Isolation and characterization of a mycovirus from Plasmopara halstedii

1992 ◽  
Vol 70 (9) ◽  
pp. 1734-1737 ◽  
Author(s):  
D. E. Mayhew ◽  
A. L. Cook ◽  
T. J. Gulya
Keyword(s):  
Molecular Weight ◽  
Key Words ◽  
Helianthus Annuus ◽  
Downy Mildew ◽  
Type A ◽  
Uv Absorbance ◽  
Plasmopara Halstedii ◽  
Isolation And Characterization ◽  
Virion Surface

A virus was isolated from Plasmopara halstedii, the causal agent of sunflower downy mildew. The virus contained two species of ssRNA with molecular weight (MW) of 1.05 × 106 and 0.53 × 106. One polypeptide with a MW of 37.5 × 103 was associated with whole virus. Purified preparations consisted of two components, the top component consisting primarily of empty shells. The A260/A280 UV absorbance ratio for the bottom component ranged from 1.25 to 1.35. The partially purified virions were isometric, approximately 32 nm in diameter, and had projections of approximately 4 nm on the virion surface. This virus may be related to the type A virus in Sclerophthora macrospora. Key words: mycovirus, downy mildew, Helianthus annuus.

scholarly journals BIOCHEMICAL AND HISTOLOGICAL CHANGES ASSOCIATED WITH DOWNY MILDEW (Plasmopara halstedii (Farl.) Berl. and de Toni) INFECTION IN SUNFLOWER (Helianthus annuus L.) / MODIFICACIONES BIOQUIMICAS E HISTOLOGICAS RELACIONADAS CON LA INFECCION DEL GIRASOL (Helianthus annuus L.) POR EL AGENTE DE MILDIU (Plasmopara halstedii (Farl.) Berl. y de Toni) / CHANGEMENTS BIOCHIMIQUES ET HISTOLOGIQUES ASSOCIÉS À L’INFECTION DU TOURNESOL (Helianthus annuus L.) PAR LE MILDIOU (Plasmopara halstedii (Farl.) Berl. et de Toni)

Helia ◽  
2000 ◽  
Vol 23 (33) ◽  
pp. 1-18
Author(s):  
B.R. Prashanth Kumar ◽  
Mahesh J. Kulkarni ◽  
B.N. Veena Rao ◽  
K. Chandrika ◽  
V.R. Balakrishna Gowda ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Helianthus Annuus ◽  
High Molecular Weight ◽  
Downy Mildew ◽  
Soluble Sugars ◽  
Molecular Weight Protein ◽  
High Molecular Weight Protein ◽  
Plasmopara Halstedii ◽  
Helianthus Annuus L ◽  
Weight Protein

SUMMARY Biochemical changes associated with downy mildew infection in sunflower revealed an increase in the total soluble proteins, 0-40% ammonium sulfate fractionated proteins and total soluble sugars. Molecular sieve chromatography of 0-40% ammonium sulphate fraction revealed for presence of a high molecular weight protein and polysaccharide in the downy mildew infected sunflower leaves. Antibodies raised against the high molecular weight protein and polysaccharide were used in the western blot/dot blot analysis. It has been shown that the downy mildew disease also induces PR proteins, which have antigenic homology with PR-S protein, a member of the PR-5 class of proteins. An increase in iPA (isopentenyl adenosine) content in the infected sample was observed. A positive correlation exists between iPA level, 0-40% protein and total sugar content. Histological studies revealed that the downy mildew fungal mycelium extensively proliferates, ramifies extensively the leaf tissue and forms a nutritional link with the host cell by producing the intracellular haustorium.

Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

1974 ◽  
Vol 31 (01) ◽  
pp. 072-085 ◽  
Author(s):  
M Kopitar ◽  
M Stegnar ◽  
B Accetto ◽  
D Lebez
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Isolation And Characterization ◽  
Ph Range ◽  
Inhibition Studies ◽  
Final Purification

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

Preparation and Characterization of the High Molecular Weight [3H]Hyaluronic Acid

1992 ◽  
Vol 57 (10) ◽  
pp. 2151-2156 ◽  
Author(s):  
Peter Chabreček ◽  
Ladislav Šoltés ◽  
Hynek Hradec ◽  
Jiří Filip ◽  
Eduard Orviský
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Methyl Bromide ◽  
High Performance ◽  
Hydrogen Atoms ◽  
Isolation And Characterization ◽  
Labelling Procedure ◽  
Enzymatic Depolymerization

