• Title/Summary/Keyword: Plant Cell Culture

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Resistance of Plants to Herbicide (제초제(除草劑)에 대한 식물(植物)의 저항성(抵抗性))

  • Kim, Kil-Ung
    • Korean Journal of Weed Science
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    • v.4 no.1
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    • pp.96-106
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    • 1984
  • Changes in weed floras and development of plant resistance to herbicides seemed to be closely related with increased and repeated use of herbicides. Herbicide use increased from 5% of the total consumption of pesticide in 1950 to 45% in 1976 in world basis. About 200 herbicides have been introduced to agriculture so as to control about 206 weed species which have been recorded important to human beings. In Korea, there was about 351 times in increased use of herbicides from 1966 to 1982. Interspecific selection by herbicide is mainly responsible for changes in weed floras and resulted in varying tolerance or susceptibility to herbicides, together with the changes of agricultural practices. The present trend toward continuous cereal cultivation throughout world will lead to type of changes in weed floras favorable to therophyte which can survive under unfavorable conditions as seeds rather than the types of geophyte which can survive unfavorable seasons as buds placed below soil surface. However, geophyte such as Sagitaria pygmaea, and Scirpus jurtcoides, and Cyperus rotundus and Cynodon dactylon in temperate warm climate become severe paddy weeds, presumably because of the removal of annual weeds by herbicides. Since differential tolerance to 2,4-D was firstly reported in Agrostis stolofera, about 30 species of weeds in 18 genera are presently known to have developed resistance to triazine herbicides. Resistance of weed biotypes to triazine herbicide is not mainly due to limited absorption and translocation or to the difference in metabolism, but is the result of biochemical changes at the site of metabolic activity, such as a loss of herbicide affinity for triazine binding site in the photosystem II complex of the chloroplast membrane. Genetical study showed that plastid resistance to triazine was wholly inherited through cytoplasmic DNA in the case of Brassica campestris. Plant tissue culture method can be utilized as an alternate mean of herbicide screening and development of resistance variants to herbicides as suggested by Chaleff and Parsons. In this purpose, one should be certain that the primary target process is operational in cell culture. Further, there are a variety of obstacles in doing this type of research, particularly development of resistance source and it's regeneration because cultured cells and whole plants represent different developmental state.

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Effect of Methyl Jasmonate on in vitro Bulblet Formation and Enlargement from Shoot Clump of Allium victorialis (산마늘 다신초 덩어리로부터 인경 형성과 비대에 미치는 methy jjasmonate의 영향)

  • Park, So-Young;Ahn, Jin-Kwon;Lee, Wi-Young;Park, Hae-Chin
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.79-82
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    • 2004
  • The effect of abscisic acid (ABA) and methyl jasmonate (MeJA) on bulblet formation from the culture of Allium victorialis var. platyphyllum Makino was studied. Shoot clumps were cultured on MS medium containing ABA (0, 0.01∼2.0mg/L) and MeJA (0.01∼5.0mg/L). ABA at low concentrations (0.01mg/L) induced shoot proliferation without bulblet formation. However, bulblet formation started on the medium containing MeJA approximately in 4-6 weeks of culture. Furthermore, 1.0mg/L MeJA resulted in bulblet formation at high frequency (100%) as compare to the control (46.1%). Cortical cells of the bulblets enlarged on medium with MeJA had dense protein-like substance in expanded and round cells when examined under the microscope. The data described here show that formation and enlargement of bulblets from Allium victorialis can be improved by addition of appropriate concentration of MeJA.

Antiviral Activity of Antibiotic Peptaibols, Chrysospemins B and D, Produced by Apiocrea sp. 14T against TMV Infection

  • Kim, Young-Ho;Yeo, Woon-Hyung;Kim, Young-Sook;Chae, Soon-Young;Kim, Kap-Sik
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.522-528
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    • 2000
  • A total of about 300 fungal isolates from forest havitats were screened for inhibitors of tobacco mosaic virus (TMV) infection using its local lesion host, Nicotiana tabacum cv. Xanthi nc. Ine of the isolates, 14T, showed a strong activity against TMV infection, and was identified as an Apiocrea sp. based on its morphological characterstics. Rice was an optimum culture medium for its fermentation, and two antiviral compounds, KGT 141 and KGT 142, were resolved from the rice culture through column chromatography, TLC, and HPLC. By NMR and FAB-MS, the two compounds were identified as chrysospermins B (KGT 141) and D (KGT 142), both of which are peptaibols with 19-mer amino acids possessing an acetylated N-terminus and a hydroxy-amino acid (tryptophanol) at the C-terminus. Both compounds showed inhibitory activities against TMV infection, but chrysospermin D showed the stronger activity than chrysospermin B. The former of $100{\;}\mu\textrm{g}/ml$ and 54.7% at $10{\;}\mu\textrm{g}/ml$, respectively. Furthermore, the chrysospermins were highly cytotoxic toward cancer cell lines of PC-3 (prostrate) and K562 (leukemia), and inhibited growth of the Gram-positive bacteria tested, especially the plant pathogenic bacterium Corynebacterium lilium. To the best of our knowledge, this is the first report on the inhibition of plant virus infection by antimicrobial peptaibols.

