• Title/Summary/Keyword: Regeneration frequency

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High frequency somatic embryogenesis through leaf explant-derived callus culture in Muscari armeniacum cv. 'Early Giant' (무스카리 'Early Giant' 잎 절편 유래 캘러스 배양을 통한 고빈도 체세포배 발생)

  • Lee, Hyang-Bun;Jeon, Su-Min;Chung, Mi-Young;Han, Jeung-Sul;Kim, Chang-Kil;Lim, Ki-Byung;Chung, Jae-Dong
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.69-74
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    • 2012
  • Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors (Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換))

  • Son, Suk Gyu;Hyu, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.164-172
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    • 1998
  • This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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The Status of Endangered Plants Distributed in the Middle Eastern Area of Korea and Evaluation of the Risk Factors (우리나라 중동부지역에 분포하는 멸종위기야생식물 현황과 위험요인 평가)

  • Kim, Young-Chul;Chae, Hyun-Hee;Hong, Bo-Ram;Oh, Hyun-Kyung;Lee, Kyeong-Hwa;Lee, Kyu-Song
    • Korean Journal of Environment and Ecology
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    • v.30 no.3
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    • pp.291-307
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    • 2016
  • Evaluation of the extinction risk of endangered plants at international, national as well as at regional levels is essential to the implementation of plans for direct conservation activities. Reports indicate that 34 endangered plants are distributed in the middle eastern area of Korea. For each endangered plant, we investigated the sites, area of extents, population size, and factors that affect population extinction. We assessed risk factors based on 10 evaluation criteria including the results from the investigation and the life traits each endangered plant has. As a result of evaluating the risk factors, these 34 endangered plants are classified into 3 groups: the first category comprises 12 endangered plants that require active and urgent conservation of habitats due to multiple risk factors; the second group has 16 endangered plants that should be able to persist with the removal of a few direct risk factors; the third category has 6 endangered plants that can persist with minimal management due to comparatively large distributed area and numerous individuals. It was found that most major risk factors in the population of endangered plants are caused by disruption of habitats and population extinction due to the increase of human habitation in the concerned areas, development and illegal harvesting. Futhermore, ecological collapse from decreasing habitats and malfunctioning mechanism of extinction and regeneration due to the changes of vegetational environment can be the other causes. From the area of the present investigations, we selected 5 regions according to the number of species and the frequency of appearance and importance of conservation measures. Also, we suggested a conservation strategy according to the regional characteristics. We suggest that the method for evaluating extinction risk of endangered plants includes distributional data and life traits of species. In addition, we underscore the necessity for understanding population dynamics and ecological niche of the each target species.

Elimination of Grapevine fleck virus from infected grapevines 'Kyoho' through meristem-tip culture of dormant buds (휴면아 경정 배양법을 통한 포도 '거봉' 에서 Grapevine fleck virus의 제거)

  • Kim, Mi Young;Cho, Kang Hee;Chun, Jae An;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.401-408
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    • 2017
  • Herein, we report the meristem-tip culture from dormant buds of grape 'Kyoho' single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2-5 leaf primordia, and 1-4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at $4^{\circ}C$). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.

Effects of 2-Mercaptoethanol on the Protoplast Formation and Osmotic Stabilizers on the Protoplast Reversion of Pyricularia oryzae Cavara (벼 도열병균(稻熱病菌)의 원형질체(原形質體) 생성(生成)에 미치는 2-Mercaptoethanol과 복귀(復歸)에 미치는 삼투압 안정제(安定劑)의 영향(影響))

  • Kim, Heung-Tae;Chung, Hoo-Sup
    • The Korean Journal of Mycology
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    • v.17 no.1
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    • pp.1-6
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    • 1989
  • The mycelia of Pyricularia oryzae were treated with enzyme solution mixture consisting of Driselase, ${\beta}-Glucuronidase$, Cellulase, and Macerozyme R-10 for the production of protoplasts. More protoplasts were formed from mycelia of race KJ 101 of P. oryzae than that of race KI 315a in the enzyme mixtures. The number of protoplasts was decreased in the untreated control three hrs after the enzyme treatment, whereas the number was increased in the treatments with 10, 50 and 100 mM 2-mercaptoethanol, respectively. The number of protoplasts increased to reach maximum at five hrs after treatment of 10 mM 2-mercaptoethanol, but the least was found in 200 mM. The protoplasts of P. oryzae in a liquid medium containing 2.5% yeast extract, and 2% dextrose reverted to the mycelia after five hrs shaking incubation at $27^{\circ}C$. Some protoplasts produced yeast like buds and the buds were developed to irregularly shaped chains of cell protruded a germ tube like hypha from the distal cell. Once in a while a germ tube like hypha protruded directly from the protoplasts. Except in the first type of reversion, other protoplasts reverted to the normal mycelia. The reversion frequency was highest on PDA with stabilizer of 0.6 M KCl. No reversion of protoplasts occurred on water agar regaardless oftreaatments.

