• Title/Summary/Keyword: shoot regeneration

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Production and Characterization of Nitrate Reductase Deficient Mutants in Petunia parviflora

  • Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.6
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    • pp.706-715
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    • 2006
  • Nitrate reductase deficient (NR) mutant lines were selected indirectly by their resistance to 100mM chlorate in cell cultures of P. parviflora. A total of 585 chlorate resistant lines were confirmed by a second passage on a high concentration of chlorate. Frequency of spontaneous mutation was $9.7{\times}10^{-7}$ in 3 month old suspension-cultured cells, and in non-selective media containing amino acids as sole nitrogen source. The frequency of mutation could be increased up to 11-fold by culture for 12 months. Out of 40 randomly selected calli, 22 were fully deficient in NR. The rest of the clones contained a decreased level of NR activity. Further characterization was carried out in 13 mutant lines which were fully deficient in NR and in 5 mutant lines containing residual (0-7.0%) NR activity, as compared to wild-type cells cultured on the same medium. The $NR^-$ mutants were tentatively classified as defective in the NR apoenzyme (nia-type; 11 mutant lines including the 5 with residual NR activity) or in the molybdenum cofactor (cnx-type; 7 mutant lines) by the XDH activity. The cnx-type could be further classified into two groups. In one group (5 mutant lines) of these, the NR activity could be partially restored by nonphysiologically high (1.0mM) molybdate in the culture medium. Both types of $NR^-$ mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They also proved to be extremely sensitive to the standard medium ($MSP_1$) containing nitrate and ammonium. Shoot regeneration was obtained only in the $NR^-$ mutants, which contained residual NR activity, but they so far have failed to grow into plants.

Developing a mass propagation technique for Aralia elata via somatic embryogenesis

  • Moon, H.K.;Lee, J.S.;Kim, T.S.
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2000.10a
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    • pp.114-115
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    • 2000
  • Aralia elata is found in mountain areas all over Korean peninsula. Aralia elata is the scientific name for Japanese angelica tree. The tree belongs to the family Araliaceae, commonly known as ginseng family. Bud sprouts from apical shoot tip of the plants are rich in flavor and thus mainly used for both folk medicine and vegetable. The stalks with apical buds are gathered in the early spring and planted in sandy soil or water in the greenhouse. The sprouting buds are then collected and sold as fresh vegetable. Although the plants have been used for food, they have been cultivated in a very small scale. In spring, local farmers just go around mountain areas to search the trees and gather the stalks as much as they get and sell them to the market. No conservation efforts have been made to stop the exploitation or to save the dwindling population. We tried to provide local farmers with the plants that may be used as an alternative to stalks from wild populations. This will bel! p conserve the wild populations. However, it is hard to propagate them either by conventional cuttings or by seed germination in a short period of time. Mass propagation using tissue culture systems have shown a great promise with several woody plants. Recently we developed a mass propagation technique via somatic embryogenesis system using mature and/or juvenile explants for Aralia elata. Several factors affecting somatic embryogenesis system including SE(somatic embryo) induction, embryogenic callus proliferation, SE germination, plant regeneration and transplanting to field frill be presented. And some problems arising for the somatic embryogenesis system will be also discussed.

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Developing a mass propagation technique for Aralia elata via somatic embryogenesis

  • Moon, H.K.;Lee, J.S.;Kim, T.S.
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2000.10b
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    • pp.16-17
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    • 2000
  • Aralia elata is found in mountain areas all over Korean peninsula. Aralia elata is the scientific name for Japanese angelica tree. The tree belongs to the family Araliaceae, commonly known as ginseng family. Bud sprouts from apical shoot tip of the plants are rich in flavor and thus mainly used for both folk medicine and vegetable. The stalks with apical buds are gathered in the early spring and planted in sandy soil or water in the greenhouse. The sprouting buds are then collected and sold as fresh vegetable. Although the plants have been used for food, they have been cultivated in a very small scale. In spring, local farmers just go around mountain areas to search the trees and gather the stalks as much as they get and sell them to the market. No conservation efforts have been made to stop the exploitation or to save the dwindling population. We tried to provide local farmers with the plants that may be used as an alternative to stalks from wild populations. This will hel! p conserve the wild populations. However, it is hard to propagate them either by conventional cuttings or by seed germination in a short period of time. Mass propagation using tissue culture systems have shown a great promise with several woody plants. Recently we developed a mass propagation technique via somatic embryogenesis system using mature and/ or juvenile explants for Aralia elata. Several factors affecting somatic embryogenesis system including SE(somatic embryo) induction, embryogenic callus proliferation, SE germination, plant regeneration and transplanting to field will be presented. And some problems arising for the somatic embryogenesis system will be also discussed.lso discussed.

