Evaluation of the presence of double-stranded RNA in Ceratocystis ulmi

1985 ◽  
Vol 63 (2) ◽  
pp. 297-300 ◽  
Author(s):  
J. G. Hoch ◽  
S. M. Tavantzis ◽  
R. J. Campana ◽  
S. L. Anagnostakis

Nineteen cultures of Ceratocystis ulmi were classified by mating type and growth rate in vitro. Eleven of the cultures contained double-stranded RNA (dsRNA) with 52% of the segments having molecular weights (MWs) ranging from 1.5 × 106 to 2.0 × 106. Eight of 11 fast-growing (aggressive) isolates contained one to five dsRNA segments with MWs ranging from 0.6 × 106 to 3.1 × 106, whereas 3 of 8 slower growing (nonaggressive) cultures had one or two dsRNA species with MWs ranging from 1.8 × 106 to 2.7 × 106. In addition, cultures with both A and B mating types were found to either contain or lack dsRNA. Thus, there was no clear association between the presence of dsRNA or specific components of dsRNA and aggressiveness or nuclear mating type in the cultures of C. ulmi examined in this study.


Author(s):  
Alejandra Mondragón-Flores ◽  
Patricia Manosalva ◽  
Salvador Ochoa-Ascencio ◽  
Marlene Díaz-Celaya ◽  
Gerardo Rodríguez-Alvarado ◽  
...  

<em>Phytophthora cinnamomi</em> is the pathogen most frequently associated with avocado root rot. In Zitácuaro, Michoacán, production has increased by 19.8%; however, there are no studies of root rot in this area. The objective of the study was to characterize the isolates obtained from avocado roots and assess the sensitivity to fungicides. Samples from 5 avocado orchards were collected, sampling 5 trees per orchard (a total of 25 samples). The samples isolated were characterized morphological and molecularly. Mating type was analyzed using reference isolates of<em> P. cinnamomi</em> A1 (isolate from camelia) and A2 (isolate from avocado). To confirm the pathogenicity, tests were performed on avocado fruits with the isolates. The sensitivity of 15 isolates to potassium phosphite and to metalaxyl-M at different concentrations was evaluated<em> in vitro</em>. In a subgroup of six isolates, it was evaluated whether there was a relationship between growth rate and potassium phosphite sensitivity. Fifteen isolates were obtained with coenocytic coraloid mycelium, chlamydospores, sporangia without papilla, ovoid to ellipsoid, with internal proliferation, heterothallic with mating type A2, with amphigynous antheridia and plerotic oospores, characteristics consistent with <em>P. cinnamomi</em>. The inoculated isolates were pathogenic on avocado fruits. The isolates were more sensitive to potassium phosphite than to metalaxyl-M, with mean EC50 values of 24.62 and 0.215 ?g mL-1 of i.a., respectively. No relationship was observed between growth rate and potassium phosphite sensitivity. It is necessary to obtain a greater number of<em> P. cinnamomi</em> isolates for virulence studies.



2019 ◽  
Vol 6 (1) ◽  
pp. 11-15
Author(s):  
Hari Krishna Saiju ◽  
Abhishesh Bajracharya ◽  
Brishav Rajbahak ◽  
Stuti Ghimire

Paulownia is a fast-growing woody tree, native to the forests of China. It belongs to the family Scrophulariaceae and is mainly used as a source of wood for furniture and musical instruments. Due to its fast-growing nature and high-quality of wood, there has been growing interest in cultivation and research of Paulownia in Nepal. Growth comparison was performed by measuring shoot length in in vitro condition. Among two species of Paulownia - Paulownia tomentosa (Thunb.) Steud and Paulownia fortuneii (Seem.) Hemsl., the growth rate of P. tomentosa was found to be 0.355 cm/week while that of P. fortuneii was found to be 0.637 cm/week in in-vitro conditions in MS medium supplemented with 0.1 mg/l NAA and 1mg/l BAP. Optimization of rooting methods was also performed, in which, sand rooting was found to be easier and more effective than in-vitro rooting. Dipping the plantlets in 1 mg/l of NAA was found to produce longer and denser roots than lower or higher concentrations during sand rooting.



