scholarly journals Effects of Culture Media on Phytophthora palmivora Growth, α-elicitin Production and Toxicity to Dendrobium

2018 ◽  
Vol 46 (2) ◽  
pp. 630-638 ◽  
Author(s):  
Apinya KHAIRUM ◽  
Oythip POOLSAWAT ◽  
Paniti PORNBUNGKERD ◽  
Akkawat THARAPREUKSAPONG ◽  
Sopone WONGKAEW ◽  
...  
Keyword(s):  
In Vitro Selection ◽  
Culture Media ◽  
Protein Band ◽  
Phytophthora Palmivora ◽  
Black Rot ◽  
Potato Dextrose Broth ◽  
Phytophthora Spp ◽  
Sds Page ◽  
Identical Amino Acid Sequence

Four culture media were evaluated for their ability to induce Phytophthora palmivora growth and produce culture filtrate (CF), and to determine a CF concentration and culture period effective for in vitro screening of black rot resistance in Dendrobium cv. ‘Earsakul’. Mycelial fresh weights of P. palmivora cultured in potato dextrose broth (PDB; the most commonly used medium for fungi), pea sucrose broth (PSB; a medium frequently used for Phytophthora spp.), and Murashige and Skoog broth (MSB; the most popular plant tissue culture medium) were found to be significantly higher than that in the newly developed modified oat meal broth (MOMB). When the total proteins of CFs were analysed with SDS-PAGE, a protein band of 10.5 kDa MW was found in CFs from all media with the highest level in PSB. LC-MS/MS analysis identified this protein as α-elicitin that had an identical amino acid sequence to the α-elicitin hibernalin of P. hibernalis and syringicin from P. syringae. The optimum conditions for in vitro selection of Dendrobium for black rot resistance using α-elicitin-containing CFs were also determined by evaluating the CF toxicity on Dendrobium protocorm-like bodies (PLBs) when cultured in all media supplemented with 0, 30, 50 and 100% CFs for seven, 14 and 21 d. The levels of PLB necrosis varied according to medium types, CF concentrations and culture periods. The maximum percentage of PLB necrosis (100%) was obtained in PSB supplemented with 50 and 100% CFs, and the severity of PLB necrosis was highest when treated with 100% CF for 14 and 21 d.

Evaluation of drought resistance-related traits in durum wheat somaclonal lines selected in vitro

2004 ◽  
Vol 44 (1) ◽  
pp. 27 ◽  
Author(s):  
M. Bajji ◽  
P. Bertin ◽  
S. Lutts ◽  
J-M. Kinet
Keyword(s):  
Durum Wheat ◽  
Somaclonal Variation ◽  
Drought Resistance ◽  
In Vitro Selection ◽  
Culture Media ◽  
Field Studies ◽  
Resistance Mechanisms ◽  
Wide Range ◽  
Triticum Durum Desf

Somaclonal variation associated with in vitro selection has been used as a source of variability to improve drought resistance of 3 durum wheat (Triticum durum Desf.) cultivars (Selbera, Sebou, and Kyperounda). In a previous study, R0 plants with improved drought resistance-related characters were regenerated after selection on culture media containing polyethylene glycol (PEG). This improvement was transmitted to the R1 progeny. The present study analysed the behaviour of the selected tissue culture-derived lines in subsequent R2, R3 and R4�generations. Differences in electrolyte leakage, chlorophyll fluorescence (Fv/Fm), stomatal conductance and days to heading were found between the parental cultivars and most of their in vitro-derived lines. The changes may differ from one cultivar to another. Many promising somaclonal lines still presented improvement for at least 3 of the 4�parameters measured comparatively to initial cultivars. Somaclonal variation thus appears to induce a wide range of modifications among individual components of drought-resistance mechanisms. These improved traits could be valuable if shown to be inherited and to give enhanced agronomic performances in future field studies.