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Transformation of PAT gene into Lettuce (Lactuca sativa L.) using Agrobacterium tumefaciens (Agrobacterium tumefaciens를 이용한 상추 (Lactuca sativa L.)의 PAT유전자 형질전환)

  • 류정아;김창길;이현숙;최경배;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.197-200
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    • 2001
  • Agrobacterium tumefaciens MP90 harboring PAT (phosphinothricin acetyltransferase) and NPTII-GUS gene were used for the genetic transformation of lettuce (Lactuca Sativa L.). Shoot regeneration from cotyledon explants were obtained from the MS medium supplemented with 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin and 500 mg.L$^{-1}$ carbenicillin after cocultivation with A. tumefaciens for 2 days. Kanamycin resistance test of transgenic plants indicated that the NPTII gene was integrated into the lettuce genome and was stably expressed. PCR and northern blot analysis indicated that bialaphos resistance gene (PAT) was stably integrated into the lettuce genome. The transgenic plant sprayed with Basta (1500x) remained healthy with continuous growth, while the control group exhibited fatality.

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In vitro propagation of endangered species, Hylotelephium ussuriense (Kom.) H. Ohba (멸종위기종 둥근잎꿩의비름 (Hylotelephium ussuriense (Kom.) H. Ohba)의 기 내 증식)

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Kim, Ji-Ah;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.38-43
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    • 2014
  • To establish the system of in vitro plant regeneration, the different explants (stem with axillary bud and stem without axillary bud) of Hylotelephium ussuriense were cultured on the Murashige and Skoog's medium containing 6-benzylaminopurine (BA) and indolebutyric acid (IBA). The adventitious shoot induction was more effective in the stem with axillary bud explants than the stem without axillary bud explants, and was the best on MS medium containing 3.0 mg/L BA and 0.01 mg/L IBA. Frequency of plantlet growth was not significantly treated on MS and sucrose. Total chlorophyll contents under ventilation treatment were higher than those in control (non-ventilation). This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Seasonal Dynamics of the Seagrass Zostera marina on the South Coast of the Korean Peninsula

  • Lee, Kun-Seop;Kang, Chang-Keun;Kim, Young-Sang
    • Journal of the korean society of oceanography
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    • v.38 no.2
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    • pp.68-79
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    • 2003
  • Although seagrasses are relatively abundant, few studies have been conducted on seagrass physiology and ecology in Korea. Zostera marina is the most abundant seagrass species, widely distributed throughout all coastal areas of the Korean peninsula. To examine seasonal dynamics and spatial variations of eelgrass, Zostera marina distributed on the coast of Korea, morphological characteristics, biomass, tissue nutrient constituents, leaf productivity and environmental factors were monitored monthly from the eelgrass beds in Kabae Bay and Kosung Bay on the south coast of the Korean peninsula from June 2001 to June 2002. Eelgrass density, biomass, morphological characteristics, leaf productivities, and tissue nutrient constituents exhibited clear seasonal variations, and these seasonal trends reflected seasonal changes in water temperature. Eelgrass shoot density and biomass at Kabae Bay site showed more obvious seasonal trends than Kosung Bay. No strong seasonality in Kosung Bay site appeared to be caused by high water temperature ($>30{\circ}C$) during summer months at this site. Despite differences in nutrient availabilities between two study sites, eelgrass biomass and leaf productivities were not significantly different between study sites, and this lack of spatial variations implies that the ambient nutrient availabilities at the present study sites are in excess of seagrass nutrient demand. Eelgrass tissue N content and sediment pore water DIN concentrations exhibited reverse relationship at the present study. This reverse relationship suggests in situ nutrient concentrations are not good indicator of nutrient availabilities, and regeneration and turnover rates of sediment nutrients are also important factors to determine nutrient availabilities at the site.