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The Structure of Vegetation in Chamaecyparis obtusa Plantations (편백인공림(人工林)의 식생구조(植生構造)에 관(關)한 연구(硏究))

  • Goo, Gwan Hyo;Lee, Kang Young
    • Journal of Korean Society of Forest Science
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    • v.80 no.4
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    • pp.393-407
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    • 1991
  • The vegetation structure within Chamaecyparis obtusa plantation was analyzed for the purpose of applying the effective forestation method for Chmaecyparis obtusa plantation, tending and regeneration in the southern districts of korea. The results were as follows ; 1. The importance percentage was high in the order of Eurya japonica, Rhus verniciflua, Chamaecyparis obtusa, Lindera erythrocarpa, Carpinus laxiflora, Styrax japonica, Viburnum dilatatum, Zanthoxylum piperitum and Smilax china among the vegetation of Chamaecyparis obtusa. Importance percentage of natural seedling of Chamaecyparis obtusa was high in lower story but gradually decreased in middle story. 2. The basal area of upper trees had a negative correlation with the density of natural seedlings in the middle and lower story, and it represents that the basal area of upper trees had some effect on the density of natural seedlings within understories. 3. The rate of the A and B class by Raunkiaer's frequency was higher in the vegetation of middle story than that of lower story. 4. By Morisita's index, the species of Chamaecyparis obtusa, Rhus verniciflua, Lindera erythrocarpa, Smilax china. Callicarpa japonica and Lindera obtusiloba were randomly distributed at lower story, but they were aggregatively distributed at middle story. At all of middle and lower story, Eurya japonica and Viburum dilatatum were randomly distributed, and Carpinus laxiflora, Zanthoxylum piperitum and Picrasma quassioides were aggregatively distributed. 5. The number of appearance species and the value of species diversity in western survey area were more than that of eastern survey area. 6. The value of species diversity at lower story was higher than that of middle story, and it represents that the number of individuals of appearance species was composed more even at lover story than middle story. 7. According to cluster analysis by similarity index, the survey areas were separated from inland and seacoast districts. 8. Judging from each stories ordination analysis by dissimilarity index, the vegetation was separated from lower and middle story, and the vegetation of lower story was more progressed succession stage than that of middle story. 9. In Chamaecyparis obtusa stands, Eurya japonica had a positive correlation with Sorbus alnifolia, Hex macropoda. Ficus erecta and Trachelospermum asiaticum, but it had a negative correlation with Zanthoxylum piperitum, Carpinus laxiflora and Parthenocissus tricuspidata. 10. In estimation of the productivity of Chamaecyparis obtusa stands, the value of SC (Conic surface) and VP (Parabolic volume) for upper trees was 94.5% and 99.63%, respectively and SC and VP of middle story was 5.49% and 0.37%, respectively. In the species of middle story, material productivity was high in order of Eurya japonica. Lindera eryhrocarpa, Rhus verniciflua. Carpinus laxiflora and Styrax japonica.

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Effects of the Aquatic Vascular Plants on the Lake Ecosystem in the Upper Stream Wetlands of the Namgang-Dam (남강댐 상류 습지에서 수생관속식물이 호소생태계에 미치는 영향)

  • Oh, Kyung-hwan;Lee, Jeong-Hwan;Kim, Cheol-Soo;Son, Sung-Gon;Lee, Pal-Hong
    • Journal of Wetlands Research
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    • v.1 no.1
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    • pp.29-44
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    • 1999
  • Vegetation structure and distribution of the vascular hydrophytes and hygrophytes, and the growth pattern, standing crop and amounts of nutrient uptake by Salix species were investigated in the upper stream wetlands of the Namgang-Dam, Chinju-city, Gyeongsangnam-do, Korea from April to November in 1997. The flora was composed of 43 hydrophytes and 241 hygrophytes, or total 284 vascular plants. The life forms of the hydrophytes were classified as 27 kinds of emergent plants, 4 floating-leaved plants, 3 free-floating plants, and 9 submersed plants. In the herb layer, the dominant species was Persicaria hyciropiper, and the ranges of the species diversity indices (H'), equitabilities, (J') and community similarity indices (CCs) were 1.59~1.89, 0.87~0.96, and 0.35~0.83, respectively. In the shrub and subtree layers, 17 kinds of Salix species were supposed to the pioneer plants at the early stage of the succession. The number of branches per main stem of Salix species was 5.0. The DBH class-frequency histograms of Salix species were the reverse J type, and the natural regeneration of the Salix community was expected. Basal area of Salix species per square meter was $24.87cm^2$. Volume of Salix species per square meter was $12,008cm^3$ and total phytomass of the Salix species was estimated as 12,894 ton. Biomass distribution of Salix species in the stem, the branch and twig, and the leaf was 64.1%, 28.1%, and 7.8%, respectively. The amounts of nitrogen and phosphorus absorbed by Salix species were 68,022 and 19,823 kg. It was recommended that application and conservation of the wetland and other counterplans are indispensable to reduce the adverse effects of water pollution and to preserve the wetland ecosystem.

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