2016 ◽  
Vol 106 (12) ◽  
pp. 1521-1529 ◽  
Author(s):  
Tamieka L. Pearce ◽  
Jason B. Scott ◽  
Frank S. Hay ◽  
Sarah J. Pethybridge

Tan spot of pyrethrum (Tanacetum cinerariifolium) is caused by the ascomycete Didymella tanaceti. To assess the evolutionary role of ascospores in the assumed asexual species, the structure and arrangement of mating-type (MAT) genes were examined. A single MAT1-1 or MAT1-2 idiomorph was identified in all isolates examined, indicating that the species is heterothallic. The idiomorphs were flanked upstream and downstream by regions encoding pyridoxamine phosphate oxidase-like and DNA lyase-like proteins, respectively. A multiplex MAT-specific polymerase chain reaction assay was developed and used to genotype 325 isolates collected within two transects in each of four fields in Tasmania, Australia. The ratio of isolates of each mating-type in each transect was consistent with a 1:1 ratio. The spatial distribution of the isolates of the two mating-types within each transect was random for all except one transect for MAT1-1 isolates, indicating that clonal patterns of each mating-type were absent. However, evidence of a reduced selection pressure on MAT1-1 isolates was observed, with a second haplotype of the MAT1-1-1 gene identified in 4.4% of MAT1-1 isolates. In vitro crosses between isolates with opposite mating-types failed to produce ascospores. Although the sexual morph could not be induced, the occurrence of both mating-types in equal frequencies suggested that a cryptic sexual mode of reproduction may occur within field populations.



2008 ◽  
Vol 76 (7) ◽  
pp. 2923-2938 ◽  
Author(s):  
Xiaorong Lin ◽  
Kirsten Nielsen ◽  
Sweta Patel ◽  
Joseph Heitman

ABSTRACT Hybridization with polyploidization is a significant biological force driving evolution. The effect of combining two distinct genomes in one organism on the virulence potential of pathogenic fungi is not clear. Cryptococcus neoformans, the most common cause of fungal infection of the central nervous system, has a bipolar mating system with a and α mating types and occurs as A (haploid), D (haploid), and AD hybrid (mostly diploid) serotypes. Diploid AD hybrids are derived either from a-α mating or from unisexual mating between haploid cells. The precise contributions of increased ploidy, the effect of hybridization between serotypes A and D, and the combination of mating types to the virulence potential of AD hybrids have remained elusive. By using in vitro and in vivo characterization of laboratory-constructed isogenic diploids and AD hybrids with all possible mating type combinations in defined genetic backgrounds, we found that higher ploidy has a minor negative effect on virulence in a murine inhalation model of cryptococcosis. The presence of both mating types a and α in AD hybrids did not affect the virulence potential, irrespective of the serotype origin. Interestingly, AD hybrids with only one mating type behaved differently, with the virulence of αADα strains similar to that of other hybrids, while aADa hybrids displayed significantly lower virulence due to negative epistatic interactions between the Aa and Da alleles of the mating type locus. This study provides insights into the impact of ploidy, mating type, and serotype on virulence and the impact of hybridization on the fitness and virulence of a eukaryotic microbial pathogen.



2020 ◽  
Author(s):  
Veronica Farmer ◽  
Göker Arpağ ◽  
Sarah Hall ◽  
Marija Zanic

ABSTRACTThe GTP-tubulin cap is widely accepted to protect microtubules against catastrophe. The GTP-cap size is thought to increase with the microtubule growth rate, presumably endowing fast-growing microtubules with enhanced stability. It is unknown what GTP-cap properties permit frequent microtubule catastrophe despite fast growth. Here, we investigate microtubules grown in vitro in the presence and absence of the microtubule polymerase XMAP215. Using EB1 as a GTP-cap marker, we find that GTP-cap size increases regardless of whether growth acceleration is achieved by increasing tubulin concentration or by XMAP215. In spite of the increased mean GTP-cap size, microtubules grown with XMAP215 display increased catastrophe frequency, in contrast to microtubules grown with more tubulin, for which catastrophe is abolished. However, microtubules polymerized with XMAP215 have large fluctuations in growth rate and EB1 intensity; display tapered and curled ends; and undergo catastrophe at faster growth rates and with higher EB1 end-localization. Our results underscore the role of growth irregularities in overall microtubule stability.



