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Jung YS Yang BK Jeong YT Islam R Kim SM Song CH 《Journal of microbiology and biotechnology》2008,18(8):1431-1438
Immunomodulating activities of water-soluble exopolysaccharides (LL-EX) obtained from submerged mycelial culture of Lentinus lepideus were studied and their effectiveness was compared with lipopolysaccharide (LPS). The influence of the LL-EX on macrophage cellular lysosomal enzyme activity was to stimulate up to 267%, 392%, and 464% at the level of 10, 50, and 100 mug/ml, respectively. When the LL-EX was further fractionated into LL-Fr.I and Fr.II by Sepharose CL-6B gel chromatography, the cellular lysosomal enzyme activity of LL-Fr.II (2.1- fold) was higher than Fr.I (1.2-fold). Moreover, both LL-Fr.I and Fr.II stimulated the cytokines IL-1beta, TNF-alpha, and IL-6 in macrophages. In mixed lymphocyte reaction, LL-Fr.I and Fr.II enhanced the splenocyte proliferation up to 1.2-fold and 1.4-fold (50 mg/ml), respectively, stimulating only T lymphocytes. The fractions of LL-EX not show any direct toxicity against human gastric adenocarcinoma cell (AGS). The molecular masses of LL-Fr.I and Fr.II were estimated to be about 1,986 kDa and 21 kDa, respectively. The total sugar and protein contents of the two fractions were 84.97% and 69.88% and 15.03% and 30.12%, respectively. The sugar and amino acid compositions of the LL-Fr.I and Fr.II were also analyzed in detail. 相似文献
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Aspects of protoplast culture and plant regeneration 总被引:1,自引:0,他引:1
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A yield of 2–4×106 protoplasts/g F.W. could be obtained when fresh cauliflower inflorescence segments were digested with 2% cellulase Onozuka R-10, 1% cellulase RS and 0.4% Macerozyme R-10 in CPW18S for 7 to 10 h. Purified protoplasts were cultured in K8p liquid and agarose medium. Although protoplasts in liquid medium divided earlier than in agarose, protoplast-derived cells in liquid culture could not avoid browning. With agarose culture, sustained division and callus formation could be achieved. After 20 days, calli were transferred onto B5 agar medium with ZT 1.5 mg l-1, BA 0.5 mg l-1 and IAA 0.1 mg l-1 for shoot formation. The frequency of bud formation varied from 56.7% for calli of 1mm in size to 5.6% for 5mm calli. The shoots formed were rooted in B5 medium containing 0.5 mg l-1 IBA, and the regenerated plants were transplanted to pots and grew normally. It took about two months from protoplasts to the regenerated plants.Abbreviations Ade
adenine
- BA
6-benzyl aminopurine
- CH
casein hydrolysate
- CM
coconut milk
- 2,4-D
2,4,-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- Gln
glutamine
- NAA
-naphthylacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- ZT
zeatin 相似文献
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J. J. Rybczyński 《Plant cell reports》1989,8(7):383-386
Explants from hypcotyls and cotyledons of Browalia speciosa were shown to regenerate plantlets.Protoplasts were isolated from etiolated cotyledon material, and, although callus was readily obtained, plantlet regeneration was not observed using numerous hormone regimes.Abbreviations M
Mannitol
- 2,4-D
Dichlorophenoxyacetic acid
- NAA
Naphthalene-acetic acid
- BAP
Benzylaminopurine
- MS medium
Murashige and Skoog (1962) medium
- UM medium
Uchimiya and Murashige (1974) medium
- COT
cotyledon
- SH
shoot
- R
root 相似文献
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Julie Samson Éric Langlois Jiyu Lei Yves Piché Robert Chênevert 《Biotechnology letters》1998,20(4):355-358
At an initial concentration of 50 mg/l in liquid culture, 2,4,6-trinitrotoluene (TNT) was totally transformed by the three following fungi: Ceratocystis coerulescens, Lentinus lepideus and Trichoderma harzianum. 2,4-Dinitrotoluene (DNT) was also degraded by C. coerulescens and T. harzianum but not by L. lepideus. Composition of the culture medium in terms of carbon and nitrogen affected the transformation of TNT and DNT into amino-intermediates. These metabolites accounted for less than one quarter of the initial concentration of aromatics. In some instances, these transient intermediates disappeared at the end of the incubation period. © Rapid Science Ltd. 1998 相似文献
