• Journal of Plant Biotechnology
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• 제36권2호
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• pp.137-143
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• 2009

An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

• Journal of Plant Biotechnology
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• 제34권2호
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• pp.161-166
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• 2007

In vitro shoot regeneration and multiple shoot induction has been obtained from the stolen node explants in Eremochloa ophiuroides (Munro) Hack. The highest number of shoots ($10.66{\pm}0.21$) was observed from initial explants after one month culture duration on Murashige and Skoog (MS) medium containing 6-benzyladenine (BA: 0.5 mg/l). First generation shoot was excised and sub-cultured on the same fresh media for further multiplication of shoots. An enhanced number of second round shoots ($15.33{\pm}0.21$) was obtained compared to the initial culture media containing BA (0.5 mg/l). The number of shoots/stolon node was higher among all the concentrations of BA than kinetin (KN). In vitro regenerated shoots were successfully rooted in the phytohormone free MS medium. Plantlets generated with roots were transferred to pots containing compound mixture of soil and kept in green house conditions. Acclimatized plants showed 100% survival rate with normal morphology in green house conditions. The present study demonstrates the effect of explant and different plant growth regulators towards in vitro response in E. ophiuroides. Moreover, the study reveals the effect of cytokinin on induction of shoot number per stolen node explant in E. ophiuroides.

• 식물조직배양학회지
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• 제24권6호
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• pp.357-360
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• 1997

쇠무릎(A. japonica) 다량증식 방법의 일환으로 액아를 이용한 multiple shoot유도를 위한 최적 조건을 조사하였다. 기내에서 증식하고 있는 식물체로부터 액아를 적출하여 NAA, 2,4-D 및 BA가 여러 농도로. 첨가된 MS 배지(3% sucrose, 0.2% gelrite)에서 6주 간 배양한 결과, shoot의 발생은 1mg/L NAA와 2mg/L BA가 첨가된 처리구에서 액아 당 평균 25.8개로 가장 많았다. 신초의 발생 빈도는 조금 낮지만 발생된 신초가 완전한 식물체로 되기 위한 크기를 고려할 때의 조합은 0.5 mg/L NAA와 1 mg/L BA가 첨가된 처리구에서 액아 당 19.7개의 신초가 발생하여 가장 좋은 결과를 얻었다. 한편 발생된 신초는 절취하여 0.1mg/L IBA가 첨가된 1/2 MS 배지에서 배양했을 때 뿌리의 발생과 신장이 가장 양호하였으며, 식물체의 생육도 왕성하였다. 발근된 식물체를 기외로 이식 후 활착시켜 쇠무릎의 액아 배양을 통한 다량증식의 가능성을 확인하였다.

• 한국작물학회지
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• 제51권spc1호
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• pp.237-241
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• 2006

국내육성 유채품종 간 반수체식물 생산성을 비교하였다. 화아로부터 분리된 소포자를 13%의 sucrose, $0.05mg/{\ell}$ BA 와 $0.5mg/{\ell}$ 의 NAA가 첨가된 NLN 배지에서 배양하였다. 반수체 배의 생산에는 유전형이 중요한 요인이였으며, 탐라유채가 가장 높은 반수체 배 생산능을 보여 화아당 176개의 반수체 배가 발생하였으나, 한라유채와 영산유채는 배 생산은 물론 세포분열도 관찰되지 않았다. 발생한 반수체 배를 NLN배지에서 현탁배양하여 Multilobe abnormal embryos를 형성시켰으며, 계속하여 생장조절제가 첨가되지 않은 MS고체 배지에 치상 배양하여 반수체식물체를 유도 하였다. 재생된 반수체 식물체는 성공적으로 순화되었다.

• Journal of Plant Biology
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• 제39권2호
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• pp.127-135
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• 1996

An efficient system of somatic embryogenesis was established for the red pepper plant (Capsicum annuum L. cv. Nokkwang) usign immature zygotic embryos. The size of the immature zygotic embryos and the concentrations of 2, 4-D and sucrose were found to be critical. Somatic embryos were induced via callus or directly from explants and regenerated into plantlets successfully. When zygotic embryos 1~2 mm long were cultured on the modified Murashige-Skoong (MS) medium supplemented with 2 mg/L 2, 4-D for 3 weeks in the dark, somatic embryos were induced directly from the apical region of zygotic embryos with the highest frequency being approximately 90%. To mature the somatic embryos, ABA and an ethylene inhibitor AgNO3 were used. The highest frequency of shoot regeneration (25% in each) resulted at 2$\mu$M ABA or 20$\mu$M AgNO3 treatment at rates 3.7 and 1.6 times control, respectively. Shoots developed mainly from the cotyledonary node on CoCl2-containing medium, and from the upper side of cotyledon on medium containing AgNO3 while the embryos on the control medium produced shoots from both the cotyledonary node and the upper region of cotyledons both at frequencies of 50%. Indirect somatic embryogenesis via callus was induced at an efficiency of approximately 10% with zygotic embryos 3~4 mm long cultured on MS medium containing 5~10 mg/L, 2, 4-D for 5~7 weeks under a continuous light condition. The plants regenerated from the somatic embryos were morphologically normal. Using scanning electron microscopy, the direct and indirect somatic embryogeneses were observed to follow the globular, heart and torpedo stages, similar to zygotic embryogenesis. Also, suspensors appeared in the early globular and ovoid-shaped late globular embryos during indirect somatic embryogenesis.

