Adaptive tolerance of Fusarium solani to benzimidazole derivatives in vitro

1973 ◽  
Vol 51 (10) ◽  
pp. 1725-1732 ◽  
Author(s):  
Lloyd T. Richardson
Keyword(s):  
Fusarium Solani ◽  
Fusarium Species ◽  
Benzimidazole Derivatives ◽  
Wild Type ◽  
Cross Tolerance ◽  
Single Exposure ◽  
Single Spore ◽  
Slow Growing ◽  
Dosage Response Curve

The growth response of Fusarium solani to benomyl or thiabendazole in the medium was found to differ from that of many other Fusarium species tested. In contrast to the steep, linear dosage–response curve typical for this genus, the curve for F. solani invariably reaches a maximum at about 1.0 μg/ml and is horizontal from there on. During a single exposure to either benomyl or thiabendazole, F. solani develops tolerance to both toxicants, but its subsequent growth on untreated medium is retarded. Cross-tolerance to other benzimidazoles is also induced. Such adapted strains generally maintain their slow growth habit and tolerance indefinitely through successive transfers to either untreated or treated medium. Occasionally a colony growing slowly on untreated medium will produce a sector which grows normally but proves to be sensitive when transferred to treated medium. Conversely, colonies of this sensitive strain, while growing extremely slowly on treated medium, more frequently produce a sector like the resistant strain. From observation of the response of a large number of single-spore isolates it is concluded that in the presence of a benzimidazole a high proportion of the wild population can mutate to the slow-growing, resistant type. Such mutants occasionally revert to the wild type.

scholarly journals EUCAST Determination of Olorofim (F901318) Susceptibility of Mold Species, Method Validation, and MICs

2018 ◽  
Vol 62 (8) ◽  
Author(s):  
Karin Meinike Jørgensen ◽  
Karen M. T. Astvad ◽  
Rasmus Krøger Hare ◽  
Maiken Cavling Arendrup
Keyword(s):  
Fusarium Solani ◽  
Susceptibility Testing ◽  
Wild Type ◽  
Preparation Methods ◽  
Content Type ◽  
Microtiter Plates ◽  
Upper Limits ◽  
Limited Variation

ABSTRACT Olorofim is a novel antifungal agent with in vitro activity against Aspergillus and some other molds. Here, we addressed technical aspects for EUCAST olorofim testing and generated contemporary MIC data. EUCAST E.Def 9.3.1 testing was performed comparing two plate preparation methods (serial dilution in medium [serial plates] versus predilution in DMSO [ISO plates]), two lots of olorofim, visual (visual-MIC) versus spectrophotometer (spec-MIC) reading, and four polystyrene plates using 34 to 53 Aspergillus isolates from five genera. Subsequently, olorofim MICs were compared to itraconazole, voriconazole, posaconazole, and amphotericin B MICs for 298 clinical mold isolates (2016 to 2017). Wild-type upper limits (WT-UL) were determined following EUCAST principles for epidemiologic cutoff value (ECOFF) setting. Olorofim median MICs comparing serial plates and ISO plates were identical (25/36 [69%]) or one dilution apart (11/36 [31%]). Interperson agreement for visual-MICs was 92% to 94%/100% for ≤1/≤2 dilutions, respectively. The visual-MIC values across tested microtiter plates and olorofim lots revealed only discrete differences (≤1 dilution lower for treated plates). No single spec-MIC criterion was applicable to all species. Olorofim MICs were low against 275 Aspergillus species isolates (modal MIC, 0.06 mg/liter; MIC range, < 0.004 to 0.25 mg/liter) and three dermatophytes (MICs 0.03 to 0.06 mg/liter). MICs against Fusarium were diverse, with full inhibition of F. proliferatum (MIC, 0.016), 50% growth inhibition of Fusarium solani at 1 to 2 mg/liter, and no inhibition of F. dimerum. Olorofim displayed potent in vitro activity against most mold isolates and was associated with limited variation in EUCAST susceptibility testing.

