Genetic and Phenotypic Diversity among Botrytis cinerea Isolates in Iran

Forty-four Botrytis cinerea isolates from different hosts and geographical regions were studied for colony morphology, mycelial growth rate at different temperatures, pathogenicity and molecular diversity. Botrytis cinerea isolates had temperature optima of 20–25°C and isolates showed variation in growth rate at different temperatures. Two morphological types were identified among tested isolates: mycelial and sclerotial. The pathogenicity of isolates was tested on grapevine leaves, and it was revealed that nine of 44 isolates were non-pathogenic and among them seven were of mycelial type. There was no correlation between mycelium growth rate and pathogenicity. Genetic diversity was investigated using nine arbitrary decaprimers. No relationship was found between molecular clusters and geographical region or sampling time; whereas isolates from the same plant host tended to cluster with each other. Seven of nine non-pathogenic isolates were separated from pathogenic ones.     

Genetic diversity and pathogenicity of the grass pathogen Xanthomonas translucens pv. graminis

Genetic diversity of Xanthomonas translucens pv. graminis (X.t.g.), the causal agent of bacterial wilt in forage grasses, was assessed using 16S rDNA sequencing, AFLP analysis and pathogenicity screening on three Italian ryegrass (Lolium multiflorum) cultivars. The aim of this in depth analysis was to provide insight into geographic variation and race specificity of X.t.g. in order to develop strategies for plant protection and resistance breeding. 16S rDNA sequencing of 29 putative X.t.g. isolates allowed to assign 28 isolates to the pv. translucens while one isolate was identified as X.t. pv. arrhenatheri. AFLP analysis and UPGMA clustering resulted in two distinct clusters with a similarity of 90% based on Nei and Li's coefficient and 306 polymorphic markers. A significant effect of geographic location of the collection sites on genetic diversity was detected by redundancy analysis using AFLP markers and the explanatory variables longitude, latitude and altitude. The two groups identified with redundancy analysis were congruent to the major clusters identified with cluster analysis. All X.t.g. isolates identified as pv. graminis were pathogenic, showing mostly moderate to high pathogenicity. However, the three cultivars used for pathogenicity testing showed significant differences in susceptibility and a significant interaction between cultivars and isolates was observed, indicating an at least partial race-specific resistance. With the information provided, targeted selection of X.t.g. isolates may allow one to efficiently address specific tasks in resistance breeding.     

Genetic correlates of in vivo viral resistance to indinavir, a human immunodeficiency virus type 1 protease inhibitor.

Indinavir (IDV) (also called CRIXIVAN, MK-639, or L-735,524) is a potent and selective inhibitor of the human immunodeficiency virus type 1 (HIV-1) protease. During early clinical trials, in which patients initiated therapy with suboptimal dosages of IDV, we monitored the emergence of viral resistance to the inhibitor by genotypic and phenotypic characterization of primary HIV-1 isolates. Development of resistance coincided with variable patterns of multiple substitutions among at least 11 protease amino acid residues. No single substitution was present in all resistant isolates, indicating that resistance evolves through multiple genetic pathways. Despite this complexity, all of 29 resistant isolates tested exhibited alteration of residues M-46 (to I or L) and/or V-82 (to A, F, or T), suggesting that screening of these residues may be useful in predicting the emergence of resistance. We also extended our previous finding that IDV-resistant viral variants exhibit various patterns of cross-resistance to a diverse panel of HIV-1 protease inhibitors. Finally, we noted an association between the number of protease amino acid substitutions and the observed level of IDV resistance. No single substitution or pair of substitutions tested gave rise to measurable viral resistance to IDV. The evolution of this resistance was found to be cumulative, indicating the need for ongoing viral replication in this process. These observations strongly suggest that therapy should be initiated with the most efficacious regimen available, both to suppress viral spread and to inhibit the replication that is required for the evolution of resistance.     

Does selection by resistant hosts trigger local adaptation in plant–pathogen systems?

Understanding the consequences of selection by host resistance on pathogen population structure provides useful insights into the dynamics of host-parasite co-evolution processes and is crucial for effective disease management through resistant cultivars. We tested general vs. local population adaptation to host cultivars, by characterizing a French collection of Phytophthora infestans (the causal organism of potato late blight) sampled during two consecutive years on cultivars exhibiting various levels of resistance. Local populations were structured by the host for virulence (qualitative pathogenicity) but also for aggressiveness (quantitative pathogenicity). All populations had a low genotypic diversity for amplified fragment length polymorphisms (AFLPs), and presumably consisted of a few closely related clonal lineages. No correlation was detected between pathogenicity traits and AFLP genotypes. The data support the hypothesis of general adaptation for aggressiveness, to which directional selection for virulence is superimposed when race-specific resistance is introduced.     

