Virulence and aggressiveness of single-zoospore isolates of Phytophthora fragariae

A system for scoring the virulence of isolates of Phytophthora fragariae based on a scale of root rot from 0 ( no symptoms ) to 5 (76-100% roots roiled) on a series of strawberry cultivars is described. Thirty-two single-zoospore isolates from one field site were compared by subjecting their root rot scores to cluster analysis and this grouped them into two major clusters equivalent to physiologic races B66–3 and B66-11, Different sub-clusters of isolates of race B66-11 produced different degrees of rotting on the same hosts. Apart from differences in virulence between the sub-clusters there was some evidence for differences in aggressiveness between isolates within sub-clusters.
Increasing inoculum concentration by over 300-fold increased rotting by c . 25% but did not alter the rankings of different isolate/host combinations. Repeated passage of isolates through cultivars of differing susceptibilities did not affect their pathogenicity.     

Virulence characteristics of apple scab (Venturia inaequalis) isolates from monoculture and mixed orchards

Isolates of Venturia inaequalis were sampled from monoculture and mixed orchards of three apple cultivars: Bramley, Cox and Worcester. In addition, single-ascospore progeny isolates were obtained from three crosses between pairs of isolates originating from the three cultivars in monoculture orchards. These field isolates and single-ascospore progenies were inoculated onto each of the three cultivars in a glasshouse. The patterns of infection showed that all three cultivars, commonly regarded as susceptible to scab, contained some specific resistance factors and that scab isolates from both mixed and monoculture orchards appeared to have co-evolved with cultivars. A much higher proportion of isolates from cv. Worcester in the mixed orchard were unable to infect any of the three cultivars than isolates from any other combination of cultivar and orchard type, but there was no other difference between isolates from mixed vs. monoculture orchards. Many isolates could infect both Bramley and Cox, or Cox and Worcester; but only a single isolate could infect both Bramley and Worcester. Results from the testing of 61 single-ascospore progeny isolates suggested that virulence towards Bramley, Cox and Worcester was controlled by at least one, two and three factors, respectively. Moreover, the exact nature of the interactions between these factors in determining virulence depended on the particular pair of isolates concerned.     

Sensitivity of cells and protoplasts of hop cultivars to cytotoxic components of culture filtrates of Verticillium albo-atrum isolates from hop

Leaf mesophyll protoplasts and cell suspension cultures obtained from resistant and susceptible cultivars of hop ( Humulus lupulus ) exhibited differential cell death responses when treated with sterile culture filtrates (CF) obtained from four isolates of Verticillium albo-atrum showing a range of virulence towards hop. The extent of cytotoxicity was correlated with both the virulence of the isolates and the resistance of the cultivars. Cytotoxicity was attributed to a low-molecular-weight, heat-labile, component of CF. Cell cultures from a susceptible and a resistant cultivar exhibited similar, growth-related, patterns of sensitivity to CF. The extent of inhibition of callus growth caused by CF treatment was related to the level of the resistance of the cultivar.     

Responses of cucumber cultivars to induction of systemic resistance against anthracnose by plant growth promoting fungi

Initial experiment on the reactions of five Japanese cultivars of cucumber toColletotrichum orbiculare infection in the greenhouse revealed that cv Suyo and Gibai were susceptible and moderately susceptible, respectively, while cv Shogoin fushinari and Sagami hanjiro were resistant to infection byC. orbiculare; cv Ochiai fushinari was moderately resistant. The ability of 16 plant growth promoting fungi (some isolates belonged to species ofPhoma and some non-sporulating isolates) isolated from zoysiagrass rhizospheres to induce systemic resistance in the above five cucumber cultivars was tested by growing plants in potting medium infested with barley grain inocula of PGPF in the greenhouse. The second true leaves of 21-day-old plants were challenge inoculated withC. orbiculare and disease assessed. Nine, out of 16 isolates, caused significant reduction of disease caused byC. orbiculare in at least two cultivars.Phoma isolates (GS8-1 and GS8-2) and non-sporulating isolates (GU21-2, GU23-3, and GU24-3) significantly reduced the disease in all the five cultivars. The disease suppression in cucumber was due to the induction of systemic resistance, since the inducer(s) and the pathogen were separated spatially and that the inducer did not colonize aerial portions. The resistance induced by certain isolates in a susceptible cultivar was less than that in a resistant cultivar. Disease suppression caused by isolate GU21-2 was similar to theC. orbiculare induced control in certain cultivars. The average rate of expansion of lesion diameter on leaves due toC. orbiculare was slower due to induction with the selected plant growth promoting fungi compared to the uninduced control plants. Roots of four cultivars were colonized by only three isolates, however, roots of one cultivar (Suyo) was colonized by five isolates suggesting the cultivar-specific root colonization ability.Abbreviations cv cultivar(s) - PGPF plant growth promoting fungal isolates - PGPR plant growth promoting rhizobacteria     