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An efficient method to produce water-soluble polysaccharides from Lentinus lepideus is described. The productivity of both endopolysaccharides (PPS) and exopolysaccharides (EPS) was compared under various culture conditions. The effect of treating their own PPS and EPS on immune cytokine production was also studied in relation to culture factors. High yield production of EPS required moderate culture temperature (25 degrees ) as well as long culture period (16-20 days). In contrast, PPS production required high culture temperature (30 degrees ) and short culture period ( days). Most of the carbon sources did not affect polysaccharides and mycelial production except for sucrose. Immune cytokine levels in the EPS treatment varied among carbon sources or culture periods. PPS did not appear to affect much on the production of cytokines, regardless of the culturing factors, except for the culture period. These results suggest that the optimal culture conditions for L. lepideus vary according to culture purposes, and different culture conditions should be used for different targets including mycelial biomass, EPS, and PPS. Whereas the immunomodulating activity of EPS appeared to be affected by culture conditions in L. lepideus, that of PPS did not. 相似文献
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Summary Regeneration of Beauveria bassiana GK2016 protoplasts demonstrated three phases: (1) enlargement, (2) formation of a chain of budding cells, and (3) development of a true germ tube from the original protoplast. Regeneration frequencies of up to 80% were obtained when using the stabilizer ammonium sulphate. Using electroporation, protoplasts were transformed to methyl 1,2-benzimidazole carbamate (MBC) resistance with the Neurospora crassa MBC-resistant -tubulin gene. A transformation efficiency of one of three transformants per 5 g vector DNA was obtained. The MBC phenotype was stable and transformants grew in the presence of 5 g MBC/ml. Southern DNA-DNA hybridization analysis demonstrated that integration of the vector into the chromosomal DNA had occurred.
Correspondence to: G. G. Khachatourians 相似文献
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Studies on stress ethylene and ethane during protoplast isolation from water-stressed and waterlogged donor plants Nicotiana tabacum L. xanthi-nc, show a correlation between ethane, but not ethylene, release and protoplast survival in vitro. Ethane release shows a high negative correlation with protoplast survival potential from donor plants subjected to both stresses. Ethylene showed a high negative correlation with protoplast survival potential in tissues from water-stressed but not from long-term waterlogged plants. The absence of correlation in the latter may be related to decreased ability to produce ethylene in hyperstressed plants.
The results are discussed in relation to the use of stress ethane release as a parameter of the physiological status of the plant. 相似文献
The results are discussed in relation to the use of stress ethane release as a parameter of the physiological status of the plant. 相似文献
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Protoplast formation, stabilization and regeneration was improved for 4 strains (erythromycin producers) of Saccharopolyspora erythraea. A modified medium was developed for protoplast regeneration. Parental and protoplast-regenerated strains exhibited changes in morphology, ultrastructure, and antibiotic production. 相似文献
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Summary Treatment ofBacillus thuringiensis andAgrobacterium tumefaciens taken from the early growth phase (8 h) with lysozyme at 1 mg/ml gave 90–99% protoplast formation and 10–12% protoplast regeneration on the minimal medium in absence of plasma expander (Bovine serum albumin). Enhanced fusion frequency was obtained when protoplasts from 8 h grown cells were used for fusion experiments. 相似文献
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A method for the isolation of plant protoplasts is presented that uses inexpensive sources of enzymes. It is suitable for student use in Biotechnology. 相似文献
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Efficient protoplast regeneration for some homofermentative lactobacilli and pediococci 总被引:2,自引:0,他引:2
Conditions for protoplast regeneration were examined for several strains of homofermentative lactobacilli and pediococci isolated from silage. Attempts to regenerate protoplasts using previously published agar regeneration media for lactobacilli were unsuccessful for most of the strains. Replacing or increasing colloidal substances in a medium containing raffinose and MgCl(2) as osmotic stabilizers enabled efficient regeneration of the protoplasts at a frequency of 10-99%. A medium containing gelatin, polyvinylpyrrolidone (PVP) and no agar was effective for Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus rhamnosus protoplasts. An agar medium containing PVP (PVP medium) was effective for Pediococcus sp. protoplasts, and addition of agarose to the PVP medium enabled regeneration of Lactobacillus casei protoplasts. A medium containing calcium alginate gel and no agar was effective for Lactobacillus curvatus protoplasts. The type of colloidal substance required for protoplast regeneration varied from species to species. This result suggested that several kinds of media may be necessary to regenerate protoplasts for all the genera of lactobacilli and pediococci. 相似文献