1984 ◽  
Vol 4 (1) ◽  
pp. 92-100 ◽  
Author(s):  
D J Thiele ◽  
E M Hannig ◽  
M J Leibowitz

The L double-stranded (ds) RNA component of Saccharomyces cerevisiae may contain up to three dsRNA species, each with a distinct sequence but with identical molecular weights. These dsRNAs have been separated from each other by denaturation and polyacrylamide gel electrophoresis. The 3' terminal sequences of the major species, LA dsRNA, were determined. Secondary structural analysis supported the presence of two stem and loop structures at the 3' terminus of the LA positive strand. In strain T132B NK-3, both the LA and LC species are virion encapsidated. Two distinct classes of virions were purified from this strain, each with a different RNA polymerase activity and with distinct protein components. The heavy virions harbored LA dsRNA, whereas the LC dsRNA species co purified with the light virion peak. Thus, LA and LC dsRNAs, when present in the same cell, may be separately encapsidated.



1981 ◽  
Vol 89 (2) ◽  
pp. 338-345 ◽  
Author(s):  
U Euteneuer ◽  
J R McIntosh

The polarity of kinetochore microtubules (MTs) has been studied in lysed PtK1 cells by polymerizing hook-shaped sheets of neurotubulin onto walls of preexisting cellular MTs in a fashion that reveals their structural polarity. Three different approaches are presented here: (a) we have screened the polarity of all MTs in a given spindle cross section taken from the region between the kinetochores and the poles, (b) we have determined the polarity of kinetochore MTs are more stable to cold-treated spindles; this approach takes advantage of the fact that kinetochore MTs are more stable to cold treatment than other spindle MTs; and (c) we have tracked bundles of kinetochore MTs from the vicinity of the pole to the outer layer of the kinetochore in cold-treated cells. In an anaphase cell, 90-95% of all MTs in an area between the kinetochores and the poles are of uniform polarity with their plus ends (i.e., fast growing ends) distal to the pole. In cold-treated cells, all bundles of kinetochore MTs show the same polarity; the plus ends of the MTs are located at the kinetochores. We therefore conclude that kinetochore MTs in both metaphase and anaphase cells have the same polarity as the aster MTs in each half-spindle. These results can be interpreted in two ways: (a) virtually all MTs are initiated at the spindle poles and some of the are "captured" by matured kinetochores using an as yet unknown mechanism to bind the plus ends of existing MTs; (b) the growth of kinetochore MTs is initiated at the kinetochore in such a way that the fast growing MT end is proximal to the kinetochore. Our data are inconsistent with previous kinetochore MT polarity determinations based on growth rate measurements in vitro. These studies used drug-treated cells from which chromosomes were isolated to serve as seeds for initiation of neurotubule polymerization. It is possible that under these conditions kinetochores will initiate MTs with a polarity opposite to the one described here.



2020 ◽  
Vol 6 (1) ◽  
pp. 14 ◽  
Author(s):  
Austin M. Perry ◽  
Aaron D. Hernday ◽  
Clarissa J. Nobile