A Japanese powdery mildew isolate with exceptionally large infection efficiency on Mlo-resistant barley

A Japanese field isolate (Race I) of Erysiphe graminis f.sp, hordei was tested on 17 barley lines carrying the mlo powdery mildew resistance gene. Race I produced many successful infections with infection type larger than or equal to 2 on six lines (M66, MC20, SRI, SR7, A tem and Totem), On the remaining 11 lines it reacted with infection type 0. Colony numbers on the Mlo-lines were between 26% and 12 9% of the numbers on a susceptible cultivar Manchuria, These numbers were larger than, or similar to those produced by isolate HL3/5, which has the highest recorded infection efficiency on Mlo-resistant cultivars. The interaction between isolates and lines was highly significant. The isolate GE3, from which HL3/5 was derived by selection, gave rise to occasional colonies corresponding to less than 03° o of the number produced on cv. Manchuria.     

Host-pathogen relationship of geographically diverse isolates of Septoria tritici and wheat cultivars

Pathogenic variability of 14 Septoria tritici isolates from different locations in the USA (California, Oregon, and Texas) was determined on seedlings of two sets of geographically diverse wheat cultivars under greenhouse conditions. Significant isolate effects, cultivar effects, and isolate × cultivar interactions were found, and a substantial amount of variation was accounted for by the interaction terms compared with the main effects of isolate and cultivar. All isolates were pathogenic on the cultivars tested but the degree of virulence on the individual cultivars varied among isolates. Linear contrasts between all homologous combinations (isolate × cultivar combination of same geographic location) and all heterologous combinations (isolate × cultivar combination of different locations) indicated that homologous combinations produced significantly more disease than heterologous combinations. The results demonstrate location-specific adaptation of S. tritici. Implications of pathogenic variability and local adaptation in S. tritici are discussed.     

A new aggressive strain of Verticillium albo-atrum in Verticillium resistant cultivars of tomato in the Netherlands

In 1991 serious losses caused byVerticillium wilt were found on two holdings in the Westland glasshouse district in the Netherlands in which theVerticillium resistant tomato cultivars Calypso and Criterium were grown in soilless systems. Isolates from diseased plants were identified asVerticillium albo-atrum.In inoculation experimentsVerticillium resistant tomato cultivars were seriously affected by the new isolates but not by a control isolate. Moneydor, a susceptible cultivar without the Ve gene, was the most seriously diseased by all isolates. The isolates from theVerticillium resistant tomato cultivars were less virulent on the susceptible cultivar than the control isolate.     

Characterization of Ascochyta isolates and susceptibility of pea cultivars to the ascochyta disease complex in Alberta

The relative virulence of 109 Ascochyta isolates collected from pea fields in Alberta from 1996 to 1998 were evaluated on 10-day-old seedlings by the excised leaf-assay technique. Twenty-eight isolates were avirulent, while the others produced lesions of various sizes on pea leaves. DNA samples from 86 isolates were amplified by the RAPD technique using PCR with single primers. One dominant genotype of Ascochyta pisi was identified throughout Alberta, but variations in virulence were not clearly differentiated by the RAPD technique. Five Ascochyta isolates, four virulent and one avirulent, were used to assess the susceptibility of 20 field pea cultivars available in Alberta, including 13 yellow types and seven green types. Based on symptom development, the yellow-type cultivars Swing, Eiffel and Delta, and the green-type cultivar Orb, were the most susceptible. Of yellow-type cultivars, Voyageur, Carneval and Montana were most resistant to A scochyta infection.     

小麦条锈菌异核新菌系的筛选及核游离试验研究

 采用生物学和细胞学方法研究小麦条锈菌的异核作用。不同小种的单孢菌系间的混合接种繁殖筛选出三个新菌系。新菌系的毒性明显不同于相应的原始菌系,新菌系夏孢于芽管的多核率较原始菌系高,这表明三个新菌系是经异核作用产生的。异核体的多核率随继代接种而降低,但有的菌系的毒性仍保持稳定。在采自陇南自然条件的条锈菌的夏孢子群体中,存在有3核和4核体,感病品种上夏孢子的多核率明显高于抗病品种。这一结果表明小麦条锈菌异核作用在感病品种上的发生机率较在抗病品种上的为高。     

Occurrence of two races of Puccinia kuehnii causing orange rust of sugarcane in Florida