Regeneration of plants from alginate-encapsulated axenic nodal segments of Paederia foetida L. - A medicinally important and vulnerable plant species

  • Behera, Biswaranjan;Behera, Shashikanta;Shasmita, Shasmita;Mohapatra, Debasish;Barik, Durga Prasad;Naik, Soumendra Kumar
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.255-263
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    • 2021
  • Paederia foetida L. is an important medicinal plant that has been used for the treatment of various gastrointestinal related ailments by different tribal communities in India. This plant is also known for its use as a food. Due to overexploitation, P. foetida has been classified as a vulnerable plant in some states of India. The propagation of P. foetida by conventional methods is easy but very slow. Synthetic seed technology offers incredible potential for in vitro propagation of threatened and commercially valuable plants, and can also facilitate the storage and exchange of axenic plant material between laboratories. However, synthetic seed production for P. foetida has not yet been reported. Thus, to the best of our knowledge, the present study is the first attempt to produce synthetic seeds of P. foetida by calcium alginate encapsulation of in vitro regenerated axenic nodal segments. Sodium alginate (3%) and CaCl2 (100 mM) were found to be the optimal materials for the preparation of ideal synthetic seeds, both in terms of morphology and germination ability. The synthetic seeds showed the best germination (formation of both shoot as well as root; 83.3%) on ½ MS medium augmented with 0.5 mg/L indole-3-acetic acid. The plantlets obtained from these synthetic seeds could be successfully acclimatized under field conditions. We also studied the storage of these synthetic seeds at low temperature and their subsequent sprouting/germination. The seeds showed a germination rate of 63.3% even after 21 days of storage at 4 ℃; thus, they could be useful for transfer and exchange of P. foetida germplasm.

Development of transgenic disease-resistance root stock for growth of watermelon.(oral)

  • S.M. Cho;Kim, J.Y.;J.E. Jung;S.J. Mun;S.J. Jung;Kim, K.S.;Kim, Y.C.;B.H. Cho
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.65.2-65
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    • 2003
  • To protect the plant against several soil-borne pathogens, we are currently constructing disease-resistant transgenic root stock for the growth of cucurbitaceae vegetable plants, watermelon and gourd. We made a watermelon cDNA library from Cladosporium cucumerinum-Infected leaves for substractive hybriazation and differential screening. We isolated the several pathogen inducible cDNA clones, such as caffeoyl-CoA-methyltransferase, LAA induced protein, receptor-like kinase homolog, hydroxyproline-rich glycoprotein, catalase, calmodulin binding protein, mitochondrial ATPase beta subunit, methyl tRNA synthetase and WRKY transcription factors. We previously obtained CaMADS in pepper and galactinol synthase ( CsGolS) in cucumber that were confirmed to be related with disease-resistance. CaMADS and CsGolS2 were transformed into the inbred line 'GO701-2' gourd, the inbred line '6-2-2' watermelon and the Kong-dye watermelon by Agrobacterium tumerfaciens LBA4404. Plant growth regulators (zeatin, BAP and IAA) were used for shoot regeneration and root induction for optimal condition. Putative transgenic plants were selected in medium containing 100mg/L kanamycin and integration of the CaMADS and CsGO/S2 into the genomic DNA were demonstrated by the PCR analysis. We isolated major soil-borne pathogens, such as Monosporascus cannonballus, Didymella bryoniae, Cladosporium cuvumerinum from the cultivation area of watermelon or root stock, and successfully established artificial inoculation method for each pathogen. This work was supported by a grant from BioGreen 21 program, Rural Development Administration, Republic of Korea.

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Effect of Medium Composition on in Vitro Shoot Regeneration from Leaves of Cassava (Manihot esculenta Crantz) Through Somatic Embryogenesis and Callus Induction (카사바 잎 절편 유래 체세포배 배양시 배지조성이 기내 식물체 재분화에 미치는 영향)

  • Young Hee Kwon;Joung Kwan Lee;Hee Kyu Kim;Kyung Ok Kim;Ju Hyoung Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.19-19
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    • 2020
  • The Cassava(Manihot esculenta Crantz) is a tropical root crop, originally from Amazonia, that provides the staple food of an estimated 800 million people worldwide. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. So we tried to optimize protocols for mass production of somatic embryo amenable to large-scale vegetative propagation of Cassava. After in vitro eight-week culture of leaves of Cassava, the medium which contained the 2,4-D, BAP and IBA showed the highest callus induction rate, embryogenesis callus formation rate and somatic embryo formation in Cassava culture. In the medium with GA3 and myo-inositol, shoots were most vigorously regenerated from somatic embryos of Cassava. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

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Optimization of in vitro lily culture system with different treatments of taurine (타우린 처리에 의한 나리 기내 식물체 생산체계 최적화)

  • Lee, Sang-Hee;Yang, Hwan-Rae;Kim, Sun Tae;Jun, Tae Hwan;Kim, Yong Chul;Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.484-489
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    • 2017
  • Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.