were only produced calli on AA medium (modified B$_{5}$+8.0 mg/L 2,4-D ＋0.2 mg/L kinetin), but callus formation was quite low (less than 1%).).

• Korean Journal of Plant Tissue Culture
• /
• v.28 no.5
• /
• pp.243-248
• /
• 2001

This study was peformed to find out the optimal conditions for the selection of transformed cells using already established transgenic plants. Several transgenic poplar (Populus alba$\times$P giandulosa) lines carrying npt II or hpt gene as a selectable marker were tested against kanamycin or hygromycin. Two culture explants, leaf discs and nodes, were compared regarding their sensitivity to the antibiotics. When leaf discs of untransformed control plants were cultured on callus inducing media in the presence of varying levels of kanamycin or hygromycin, only those cultured on the media containing lower than 50 mg/L kanamycin or 2 mg/L hygromycin formed callus. However, much higher concentration of kanamycin was needed to suppress the growth of axillary buds of untransformed plants. On the other hand, hygromycin at the concentration of 5 mg/L effectively suppressed shoot growth of untransformed plants. Root induction from untransformed plants could also be suppressed at the concentration of 50 mg/L kanamycin or 5 mg/L hygromycin. The transgenic plants showed resistance to 100 mg/L kanamycin or 50 mg/L hygromycin in the growth of callus, shoots, and roots. Hygromycin appeared to be more efficient in selecting untransformed cells than kanamycin.

• Korean Journal of Plant Resources
• /
• v.18 no.3
• /
• pp.382-389
• /
• 2005

This study was conducted to establish the system of effective in vitro propagation by various explant sources culture of Bippeastrum hybridum Hort, 'Dazzler'. We tested the effects of optimal explant source, plant growth regulators on bulblet formation and plant regeneration. Callus was readily produced on the different tissues excised from floral buds whereas, bulbs and shoots were formed only on pedicel explants as compared with anthers, styles and ovaries. Pedicel is the best optimal explant for in vitro propagation. Two distinct pathways, organogenesis through callus and direct bulblet formation, could be recognized in pedicel culture. Up to the $80-100\%$ of bulblet formation and shoot organogenesis from the pedicel in fifteen days before anthesis were effectively induced by MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA. Plantlet regeneration was successfully achieved from pedicel-derived callus, via shoot bud induction or direct bulblet formation. The bulblets with blooming flower were produced within 2 years.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2018.04a
• /
• pp.54-54
• /
• 2018

억새(Miscanthus spp.)는 우리나라 등 동아시아가 원산이며 바이오매스량이 많아 바이오에너지 원료작물로서 잠재성이 크기 때문에 2세대 바이오에탄올 생산 원료로 주목을 받고 있고, 축산깔개 및 토양개량제 등으로도 이용되고 있다. 미국과 유럽 등에서는 생태계 교란 방지를 위해 4배체 물억새(M. sacchariflorus)와 2배체 참억새(M. sinensis)의 종간 교잡 이질 3배체인 불임성 억새(M. x giganteous)을 주로 재배하고 있으나, 단일 유전형의 품종으로 병해충과 자연재해에 취약하여 다양한 억새 품종의 개발이 시급한 실정이다. 본 연구는 억새를 재료로 하여 반수체 및 배수체 확보를 통한 배수성 별 특성 평가와 함께 기내배양을 통해 탈분화 및 재분화 시스템을 구축하여 억새의 육종 효율을 높이기 위해서 실시하였다. 억새 종자로부터 캘러스의 유도는 MS배지와 N6배지에 1mg/L 2.4-D를 첨가하였을 때 비배발생 캘러스(nonembryogenic callus)가 유도되었고, N6배지에 3~5 mg/L 2,4-D를 첨가하였을 때 배발생 캘러스(embryogenic callus)가 발생하였다. MS배지보다는 N6배지에서 캘러스 유도율이 높았으며, 식물생장조절제로 2,4-D와 BA 조합처리 보다 2,4-D 단용 처리하였을 때 캘러스 유도율이 더 높았다. 억새 종에 따른 캘러스 유도율은 물억새가 25.2~49.3%, 참억새는 30.3~52.0%였고 거대억새는 62.6~81.1%로 나타났다. 억새 신초 및 줄기로부터의 캘러스 유도율은 물억새가 4.4~17.2%, 참억새는 1.8~7.7%, 거대억새는 15.3~19.9%로 나타나 종자에 비해 매우 낮았다. 미성숙화기로부터의 캘러스 유도는 억새 종에 따른 차이가 없었으며, 3mg/L 2,4-D 첨가 배지에서 캘러스 유도율이 비교적 낮았고(60~80%), 1mg/L와 5mg/L의 2,4-D가 첨가된 배지에서 캘러스 유도율이 높게(90~95%) 나타났다.

• Journal of Plant Biotechnology
• /
• v.32 no.4
• /
• pp.301-306
• /
• 2005

Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

• Korean Journal of Plant Tissue Culture
• /
• v.26 no.2
• /
• pp.127-132
• /
• 1999

Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.

• Korean Journal of Medicinal Crop Science
• /
• v.2 no.3
• /
• pp.219-226
• /
• 1994

Immature ovule of intergeneric $F_1$ hybrid between Platycodon grandiflorum x Codonopsis lanceolata for producing embryogenic callus. somatic embryos and plant regeneration were cultured in vitro on various medium as well as MT(Murashige Tucker)medium treated with different concentration of plant growth regulators. Embryogenic callus induction was highest in the treatment of NAA 0.5 $mg/{\ell}$ and zeatin 0.01 $mg/{\ell}$ added on MT medium, whereas it was lower in treatments with auxins alone. MT medium were more effective in production of somatic embryos from incubated embryogenic callus. Most favorable plant growgh regulator for producing somatic embryos was 2. 4-D 0.5 $mg/{\ell}$ and zeation. BAP 0.01$mg/{\ell}$, but hormone-free and auxins alone were less effective. NAA 0.01$mg/{\ell}$ added with zeation 0.5 $mg/{\ell}$ was effective as high as NAA 0.01 $mg/{\ell}$ alone in normal plant regeneration from somatic embryo.