Isolation and Identification of Eutypa lata from Pistacia vera in Greece

The fungus Eutypa lata (syn. E. armeniacae), known as the causal agent of the death of many different woody plants, was found on dead branches of pistachio (Pistacia vera L.) in Greece. Isolations from diseased branches yielded consistently typical colonies of the asexual stage of the fungus (Libertella blepharis, syn. Cytosporina sp.), which proved to be undistinguishable from other cultures of the pathogen obtained from 15 different hosts. Furthermore, all isolates from pistachio tested for pathogenicity on apricot were pathogenic and yielded characteristic cankers.     

Host specificity,pathogenicity and the presence of virulence genes in Iranian strains of Pseudomonas syringae pv. syringae from different hosts

Pseudomonas syringae pv. syringae (Pss) strains were isolated from almond, apricot, peach, pear, sweet cheery and wheat in Kohgiluye and Boyer-Ahmad, Kordestan, Fras and Chaharmahal and Bakhtiari provinces of Iran. The strains were examined for host specificity, the presence of virulence genes and pathogenicity on different hosts. After inoculation of isolates, in compatible reactions bacterial populations increased within six days of inoculation and final cell numbers increased several-fold over initial inoculum levels, but in incompatible reactions, bacterial populations declined within four days of inoculation. Almond, sweet cherry and wheat isolates induced progressive necrotic symptoms on almond leaves and stems. Apricot, peach and sweet cherry isolates induced necrotic lesions when inoculated on apricot leaves. On pear leaves and stems, only the pear isolate incited pathogenic reaction and isolates from other hosts did not. The syrB gene was detected in all of the tested isolates. Almond and pear isolates did not have the syrD gene. The sypA gene was detected in the almond, peach, pear and sweet cherry isolates while the sypB gene was detected in the apricot, peach, sweet cherry and wheat isolates. Almond, apricot, pear and wheat isolates gave negative results for the detection of nit gene. The gene Ach, was detected only in the peach isolate and gene hrmA, was detected only in the wheat isolate. This study indicates that host specificity exists among different Pss strains, and genes responsible for syringomycin and syringopeptin production contribute to the virulence of Pss strains.     

Intraspecies diversity of Macrophomina phaseolina in Iran

To study the pathogenic and genetic diversity of the Macrophomina phaseolina in Iran, 52 isolates of the fungus were isolated from 24 host plants across the 14 Iranian provinces. All isolates were confirmed to the species based on the species-specific primers. The aggressiveness of M. phaseolina isolates was evaluated on the common bean. Based on the pathogenicity tests, M. phaseolina isolates from the different hosts displayed different levels of aggressiveness on the common beans. The results showed that there was significant variation in the aggressiveness of the pathogen; however, there was no distinct pattern of differentiation based on the host or geographical origin linked to the virulence of the isolates, as frequently theisolates from the same host or geographical origin had different levels of aggressiveness. Inter-simple sequence repeat (ISSR) markers were used to assess the genetic diversity of the fungus. The unweighted pair-group method, using arithmetic mean clustering of data, showed that isolates did not clearly differentiate to the specific group according to the host or geographical origins; however, usually the isolates from the same host or the same geographical origin tend to group nearly. Our results did not show a correlation between the genetic diversity based on the ISSR and pathogenic patterns on common bean in the greenhouse. Similar to the M. phaseolina populations in the other countries, the Iranian isolates were highly diverse based on the pathogenic and genotypic characteristics.     

Total soluble protein profiles of Beauveria bassiana and their relationship with virulence against Helicoverpa armigera

Intracellular total soluble proteins of Beauveria bassiana are believed to play an important role in virulence against insect hosts. Thirty B. bassiana isolates collected from different geographical regions and host ranges were characterised by total soluble proteins present in cells, using the SDS–PAGE technique to differentiate the isolates based on virulence and host insect origin. In vitro analysis of total soluble protein profiles of 30 isolates was studied to understand the relationship of isolates with their host of origin and virulence against Helicoverpa armigera. There was a positive relationship between virulence and host origin. All the non-virulent isolates are grouped together. Similarly, highly virulent isolates against H. armigera were grouped together. The relationship between total soluble proteins and pathogenicity was positively correlated. Thirty isolates shared only 22% similarity in their protein profiles.     