Production of tumours and roots by Ephedra following Agrobacterium rhizogenes infection

1994 ◽  
Vol 72 (2) ◽  
pp. 203-207 ◽  
Author(s):  
Niamh A. O'Dowd ◽  
David H. S. Richardson
Keyword(s):  
Growth Rate ◽  
Plant Growth ◽  
Agrobacterium Rhizogenes ◽  
Dry Weight ◽  
Wild Type ◽  
Bacterial Strains ◽  
Heat Treated ◽  
Slow Growing

This paper contains the first report that stems of the Gnetophyte Ephedra respond to infection by Agrobacterium rhizogenes by producing roots and tumours in vivo and in vitro. Of the bacterial strains employed, the wild-type Ar2629 gave the maximum response, and strain LBA9402 was also effective. In no case did heat-treated A. rhizogenes produce tumours or roots. Excised tumour tissues were cultured for more than 2 years in the absence of exogenous plant-growth regulators without any deterioration in growth rate. In vivo tumours of Ephedra fragilis and Ephedra minima contained up to 0.3% dry weight l-ephedrine, and slow-growing in vitro cultured tumours of E. fragilis contained up to 0.01% l-ephedrine, but alkaloid was not detected in faster growing isolates. Key words: Agrobacterium rhizogenes, alkaloid, Ephedra, l-ephedrine, Gnetophytes, gymnosperm, tumours.

In vitro variation in pathogenicity of Fusarium species causing head blight in relation to their isolation from barley head

2021 ◽  
Author(s):  
Nachaat Sakr
Keyword(s):  
Quantitative Resistance ◽  
Fusarium Species ◽  
Fusarium Verticillioides ◽  
Fusarium Culmorum ◽  
Head Blight ◽  
Single Spore ◽  
Barley Cultivars ◽  
Growing Seasons ◽  
Depth Study

Abstract Till now, no published study is available on the variation in pathogenicity of Fusarium head blight (FHB) pathogens in relation to their isolation origin in barley head. To end this, two barley cultivars of contrasting quantitative resistance were artificially infected by four FHB species under field conditions over two consecutive growing seasons. Then, pathogenicity tests were conducted under in vitro conditions on single-spore cultures originated from both kernels and glumes in the heads. Different pathogenicity was detected among Fusarium species originated from both kernels and glumes, indicating that the same isolate from glumes and kernels differs in pathogenicity on leaves/seedlings. Isolates of Fusarium culmorum and Fusarium verticillioides originated from infected kernels had shorter latent periods and higher area under disease progress curves compared to isolates originated from glumes, and the reverse was observed for the Fusarium equiseti isolate. In the case of Fusarium solani, isolates originated from kernels or from glumes were equally pathogenic. Primarily findings in this first in-depth study have implications for breeding programs relied principally on actual quantification of pathogenicity in Fusarium species present in a given environment. The sampling of fungi should take into account the presence of Fusarium species of interest on kernels or glumes.

scholarly journals First Report of Fusarium Crown and Root Rot Caused by Fusarium solani on St. John's-Wort in Argentina

Plant Disease ◽  
2004 ◽  
Vol 88 (9) ◽  
pp. 1050-1050 ◽  
Author(s):  
S. Gaetán ◽  
M. Madia ◽  
R. Cepeda
Keyword(s):  
Fusarium Solani ◽  
Root Rot ◽  
Fusarium Species ◽  
Buenos Aires Province ◽  
Single Spore ◽  
Susceptible Host ◽  
First Report ◽  
St John’S Wort ◽  
St John's Wort ◽  
Crown And Root Rot

