• 식물조직배양학회지
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• 제27권3호
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• pp.191-195
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• 2000

사과나무 교배잡종 (P.16 $\times$ M. prunifolia)의 자엽과 배축 조직을 배양하여 신초의 재분화에 미치는 2,4-D, NAA 키네틴, BA, thidiazuron의 처리효과를 조사하였다. 자엽조직은 1.0 mg/L＋ NAA +2.0 mg/L BA 처리구에서, 배축조직은 0.5 mg/L NAA+0.5mg/L BA처리구에서 가장 많은 수의 신초가 분화하였으며, BA 무첨가구에서는 신초의 재분화가 전혀 이루어지지 않았다. 자엽조직보다는 배축조직을 배양하는 것이 더 효과적이었으며 특히, 배축의 상위부분을 치상하였을때 신초 재분화가 더 양호하였다. 재분화된 신초를 1/2 MS에 1.0mg/L NAA를 첨가한 발근배지에 계대배양한 결과 발근이 양호하였으며 정상적인 식물체로 생장하였다.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.101-107
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• 2010

An efficient plant regeneration system has been developed for figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), which is exclusively used as a rootstock for cucumber. The protocol is based on results obtained from a series of culture experiments involving different parts of the cotyledons and various media. The culture of cotyledon explants was critical for the enhancement of shoot regeneration frequency. The lower parts of the cotyledon excised at the plumule base were found to display a markedly enhanced production of adventitious shoots compared to other cotyledon regions. Culture in silver nitrate-supplemented Murashige and Skoog (MS) medium was not beneficial for shoot regeneration and suppressed root regeneration. Efficient shoot regeneration was obtained on MS medium containing 1.0 $mg\;l^{-1}$ zeatin and 0.1 $mg\;l^{-1}$ indole-3-acetic acid. Regenerated shoots successfully elongated and rooted in medium containing 0.1 $mg\;l^{-1}$ 1-naphthalene-acetic acid after 10-15 days of subculturing. The plantlets were satisfactorily acclimatized in a greenhouse and grew into normal plants without any morphological alterations.

• 식물조직배양학회지
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• 제25권5호
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• pp.301-304
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• 1998

자엽으로 부터 신초 유기율은 MS기본배지와 MS기본배지에 IAA 0.25mg/L 또는 0.50mg/L과 zeatin 2.0mg/L 또는 4.0mg/L를 단독 및 조합 처리한 배지중에서 M배지에 IAA 0.25mg/L + zeatin 2.0mg/L 첨가된 배지가 63%로 가장 양호하였다. 신초 유기시기는 품종 및 배지에 따라 치상후 9~25일의 차이가 있었는데 MS배지 + zeatin 2.0mg/L + IAA 0.25 mg/L배지에서 영양재래, 풋고추, 342, Kakovskij-A-35, Gris I-A-1가 빠르게 유기 되었으며, MS배지 + BA 2.0mg/L + IAA 1.0mg/L 배지에서 COO485 Long Hot P5-3, Gris I-A-1에서 유기가 빨았다. 신초 유기율도 품종 및 배지에 따라 달랐는데 MS 배지 + zeatin 2.0mg/L + IAA 0.25mg/L 배지에서는 621, 영양재래, Nikko(일광), 적색물고추, Ch-6Num-216, 풋고추, Kakovskij-A-35가, MS배지 + BA 2.0mg/L + IAA 1.0mg/L 배지에서는 Fresno Chile, PI 169126, 적색물고추, PI 297438가 90%이상으로 높았다.

• Journal of Plant Biology
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• 제17권4호
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• pp.171-174
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• 1974

Cotyledon of Panax ginseng was cultured in the growth regulator-free Knudson C medium comprising only several kinds of mineral salts and sucrose. Shoot primordium or callus developed profusely from the cotyledonary tissue and finally plantlets were produced directly from the shoot primordium or indirectly through callus. Microscopic examination revealed that the epidermal cell as well as the mesophyll cell of the cotyledon became meristematic and divided, changing into multinucleate cell or multicellular body, eventually developing into either a shoot primordium or callus.