Pathogenicity to Wheat of Isolates of Pseudocercosporella herpotrichoides (Fron) Deighton from Five Graminaceous Hosts

Eleven isolates of Pseudocercosporella herpotrichoides were obtained from field material of the Gramineae Agropyron repens, Alopecurus myosuroides, Apera spica-venti, Hordeum vulgare and Triticum aestivum. Four of these isolates belonged to the variety acuformis and 7 isolates to the variety herpotrichoides of the fungus. In a greenhouse experiment, winter wheat was inoculated with these isolates. All isolates were pathogenic on wheat. They differed in virulence, but these differences could not be related to the nature of their original hosts nor to their classification as variety acuformis or herpotrichoides.     

Sarcodontia pachyodon: a Canker and White‐rot Agent of Plane‐trees

Some lignivorous hymenomycete fungi are capable of causing both cankers and decay in stemwood of adult trees. Recently in Tuscany (Italy), Platanus x acerifolia trees were found colonized by Sarcodontia pachydon (Polyporales, Meruliaceae), a fungus associated with white rot and stem cankers on different host tree species. Because the relationship S. pachyodon‐plane‐tree was only preliminary studied, we decided to investigate whether isolates obtained from this host are distinct from those commonly collected from oaks. For this purpose, isolates obtained from plane‐tree and from holm oak (Quercus ilex) were compared by in vitro test and molecular markers. Results showed that fungal isolates did not differ in growth nor in wood degradation, also molecular tests revealed relative similarity among fungal samples.     

Reproductive Fitness and Random Amplified Polymorphic DNA Variation among Isolates of Pratylenchus vulnus

The reproductive fitness of seven isolates of Pratylenchus vulnus from different geographical areas and hosts was assessed in monoxenic cultures (carrot), and greenhouse cultures (plum, sour orange, and quince). The genetic makeup of the different isolates was compared by Random Amplified Polymorphic DNA (RAPD-PCR). The apple (PvAP-S) and apricot (PvAT-F) isolates reproduced less in monoxenic cultures than the rose (PvRO-S) and walnut (PvWA-A and PvWA-U) isolates. On plum, the rose isolate (PvRO-S) reproduced better than the apple (PvAP-S) and walnut isolate from the United States (PvWA-U). On sour orange, the apple (PvAP-S), unknown origin (PvU-UK), and walnut isolate from Argentina (PvWA-A) multiplied well, whereas the walnut isolate from the United States (PvWA-U), apricot (PvAT-F), and rose (PvRO-S) did not. On quince, the apple (PvAP-S) and walnut (PvWA-U) isolates showed a higher reproduction than the one from unknown origin (PvU-UK). RAPD-PCR patterns among the seven P. vulnus isolates were similar, although high intraspecific varibility was detected. Very few bands of P. neglectus were shared by any population of P. vulnus. A high degree of similarity was found among the patterns corresponding to the rose (PvRO-S), apple (PvAP-S), walnut from the United States (PvWA-U), and unknown origin (PvUK-U) isolates. The apricot isolate (PvAT-F) was the most dissimilar among the seven isolates. No correlation could be established between the genetic variation of P. vulnus detected by RAPD-PCR and reproductive fitness. Results demonstrate high genetic varibility between geographically separated populations of P. vulnus.     

Molecular genotyping of Sclerotinia sclerotiorum isolates from different regions and host plants in Iran

Genetic variation among Sclerotinia sclerotiorum isolates from different regions and host plants were investigated using pathogenicity test, mycelial compatibility groups (MCGs) and molecular markers. Six MCGs were identified and significant differences of virulence variability were observed within and among MCGs. Cluster analysis of combined repetitive sequence-based polymerase chain reaction and randomly amplified polymorphic DNA data discriminated 12 isolates into 11 genotypes, indicating high level of genetic polymorphism among tested isolates. Twelve isolates clustered into four major groups corresponding to their hosts andgeographical region. The variability found within closely related isolates of S.sclerotiorum indicated that such morphological and molecular markers are useful in population studies of this pathogen.     

