scholarly journals First Report of Damping-Off of Durum Wheat Caused by Arthrinium sacchari in the Semi-Arid Saskatchewan Fields

Plant Disease ◽  
2007 ◽  
Vol 91 (4) ◽  
pp. 469-469 ◽  
Author(s):  
D. C. Mavragani ◽  
L. Abdellatif ◽  
B. McConkey ◽  
C. Hamel ◽  
V. Vujanovic
Keyword(s):  
Durum Wheat ◽  
Search Algorithm ◽  
Plant Biotechnology ◽  
In Vitro Assay ◽  
Damping Off ◽  
Fusarium Spp ◽  
Vitro Assay ◽  
First Report ◽  
Semi Arid

Durum wheat (Triticum durum Desf.) is an important crop in western Canada. In 2005, Arthrinium sacchari was frequently isolated from soil and durum wheat plants of the semi-arid fields of Swift Current, Saskatchewan, Canada (50°16′N, 107°44′W). The susceptibility of durum wheat to damping-off caused by this fungus was evaluated. To our knowledge, this is the first report of A. sacchari in North America (1) and the first mention of its association with durum wheat. DNA was extracted (MoBio Isolation Kit, Carlsbad, CA) from 2-week-old A. sacchari isolates (FBC.3, FBC.45, and FBC.143) grown on PDA. The internal transcribed spacer (ITS) of the rDNA was amplified from each isolate and sequenced (Plant Biotechnology Institute, Saskatoon, SK, Canada) and similarity analyses were performed using the BLAST search algorithm in GenBank. All three sequences (Accession Nos. EF076710, EF076711, and EF076712A) showed 99% similarity with A. sacchari (Accession No. AF393679). An in vitro assay was performed by placing 1-cm2 agar plugs containing mycelia of A. sacchari (FBC.3, FBC.45, and FBC.143) onto surface-sterilized durum. Surface-sterilized seeds inoculated in the same way with Fusarium graminearum or F. avenaceum were used as negative controls, and noninoculated surface-sterilized seeds were used as a positive control. A second in vitro assay involved inoculating the same isolates onto seeds placed in sterilized sandy soil. In both assays, 10 seeds per petri plate and three plates per treatment were used and plates were incubated at 21°C for 1 week in darkness. All experiments were performed twice. On PDA, preemergence damping-off was found in 60% of A. sacchari FBC.3, 55% of A. sacchari FBC.45 and FBC.143, 50% of F. avenaceum, and 58% of F. graminearum inoculated seeds. In sterilized soil, the incidence of preemergence damping-off ranged from 43 to 30%. Subsequent incubation over a period of 3 weeks resulted in 100% postemergence damping-off in A. sacchari FBC.45 and FBC.3 as well as in both Fusarium spp. inoculated controls, 60% postemergence damping-off in A. sacchari FBC.143, and no damping-off in the noninoculated control. Arthrinium and Fusarium spp. were reisolated only from symptomatic plants, satisfying Koch's postulates. In conclusion, durum wheat is highly susceptible to damping-off caused by A. sacchari, showing characteristic dark brown or violet lesions in infected tissues. A. sacchari was previously reported to be present in South America and eastern Asia. In China, it is considered an important mycotoxigenic species (2). Thus, infection of durum wheat crops with A. sacchari could pose a significant threat to North American wheat production. References: (1) D. F. Farr et al. Fungal Databases. Systematic Botany and Mycology Laboratory. Online publication. ARS, USDA, 2006. (2) X. J. Liu et al. Acta Mycol. Sinica 7:221, 1988.

scholarly journals The Effect of Various pH Medium on the Secondary Metabollites Production from Trichoderma harzianum T10 to Control Damping Off on Cucumber Seedlings

2020 ◽  
Vol 3 (2) ◽  
pp. 65
Author(s):  
Nur Chalimah ◽  
Loekas Soesanto ◽  
Woro Sri Suharti
Keyword(s):  
Secondary Metabolites ◽  
Trichoderma Harzianum ◽  
Disease Incidence ◽  
In Vitro Assay ◽  
Cucumber Seedling ◽  
Damping Off ◽  
In Planta ◽  
Vitro Assay ◽  
Cucumber Seedlings