• 식물조직배양학회지
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• 제26권1호
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• pp.59-63
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• 1999

국화 '남전'의 기내배양된 엽절편으로부터 신초 기관형성을 통한 효율적인 식물체 재생 체계를 확립하였다. 엽절편으로부터의 부정지 형성은 생장조절제 조성, 잎의 연령, 광조건, 배양용기당 절편체 수 및 치상방향에 의해 많은 영향을 받는 것으로 나타났다. 효율적인 부정지 형성을 위해서는 MS배지에 생장조절제 BA 0.5 ㎎/L 와 NAA 2.0 ㎎/L를 첨가하는 것이 가장 좋았으며, 엽절편체는 완전히 전개된 신초 상부의 1∼2번째 잎을 채취하여 4면을 절단한 다음 잎의 향축면이 배지에 닿도록 하여 Petri-dish당 8개의 엽절편을 치상하여 배양초기 1주간 암조건에서 배양하다가 명조건으로 옮겨주는 것이 가장 효과적이었다. 이와 같은 조건에서 엽절편체로부터 신초 재생률 100%, 절편체당 신초수 13.7개의 양호한 재생 이 이루어졌다. 재생된 신초는 NAA 0.1 ㎎/L를 첨가한 MS배지에서 성공적으로 발근이 이루어져 소식물체가 얻어졌다. 엽절편체로부터 얻어진 소식물체를 인공토양 (Vermiculite: Perlite=1:1)에서 활착시켜 온실에서 재배하였는데, 외견상 정상적으로 개화하였다.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

• 한국자원식물학회지
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• 제19권3호
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• pp.427-431
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• 2006

In vitro high-frequency plant regeneration of Muscari comosum var. plumosum through somatic embryogenesis was obtained via two developmental pathways: direct embryos and multiple shoots regenerated from embryogenic callus. Flower bud with pedicel, receptacle, petal and ovary wall, floral stalk and leaf as explants were cultured in MS medium supplemented with various plant growth regulators. Embryos formed directly from pedicel, receptacle and floral stalk. Depending on explant sources, the optimal medium was MS medium supplemented with 0.2 mg/L IBA and 0.3 mg/L BA, 3.0 mg/L IBA and 3.0 mg/L BA, and MS-free medium for pedicel, receptacle, and floral stalk, respectively. Multiple shoots regenerated from embryogenic cal]i which was initiated from petal, ovary and leaf were observed in MS medium with different concentrations and combinations of hormone. The most suitable medium for each type of explant was 3.0 mg/L IBA and 3.0 mg/L BA(petal and ovary) and 5.0 mg/L IBA and 5.0 mg/L BA (leaf) Furthermore, the combination of 0.1 mg/L 2,4-D and 1.0 mg/L BA was also good for all sources of explants not only for direct embryo formation, but also, for embryogenic callus induction.

• 식물조직배양학회지
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• 제24권5호
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• pp.263-268
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• 1997

다래(A. arguta)의 절간조직을 절편체로하여 식물생장조절제의 종류와 광조건이 shoot 형성에 미치는 영향을 조사하였으며 아울러 기내발근을 통하여 다래로부터 완전한 식물체를 재분화시킬 수 있는 실험체제를 확립하였다. 절간절편체는 3.0 mg/L의 zeatin이 단독첨가된 배지에 암배양한 경우 62.5%로 높은 shoot 형성률을 나타내었다. 0.5mg/L NAA가 포함된 WPM 배지에 4주 동안 배양하여 발근시킨 식물체는 vermiculite와 perlite 가 1:1(v/v)로 혼합된 배양토에 1주간 순화시킨 후 일반토양에 이식한 결과, 완전한 식물체로 생장시킬 수 있었다. 다래의 절간절편체를 shoot 유도배지에 배양하여 shoot의 형성과정을 발달단계별로 조직학적인 측면에서 관찰한 결과, 배양초기에는 신장되거나 모양이 다양한 무정형의 세포가 대부분이었으나 배양이 진전됨에 따라 세포의 크기가 작고 세포질이 충만한 분열조직군이 형성되면서 이들로부터 장차 shoot로 발달될 shoot primordia가 형성되는 것을 관찰할 수 있었다.

• 한국약용작물학회지
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• 제14권1호
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• pp.31-35
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• 2006

Agrobacterium rhizogenes ATCC15834에 의해 형질전환된 지황 (Rehmannia glutinosa)의 모상근으로부터 유식물체를 재분화 시키기 위한 최적 조건을 탐색하였다. 모상근 절편으로부터 유식물체의 재분화는 0.5 mg/l BA가 첨가된 SH배지 (3% sucrose, pH 5.8)에서 배양 5주 후 이루어졌다. 이와 같은 유식물체는 식물생장조절물질이 포함되지 않은 기본배지로 옮겼을 때 발근이 이루어졌다. 형질전환된 모상근 유래 식물체는 잎으로부터 직접 재분화된 식물체와 비교했을 때 잎이 오므라드는 경향을 보였고 줄기는 절간이 짧았으며, 뿌리는 가늘고 수량이 많은 특징을 보였다. 생체량 비교에서 형질전환식물체는 식물 하나의 무게가 1.28 g (생중량)으로 대조구인 잎 유래 재분화식물체 (0.53 g)에 비해 높았으며, 지표물질인 catalpol의 함량에 있어서도 차이를 보여주었다.