Pisatin demethylase genes are on dispensable chromosomes while genes for pathogenicity on carrot and ripe tomato are on other chromosomes in Nectria haematococca

Studies on the wide-host-range fungus Nectria haematococca MP VI have shown a linkage between virulence on pea and five of nine PDA genes that encode the ability to detoxify the pea phytoalexin, pisatin. Most of the PDA genes are on chromosomes of approximately 1.6 megabases (Mb) and two of these genes, PDA1-2 and PDA6-1, have been demonstrated to reside on approximately 1.6-Mb chromosomes that can be lost during meiosis. Prior studies also have shown that the dispensable chromosome carrying PDA6-1 contains a gene (MAK1) necessary for maximum virulence on chickpea. The present study evaluated whether the other approximately 1.6-Mb chromosomes that carry PDA genes also are dispensable, their relationship to each other, and whether they contain genes for pathogenicity on hosts other than pea or chickpea. DNA from the PDA1-1 chromosome (associated with virulence on pea) and the PDA6-1 chromosome (associated with virulence on chickpea) were used to probe blots of contour-clamped homogeneous electric field (CHEF) gels of isolates carrying different PDA genes and genetically related Pda- isolates. All of the approximately 1.6-Mb PDA-bearing chromosomes hybridized with both probes, indicating that they share significant similarity. Genetically related Pda-progeny lacked chromosomes of approximately 1.6 Mb and there was no significant hybridization of any chromosomes to the PDA1-1 and PDA6-1 chromosome probes. When isolates carrying different PDA genes and related Pda- isolates were tested for virulence on carrot and ripe tomato, there was no significant difference in lesion sizes produced by Pda+ and Pda- isolates, indicating that genes for pathogenicity on these hosts are not on the PDA-containing chromosomes. These results support the hypothesis that the chromosomes carrying PDA genes are dispensable and carry host-specific virulence genes while genes for pathogenicity on other hosts are carried on other chromosomes.     

Pectic zymogram variation and pathogenicity of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-4 to bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) isolates in Isfahn,Iran

Rhizoctonia disease, caused by Rhizoctonia solani is one of the most important fungal diseases in bean fields in Isfahan, Iran. Bean plants showing stem and root cankers were collected and Rhizoctonia-like fungi obtained from the samples were identified by anastomosis. Pure cultures of bean isolates of R. solani were identified as AG-4. There were also AG-4 isolates from tomato, potato, cucumber, alfalfa and sugar beet in the areas sampled. A total of 163 isolates of R. solani AG-4 originating from stem and root cankers of beans were examined using pectic zymogram electrophoresis. Polygalacturonase (PG) and pectin estrase isozymes were observed in all AG-4 isolates tested. One (PG) and one pectic esterase (PE) band was found in common between all isolates examined. The electrophoretic patterns were grouped into seven zymogram groups (ZGs) according to the diagnostic PG and PE bands. One ZG occurred in a high frequency throughout the areas sampled. A pathogenicity test was conducted and representative isolates of each ZG were used to inoculate healthy bean plants. The results showed that each ZG caused different symptoms with varying severity. Isolates belonging to two ZGs were highly pathogenic causing root, stem and hypocotyl cankers whereas isolates of the other ZGs produced weak or no symptoms.     

Pathogenicity studies on isolates of Leptosphaeria maculans from brassica seed production crops in south-east England

Out of 117 isolates of Leptosphaeria maculans collected from a range of brassica seed crops in south-east England, 26 were virulent when tested on cabbage cv. January King. Of these, 24 were isolated from oil-seed rape (Brassica napus) and the remainder from a swede (B. napus) and a cabbage (B. oleracea) seed crop. Virulent strains were derived from infected stems and from single ascospores of the sexual state present on diseased stubble. In glasshouse tests virulent isolates were not host-specific and caused severe cankers on cabbage, kale, swede, turnip and oil-seed rape; under field conditions, ascospores originating from diseased oil-seed rape stubble produced severe cankers and high levels of seed-borne infection in eight horticultural and vegetable brassica hosts (B. napus, B. campestris and B. oleracea). Virulent and non-virulent types could be distinguished by certain cultural characteristics. On nutrient agar virulent types grew slowly, irregularly and quickly staled whereas non-virulent types grew rapidly, regularly and did not stale. The latter also produced a yellow-brown pigment in liquid culture. The potential for cross-infection of virulent strains originating from oil-seed rape to other brassica seed crops has serious implications for disease incidence on forage and vegetable brassica seed crops.     