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Relationships Between Pathogenicty and Activities of Polygalacturonase, Laccase, and ${\beta}$-Glucosidase Produced by Botrytis cinerea (Botrytis cinerea 균주들이 생산하는 Polygalacturonase, Laccase, ${\beta}$-glucosidase의 균주 간 화성 및 병원성과의 상관관계)

  • Kim, Jong-Jin;Kim, Jae-Won;Lee, Chang-Won;Chung, Young-Ryun
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.225-231
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    • 1997
  • Activities of polygalacturonase, laccase, and intra- and extra-cellular $\beta$-glucosidase produced by 20 Botrytis cinerea isolates in liquid culture media containing cucumber cell was as a carbon source were measured and their relationships to the pathogenicity were analyzed. No significant correlations between these enzyme activities and the pathogenicity of B. cinerea were found. Mycelial growth rate on Bayendamm media, however, was higthly correlated with the pathogenicity (r=0.522) anong these isolates. Immuno-blot analysis of the culture filtrate using antibody against against exo-polygalacturonase revealed that only one band with molecular weight of 66 kDa was detected amone 34 tested isolates. It appears that these enzymes may not be primary factors in dermining the pathogenicity of B. cinerea.

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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.

Comparison of Frequency Embryogenesis through Microspore Culture of Domestic Cultivars in Brassica napus L. (소포자 배양에 의한 반수체 식물유도 효율이 높은 국내 유채 품종 선발)

  • Park, Yoon-Jung;Kim, Kwang-Soo;Jang, Young-Seok;Kim, Chul-Woo;Bang, Jin-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.237-241
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    • 2006
  • This experiment was carried out comparison with haploid plants productivity by microspore culture among domestic cultivars of Brassica napus L. Isolated microspore from flower buds were cultured on NLN medium supplemented with 13% sucrose, $0.05mg/{\ell}$ BA and $0.5mg/{\ell}$ NAA. Genotype was important factor in haploid embryo productivity 'Tamlayuchae' showed the highest haploid embryo production frequency (176 embryos formed from 1 flower bud). But, 'Hallayuchae' and 'Youngsanyuchae' were not generated embryo even cell division. When suspension culture on NLN liquid medium at 100 rpm, embryos were developed multilobe abnormal embryo cluster. Multilobe abnormal embryos on MS medium basal solid medium were regenerated multiple shoots. Regenerated haploid plant with well developed shoots and roots on MS basal medium were successfully transferred to pots.

Isolation and Culture of In Vitro Cultured Populus alba×P. grandidentata Protoplasts (Populus alba×P. grandidentata 조직배양(組織培養) 식물체(植物體) 원형질의 분리(分離)와 배양(培養))

  • Chun, Young Woo
    • Journal of Korean Society of Forest Science
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    • v.71 no.1
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    • pp.45-49
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    • 1985
  • Protoplast-source meterial and enzyme strength had a significant influence on protoplast yield from hybrid poplar, Populus alba ${\times}$ P. grandidentata. The yield of protoplasts from in vitro culture of 1 month-old plantlets was more than that from greenhouse grown 4 month-old stock plant. In vitro cultured plantlets regulary produced more viable protoplasts with E-I enzyme solution (0.5% cellulase and 0.1% macerase) than those with E-II enzyme solution (1.0% cellulase and 0.2% macerase) after overnight incubation. The mean yield of protoplasts from in vitro cultured plantlets was $4{\times}10^6$ with E-I enzyme solution. Cell division was observed in these protoplast cultures after 7-10 days. Protoplast-derived hybrid poplar cells survived over 3 weeks in culture and some continuous cell divisions were evident. Other aspects associated with protoplasts from in vitro cultured plantlet are also discussed.

