• Title/Summary/Keyword: 기내배양

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Analysis of UreB Protein Synthesis from Transgenic Lily Pollen (형질전환 백합화분을 이용한 UreB단백질의 발현분석)

  • 박희성;박인혜
    • KSBB Journal
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    • v.17 no.6
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    • pp.577-581
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    • 2002
  • In an attempt to produce recombinant proteins using the pollen enriched in some plant species, a 1.7 kb DNA encoding urease subunit B (UreB) amplified by PCR from Helicobacter pylori urease gene cluster in pH808 plasmid was cloned to be expressed under CaMV35S promoter in lily (Lilium longiflorum) pollen tubes elongated in vitro. Lily pollen at early germinating stage was transformed with the ureB DNA using Agrobacterium via vacuum infiltration and, incubated for a full pollen tube growth 16 - 24 h in the dark in the presence of kanamycin. DNA integration and expression in the transgenic pollen were analyzed by the standard molecular techniques and the results suggest that the pollen in vitro may be employed as a protein factory in a disposable fashion.

Effects of 2,4-D Treatment on the Callus Induction of Polygonatum stenophyllum Maxim. (2,4-D처리가 층층둥굴레(Polygonatum stenophyllum Maxim.) 캘러스 유도에 미치는 영향)

  • Park, Min Wan;Bae, Kee Hwa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.32-32
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    • 2018
  • 층층둥굴레(Polygonatum stenophyllum Maxim.)는 백합과(Liliaceae) 둥굴레속(Polygonatum)에 속하는 다년생초본이다. 둥굴레속의 근경은 점액질이 풍부하고 전분질, 아미노산, 알카로이드 등의 성분을 다량 함유하고 있어, 식품학적으로도 우수한 것으로 보고되었다. 그리고 새순과 여린줄기는 식약청 식품원재료서비스에 등록된 식용가능 식물자원이다. 본 연구에 사용된 층층둥굴레는 경기도 황학산수목원에서 2년, 5년생과 종자를 분양받아 종자는 기내에서 발아시킨 후 실험에 사용하였다. 야생의 개체를 무균적으로 기내도입하기 위해 차아염소산나트륨의 농도를 0~1.5%까지 달리하여 멸균한 결과, 유의적 차이는 없으나 1~1.5%처리가 비오염율이 20% 내외로 조사되었다. 층층둥굴레 캘러스 유도에 미치는 옥신류 호르몬의 영향을 알아보기 위해 멸균된 근경을 절단하고 옥신류 호르몬의 종류와 농도가 각각 다르게 처리된 MS배지에 치상하여 배양한 결과, 2,4-D 0.5mg/L 처리구에서 캘러스 유도율이 87%로 가장 높게 조사 되었다. 이러한 결과는 층층둥굴레의 기내배양 방법을 통해 효과적으로 모본과 동일한 층층둥굴레의 대량생산 체계를 만들 수 있는 참고자료로 활용될 수 있고 더불어 관련 산업계의 소재다양화 측면에서 유용하게 사용될 것으로 보여진다.

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In vitro propagation and multiple shoot induction of Rhodiola rosea L. by axillary bud culture (홍경천(Rhodiola rosea L.)의 액아배양을 통한 다신초 유도 및 기내 대량증식)

  • Bae, Kee-Hwa;Ko, Myung-Suk;Kim, Nam-Young;Song, Jae-Mo;Song, Gwan-Pil
    • Journal of Plant Biotechnology
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    • v.39 no.2
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    • pp.114-120
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    • 2012
  • An efficient in vitro propagation was established by using axillary bud explants of roseroot (Rhodiola rosea L.), which has been known as a medicinal plant in East Asia. Among various media tested, MS medium supplemented with 1.0 mg/L BA and 1.0 mg/L $GA_3$ was found to be the best for multiple shoot formation (15 axillary shoots per axillary bud). In addition 1/2MS medium containing 50 g/L sucrose was best for shoot elongation (7.8 cm) and increasing total chlorophyll contents (8.64 mg/g) best. Maximum number of roots (17.7 roots per explant) was observed on the medium without plant growth regulators. Propagated plants were successfully acclimatized to ex vitro conditions, with a survival frequency of 97% after 12 weeks. Most rooted shoots grew well and produced viable seeds when grown in vitro culture conditions. Therefore, R. rosea can be effectively propagated in vitro by the system we developed in this study.