Pinewood Nematode Species Complex: Interbreeding Potential and Chromosome Number

Interbreeding potential, chromosome number, and host range were compared among several isolates and species of Bursaphelenchus from diverse geographic areas. Some isolates from North America, Japan, and France had a wide-ranging interbreeding potential, whereas others were restricted in their potential to hybridize with other isolates. Although interbreeding occurred in the laboratory between some "M" and "R" forms of B. xylophilus, interbreeding of B. xylophilus and B. mucronatus was rare. The hybrids had the pathogenicity of the parent with the broader host range. This fact suggests that virulence may be inherited as a dominant character or that increased virulence may have resulted from differences in hybrid vigor. The haploid chromosome number of the different isolates separated the isolates into three groups and distinguished B. xylophilus from B. mucronatus. The findings suggest that the pinewood nematode species complex consists of sibling species that have evolved by reproductive isolation, that the French isolate is a new species, and that B. xylophilus and B. mucronatus have evolved from a common ancestor.     

Identification of sources of Escherichia coli in South Carolina estuaries using antibiotic resistance analysis

Fecal pollution from nonhuman (pets, livestock or wildlife) and human sources is often one of the major factors associated with urbanization that contribute to the degradation of water quality. Methods to differentiate animal from human sources of fecal coliform contamination could assist resource managers in developing strategies to protect shellfish harvesting areas and recreational waters. In this study, surface water samples were collected from both a developed and an undeveloped watershed in coastal South Carolina. Influent and effluent samples from several wastewater treatment plants (WWTPs) in the same area were also collected. Most Probable Numbers (MPNs) of fecal coliforms were determined for all samples. Escherichia coli isolates were analyzed for antibiotic resistance (AR) to 10 antibiotics. Then, AR indices (no. of resistant/total no. of antibiotics tested), were calculated for each isolate and site. Results indicated that MPNs from the WWTP samples were significantly higher than those from the developed watershed which were significantly higher than those from the undeveloped watershed (p<0.0001). The AR analyses suggested that there was a trend toward increased antibiotic resistance in samples for the urbanized Broad Creek (BC) watershed. In the Okatee River (OR), E. coli isolates from three sites (20%) showed resistance to a single antibiotic (penicillin) but in BC, isolates from seven sites (47%) were resistant to multiple antibiotics, and the predominant resistance pattern was chlortetracycline-oxytetracycline-tetracycline. Raw sewage isolates from most WWTPs contained E. coli that exhibited resistance to multiple antibiotics. Cluster analysis indicated that all resistant OR sites had antibiotic resistant isolates that matched AR patterns found in isolates from WWTPs. Similarly, six of the seven sites in BC had AR patterns that matched with resistance patterns in WWTPs. These results suggest that AR testing may be a useful tool for differentiating E. coli from human and wildlife sources. Further testing of bacterial isolates from known animal sources is necessary to better assess the utility of this approach.     

Population Structure of Puccinia recondita in Western Europe During 1995, as Assessed by Variability in Pathogenicity and Molecular Markers

The population structure of Puccinia recondita f. sp. tritici (Prt) in western Europe was examined by assessing variability in pathogenicity and in randomly amplified polymorphic DNA (RAPD) among 61 single uredinial isolates. The isolates were chosen to represent pathotypes detected in a previous survey of pathogenic variability in the fungus in western Europe in 1995. Thirty‐five pathotypes were identified by assessing infection types produced by the 61 isolates on 24 differential lines, each with a single gene for resistance to Prt. In contrast, only 18 RAPD phenotypes were identified by scoring 19 polymorphic RAPD bands generated with eight RAPD primers. When analysed by cluster and bootstrap analyses, the pathogenicity and RAPD results revealed little evidence for robust distinct clusters among the isolates. Multiple isolates of several pathotypes collected from widely separated locations such as Belgium, Germany, France, Italy and Switzerland had the same RAPD phenotype, providing evidence of clonal migration over considerable distances in western Europe. Some variability (one or two band differences) was observed in RAPD phenotype within several pathotypes, indicating the possible occurrence of genetic changes independent of pathogenicity, and/or the independent development of pathotypes with different genetic backgrounds. Two groups of isolates identified in the 1995 survey, differentiated by pathogenicity for genes Lr3a, Lr3bg, Lr3ka and Lr30, were not distinguished by RAPD phenotype, indicating that the groups probably do not constitute separate lineages within the pathogen population. Little correlation was apparent between the polymorphisms observed in pathogenicity and RAPD phenotypes. The similarity in the genetic backgrounds of the isolates, as assessed by RAPD markers, suggest that the observed differences in pathogenicity may have arisen by selection for specific virulences corresponding to genes for resistance in wheat cultivars grown in the region. Three isolates of pathotype 3, restricted in its distribution to southern France during 1995, were distinct from all other isolates in RAPD phenotype. Circumstantial evidence suggests that this pathotype originated from northern Africa, and that it belongs to a group of leaf rust pathogens specialized to durum wheats.     