Biofilms, structured and densely packed communities of microbial cells attached to surfaces, are considered to be the natural growth state for a vast majority of microorganisms. The ability to form biofilms is an important virulence factor for most pathogens, including the opportunistic human fungal pathogen Candida albicans. C. albicans is one of the most prevalent fungal species of the human microbiota that asymptomatically colonizes healthy individuals. However, C. albicans can also cause severe and life-threatening infections when host conditions permit (e.g., through alterations in the host immune system, pH, and resident microbiota). Like many other pathogens, this ability to cause infections depends, in part, on the ability to form biofilms. Once formed, C. albicans biofilms are often resistant to antifungal agents and the host immune response, and can act as reservoirs to maintain persistent infections as well as to seed new infections in a host. The majority of C. albicans clinical isolates are heterozygous (a/α) at the mating type-like (MTL) locus, which defines Candida mating types, and are capable of forming robust biofilms when cultured in vitro. These “conventional” biofilms, formed by MTL-heterozygous (a/α) cells, have been the primary focus of C. albicans biofilm research to date. Recent work in the field, however, has uncovered novel mechanisms through which biofilms are generated by C. albicans cells that are homozygous or hemizygous (a/a, a/Δ, α/α, or α/Δ) at the MTL locus. In these studies, the addition of pheromones of the opposite mating type can induce the formation of specialized “sexual” biofilms, either through the addition of synthetic peptide pheromones to the culture, or in response to co-culturing of cells of the opposite mating types. Although sexual biofilms are generally less robust than conventional biofilms, they could serve as a protective niche to support genetic exchange between mating-competent cells, and thus may represent an adaptive mechanism to increase population diversity in dynamic environments. Although conventional and sexual biofilms appear functionally distinct, both types of biofilms are structurally similar, containing yeast, pseudohyphal, and hyphal cells surrounded by an extracellular matrix. Despite their structural similarities, conventional and sexual biofilms appear to be governed by distinct transcriptional networks and signaling pathways, suggesting that they may be adapted for, and responsive to, distinct environmental conditions. Here we review sexual biofilms and compare and contrast them to conventional biofilms of C. albicans.



1998 ◽  
Vol 76 (8) ◽  
pp. 1412-1417 ◽  
Author(s):  
Emma T Steenkamp ◽  
Brenda D Wingfield ◽  
Wijnand J Swart ◽  
Michael J Wingfield

Sphaeropsis sapinea (Fr.) Dyko & Sutton is an opportunistic pathogen of various Pinus spp., causing severe shoot blight and dieback. Some isolates of S. sapinea display characteristics such as reduced virulence, reduced growth rate, lack of pigmentation, altered colony morphology, and suppressed conidiation. South African isolates of S. sapinea displaying a range of growth patterns, including reduced growth rate and atypical morphology, were screened for the presence of double-stranded RNA (dsRNA). They were also tested for relative virulence in pathogenicity tests. Double-stranded RNA was isolated by means of phenol extraction and cellulose chromatographic purification. A single species of dsRNA (± 4.3 kilobase pairs in size) was obtained from two slow-growing isolates and two isolates with more regular growth. The virulence of these dsRNA-containing isolates was tested on mature Pinus patula Schlecht. et Cham. trees. Although reduced virulence was positively correlated with slower growth in vitro, the presence of dsRNA could not be linked to either of these characteristics.Key words: double-stranded RNA, hypovirulence, Sphaeropsis sapinea.



1984 ◽  
Vol 4 (1) ◽  
pp. 92-100
Author(s):  
D J Thiele ◽  
E M Hannig ◽  
M J Leibowitz

The L double-stranded (ds) RNA component of Saccharomyces cerevisiae may contain up to three dsRNA species, each with a distinct sequence but with identical molecular weights. These dsRNAs have been separated from each other by denaturation and polyacrylamide gel electrophoresis. The 3' terminal sequences of the major species, LA dsRNA, were determined. Secondary structural analysis supported the presence of two stem and loop structures at the 3' terminus of the LA positive strand. In strain T132B NK-3, both the LA and LC species are virion encapsidated. Two distinct classes of virions were purified from this strain, each with a different RNA polymerase activity and with distinct protein components. The heavy virions harbored LA dsRNA, whereas the LC dsRNA species co purified with the light virion peak. Thus, LA and LC dsRNAs, when present in the same cell, may be separately encapsidated.



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