Appropriate in Vitro Culture Conditions of Growing Medium for New Ever-bearing Strawberry 'Goha' (사계성 신품종 딸기 '고하'의 기내배양을 위한 배지의 적정 조건)

  • Lee, Jong-Nam;Kim, Hye-Jin;Kim, Ki-Deog;Kwon, Young-Seok;Im, Ju-Sung;Yeoung, Young-Rok;Lim, Hak-Tae
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1051-1056
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    • 2010
  • This study was carried out to determine suitable in vitro culture conditions of new ever-bearing strawberry ($Fragaria$ $ananassa$ Duch.), 'Goha'. Four-week old plantlets which were derived from the meristem culture were used in this study. Three different culture media including MS, Gamborg B5 and White medium were used for the plant culture and the medium concentrations were at the 5 levels of $1/3{\times}$, $1/2{\times}$, $1{\times}$, $2{\times}$ and $3{\times}$. Sucrose content ranged at four levels of 1, 3, 5 and 8% (w/v). Crown diameter on the MS medium was thicker (2.1 mm) than in other media. Fresh weight on the MS medium was 482 mg, which was heavier than 88 mg or 260 mg of Gamborg B5 or White medium, respectively. Particularly, 1/2MS medium was found to have higher growth rate than these in other treatments (fresh weight, dry weight and D/F rate). Shoot length in the treatment of 1% sucrose concentration was 3.6 cm which was the longest. Shoot length was in inverse proportion to the increasing concentration of sucrose. Fresh weight was increased up to 3% sucrose concentration, but decreased above 5% sucrose concentration. From the results, we found that the best condition for in vitro culture of new ever-bearing strawberry 'Goha' was 1/2MS medium supplemented with 1% sucrose concentration.

In Vitro Plant Regeneration of Siberian Wildrye Grass from Mature Seed-derived Callus (Siberian Wildrye Grass의 성숙종자 유래의 캘러스로부터 식물체 재분화)

  • Lee, Ki-Won;Chinzorig, Ochirbat;Choi, Gi-Jun;Yoon, Sei-Hyung;Kim, Ki-Yong;Ji, Hee-Chung;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.3
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    • pp.217-222
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    • 2011
  • Success in molecular breeding for better adapted varieties to environmental stresses depend upon the concerted efforts by various research including tissue culture, transformation, genetics and breeding. In order to optimize tissue culture conditions of Siberian wildrye grass, the effects of plant growth regulators on callus induction and plant regeneration were investigated with mature seeds. The highest callus induction frequency was observed when the mature seeds were cultured on MS medium supplemented with 5 mg/L 2,4-D. The highest plant regeneration frequency was observed when callus was transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots were transplanted to the soil. A short tissue culture period and regeneration system would be beneficial for molecular breeding of Siberian wildrye grass by the production of transgenic plant.

In vitro propagation of a rare and endangered species, Echinosophora koreensis Nakai, by axillary bud culture (희귀 및 멸종위기 수종 개느삼의 액아배양을 통한 기내번식)

  • Moon, Heung-Kyu;Kim, Yong-Wook
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.229-234
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    • 2008
  • An efficient micropropagation was established by using axillary bud explants from two-year-old tree(Echinosphorea koreensis Nakai), which has been known as a rare and endangered species. Among various basal media tested, DKW medium was shown to be the best for axillary shoot elongation. The addition of both BA and TDZ to the medium induced 6 to 10 shoots per explant during eight weeks of culture, without showing any abnormal morphology at the shoot proliferation stage. However, high concentration of TDZ(>0.05 mg/L) appeared to cause hyperhydration on either leaf or shoot at the later developmental stage. Approximately 20% of shoots produced roots by the addition of 1.0 mg/L NAA but not by IBA($0.2{\sim}1.0$ mg/L). Ex vitro micro-cuttings were better source for root induction; up to 58.6% of the micro-cuttings rooted when 100 mg/L IBA was applied to the soil(vermiculite). More than 90% of plantlets with roots were successfully acclimatized and grew normally in the field. Therefore, we suggest that this endangered tree species can be effectively micropropagated by axillary bud culture system developed in this study.