海南省潜伏香蕉果实的炭疽菌对特克多的抗药性

对来自海南省13个地区的67个Colletotrichum musae（Berk. & Curt.）Arx菌株进行抗药性检测,所测菌株均表现出敏感,即未产生特克多抗药性。室内采用高浓度特克多和90～95%致死剂量紫外光进行抗性诱导,获得的抗性菌株均能在1000g/ml特克多的PDA培养基上生长,但EC50值相差很大,最高达130g/ml,而最低为0.87g/ml。抗性菌株对多菌灵和甲基托布津均表现正交互抗药性。连续无毒培养10代后,所有菌株仍可在5g/ml特克多的培养基上生长,表现抗性遗传稳定。抗性菌株的产孢能力和致病力与敏感菌相比并没有下降,表现很高的适应能力。     

BREEDING PEARS FOR RESISTANCE TO THE PEAR SCAB FUNGUS, VENTURIA PIRINA ADERH.

Three clones of Venturia pirina have been tested for their pathogenicity towards forty varieties of cultivated pear. From observations on the reactions of these forty varieties, twelve were selected for differentiating the pathogenicity of sixteen clones of the fungus.
These tests differentiated four main groups of biotypes of V. pirina. Clones isolated from Conference showed wide differences in range of pathogenicity. One isolate from Conference was shown to parasitize six of the most widely grown pears in England: other Conference biotypes had a narrow range.
No variety of pear was found immune to all the isolates. Beurré Giffard was lightly infected by an isolate from Durondeau, but was resistant to all the other isolates tested.     

Host Range of Australian Phoma ligulicola var. inoxydablis Isolates from Pyrethrum

Ray blight, caused by Phoma ligulicola var. inoxydablis is one of the most damaging diseases of pyrethrum ( Tanacetum cinerariifolium [Trevir.] Sch. Bip.) in Australia. The pathogenicity of P. ligulicola var. inoxydablis to a range of ornamental and other plant species was tested to determine potential sources of inoculum into pyrethrum crops. Differences were identified in the host range of P. ligulicola var. inoxydablis isolates in this study in comparison with isolates reported from garden chrysanthemum ( Chrysanthemum morifolium L.), most likely to be P. ligulicola var. ligulicola . Australian P. ligulicola var. inoxydablis isolates were unable to infect and cause disease following repeated inoculation to zinnia ( Zinnia elegans L.), sunflower ( Helianthus annuus L.), dahlia ( Dahlia variabilis Desf.), and several cultivars of crisphead lettuce ( Lactuca sativa L.). French marigold ( Tagetes patula L.) was recorded as a susceptible host for this pathogen for the first time. Moreover, the susceptibility of annual chrysanthemum ( Chrysanthemum carinatum L.) to infection by P. ligulicola var. inoxydablis was confirmed. Implications for disease management in Tasmanian pyrethrum fields are discussed.     

不同寄主来源的灰葡萄孢对番茄的致病力分化研究

Eighteen isolates of Botrytis cinerea were obtained from the diseased plant tissue collected in Hefei, Bengbu, Changfeng and Hexian in Anhui province, by means of tissue isolating method. The pathogenicity of the isolates of B. cinerea from different hosts to the fruits and leaves of tomato were investigated by applying wound inoculation with mycelial blocks. The results showed that all of the tested isolates caused grey mould on tomato fruits, but there was significant difference in the average diameters of the lesions caused by different isolates, suggesting that there was significant differentiation in pathogenicity of B. cinerea strains to tomato fruits among isolates. According to the average diameters of the lesions on tomato fruits, the pathogenicity of the all isolates was classified into three categories: strong, intermediate and weak. In general, the isolates from tomato were more strongly pathogenic to tomato fruits than the isolates from strawberry, grape and capsicum. However, there was difference in pathogenicity among the different isolates from the same host, and the pathogenicity difference was not obviously related to the localities of isolates. After inoculating of tomato leaves, all of the tested isolates except CF3 caused grey mould on tomato leaves, but there was significant difference in the average diameters of the lesions caused by different isolates; and the difference in pathogenicity to tomato leaves was not obviously related to the host and locality of isolates.     