Orange rust of sugarcane caused by Puccinia kuehnii was first reported in Florida in 2007. Since then, several sugarcane cultivars that were resistant during the initial epidemics became susceptible within a few years. These shifts in resistance were attributed to the evolution of the pathogen and appearance of new races. To study the variation in virulence of P. kuehnii, healthy leaf pieces of sugarcane cultivars susceptible to orange rust were brush inoculated with isolates of P. kuehnii collected from susceptible cultivars in the field. After inoculation, leaf pieces were placed in an incubator and disease severity based on the number of rust uredinia was determined 2 weeks postinoculation. Isolates of P. kuehnii collected from sugarcane cultivar CP 89-2143, which only showed severe symptoms of orange rust starting in 2011–2012, produced 300%–500% more uredinia on CP 89-2143 than the isolates collected from cultivar CL 85-1040 that has been susceptible since 2007. Sugarcane cultivar CL 85-1040 exhibited high and equivalent numbers of uredinia regardless of the inoculated isolate of the pathogen. These data support the occurrence of pathogenic specialization within P. kuehnii and the existence of at least two races of this pathogen in Florida. Analysis of amplified fragment-length polymorphism among isolates of P. kuehnii from cultivars CP 89-2143 and CL 85-1040 differing in resistance to orange rust revealed genetic variation among rust uredinia. However, this variation was not associated with a specific sugarcane cultivar, suggesting that pathogenic variation was not linked to major, but rather to small genetic changes within the genome of P. kuehnii.     

Identification of an Avirulence Gene in the Fungus Magnaporthe grisea Corresponding to a Resistance Gene at the Pik Locus

ABSTRACT A rice isolate of Magnaporthe grisea collected from China was avirulent on rice cvs. Hattan 3 and 13 other Japanese rice cultivars. The rice cv. Hattan 3 is susceptible to almost all Japanese blast fungus isolates from rice. The genetic basis of avirulence in the Chinese isolate on Japanese rice cultivars was studied using a cross between the Chinese isolate and a laboratory isolate. The segregation of avirulence or virulence was studied in 185 progeny from the cross, and monogenic control was demonstrated for avirulence to the 14 rice cultivars. The resistance gene that corresponds to the avirulence gene (Avr-Hattan 3) is thought to be located at the Pik locus. Resistance and susceptibility in response to the Chinese isolate in F(3) lines of a cross of resistant and susceptible rice cultivars were very similar to the Pik tester isolate, Ken54-20. Random amplified polymorphic DNA markers and restriction fragment length polymorphism markers from genetic maps of the fungus were used to construct a partial genetic map of Avr-Hattan 3. We obtained several flanking markers and one co-segregated marker of Avr-Hattan 3 in the 144 mapping population.     

Spatial-Temporal and Quantitative Analysis of Growth and EPS I Production by Ralstonia solanacearum in Resistant and Susceptible Tomato Cultivars

ABSTRACT One susceptible and two resistant cultivars of tomato were tested for differences in infection by Ralstonia solanacearum and for the subsequent multiplication, colonization, and production of the wilt-inducing virulence factor, exopolysaccharide I (EPS I). Bacterial ingress into the taproot was fastest in the susceptible cv. Marion, followed by the resistant cvs. L285 (fivefold slower) and Hawaii 7996 (15-fold slower). Once inside the taproot, R. solanacearum colonized, to some extent, almost all regions of the resistant and susceptible plants. However, colonization occurred sooner in the susceptible than in the resistant cultivars, as measured by viablecell counts of bacteria in the midstems. Rates of multiplication and maximum bacterial cell densities were also greater in the susceptible than in the resistant cultivars. Growth experiments utilizing xylem fluid from infected and uninfected plants indicated that neither antimicrobial activities nor reduced levels of growth-supporting nutrients in the xylem fluids were responsible for the reduced bacterial multiplication in the resistant cultivars. Quantification of EPS I in the infected plants, using an enzyme-linked immunosorbent assay, revealed that the bacterial populations in the susceptible cultivar produced greater amounts of EPS I per plant than those in the resistant cultivars. Immunofluorescence microscopy using antibodies against either EPS I or R. solanacearum cells revealed that bacteria and EPS I were distributed throughout the vascular bundles and intercellular spaces of the pith in the susceptible cultivar, whereas in the resistant cultivars, bacteria and EPS I were restricted to the vascular tissues.     

Evaluation of antisera raised against Phytophthora fragariae for detecting the red core disease of strawberries by enzyme-linked immunosorbent assay (ELISA)

Antiserum was raised against pooled mycelial suspensions from five isolates (designated Pf 1, Pf 2, Pf 3, Pf 10 and Pf 11) representing five physiologic races of Phytophthora fragariae. In enzyme-linked immunosorbent assay (ELISA), this antiserum detected homologous soluble antigens at protein concentrations as low as 2 ng/ml.
Fungal antigens could also be detected in extracts of strawberry plants infected with P. fragariae. Root extracts prepared from the alpine strawberry Fragaria vesca and F. ananassa cv. Cambridge Favourite infected with any of the five isolates studied produced strong reactions in ELISA. In F. vesca , ELISA-positive material was detectable 6-8 days after inoculation before macroscopic symptoms appeared. The cultivar Red Gauntlet, which is resistant to Pf 1, 2 and 3 but susceptible to Pf 10 and 11, reflected this differential response in ELISA; the absorbance produced by extracts of plants infected with virulent isolates was significantly higher than that obtained with the corresponding extracts of plants inoculated with a virulent isolates. The recently introduced cultivars Hapil, Ostara and Providence were found to be susceptible to all isolates in this study: the corresponding root extracts were also positive in ELISA.
The antiserum also detected P cactorum infections. Nevertheless, the ELISA test described should prove valuable in screening certified strawberry stocks.     