Identification of thecrygene inBacillus thuringiensisstrain WZ-9 and its toxicity againstHenosepilachna vigintioctomaculata

2008 ◽  
Vol 5 (3) ◽  
pp. 245-250 ◽  
Author(s):  
Song Ping ◽  
Wang Qin-Ying ◽  
Wu Hui-Xian ◽  
Lu Xiu-Jun ◽  
Wang Yong
Keyword(s):  
Amino Acid ◽  
Culture Media ◽  
Stationary Phases ◽  
Protein Band ◽  
Growth Cycle ◽  
Amino Acid Residues ◽  
Reading Frame ◽  
Gene Sequence Analysis ◽  
Sds Page ◽  
Encoding Gene

AbstractBacillus thuringiensisstrain WZ-9, isolated from soil in Hebei province, China, was effective againstHenosepilachna vigintioctomaculatalarvae. The strain presented bipyramidal crystals with a protein band of 130 kDa in SDS–PAGE. The pH changes of the culture media showed important fluctuations during the 24 h growth cycle. The pH varied less in log and stationary phases than it did in the exponential phase. Bioassay results showed that the WZ-9 strain was only harmful to larvae ofH. vigintioctomaculataand not to either adults ofH. vigintioctomaculataor other several lepidopteran and coleopteran insects. LC50to second-instar larvae ofH. vigintioctomaculatawas 2.95×107cells/ml after 72 h. Genotypic investigations showed that this strain possessed thecry7gene. Sequence analysis demonstrated that the encoding gene contained an open reading frame (ORF) of 3414 bp and encoded 1138 amino acid residues. The deduced amino acid sequence was 99.65% identical to that of the reported Cry7Ab2 sequences. This gene was designated by the Bt δ-endotoxin nomenclature committee as Cry7Ab3 with accession number BI 1015188 in the GenBank database.

scholarly journals Comparison of Four Systems to Test the Tolerance of ‘Fortune’ Mandarin Tissue Cultured Plants to Alternaria alternata

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1321
Author(s):  
Margarita Pérez-Jiménez ◽  
Olaya Pérez-Tornero
Keyword(s):  
Culture Filtrate ◽  
Alternaria Alternata ◽  
In Vitro Selection ◽  
Culture Media ◽  
Severe Disease ◽  
Mutation Breeding ◽  
Brown Spot ◽  
Fungus Culture ◽  
Alternaria Brown Spot

Alternaria brown spot is a severe disease that affects leaves and fruits on susceptible mandarin and mandarin-like cultivars, and is produced by Alternaria alternata. Consequently, there is an urge to obtain new cultivars resistant to A. alternata, and mutation breeding together with tissue culture can help shorten the process. However, a protocol for the in vitro selection of resistant citrus genotypes is lacking. In this study, four methods to evaluate the sensitivity to Alternaria of mandarin ‘Fortune’ explants in in vitro culture were tested. The four tested systems consisted of: (1) the addition of the mycotoxin, produced by A. alternata in ‘Fortune’, to the propagation culture media, (2) the addition of the A. alternata culture filtrate to the propagation culture media, (3) the application of the mycotoxin to the intact shoot leaves, and (4) the application of the mycotoxin to the previously excised and wounded leaves. After analyzing the results, only the addition of the A. alternata culture filtrate to the culture media and the application of the mycotoxin to the wounded leaves produced symptoms of infection. However, the addition of the fungus culture filtrate to the culture media produced results, which might indicate that, in addition to the mycotoxin, many other unknown elements that can affect the plant growth and behavior could be found in the fungus culture filtrate. Therefore, the application of the toxin to the excised and wounded leaves seems to be the most reliable method to analyze sensitivity to Alternaria of ‘Fortune’ explants cultured in vitro.