Characterization of the Bacteroides fragilis pathogenicity island in human blood culture isolates

Bacteroides fragilis is an important anaerobic pathogen accounting for up to 10% of bacteremias in adult patients. Enterotoxin producing B. fragilis (ETBF) strains have been identified as enteric pathogens of children and adults. In order to further characterize the B. fragilis pathogenicity island (BfPAI) and using PCR assays for bft- and mpII-metalloprotease genes, we determined the frequency of B. fragilis strains with pattern I (containing the BfPAI and its flanking region), pattern II (lacking both the BfPAI and the flanking region), and pattern III (lacking the BfPAI but containing the flanking region) in 63 blood culture isolates. The results were compared to 197 B. fragilis isolates from different clinical sources. We found 19% of blood culture isolates were pattern I (ETBF), 43% were pattern II (NTBF) and 38% were pattern III (NTBF). Comparatively, B. fragilis isolates from other clinical sources were 10% pattern I, 47% pattern II and 43% pattern III. This suggests that the pathogenicity island and the flanking elements may be general virulence factors of B. fragilis.     

Genotypic Diversity and Virulence Characteristics of Clinical and Environmental Vibrio vulnificus Isolates from the Baltic Sea Region

The genetic diversity of Vibrio vulnificus isolates from clinical and environmental sources originating from the Baltic Sea region was evaluated by multilocus sequence typing (MLST), and possible relationships between MLST clusters, potential genotypic and phenotypic traits associated with pathogenicity, and source of isolation were investigated. The studied traits included genotyping of polymorphic loci (16S rRNA, vcg, and pilF), presence/absence of potential virulence genes, including nanA, nab, and genes of pathogenicity regions, metabolic features, hemolytic activity, resistance to human serum, and cytotoxicity to human intestinal cells. MLST generated 35 (27 new) sequence types and divided the 53 isolates (including four reference strains) into two main clusters, with cluster I containing biotype 1 and 2 isolates of mainly environmental origin and cluster II containing biotype 1 isolates of mainly clinical origin. Cluster II isolates were further subdivided into two branches. Branch IIB included isolates from recent cases of wound infections that were acquired at the German Baltic Sea coastline between 2010 and 2011 and isolates from seawater samples of the same regions isolated between 1994 and 2010. Comparing the MLST data with the results of genotyping and phenotyping showed that strains of MLST cluster II possess a number of additional pathogenicity-associated traits compared to cluster I strains. Rapid microbiological methods such as matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry combined with typing of selected virulence-associated traits (e.g., serum resistance, mannitol fermentation, nanA, and pathogenicity region XII) could be used for risk assessment purposes regarding V. vulnificus strains isolated from the Baltic Sea region.     

Changes of Genotype,Sensitivity and Aggressiveness in Phytophthora infestans Isolates Collected in European Countries in 1997, 2006 and 2007

A total of 241 isolates of Phytophthora infestans were collected in 1997, 2006 and 2007 in eight European countries and characterized with molecular markers (simple sequence repeats, SSR genotypes) and phenotypic traits such as sensitivity to fungicides, mating type and aggressiveness. The mating type distribution changed from mainly A1 in 1997 to a majority of A2 in 2007. No resistant isolates were detected for fluazinam and mandipropamid, whereas the proportion of isolates resistant to mefenoxam (MFX) was high and increased over the years. There was no genetic link between mating type and MFX resistance. Aggressiveness (product between lesion expansion and sporulation capacity) was slightly higher for MFX‐resistant compared to sensitive isolates and for isolates collected later compared to earlier in the same season. It was about equally high for A1 and A2 types, and for French isolates in 1997 and British isolates in 2007, but lower for French isolates in 2007. Six different SSR genotype families were distinguished. In 1997, populations were dominated by genotype families I and III/IV, which significantly declined in 2007 being largely displaced by genotype families II (‘blue 13’ type) and V, which are by coincidence mainly A2 MFX resistant and A1 MFX sensitive, respectively. However, mating type and MFX resistance were genetically not linked to SSR genotypes.     

云南两个县陆稻稻瘟病菌稻巨座壳交配型测定及致病性分析

2014年,自云南省沧源县及耿马县陆稻地方品种上分离99个稻瘟病菌稻巨座壳单孢菌株,采用4个已知交配型的标准菌株对其进行育性和交配型测定.结果 表明,两地稻巨座壳菌株具较高的育性,平均可交配率高达90.8％,且可育菌株中,MAT1-1和MAT1-2菌株分别占60.9％和39.1％;分别随机对沧源县南撒村和班考村同一田块...     