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Fermentation Process Development of Recombinant Hansenula polymorpha for Gamma-Linolenic Acid Production

  • Khongto, B.;Laoteng, K.;Tongta, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1555-1562
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    • 2010
  • Development of the strain and the fermentation process of Hansenula polymorpha was implemented for the production of ${\gamma}$-linolenic acid ($GLA,\;C18:3{\Delta}^{6,9,12}$), an n-6 polyunsaturated fatty acid (PUFA) that has been reported to possess a number of health benefits. The mutated ${\Delta}^6$-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without the utilization of methanol, a high-cell-density culture of the yeast recombinant carrying the ${\Delta}^6$-desaturase gene was then achieved by fed-batch fermentation under glycerol-limited conditions. As a result, high levels of the ${\Delta}^6$-desaturated products, octadecadienoic acid ($C18:2{\Delta}^{6,9}$), GLA, and stearidonic acid ($C18:4{\Delta}^{6,9,12,15}$), were accumulated under the derepression conditions. The GLA production was also optimized by adjusting the specific growth rate. The results show that the specific growth rate affected both the lipid content and the fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates tested, the highest GLA concentration of 697 mg/l was obtained in the culture with a specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor ${\Delta}^6$-desaturase gene was similar to that of blackcurrant oil, with both containing similar proportions of n-3 and n-6 essential fatty acids.

Enhanced Production of hCTLA4Ig by Adding Sodium Butyrate and Sodium Pyruvate (Sodium butyrate와 sodium pyruvate 첨가에 의한 hCTLA4Ig 생산성 증대)

  • Yoo, Mi-Hee;Kim, Soo-Jin;Kwon, Jun-Young;Nam, Hyung-Jin;Kim, Dong-Il
    • KSBB Journal
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    • v.26 no.5
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    • pp.386-392
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    • 2011
  • Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig), an immunosuppressive agent, was expressed in transgenic rice cells using RAmy3D promoter and RAmy1A signal peptide for the inducible production and secretion into culture media by sugar depletion. In this study, sodium butyrate was used as a small molecular enhancer (SME) to enhance the production of hCTLA4Ig in transgenic rice cell suspension cultures. When 1 mM sodium butyrate was added in sugar-free media, relative viability was not reduced, while the productivity was improved 1.3-fold. In addition, by supplementing 87 mM sodium pyruvate as an alternative energy source during the production phase, death rate of the cells was decreased. When sodium pyruvate was not added, most cells became dead at day 6. However, by adding sodium pyruvate, 18% of viability can be maintained until day 10 and the production of hCTLA4Ig was enhanced 1.4-fold. When the combination of sodium pyruvate and sodium butyrate at optimum concentrations was added, the highest viability and hCTLA4Ig production could be obtained. The highest level of hCTLA4Ig reached up to 35 mg/L at day 10.

Plant Cell-Wall Degradation and Glycanase Activity of the Rumen Anaerobic Fungus Neocallimastix frontalis MCH3 Grown on Various Forages

  • Fujino, Y.;Ushida, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.5
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    • pp.752-757
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    • 1999
  • Studies were made of digestion of timothy (Pheleum pretense) hay, tall fescue (Festuca elatior) hay, and rice (Oryza sativa) straw in pure cultures of rumen anaerobic fungus, Neocallimastix frontails MCH3. The fungus was inoculated on ground forages (1%, w/v) in an anaerobic medium and incubated at $39^{\circ}C$. Incubation was continued for 24, 48, 72 and 96 h. The losses of dry matter, xylose and glucose of forage during incubation were determined at the end of these incubation periods. Xylose and glucose were considered to be released from xylan and cellulose, respectively. The digested xylan to digested cellulose (X/C) ratios of the substrate were calculated. Xylanase and carboxymethyl cellulose (CMCase) of culture supernatant and residual substrate was measured at the same time. The X/C ratios in the cultures on timothy hay and rice straw were greater than 0.5 in the first 24-h incubation period. The values were smaller than 0.3 in tall fesque. The ratio of xylanase activity to that of CMCase in the first 24-h incubation period correlated well with the traits in X/C ratio. However xylanase activity was still superior to CMCase in the following incubation period (48 to 96 h), although the glucose (designated as cellulose) was more intensively digested than xylose (designated as xylan). The production of these polysaccharidases appeared to correlate with substrate cell-wall sugar composition, xylose to glucose ratios, at the beginning of fast growing period.