Molecular characterization of Dendrobium ‘Earsakul’ mutants from in vitro selection for black rot resistance

2017 ◽  
Vol 19 (02) ◽  
pp. 130-134
Author(s):  
T.P. Tantasawat ◽  
A. Khairum ◽  
A. Tharapreuksapong ◽  
O. Poolsawat ◽  
P.A. Tantasawat
Keyword(s):  
Molecular Characterization ◽  
In Vitro Selection ◽  
Black Rot ◽  
Selection For

scholarly journals Biotechnology of biomass production in vitro of fungi isolated from Pinus

2020 ◽  
Vol 9 (10) ◽  
pp. e7809109080
Author(s):  
Paula Francislaine Moura ◽  
Celso Garcia Auer ◽  
Katlin Suellen Rech ◽  
Camila Freitas de Oliveira ◽  
Cristiane da Silva Paula de Oliveira ◽  
...  
Keyword(s):  
Biomass Production ◽  
Large Scale ◽  
Culture Media ◽  
In Vitro Cultivation ◽  
Malt Extract ◽  
Scale Production ◽  
Potato Dextrose Broth ◽  
Mechanical Stirring ◽  
Large Scale Production

Fungi are organisms capable of synthesizing metabolites of industrial interest and the standardization of biomass production for the extraction of these compounds has biotechnological applications. The objective of this work was to optimize the in vitro cultivation process for fungi isolated from Pinus sp., standardizing the best conditions for the production of biomass, contributing to its large scale production. Therefore, the conditions of in vitro cultivation of the fungi Botrytis cinerea, Rhizoctonia sp. and Suillus sp., were evaluated based on the maximum production of dry biomass (PBS), varying temperature, medium and cultivation time. The fungi were grown in glass flasks with liquid culture media, in a BOD chamber, without mechanical stirring. Potato-dextrose broth - PD broth (PD), Czapek - CZ broth (CZ) and Malt Extract - EM broth (EM) were evaluated at temperatures ranging from 8 to 32 ºC and incubation times from 7 to 35 days. PD broth showed better results for fungi B.cinerea and Rhizoctonia sp., when compared to CZ and EM broths, in PBS, while Suillus sp. showed better development in EM broth. The best growth temperature based on PBS was 12 ºC and 16 ºC, with 28 and 35 days of cultivation.

scholarly journals In vitro expression demonstrates impaired secretion of the γAsn319, Asp320 deletion variant fibrinogen

2005 ◽  
Vol 94 (07) ◽  
pp. 53-59 ◽  
Author(s):  
Satomi Kani ◽  
Fumiko Terasawa ◽  
Susan T. Lord ◽  
Minoru Tozuka ◽  
Hiroyoshi Ota ◽  
...  
Keyword(s):  
Culture Media ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Fibrinogen Concentration ◽  
Ovary Cells ◽  
Cell Lysates ◽  
Sds Page ◽  
The Media ◽  
Kinetics Of

SummaryThe hypodysfibrinogenemia Otsu is caused by the two-residue deletion,γAsn319 and γAsp320. Analysis of plasma or purified fibrinogen from the heterozygous propositus revealed that the amount of variant γ-chain was lower than that of normal γ-chain. In order to examine the basis for this difference, we transfected Chinese hamster ovary cells and established stable cell lines that expressed both chains, γΔ/γN, only the normal chain, γN, and only the variant chain, γΔ. We measured fibrinogen concentration of confluent cultures by ELISA. We found the ratios of the concentrations in the media to the concentrations in the cell lysates of γΔ, γΔ/γN, and γN-cells were 0.42, 0.60, and 1.00, respectively. We measured the concentrations of the γΔ and γN chains by densitometric analysis of samples following separation by SDS-PAGE and found the fraction of γΔ-chains in cell lysates was always greater than the fraction in the respective culture media. We examined the kinetics of fibrinogen synthesis, assembly and secretion in pulse-chase experiments, and found that the γΔ-chain was assembled into intact fibrinogen at a rate similar to assembly of the γN-chain into normal fibrinogen, but was secreted into the medium at a slightly slower rate than normal fibrinogen. Considered together, these experiments indicate secretion of the variant fibrinogen was slightly impaired. These results suggest that the reduced level of γΔ319,320 fibrinogen in the plasma of the Otsu patient arises from modestly impaired secretion of this variant fibrinogen.

Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

1994 ◽  
Vol 52 ◽  
pp. 270-271
Author(s):  
Henry H. Eichelberger ◽  
John G. Baust ◽  
Robert G. Van Buskirk
Keyword(s):  
Cold Storage ◽  
Structural Integrity ◽  
Culture Media ◽  
Cell Structure ◽  
Natural State ◽  
Sodium Cacodylate ◽  
Ruthenium Tetroxide ◽  
Cacodylate Buffer ◽  
Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

The Anticoagulant Properties of a Modified Form of Protein S

1988 ◽  
Vol 60 (02) ◽  
pp. 298-304 ◽  
Author(s):  
C A Mitchell ◽  
S M Kelemen ◽  
H H Salem
Keyword(s):  
Monoclonal Antibody ◽  
Anticoagulant Activity ◽  
Activated Protein C ◽  
Factor Xa ◽  
Protein S ◽  
Human Neutrophil Elastase ◽  
Factor X ◽  
Sds Page

SummaryProtein S (PS) is a vitamin K-dependent anticoagulant that acts as a cofactor to activated protein C (APC). To date PS has not been shown to possess anticoagulant activity in the absence of APC.In this study, we have developed monoclonal antibody to protein S and used to purify the protein to homogeneity from plasma. Affinity purified protein S (PSM), although identical to the conventionally purified protein as judged by SDS-PAGE, had significant anticoagulant activity in the absence of APC when measured in a factor Xa recalcification time. Using SDS-PAGE we have demonstrated that prothrombin cleavage by factor X awas inhibited in the presence of PSM. Kinetic analysis of the reaction revealed that PSM competitively inhibited factor X amediated cleavage of prothrombin. PS preincubated with the monoclonal antibody, acquired similar anticoagulant properties. These results suggest that the interaction of the monoclonal antibody with PS results in an alteration in the protein exposing sites that mediate the observed anticoagulant effect. Support that the protein was altered was derived from the observation that PSM was eight fold more sensitive to cleavage by thrombin and human neutrophil elastase than conventionally purified protein S.These observations suggest that PS can be modified in vitro to a protein with APC-independent anticoagulant activity and raise the possibility that a similar alteration could occur in vivo through the binding protein S to a cellular or plasma protein.

In Vitro Stability of a Tissue-Type Plasminogen Activator Mutant, BM 06.022, in Human Plasma

1994 ◽  
Vol 72 (06) ◽  
pp. 906-911 ◽  
Author(s):  
D C Rijken ◽  
E Groeneveld ◽  
M M Barrett-Bergshoeff
Keyword(s):  
Plasminogen Activator ◽  
Half Life ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tissue Type ◽  
Single Chain ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator ◽  
Sds Page ◽  
Chain Form

SummaryBM 06.022 is a non-glycosylated mutant of human tissue-type plasminogen activator (t-PA) comprising only the kringle-2 and proteinase domains. The in vivo half-life of BM 06.022 antigen is 4- to 5-fold longer than that of t-PA antigen. The in vitro half-life of the activity of BM 06.022 at therapeutic concentrations in plasma is shorter than that of t-PA. In this study the inactivation of BM 06.022 in plasma was further investigated.Varying concentrations of BM 06.022 were incubated in plasma for 0-150 min. Activity assays on serial samples showed a dose-dependent decline of BM 06.022 activity with a half-life from 72 min at 0.3 μg/ml to 38 min at 10 μg/ml. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by fibrin autography showed the generation of several BM 06.022-complexes. These complexes could be completely precipitated with antibodies against Cl-inactivator, α2-antiplasmin and α1-antitrypsin.During the incubation of BM 06.022 in plasma, plasmin was generated dose-dependently as revealed by varying degrees of a2-anti-plasmin consumption and fibrinogen degradation. SDS-PAGE and immunoblotting showed that single-chain BM 06.022 was rapidly (i. e. within 45 min) converted into its two-chain form at concentrations of 5 μg/ml BM 06.022 and higher.In conclusion, BM 06.022 at therapeutic concentrations in plasma was inactivated by Cl-inactivator, a2-antiplasmin and a j-antitrypsin. The half-life of the activity decreased at increasing BM 06.022 concentrations, probably as a result of the generation of two-chain BM 06.022 which may be inactivated faster than the single-chain form.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

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