Two methods for the preparation of high molecular weight [3H]hyaluronic acid were investigated. In the first one, hydrogen atoms in the molecule were replaced by tritium. This isotopic substitution was performed in aqueous solution using Pd/CaCO3 as the catalyst. In the second method, the high molecular weight hyaluronic acid was alkylated with [3H]methyl bromide in liquid ammonia at a temperature of -33.5 °C. High-performance gel permeation chromatographic separation method was used for the isolation and characterization of the high molecular weight [3H]hyaluronic acid. Molecular weight parameters for the labelled biopolymers were Mw = 128 kDa, Mw/Mn = 1.88 (first method) and Mw = 268 kDa, Mw/Mn = 1.55 (second method). The high molecular weight [3H]hyaluronic acid having Mw = 268 kDa was degraded further by specific hyaluronidase. Products of the enzymatic depolymerization were observed to be identical for both, labelled and cold biopolymer. This finding indicates that the described labelling procedure using [3H]methyl bromide does not induce any major structural rearrangements in the molecule.

scholarly journals Updated Characterization of Races of Plasmopara halstedii and Entomopathogenic Fungi as Endophytes of Sunflower Plants in Axenic Culture

Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 268
Author(s):  
Pedro Miranda-Fuentes ◽  
Ana B. García-Carneros ◽  
Leire Molinero-Ruiz
Keyword(s):  
Plant Growth ◽  
Downy Mildew ◽  
Entomopathogenic Fungi ◽  
Genetic Resistance ◽  
Axenic Culture ◽  
Plasmopara Halstedii ◽  
Successful Establishment ◽  
Axenic Conditions ◽  
Highly Virulent

The management of downy mildew (Plasmopara halstedii) in sunflower, is heavily dependent on genetic resistance, whilst entomopathogenic fungi (EF) can reduce other sunflower diseases. In this work, we characterized P. halstedii from Spain and other countries collected in the past few years. Twenty-three races were identified (the most frequent in Spain being 310, 304, 705 and 715), with an increasing proportion of highly virulent races. Five isolates from countries other than Spain overcame the resistance in RHA-340. In addition, we assessed the efficacy of five EF against downy mildew and their effects on sunflower growth in axenic conditions. None of the entomopathogens reduced disease severity, nor did they have any effect on plant growth when applied together with P. halstedii. In contrast, three EF reduced some of the plant growth variables in the absence of the pathogen. Microbiological and molecular diagnostics suggest that the axenic system and the short experimental time used in this study did not favor the successful establishment of EF in the plants or their potential biocontrol effect. Our results show a shift in P. halstedii racial patterns and suggest that soil as a growth substrate and long infection times are needed for EF effectiveness against downy mildew.

Isolation and Characterization of Undenatured Chlorogenic Acid Free Sunflower (Helianthus annuus) Proteins

2002 ◽  
Vol 50 (6) ◽  
pp. 1713-1719 ◽  
Author(s):  
Sergio González-Pérez ◽  
Karin B. Merck ◽  
Johan M. Vereijken ◽  
Gerrit A. van Koningsveld ◽  
Harry Gruppen ◽  
...  
Keyword(s):  
Chlorogenic Acid ◽  
Helianthus Annuus ◽  
Isolation And Characterization

scholarly journals Characterization of a Novel Fructosyltransferase from Lactobacillus reuteri That Synthesizes High-Molecular-Weight Inulin and Inulin Oligosaccharides

2002 ◽  
Vol 68 (9) ◽  
pp. 4390-4398 ◽  
Author(s):  
S. A. F. T. van Hijum ◽  
G. H. van Geel-Schutten ◽  
H. Rahaoui ◽  
M. J. E. C. van der Maarel ◽  
L. Dijkhuizen
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Lactobacillus Reuteri ◽  
Bacterial Strains ◽  
Potential Health ◽  
Isolation And Characterization ◽  
A Cell ◽  
Encoding Gene ◽  
First Time

ABSTRACT Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli. When incubated with sucrose, the purified recombinant FTF enzyme produced large amounts of fructo-oligosaccharides (FOS) with β-(2→1)-linked fructosyl units, plus a high-molecular-weight fructan polymer (>107) with β-(2→1) linkages (an inulin). FOS, but not inulin, was found in supernatants of L. reuteri strain 121 cultures grown on medium containing sucrose. Bacterial inulin production has been reported for only Streptococcus mutans strains. FOS production has been reported for a few bacterial strains. This paper reports the first-time isolation and molecular characterization of (i) a Lactobacillus ftf gene, (ii) an inulosucrase associated with a generally regarded as safe bacterium, (iii) an FTF enzyme synthesizing both a high molecular weight inulin and FOS, and (iv) an FTF protein containing a cell wall-anchoring LPXTG motif. The biological relevance and potential health benefits of an inulosucrase associated with an L. reuteri strain remain to be established.

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