In vitro propagation of oil palm (Elaeis guineensis Jacq.) clones through somatic embryogenesis and analysis of somaclonal variation by RAPD (체세포배발생을 통한 오일팜나무(Elaeis guineensis Jacq.) 클론의 기내증식 및 RAPD를 이용한 체세포변이의 검정)

  • Ahn, In-Suk;Park, Hye-Rim;Son, Sung-Ho
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.196-204
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    • 2012
  • This study was carried out to develop reliable systems for somatic embryogenesis in oil palm tree (Elaeis guineensis Jacq.), and to verify the somaclonal variants by RAPD analysis. Embryogenic callus was induced successfully on modified half-strength MS medium containing $NaH_2PO_4{\cdot}2H_2O$ and casein. Embryogenic callus was further developed to somatic embryo mass (SEM), which is very hard and bonded tightly each other. Plantlets were proliferated when SEM was cultured on modified MS medium containing half strength $NH_4NO_3$, casein and L-ascorbic acid. Plantlets were transplanted into pots containing artificial soils. When RAPD analysis was conducted using randomly selected 95 in vitro plantlets and 19 random primers, somaclonal variation was detected using BNR35 primer. There was missing band around 1 kb in #22, #28, #35, and #77 plantlets. In addition, bands obtained from #28, #35, and #77 was much stronger than other normal bands. The blast results at NCBI revealed that somaclonal variation observed in this study was related to chloroplast genome of oil palm. The results also revealed that oil palm reproduction system through somatic embryogenesis is quite reliable and early detection of somaclonal variants seem to be possible at in vitro stage by RAPD analysis.

Several Factors Affecting on In vitro Culture of Prothallus and Ex Vitro Sporophyte Formation from Prothallus of Dyropteris varia (L.) O. Kuntze (족제비고사리 전엽체의 기내배양 및 기외 포자체 형성에 미치는 제요인)

  • Jeong Jin-A;Lee Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.252-258
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    • 2006
  • The most effective conditions of In vitro culture and ex vitro sporophyte formation from prothallus were studied for mass propagation of Dyropteris varia. The most effective medium of prothallus proliferation was Murashige and Skoog's basal medium supplemented with 10:50mM of $NH_4^+:NO_3^-$ and 2% sucrose. The optimum pH level was 5.8 and prothallus growth was promoted on medium containing $0.6{\sim}0.8%$ agar. Almost of the tested growth regulators (NAA, IAA, 2,4-D, BAP, kinetin and 2ip) were inhibitory in prothallus proliferation as the concentration of growth regulators became higher. The highest number of sporophytes was obtained by transplanting prothallus on compost only than on any other soil compositions. Sporophyte formation was promoted remarkably by soaking prothallus with $100{\mu}M\;GA_3$ for 3 hours.

Acclimatization and Growth Characteristics of Plantlets of Eleutherococcus senticosus Maxim Cultured by Bioreactor (생물반응기에서 배양한 가시오갈피 유식물체의 순화 및 생육특성)

  • Li, Cheng-Hao;Lim, Jung-Dae;Kim, Myong-Jo;Kim, Na-Young;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.133-137
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    • 2005
  • Experiments were conducted to find the optimal acclimatization conditions for Eleutherococcus senticosus plantlets regenerated from bioreactor cultured somatic embryos, various acclimatizing conditions were compared regarding both survival rate and growth of the plantlets. Among the various temperature and artificial soil tested, the highest survival rate (88%) was observed when plantlets were acclimatized in Klasmann bed soil at $10^{\circ}C$ When in vitro plantlets directly transplanted to field environment, shading treatment was necessary and 50% shading was more effective than 30% shading. Transplanting season were also important for successful acclimatization of in vitro cultured plantlets, transplanted on March 20 with 50% shading exhibited the best survival rate and further growth.

Rapid Micropropagation by Stem Node Culture of Japanese Yew (주목의 줄기절간 조직배양에 의한 급속 대량증식)

  • 선정훈
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.335-337
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    • 1995
  • The effect of plant growth regulators on proliferation of shoot from stem node culture of Japanese yew (Taxus cuspidata Sieb. et Zucc.) was studied using Quoirin and Lepoivre (1977) medium. Among the cytokinin tested, BAP, kinetin, and thidiazuron at various concentrations had no effect on shoot multiplication However when zeatin at 5$\times$10$^{-5}$ M was added to the medium, an average of 6 shoots were regenerated per explant after 8 weeks of culture. The ratio of rooting ex vitro was remarkably increased up to 34% by dipping the basal end in 0.5 to 1.0% IBA on talc compared with 3% in vitro rooting. Rooted plantlets were acclimated in greenhouse conditions for one month and successfully transplanted to the field.

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