Since 2001, 15 to18% of commercial plantings of the medicinal plant St. John's-wort (Hypericum perforatum L.) in Buenos Aires Province, Argentina were affected by a new disease. Disease symptoms of crown and root rot, wilting, chlorosis, and necrosis of the leaves appeared in circular-to-irregular shaped sectors of 12- to 14-month-old plants. Symptoms began with foliage turning yellow followed by an irregular, brown necrosis of the leaf margins. Lesions coalesced to form large necrotic areas causing a severe defoliation of the basal and upper leaves. A soft rot affected the crown and roots causing a complete maceration of these tissues. Infected plants broke off easily because the crown region and the roots were destroyed. As the disease developed, a dark brown discoloration girdled the stems that progressed above the soil line to the apex. The infected stems became dry and breakable. Finally, the affected plants died. Segments (1 cm long) were taken from roots and rotted crowns of diseased plants, dipped in 70% ethanol, surface sterilized with NaOCl (1%) for 1 min, and rinsed in sterile water. Each segment was blotted dry and placed on potato dextrose agar. Plates were incubated in the dark at 26°C for 4 to 7 days. The predominate fungus isolated from the diseased tissue was identified as Fusarium solani (Mart.) Sacc. (1). Koch's postulates were completed by dipping the roots of seedlings in a 2 × 106 conidia per ml suspension of a single spore isolate for 45 min. Plants were repotted (20 inoculated and 10 controls) in a sterilized soil mix (soil/sand 2:1) and held in the greenhouse at 23 to 26°C. Characteristic symptoms identical to the original developed on 90% of inoculated plants within 2 weeks after inoculation. Symptoms included wilt and collapse, crown and root rot, and death of the plants. The fungus was recovered from symptomatic tissues. Control plants dipped into distilled water remained healthy. The experiment was repeated, and the results were identical to the first inoculations. To our knowledge, this is the first report of St. John's-wort as a susceptible host of F. solani. Reference: (1) P. E. Nelson et al. Fusarium species. An Illustrated Manual for Identification. Pennsylvania State University Press, University Park, 1983.

scholarly journals First Report of Zucchini Collapse by Fusarium solani f. sp. cucurbitae Race 1 and Plectosporium tabacinum in Italy

Plant Disease ◽  
2007 ◽  
Vol 91 (3) ◽  
pp. 325-325 ◽  
Author(s):  
S. Vitale ◽  
M. Maccaroni ◽  
A. Belisario
Keyword(s):  
Fusarium Solani ◽  
Fusarium Species ◽  
Morphological Characteristics ◽  
Its Region ◽  
Conidial Suspension ◽  
Single Spore ◽  
Running Water ◽  
First Report ◽  
Race 1 ◽  
Rot Disease

Zucchini plant collapse has been often associated with Fusarium solani f. sp. cucurbitae race 1, which is the causal agent of Fusarium crown and foot rot disease of cucurbits. In Italy, F. solani f. sp. cucurbitae race 1 has been reported on zucchini (Cucurbita pepo) in a greenhouse in the Tuscany Region (4). In spring 2005, a severe outbreak was observed on zucchini in a vast area of cultivation in the province of Venice. Isolations from necrotic vessels gave more than 20 single-spore cultures. On the basis of morphological characteristics, they were identified as F. solani (2) and Plectosporium tabacinum (3). The internal transcribed spacer (ITS) region of rDNA was amplified and sequenced. A fragment of 454 and 531 bp was 99% homologous with sequence PSU66732 and AF150472 of F. solani f. sp. cucurbitae race 1 and P. tabacinum, respectively, in the NCBI database. The nucleotide sequences have been assigned Accession No. AM408782 for F. solani f. sp. cucurbitae race 1 and AM408781 for P. tabacinum. Pathogenicity tests were conducted with four isolates of each species on 15-day-old zucchini plants and on fruit. Plants were inoculated by dipping the roots in a conidial suspension of 106 spores ml-1 for 10 min. Control plants were dipped in sterile water. Five replicates for the inoculated and control plants were used. All plants were maintained in a greenhouse at approximately 24°C. After 14 days, inoculations with F. solani f. sp. cucurbitae race 1 gave symptoms of a cortical rot at the base of the stem with a progressive yellows and wilting of leaves, while plants inoculated with P. tabacinum displayed a moderate wilting. Fruit were washed under running water, disinfected with a solution of 3% sodium hypochlorite and 5% ethanol for 1 min, and inoculated with 6-mm-diameter mycelial plugs cut from the margin of 10-day-old cultures grown on PDA. Plugs were inserted into holes (approximately 2 mm deep) made with a sterile 7-mm-diameter cork borer. Five replicates per isolate were used. Fruit were kept at room temperature (22 to 24°C) in a moist chamber. All isolates induced symptoms of fruit rotting 10 days after inoculation. All controls remained healthy. The colonies reisolated from the inoculated plants and fruit were morphologically identical to the original isolates. The results obtained proved that F. solani f. sp. cucurbitae race 1 can be considered the major pathogen in zucchini collapse, at the same time P. tabacinum may play a role in this syndrome as reported for other cucurbits (1). To our knowledge, this is the first report of zucchini plant collapse caused by F. solani f. sp. cucurbitae race 1 and P. tabacinum, and the first report of P. tabacinum on zucchini in Italy. References: (1) V. J. Garcia-Jimenez et al. EPPO Bull. 30:169, 2000. (2) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University, University Park, 1983. (3) M. E. Palm et al. Mycologia 87:397, 1995. (4) G. Vannacci and P. Gambogi. Phytopathol. Mediterr. 19:103, 1980.