Analysis of the structure of the AVR1-CO39 avirulence locus in virulent rice-infecting isolates of Magnaporthe grisea

The AVR1-CO39 gene that came from a Magnaporthe grisea isolate from weeping lovegrass controls avirulence on the rice cultivar CO39. AVR1-CO39 was not present in the genome of the rice-infecting M. grisea isolate Guyll from French Guyana, suggesting that the gene had been deleted. Molecular analysis of the deletion breakpoints in the AVR1-CO39 locus revealed the presence of a truncated copy of a previously unknown retrotransposon at the left-hand border. At the right-hand border was a truncated copy of another repetitive element that is present at multiple locations in the genome of Guyll. The structures of avr1-CO39 loci were further examined in 45 rice-infecting isolates collected in Brazil, China, Japan, India, Indonesia, Mali, and the Philippines. Most isolates showed no hybridization signal with the AVR1-CO39 probe and had the same locus structure as Guyll. Some isolates from Japan showed a signal with the AVR1-CO39 probe, but the region specifying avirulence activity was rearranged. These findings suggest that widespread virulence to 'CO39' among rice-infecting M. grisea isolates is due to ancestral rearrangements at the AVR1-CO39 locus that may have occurred early in the evolution of pathogenicity to rice.     

Nitrogen utilization in bacterial isolates from the equine cecum

A total of 114 bacterial isolates were obtained from the cecal contents of two mature cecally fistulated horses on a habitat-simulating medium containing 40% energy-depleted cecal fluid. Of these isolates, 108 were maintained in pure cultures and were tentatively grouped on the basis of cell morphology and physiological characteristics. Gram-negative rods (50.9%), gram-positive rods (22.8%), and gram-positive cocci (21.9%) represented the largest groups isolated from these animals. Fifty isolates were tested for their ability to grow in media containing urea, ammonia, peptones, or amino acids as sole nitrogen sources. None of the isolates had a unique requirement for urea or ammonia since nitrogen derived from peptones, amino acids, or both supported growth as well as did ammonia or urea in a low nitrogen medium. Of the cecal isolates, 18% were able to use urea for growth, and 20.5% were able to grow with ammonia as the sole nitrogen source. All organisms grew in the experimental media containing peptones as the sole nitrogen source. Urease activity was detected in only 2 of 114 isolates tested. The inability of isolates to use urea or ammonia as nitrogen sources may have been a reflection of growth conditions in the habitat-stimulating medium used for isolation, but it could also suggest that many cecal bacteria require nitrogen sources other then ammonia or urea for growth.     

Studies of two Frankia strains isolated from Trevoa trinervis Miers

The Frankia strains TtI 11 and TtI 12 isolated from T. trinervis Miers were characterized regarding their carbon source utilization, intrinsic antibiotic resistance, infectivity, and effectivity on the original host. Both strains grew on BAP medium supplemented with glucose, maltose, and sucrose, but differed in their ability to use other carbon sources such as propionate, pyruvate, acetate, succinate, citrate, and mannitol. The isolates were sensitive to five of the twelve antibiotics tested at 1 μg mL⁻¹ concentration: chloramphenicol, tobramycin, eritromycin, streptomycin, and rifampicin. They exhibited a variable degree of resistance at 1 μg mL⁻¹ concentraction to penicillin G, 4-fluorouracil, oleandomycin, and lincomycin. Both isolates were able to infect and nodulate the original host plant, and thus represent the first reported infective and effective microsymbionts for T. trinervis Miers, a rhamnaceous actinorhizal host. R O D Dixon Section editor     

Virulence of Vibrio vulnificus strains from marine environments

Vibrio vulnificus strains isolated from geographically diverse marine sources were compared with clinical isolates for phenotype and in vitro and in vivo production of virulence factors. There were no differences between environmental and clinical strains on the basis of biochemical characteristics or antimicrobial susceptibility patterns. Cytolysin and cytotoxin titers produced by environmental strains were generally comparable to those of clinical strains. Of 29 environmental isolates tested, 25 were pathogenic for mice. These data show that environmental V. vulnificus strains are phenotypically indistinguishable from clinical isolates and that approximately 90% of the environmental strains tested produced in vitro virulence factors and in vivo pathogenicity for mice comparable to those produced by clinical V. vulnificus isolates.