• 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 Biology
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• 제33권4호
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• pp.253-257
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• 1990

Callus-inducing ability of Alnus hirsuta was examined by culturing various tissues (leaf, hypocotyl, cotyledon and seed) on NT (Nagata & Takebe) medium, supplemented with 2.5$\mu$M 2,4-D. Leaf-originated callus was cultured on media varying in auxin (IBA and NAA) and cytokinin (BAP) concentrations to examine the effects of auxin and cytokinin on callus growth. Maximum growth was obtained at 10 $\mu$M IBA+10$\mu$M BAP and 10$\mu$M NAA without cytokinin. Cell suspensions established from cotyledon-originated callus yielded viable protoplasts after incubation for 16-18 hours in an enzyme mixture (1% (w/v) Onozuka R-10 0.5% (w/v) Macerozyme, CPW salts and 13% (w/v) mannitol, pH 5.8). Protoplasts were cultured on NT medium, supplemented with glucose, hormones and coconut milk. After 6 weeks of culture, protoplasts sustained cell divisions to form microcallus, which showed various colors from red to white.

• 한국식물병리학회지
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• 제11권4호
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• pp.292-297
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• 1995

We developed an in vitro assay method for evaluating plant growth promotion and providing an evidence that the growth promotion is rendered by growth enhancing factors. The amendment of culture filtrates of Trichoderma harzianum T95 and Gliocladium virens G872 and G872B in Murashige and Skoog (MS) agar medium enhanced the cotyledon growth of cucumber in terms of fresh weight and primary leaf area of cucumber cotyledon cuttings, of which the treatment of G. virens G872B was the most effective. The mycelial culture filtrate of G872B was more effective in the growth promotion than its conidial germling filtrate. The addition of 1% sucrose in MS mineral medium with 0.1% culture filtrates of the antagonists (T95 and G872B) was optimum for enhancing the effect of the filtrates on the growth of cotyledon cuttings in vitro. When cucumber seeds treated with G872B, Pseudomonas putida Pf3 or the G872B-Pf3 mixture were planted in a greenhouse, the rate of seed germination, biomass of shoot and root, and yield of cucumber fruits were increased, especially by G872B or the G872B-Pf3 mixture. Correspondingly, cucumber fruit yields in early to middle-cycles of harvest were significantly greater in the plots of G872B than the control and Pf3-treated plots, and the final yield was highest in the plots of the G872B-Pf3 mixture applications.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권1호
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• pp.51-55
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• 2001

This study was conducted to establish a plant cell culture system for the production of medically important secondary metabolites from Xanthium strumarium. The effects of plant growth regulators including NAA, 2,4-D, kinetin, and ABA were examined in terms of callus induction, maintenance of callus and suspension cultures. It was shown that callus was induced upon treatment with NAA while embryo was induced after treatment with 2,4-D. Callus formation was further improved by treatment with ABA and NAA. The level of callusing increased by 17-29% for the seed case, cotyledon, leaf, and hypocotyl and by 96% in the case of the root. Suspension cell lines were established using calli produced from cotyledon, hypocotyl and root and cultured at 25$\^{C}$ under light conditions. The cells grew up to 15g/L with NAA 2ppm, BA 2ppm, and ABA 1ppm treatment. Supernatants of suspension cultures of cell lines derived from coyledon and hypocotyl produced some distinctive secondary metabolites, one of which was identified as 8-epi-tomentosin, which belongs to the xanthanolides. The amounts of 8-epi-tomentosin produced by the cotyledon- and hypocotylderived cell lines were 13.4mg/L and 11.0mg/L, respectively.

• 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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• 제22권3호
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• pp.157-164
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• 1995

온전한 접합자배 또는 자엽절편을 오옥신(2,4-D, IAA)과 사이토카이닌(BA, kinetin)이 첨가된 배지에 배양하였다. 접합자배를 온전한 상태로 배양했을 경우, 오옥신은 발아를 억제하였으나 사이토카이닌은 억제하지 않았고, 체세포배의 발생은 발아되지 못하는 배에서만 유도되었다. 자엽절편을 배양한 경우, 기본배지에서 체세포배발생률이 가장 높았다. 오옥신을 첨가한 배지에서는 체세포배가 자엽의 표면에서 산재하여 발생되었는데 비해서 기본배지에서는 자엽의 기부에서만 발생되었다. 2,4-D를 첨가한 배지에서는 체세포배가 자엽의 표피 및 하표피를 포함한 다수의 세포로부터 기원되어 다배로 발생되었다. 이 체세포배를 같은 배지에 배양했을 때 일차배의 표면으로부터 이차배가 발생되었는데 이들의 경우는 주로 표피 또는 하표피의 단세포로부터 유래되었음을 조직학적으로 밝혔다.