• Journal of Plant Biotechnology
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• 제33권4호
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• pp.263-270
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• 2006

Pongamia pinnata Pierre is a tree legume, having potential in production of raw material for biodiesel. A protocol for in wk propagation of this plant was standardized using seedling explants. Growth regulators (GR) including gibberellic acid $(GA_3),\;N^6-benzylaminopurine(BA)$, thidiazuron (TDZ), and Adenine sulphate (Ads) were tested for optimum germination of seeds. Removal of seed coat prior to germination, controlled fungal growth partially but enhanced bacterial growth. Antibiotic cefotaxime was ineffective in controlling bacterial contamination. Seedling derived nodal explants and cotyledon nodes with attached cotyledons were excised and cultured for induction of shoots. Optimum sprouting and multiplication of shoot buds were obtained in MS medium supplemented with $8.88{\mu}M$ BA. These buds differentiated and rooted on medium devoid of GR. Optimum growth of Pongamia seedling was obtained in cotton plugged culture vessels. Reculturing of the cotyledon node explants produced more shoots from the same site. This process of removing shoots and reculturing of cotyledon node was followed for eight passages yielding 4 to 8 shoots in each cycle. The shoots (75%) rooted on half strength MS basal medium supplemented with 0.22% charcoal. All plants survived on transfer to soil. This is the first report on in vitro regeneration of Pongamia pinnata. This report demonstrates the possibility of coupling more than one parameter in single experiment to hasten the process of standardization. The process of cycling the nodal explant repeatedly for production of large number of shoots from single meristem may find application in genetic transformation experiments wherein meristems are used for transformation.

• 식물조직배양학회지
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• 제22권5호
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• pp.267-271
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• 1995

배추의 기내재분화율을 향상시키기 위하여 자엽을 치상재료로 하여 에틸렌 억제제인 AgNO$_3$와 STS를 첨가하여 그 영향을 조사하였다. AgNO$_3$의 경우에는 100 $\mu$M이 첨가되었을 때 STS에서는 5 $\mu$M이 첨가되었을 때, 무처리구의 4%, 2%에 비해 각각 가장 높은 22%와 15.3%의 재분화율을 보였다. 자엽연령에 대해서는 파종 후 3일 지난 자엽을 사용하였을 때 58%의 재분화율을 나타내었다. 배양용기에 따른 효과는 1회용 페트리디쉬에서 가장 높았으나 처리간에 뚜렷한 차이는 없었다. 뿌리유기 조건은 생장조절제가 포함되지 않은 처리와 NAA가 0.1 mg/L 농도로 첨가된 배지에서 가장 좋은 결과를 얻었으며 그 형태 및 밀도, 활력에서 가장 우수하였다. 기내에서 뿌리가 형성된 식물체를 4$^{\circ}C$에서 4주간 처리하여 추대를 유기하였고 형태적으로 정상적인 화기를 볼 수 있었으며 이들로부터 종자를 얻을 수 있었다.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.15-19
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• 2006

An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.25-37
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• 2004

Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).

• Journal of Plant Biotechnology
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• 제5권1호
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• pp.51-57
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• 2003

Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increased the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Salt tolerant mutants were obtained in rice the variety, 'Hawsungbyeo', through in vitro mutagenesis of in vitro cultured anther-derived calli. Various doses (30, 50, 70 and 90 Gy) of gamma ray were applied to investigate the effect of radiation on callus formation on medium containing 1% NaCl, green plant regeneration, frequency of selected doubled haploid mutants and of the salt tolerant screen. It was demonstrated that the dose of 30 and 50 Gy gamma rays had significant effects on callus formation, regeneration and selection of salt tolerance. No tolerant lines were obtained from non-mutagenized cultures. From gamma ray irradiated cultures, five tolerant lines ($M_2$generation) at germination stage and 13 tolerant lines ($M_3$genoration) at seedling stage were obtained. The frequency of salt tolerant mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve salt tolerance.