Damping-off is one of the main diseases in cucumber seedlings caused by Pythium sp. Secondary metabolites of Trichoderma harzianum T10 can conduct the control of the disease. The pH of the medium influences the production of secondary metabolites. The research aimed to determine the effective pH medium on production of T. harzianum T10 secondary metabolites, and the effect of the T. harzianum T10 secondary metabolites application in damping-off disease control also to the growth of cucumber seedling. The research was consist of two steps; 1) in vitro assay with various pH levels 5; 3; 3.5; 4; 4.5; 5.5; 6; 6.5; and 7, 2) In planta treatments consisted of control, fungicide (Mancozeb), secondary metabolites in pH 5 and 5.5 with the concentration of 5, 10 and 15% each. The research showed that: 1) the effective pH medium for the production of T. harzianum T10 secondary metabolites was 5 and 5.5. 2) application of the T. harzianum T10 secondary metabolites on pH 5 and 5.5 with a concentration of 5, 10, and 15% could decrease the disease incidence and support cucumber seedling growth.

In Vitro Assay of Platelet Adhesiveness with Washed and Tanned Human Red Cells

1968 ◽  
Vol 20 (03/04) ◽  
pp. 384-396 ◽  
Author(s):  
G Zbinden ◽  
S Tomlin
Keyword(s):  
Platelet Adhesion ◽  
Sodium Citrate ◽  
Blood Platelets ◽  
In Vitro Assay ◽  
Red Cells ◽  
Platelet Adhesiveness ◽  
Vitro Assay ◽  
In Vitro System ◽  
Human Red Cells

SummaryAn in vitro system is described in which adhesion of blood platelets to washed and tannic acid-treated red cells was assayed quantitatively by microscopic observation. ADP, epinephrine and TAME produced a reversible increase in platelet adhesiveness which was antagonized by AMP. With Evans blue, polyanetholsulfonate, phthalanilide NSC 38280, thrombin and heparin at concentrations above 1-4 u/ml the increase was irreversible. The ADP-induced increase in adhesiveness was inhibited by sodium citrate, EDTA, AMP, ATP and N-ethylmaleimide. EDTA, AMP and the SH-blocker N-ethylmaleimide also reduced spontaneous platelet adhesion to red cells. No significant effects were observed with adenosine, phenprocoumon, 5-HT, phthalanilide NSC 57155, various estrogens, progestogens and fatty acids, acetylsalicylic acid and similarly acting agents, hydroxylamine, glucose and KCN. The method may be useful for the screening of thrombogenic and antithrombotic properties of drugs.

Angiogenic Effect of Pravastatin Alone and with Sera from Healthy and Complicated Pregnancies Studied by in vitro Vasculogenesis/Angiogenesis Assay

2021 ◽  
pp. 1-9
Author(s):  
Anita Virtanen ◽  
Outi Huttala ◽  
Kati Tihtonen ◽  
Tarja Toimela ◽  
Tuula Heinonen ◽  
...  
Keyword(s):  
Direct Effect ◽  
Late Onset ◽  
Maternal Serum ◽  
In Vitro Assay ◽  
Serum Samples ◽  
Therapeutic Concentration ◽  
Tubule Formation ◽  
Vitro Assay ◽  
Angiogenesis Assay

<b><i>Objective:</i></b> To determine the direct effect of pravastatin on angiogenesis and to study the interaction between pravastatin and maternal sera from women with early- or late-onset pre-eclampsia (PE), intrauterine growth restriction, or healthy pregnancy. <b><i>Methods:</i></b> We collected 5 maternal serum samples from each group. The effect of pravastatin on angiogenesis was assessed with and without maternal sera by quantifying tubule formation in a human-based in vitro assay. Pravastatin was added at 20, 1,000, and 8,000 ng/mL concentrations. Concentrations of angiogenic and inflammatory biomarkers in serum and in test medium after supplementation of serum alone and with pravastatin (1,000 ng/mL) were measured. <b><i>Results:</i></b> Therapeutic concentration of pravastatin (20 ng/mL) did not have significant direct effect on angiogenesis, but the highest concentrations inhibited angiogenesis. Pravastatin did not change the levels of biomarkers in the test media. There were no changes in angiogenesis when therapeutic dose of pravastatin was added with maternal sera, but there was a trend to wide individual variation towards enhanced angiogenesis, particularly in the early-onset PE group. <b><i>Conclusions:</i></b> At therapeutic concentration, pravastatin alone or with maternal sera has no significant effect on angiogenesis, but at high concentrations the effect seems to be anti-angiogenic estimated by in vitro assay.