Identification of Xanthomonas campestris pv. Juglandis from (Persian) English Walnut Nursery Trees in South Africa

Bacterial isolates producing yellowish colonies on Nutrient Agar were recovered from symptoms of suspect walnut blight disease on leaves of nursery trees in the southwestern Cape Province of South Africa. The isolates were identified by pathogenicity tests on leaves of walnut and plum trees in the greenhouse. Fifteen isolates from four cultivars at two nurseries produced typical lesions of blight on walnut and one isolate. typical lesions of bacterial spot disease on plum leaves. Cluster analysis was done on 28 characteristics recorded from colony growth. colour. form. and elevation on four different culture media, and starch hydrolysis on a semi-selective medium for the isolation of Xanthomonas campestris pv. juglandis. Total DNA of the isolates was digested with restriction endonuclease Spel and resolved by contour-clamped homogeneous electric field (CHEF) electrophoresis. Two phenotypic clusters were distinguished among the 15 South African and one reference strain of X.c.pv. juglandis at the 54%S_sm level. The isolate which induced disease symptoms on plum grouped with reference strains of Xanthomonas campestris pv. pruni in a third cluster. Two-thirds of the isolates were not characterized on the semi-selective medium for X.c. pv. juglandis. DNA restriction fragment banding patterns were similar for most isolates of X.c.juglandis in the same phenotypic cluster. However, DNA banding patterns were non-distinct for some isolates with similar phenotypic characters. Phenotypic characteristics and DNA restriction fragment banding patterns of the isolates were not correlated with geographical origin or cultivar specificity.     

Traits associated with virulence to the aphid Macrosiphoniella sanborni in eighteen isolates of Verticillium lecanii

Eighteen isolates of the entomopathogenic fungus Verticillium lecanii from various world-wide locations, from insect hosts and soil were bioassayed against the aphid Macrosiphoniella sanborni in the laboratory. Virulence ranged from an isolate which achieved 100% mortality and LT 50 value (adults) of 3 days ± 0· 2 at 24 ° C, compared with the least virulent isolates causing less than 10% mortality over 14 days (when treated with an inoculum of 1 × 10⁶ conidiospores/ml). All isolates produced extracellular protease and lipase, irrespective of their virulence. A number of traits were frequently associated with the expression of virulence including fast germination, high sporulation rate, an absence of extracellular amylase activity and high extracellular chitinase activities. Large spore size was not strongly associated with virulence. There were exceptions in each variate studied, suggesting that overall expression of virulence is a result of the total complex of these and other traits still to be determined.     

Variation in two components of pathogenicity and its relationship with genetic diversity in Plasmopara halstedii (sunflower downy mildew)

The specificity of the two components of pathogenicity: virulence and aggressiveness and its relationship with genetic variability were analysed in a local Plasmopara halstedii (sunflower downy mildew) population. Pathogenic and molecular analyses were carried out on seven isolates including five progeny isolates of five races arising from two parental races 100 and 710. P. halstedii isolates showed significant differences for all aggressiveness criteria and important genetic variations. Three cases of relationship between virulence and aggressiveness for progeny isolates as compared with parental ones were found as positive, negative or uncorrelated. For solving the specificity of these cases, relationship between the two components of pathogenicity among the isolates of three different races localised in the same genetic clade was positive. The hypothesis explaining these cases is discussed.     

Variation in pathogenicity among field isolates of Phoma exigua var. foveata

Experiments carried out using a point inoculation method to infect potato tubers with Phoma exigua var. foveata demonstrated considerable variation in pathogenicity among field isolates. This variation was unlikely to be due to differential reductions in isolate pathogenicities during axenic culture. However, fresh field isolates generally produced larger lesions than stored isolates. An investigation of the distribution of pathogenicity variation in the fungus revealed that differences among isolates from different lesions taken from the same potato stock were greater than those between stocks, but pathogenicity variation within each lesion isolate was small relative to that between isolates. The importance of using isolates with a high level of pathogenicity, hence recent field isolates, in studies of this pathogen is stressed.     

Comparative Analysis of Pathogenicity and Phylogenetic Relationship in Magnaporthe grisea Species Complex

Outbreaks of rice blast have been a threat to the global production of rice. Members of the Magnaporthe grisea species complex cause blast disease on a wide range of gramineous hosts, including cultivated rice and other grass species. Recently, based on phylogenetic analyses and mating tests, isolates from crabgrass were separated from the species complex and named M. grisea. Then other isolates from grasses including rice were named as M. oryzae. Here, we collected 103 isolates from 11 different species of grasses in Korea and analyzed their phylogenetic relationships and pathogenicity. Phylogenetic analyses of multilocus sequences and DNA fingerprinting revealed that the haplotypes of most isolates were associated with their hosts. However, six isolates had different haplotypes from the expectation, suggesting potential host shift in nature. Results of pathogenicity tests demonstrated that 42 isolates from crabgrass and 19 isolates from rice and other grasses showed cross-infectivity on rice and crabgrass, respectively. Interestingly, we also found that the isolates from rice had a distinct deletion in the calmodulin that can be used as a probe.     