• Journal of Plant Biotechnology
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• 제40권2호
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• pp.65-71
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• 2013

본 연구의 목적은 희귀 및 멸종위기식물인 솔나리(Lilium cernum Komarov.) 기내 종자발아체의 인편으로부터 캘러스 유래의 식물체 재분화 조건을 확립하고자 하였다. 식물생장조절물질인 kinetin과 NAA 또는 2,4-D를 조합한 MS배지에 인편절편을 배양하였다. Kinetin과 NAA, 2,4-D를 혼합하여 처리한 모든 처리구에서 95% 이상의 캘러스 형성율을 보였으며, $3.0mg{\cdot}L^{-1}$의 kinetin과 $1.0mg{\cdot}L^{-1}$의 NAA를 조합처리한 MS배지에서 캘러스의 무게가 226 mg으로 가장 무거웠고, 캘러스 크기도 가장 우수하였다. 솔나리의 인편유래 캘러스로부터 자구형성에 미치는 BA와 NAA의 효과를 알아보기 위해 BA와 NAA를 조합한 MS배지에 캘러스를 배양하였다. 그 결과, $2.0mg{\cdot}L^{-1}$ BA와 $1.0mg{\cdot}L^{-1}$ NAA 혼합한 MS배지에서 78%의 높은 자구유도율을 보였다. MS배지 및 sucrose의 농도가 기내 건전 식물체 생산에 미치는 영향을 조사하기 위해 실험한 결과, 유의적 차이는 없이 모든 대조구와 실험구에서 건전한 유식물체를 생산할 수 있었고, 엽록소의 함량은 sucrose의 농도가 증가할수록 높아짐을 확인하였다.

• Journal of Plant Biotechnology
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• 제43권2호
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• pp.255-260
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• 2016

This research was conducted to improve the cold tolerance of the gerbera cv. Gold Eye by introduction of the Arabidopsis $Ca^{2+}/H^+$ antiporter gene (CAX1) via Agrobacterium-mediated transformation. Prior to genetic transformation, we optimized a combination of plant growth regulators; $1.0mgl^{-1}$ 6-Benzyladenine (BA) and $0.1mgl^{-1}$3-indole-acetic acid (IAA) were found to lead to proper in vitro shoot regeneration from petiole explants. In addition, $50mgl^{-1}$ kanamycin was determined to be the minimal concentration useful for selection of putative transgenic plants. In this study, transgenic gerbera expressing the Arabidopsis $Ca^{2+}/H^+$ antiporter gene (CAX1) were obtained using the optimized concentrations. We expect that introduction of the gene to the cultivar will improve cold tolerance, which will be important in the winter months.

• Journal of Plant Biotechnology
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• 제44권2호
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• pp.164-170
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• 2017

A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.

• 화훼연구
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• 제17권2호
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• pp.93-100
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• 2009

본 연구는 청나래고사리의 기내 포자체 유묘를 이용한 효율적인 대량생산 방법을 개발하기 위하여 시행되었다. 유묘의 엽신, 엽병, 근경을 곱게 다져 활성탄 0.1%를 첨가한 1/2MS배지에 배양한 결과, 근경의 절편에서만 포자체 유묘가 생산되었다. 곱게 다진 근경 절편은 1/2MS 배지에서 포자체 재생이 가장 왕성하였으며, sucrose 농도를 2%로 조절하고 $NaH_2PO_4$ $50mgL^{-1}$을 첨가할 경우 포자체 재생이 더욱 촉진되었다. Kinetin과 BA를 단용 또는 NAA, IBA와 각각 혼용하여 배양한 결과, kinetin $1{\mu}M$ 단용 처리구에서 포자체 재생이 가장 왕성하였다. BA 첨가구에서는 분열조직의 증식이 왕성하였으나. 분열조직이 포자체로 분화되지 못하는 특징을 보였다. 변형 1/2MS 배지(sucrose 2%, $NaH_2PO_4$ $50mgL^{-1}$, kinetin $1{\mu}M$, pH 5.8, agar 0.8%)에 활성탄 0 또는 0.1%를 첨가하여 고체, 액체 정체, 액체 진탕배양한 결과, 포자체 재생은 액체 진탕배양시 가장 왕성하였으며, 활성탄의 첨가는 포자체 재생을 촉진하였다.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2003년도 추계학술대회 발표논문 초록집
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• pp.163-163
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• 2003