In Vitro Assay for Single-cell Characterization of Impaired Deformability in Red Blood Cells under Recurrent Episodes of Hypoxia

Lab on a Chip ◽  
2021 ◽  
Author(s):  
YUHAO QIANG ◽  
Jia Liu ◽  
Ming Dao ◽  
E Du
Keyword(s):  
Shear Stress ◽  
Red Blood Cells ◽  
Single Cell ◽  
Blood Cells ◽  
Blood Circulation ◽  
In Vitro Assay ◽  
Vitro Assay ◽  
Cyclic Shear

Red blood cells (RBCs) are subjected to recurrent changes in shear stress and oxygen tension during blood circulation. The cyclic shear stress has been identified as an important factor that...

Purification of Chalcone Synthase from Tulip Anthers and Comparison with the Synthase from Cosmos Petals

1981 ◽  
Vol 36 (1-2) ◽  
pp. 30-34 ◽  
Author(s):  
Rainer Sütfeld ◽  
Rolf Wiermann
Keyword(s):  
Chalcone Synthase ◽  
In Vitro Assay ◽  
Flavonoid Biosynthesis ◽  
Chalcone Isomerase ◽  
Gel Chromatography ◽  
Purification Procedure ◽  
Reaction Products ◽  
Complete Elimination ◽  
Vitro Assay

Abstract Chalcone synthase was isolated from both anthers of Tulipa cv. “Apeldoorn” and petals of Cosmos sulphureus Cav. After certain prepurification steps, the enzymes were further purified using gel chromatography on Sephadex G-200 followed by repeated hydroxylapatite absorption chromatography. Both the enzymes showed the same chromatographic properties. After gel chromatography as well as after the first hydroxylapatite fractionation, the reaction products appeared as flavanones. However, after the second hydroxylapatite step, production of chalcones was observed. Like the enzyme from tulip anthers, the synthase from Cosmos petals produced the correspondingly substituted chalcones when p-coumaroyl-CoA, caffeoyl-CoA and feruloyl-CoA, respectively, were used as substractes. In both the cases, the ratios of the different chalcones produced were found to be about the same. The appearance of chalcone synthesis in this in vitro assay is caused by the complete elimination of chalcone isomerase in the purification procedure. The importance of the isomerase for flavonoid biosynthesis, particularly in plant systems which are accumulating chalcones, is discussed.

scholarly journals In Vitro Characterization of the Enzyme Properties of the Phospholipid N-Methyltransferase PmtA from Agrobacterium tumefaciens

2009 ◽  
Vol 191 (7) ◽  
pp. 2033-2041 ◽  
Author(s):  
Meriyem Aktas ◽  
Franz Narberhaus
Keyword(s):  
Agrobacterium Tumefaciens ◽  
In Vitro Assay ◽  
Enzyme Properties ◽  
Vitro Assay ◽  
Plant Infection ◽  
In Vitro Characterization ◽  
End Products ◽  
And Control

ABSTRACT Agrobacterium tumefaciens requires phosphatidylcholine (PC) in its membranes for plant infection. The phospholipid N-methyltransferase PmtA catalyzes all three transmethylation reactions of phosphatidylethanolamine (PE) to PC via the intermediates monomethylphosphatidylethanolamine (MMPE) and dimethylphosphatidylethanolamine (DMPE). The enzyme uses S-adenosylmethionine (SAM) as the methyl donor, converting it to S-adenosylhomocysteine (SAH). Little is known about the activity of bacterial Pmt enzymes, since PC biosynthesis in prokaryotes is rare. In this article, we present the purification and in vitro characterization of A. tumefaciens PmtA, which is a monomeric protein. It binds to PE, the intermediates MMPE and DMPE, the end product PC, and phosphatidylglycerol (PG) and phosphatidylinositol. Binding of the phospholipid substrates precedes binding of SAM. We used a coupled in vitro assay system to demonstrate the enzymatic activity of PmtA and to show that PmtA is inhibited by the end products PC and SAH and the antibiotic sinefungin. The presence of PG stimulates PmtA activity. Our study provides insights into the catalysis and control of a bacterial phospholipid N-methyltransferase.

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