Morphological,cultural and pathogenic characteristics of Coniothyrium zuluense isolates from different plantation regions in South Africa

Coniothyrium canker caused by Coniothyrium zuluense, is a serious stem canker disease of Eucalyptus species in sub-tropical regions of South Africa. This disease is typified by necrotic bark lesions that coalesce to form large kino-impregnated cankers along the stems of trees. The strategy currently used to manage Coniothyrium canker in plantations is to deploy Eucalyptus species or clones that are resistant to the disease. Considerable success has already been achieved in this regard, but the long-term durability of resistance is of concern. Thus, forest managers are interested in the genetic diversity of the pathogen and its potential to overcome disease resistance in planting stock. In this study, 344 isolates of C. zuluense from different plantation regions in South Africa were compared on the basis of colony colour, conidial morphology, growth characteristics on agar and pathogenicity to a susceptible E. grandis clone. Conidia of all C. zuluense isolates measured were similar in size and shape. The fungus is slow growing in culture, which is indicative of its apparent biotrophic habit, with optimum growth observed at 30 °C. Isolates of C. zuluense displayed considerable variation in colony colour and pathogenicity in inoculation trials. Variation in morphology and pathogenicity amongst isolates suggests that C. zuluense has been present in South Africa for an extended period of time, or that it is changing rapidly due to strong directional selection pressures. This revised version was published online in June 2006 with corrections to the Cover Date.     

A serious canker disease of Eucalyptus in South Africa caused by a new species of Coniothyrium

Eucalyptus spp. are being propagated extensively as exotics in plantations in South Africa, and many other parts of the world. In South Africa, a number of diseases result in serious losses to this resource. This paper describes a new and very damaging stem canker disease, which has recently appeared on plantation-grown eucalyptus in South Africa. The disease, first noted in an isolated location in Zululand is now common in other parts of the country, and is typified by discrete necrotic lesions on stems. These lesions coalesce to form large, gum-impregnated cankers and malformed stems. The causal agent of the disease, as inferred from pathogenicity tests, is a new species of Coniothyrium described here as C. zuluense. This fungus is a serious impediment to eucalypt propagation in South Africa, and is most likely a threat to similar forest industries elsewhere in the world.     

Strain Dependent Variation of Immune Responses to A. fumigatus: Definition of Pathogenic Species

For over a century microbiologists and immunologist have categorized microorganisms as pathogenic or non-pathogenic species or genera. This definition, clearly relevant at the strain and species level for most bacteria, where differences in virulence between strains of a particular species are well known, has never been probed at the strain level in fungal species. Here, we tested the immune reactivity and the pathogenic potential of a collection of strains from Aspergillus spp, a fungus that is generally considered pathogenic in immuno-compromised hosts. Our results show a wide strain-dependent variation of the immune response elicited indicating that different isolates possess diverse virulence and infectivity. Thus, the definition of markers of inflammation or pathogenicity cannot be generalized. The profound understanding of the molecular mechanisms subtending the different immune responses will result solely from the comparative study of strains with extremely diverse properties.     

Detection of double stranded RNA in phytopathogenic <Emphasis Type="Italic">Macrophomina phaseolina</Emphasis> causing charcoal rot in <Emphasis Type="Italic">Cyamopsis tetragonoloba</Emphasis>

One hundred one isolates of Macrophomina phaseolina from various hosts and eco-geographical locations were employed for elucidating relationships among genetic diversity and virulence. Highly pathogenic, moderately pathogenic, and hypovirulent cluster bean specific isolates were identified. In order to correlate respective phenotypes of plant pathogenic fungus multiple and complex patterns of dsRNA elements were analyzed. Double-stranded ribonucleic acids (dsRNA) are ubiquitous in all major groups and most of them have vast potential as biological control agents for fungi. Rate of virulence and its further association could ascertain by host plant and their fungal genotypes. Variability of the fungal genotypes decides the link between the complexity of dsRNA with different variants and the change in virulence pattern. Double-stranded RNA was identified in approximately 21.7% of M. phaseolina isolates from charcoal rot infected cluster bean varieties. After recurrent laboratory transfer on culture media, the preponderance of the isolates harboring dsRNAs developed degenerate culture phenotypes and showed reduced virulence (hypovirulence) to cluster bean. Macrophomina has successfully showed diversified and reproducible banding profile in dsRNA containing/free isolates. This is the first report of hypovirulence and detection of dsRNA in Macrophomina phaseolina isolates of cluster bean origin.