PENGEMBANGAN TEKNOLOGI SELEKSI IN VITRO BERULANG (REPEAT CYCLING–IN VITRO SELECTION) TOLERAN STRES KEKERINGAN UNTUK PERBAIKAN MUTU GENETIK DAN PRODUKSI TANAMAN KEDELAI BERMIKORIZA

2017 ◽  
Vol 1 (1) ◽  
pp. 74-84
Author(s):  
Ahmad Riduan ◽  
Rainiyati Rainiyati ◽  
Yulia Alia
Keyword(s):  
Arbuscular Mycorrhizae ◽  
In Vitro Selection ◽  
Soil Samples ◽  
Single Spore ◽  
Isolation And Characterization ◽  
Single Spore Culture ◽  
Spore Culture ◽  
Glomus Sp

Every plant rhizospheres in any ecosystem there are various living microorganisms including Arbuscular Mycorrhizae Fungi (AMF).  An isolation and characterization is required to investigate the species or type of the AMF. This research was aimed at studying the isolation and characterization of AMF sporulation in soybean rhizospheres in Jambi Province. The results of evaluation on soil samples before trapping showed that there are spores from three genus of AMF twelve types Glomus , two types Acaulospora and one type of Enthrophospora.  Following single spore culture in soybean rhizosphere, 5 spore types were obtained:  Glomus sp-1, Glomus sp-4, Glomus sp-7, Glomus sp-8 Glomus sp-10.

The URNA Genes of Herpesvirus Saimiri (Strain C488) Are Dispensable for Transformation of Human T Cells In Vitro

1999 ◽  
Vol 73 (12) ◽  
pp. 10551-10555 ◽  
Author(s):  
Armin Ensser ◽  
André Pfinder ◽  
Ingrid Müller-Fleckenstein ◽  
Bernhard Fleckenstein
Keyword(s):  
Cell Culture ◽  
Herpesvirus Saimiri ◽  
Wild Type Virus ◽  
Type Virus ◽  
Wild Type ◽  
Human T Cell ◽  
Human T Cells ◽  
Small Nuclear Rnas ◽  
T Cell Lines

ABSTRACT The herpesvirus saimiri strain C488 genome contains five genes for small nuclear RNAs, termed herpesvirus saimiri URNAs (or HSURs). Using a cosmid-based approach, all HSURs were precisely deleted from the genome. The mutant virus replicated at levels that were similar to those of wild-type viruses in OMK cells. Although the HSURs are expressed in wild-type virus-transformed human T-cell lines, the deletion does not affect viral transformation in cell culture.

scholarly journals Mutations in fbiD (Rv2983) as a Novel Determinant of Resistance to Pretomanid and Delamanid in Mycobacterium tuberculosis

2020 ◽  
Vol 65 (1) ◽  
pp. e01948-20
Author(s):  
Dalin Rifat ◽  
Si-Yang Li ◽  
Thomas Ioerger ◽  
Keshav Shah ◽  
Jean-Philippe Lanoix ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Clinical Evidence ◽  
Clinical Samples ◽  
Loss Of Function ◽  
Wild Type ◽  
Content Type ◽  
Solid Media ◽  
High Level ◽  
Level Resistance