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M. C. Dornelas F. C. A. Tavares J. C. de Oliviera M. L. C. Vieira 《Plant cell reports》1995,15(1-2):106-110
Summary Protoplasts were isolated from leaf explants ofPassiflora edulis var.flavicarpa (the yellow passion fruit) and from cell suspensions of fivePassiflora species. Chemical fusion was performed using polyethylene glycol and the microcolonies obtained were transferred to growth medium to produce calli. Electrophoresis of soluble proteins and analysis of isoenzymes from calli produced from the fusion experiments were performed to select somatic hybrids. Specific polypeptide bands allowed the identification of somatic hybrids betweenP. edulis var.flavicarpa (+)P. alata, P. edulis var.flavicarpa (+)P. amethystina, P. edulis var.flavicarpa (+)P. cincinnata, P. edulis var.flavicarpa (+)P. giberti andP. edulis var.flavicarpa (+)P. coccinea. An average of 3 to 5% hybrid calli were obtained. With the exception of theP. edulis var.flavicarpa (+)P. coccinea, whole plants were recovered from all hybrids. These somatic hybrids showed 4n=36 chromosomes, which represents a further evidence of their hybridity. 相似文献
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M. Victoria Elorza Hortensia Rico D. Gozalbo R. Sentandreu 《Antonie van Leeuwenhoek》1983,49(4-5):457-469
The transition of blastospores to the mycelial phase in Candida albicans was induced after the blastospores were kept at 4 degrees C for several hours and then transferred to a fresh medium prewarmed at 37 degrees C. Glucan was the most abundant polymer in the wall in the two morphogenetic forms but the amount of chitin was higher in the mycelial form than in blastospores. Efficient protoplasting required reducing agents and proteases together with beta-glucanases (zymolyase). Protein synthesis in regenerating protoplasts was initiated after about 30 min. Chitin synthetase, initially very low, was incorporated in important amounts into cell membranes mainly in a zymogenic state. After a few hours chitin was the most abundant polymer found in the aberrant wall of the regenerating protoplast. 相似文献
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Paul F. Hamlyn Rosemary E. Bradshaw Fiona M. Mellon Claro M. Santiago Jane M. Wilson John F. Peberdy 《Enzyme and microbial technology》1981,3(4):321-325
Several commercial polysaccharases have been compared for their ability to liberate protoplasts from fungi. These enzymes were found to contain side activities capable of hydrolysing fungal cell walls. Protoplasts have been commonly isolated from fungi using enzyme systems prepared by workers in their own laboratories. However, these procedures are time consuming and considerable variation may be found between different batches of enzyme. The present study shows that high yields of protoplasts can be prepared from a variety of fungi using relatively cheap commercial enzymes. The yields obtained were normally as good as or better than those previously produced. 相似文献
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The methods for preparation and regeneration of protoplasts were tested with respect to the strains of F. coccineum markedly differing in their capacity for antibiotic production, sporulation and the growth rate. It was found that the substrate used for the culture growth had a significant effect on the cell wall and sensitivity of the mycelium to lytic enzymes. An enzyme from Hellix pomatia and its combination with lysozyme were used for lysing the culture. The cytological investigation of the time course of the culture lysis revealed a stage-by-stage pattern of protoplast formation by means of fragmentation of the hyphal contents till a ball was formed. Two to 4 protoplasts differing in their size and structure were formed within a cell. The pH value and osmotically stabilizing component had some effect on the rate of protoplast formation. Highly productive strains were characterized by formation of protoplasts heterogenous in their size and by decreased frequency of regenerations. The enzyme-free protoplasts preserved their viability and capacity for germination in osmotically stabilizing media for 72-96 hours of storage at 4 degrees C. On solid media the regeneration frequency reached 38 per cent. The regenerated cells formed colonies morphologically similar to those of the intact culture. 相似文献