ABSTRACTThe nitroimidazole prodrugs delamanid and pretomanid comprise one of only two new antimicrobial classes approved to treat tuberculosis (TB) in 50 years. Prior in vitro studies suggest a relatively low barrier to nitroimidazole resistance in Mycobacterium tuberculosis, but clinical evidence is limited to date. We selected pretomanid-resistant M. tuberculosis mutants in two mouse models of TB using a range of pretomanid doses. The frequency of spontaneous resistance was approximately 10−5 CFU. Whole-genome sequencing of 161 resistant isolates from 47 mice revealed 99 unique mutations, of which 91% occurred in 1 of 5 genes previously associated with nitroimidazole activation and resistance, namely, fbiC (56%), fbiA (15%), ddn (12%), fgd (4%), and fbiB (4%). Nearly all mutations were unique to a single mouse and not previously identified. The remaining 9% of resistant mutants harbored mutations in Rv2983 (fbiD), a gene not previously associated with nitroimidazole resistance but recently shown to be a guanylyltransferase necessary for cofactor F420 synthesis. Most mutants exhibited high-level resistance to pretomanid and delamanid, although Rv2983 and fbiB mutants exhibited high-level pretomanid resistance but relatively small changes in delamanid susceptibility. Complementing an Rv2983 mutant with wild-type Rv2983 restored susceptibility to pretomanid and delamanid. By quantifying intracellular F420 and its precursor Fo in overexpressing and loss-of-function mutants, we provide further evidence that Rv2983 is necessary for F420 biosynthesis. Finally, Rv2983 mutants and other F420H2-deficient mutants displayed hypersusceptibility to some antibiotics and to concentrations of malachite green found in solid media used to isolate and propagate mycobacteria from clinical samples.

scholarly journals InsP7is a small-molecule regulator of NUDT3-mediated mRNA decapping and processing-body dynamics

2020 ◽  
Vol 117 (32) ◽  
pp. 19245-19253 ◽  
Author(s):  
Soumyadip Sahu ◽  
Zhenzhen Wang ◽  
Xinfu Jiao ◽  
Chunfang Gu ◽  
Nikolaus Jork ◽  
...  
Keyword(s):  
Stress Responses ◽  
Messenger Rna ◽  
Control Process ◽  
Stem Cell Differentiation ◽  
Wild Type ◽  
Inositol Pyrophosphate ◽  
Body Dynamics ◽  
Processing Body ◽  
The Stability

Regulation of enzymatic 5′ decapping of messenger RNA (mRNA), which normally commits transcripts to their destruction, has the capacity to dynamically reshape the transcriptome. For example, protection from 5′ decapping promotes accumulation of mRNAs into processing (P) bodies—membraneless, biomolecular condensates. Such compartmentalization of mRNAs temporarily removes them from the translatable pool; these repressed transcripts are stabilized and stored until P-body dissolution permits transcript reentry into the cytosol. Here, we describe regulation of mRNA stability and P-body dynamics by the inositol pyrophosphate signaling molecule 5-InsP7(5-diphosphoinositol pentakisphosphate). First, we demonstrate 5-InsP7inhibits decapping by recombinant NUDT3 (Nudix [nucleoside diphosphate linked moiety X]-type hydrolase 3) in vitro. Next, in intact HEK293 and HCT116 cells, we monitored the stability of a cadre of NUDT3 mRNA substrates following CRISPR-Cas9 knockout ofPPIP5Ks(diphosphoinositol pentakisphosphate 5-kinases type 1 and 2, i.e.,PPIP5KKO), which elevates cellular 5-InsP7levels by two- to threefold (i.e., within the physiological rheostatic range). ThePPIP5KKO cells exhibited elevated levels of NUDT3 mRNA substrates and increased P-body abundance. Pharmacological and genetic attenuation of 5-InsP7synthesis in the KO background reverted both NUDT3 mRNA substrate levels and P-body counts to those of wild-type cells. Furthermore, liposomal delivery of a metabolically resistant 5-InsP7analog into wild-type cells elevated levels of NUDT3 mRNA substrates and raised P-body abundance. In the context that cellular 5-InsP7levels normally fluctuate in response to changes in the bioenergetic environment, regulation of mRNA structure by this inositol pyrophosphate represents an epitranscriptomic control process. The associated impact on P-body dynamics has relevance to regulation of stem cell differentiation, stress responses, and, potentially, amelioration of neurodegenerative diseases and aging.

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