Geographic location and year determine virulence,and year determines genetic change,in populations of Neopseudocercosporella capsellae

White leaf spot (Neopseudocercosporella capsellae) can be severe and problematic worldwide across both horticultural and oilseed Brassicaceae, including susceptible rapeseed. In this study, 82 isolates from 2015 and 106 isolates from across Australia in 2016 were first assessed for their virulence against three different rapeseed (Brassica napus) cultivars. For both years there were significant (P < 0.001) differences. Also, there were significant (all P < 0.001) differences between isolates in each year, and between cultivars. For 2016 isolates, there were also significant differences (P < 0.001) between isolates across three different Australian states, and a significant interaction (P < 0.001) between isolates with cultivars. Of the three Australian states, isolates from Victoria were most virulent. Among tested cultivars, cv. Scoop was most susceptible. Subsequently, phylogenetic analysis of 114 of these same 2015 and 2016 isolates showed current isolates clustered separately from the majority of 2005 N. capsellae isolates collected from Western Australia a decade earlier, confirming significant genetic change within N. capsellae populations over the past decade. However, isolate clusters showed no association with geographical location. The results suggest that phylogenetic association among 2005 and 2015–2016 N. capsellae isolates is complementary with pathogenicity variations explained by geographically different N. capsellae pathogen populations. Neopseudocercosporella capsellae populations are evolving rapidly, challenging management through host resistance at a time of increasing incidence and severity of white leaf spot disease over the past decade. The outcome is well illustrated by cv. Scoop, previously resistant to 2005 isolates but moderately susceptible to 2015 and highly susceptible to 2016 isolates.     

Towards the Development of a Novel In Vitro Strategy for Early Screening of Fusarium Ear Blight Resistance in Adult Winter Wheat Plants

A novel in vitro bioassay is described for screening Fusarium ear blight (FEB) resistance in adult winter wheat plants. Seven winter wheat cultivars were assessed for components of partial disease resistance as 28 day-old detached leaf segments in the laboratory using isolates of Microdochium nivale var. nivale and M. nivale var. majus. Results were compared with disease data obtained at anthesis using the same cultivars as whole plants and the same isolates under glasshouse conditions. Significant cultivar differences were observed using detached leaves, with cv. Avalon (a Fusarium culmorum ear susceptible cultivar) having the shortest leaf incubation period, greatest leaf lesion development and shortest leaf latent period compared to cv. Spark (a Fusarium culmorum ear resistant cultivar), which had the longest leaf incubation period, least leaf lesion development and longest leaf latent period. Using whole plants, cv. Avalon had the shortest ear incubation period and greatest ear disease severity, whilst cv. Spark had the longest incubation period and least ear disease severity. Overall, cultivars of intermediate F. culmorum ear resistance expressed intermediate responses to M. nivale isolates, using both detached leaves and whole plants. Significant correlations were found with ear disease severity and ear incubation period in whole plants and components of partial disease resistance in detached leaves, with significant correlations obtained between leaf incubation period and ear disease parameters using the M. nivale var. nivale isolate. In addition, leaf latent period and leaf lesion size showed significant correlations with whole plant reactions using M. nivale var. nivale and var. majus isolates. The in vitro screening of cultivars as detached leaves using M. nivale isolates may offer a real possibility of a rapid bioassay for the early screening of FEB resistance in wheat and other cereals.     

Genetic Variation for Virulence and Resistance in the Wheat-Mycosphaerella graminicola Pathosystem III. Comparative Seedling and Adult Plant Experiments

ABSTRACT Fourteen Dutch Mycosphaerella graminicola isolates were studied for their virulence to 22 wheat cultivars in the seedling stage in an experiment set up as a completely randomized block design with three repetitions. Isolate x cultivar interactions were highly significant. Cluster analyses were applied to select three isolates with significantly different virulence characteristics for both disease parameters. These were retested in the seedling stage and used to inoculate two field experiments that were planted according to a split plot design in 1992 and 1995. Overhead inoculations were conducted after flowering to avoid the effects of plant height; hence, these experiments were intended as monocyclic tests for virulence differences between the isolates. Significant isolate x cultivar interactions were detected in each experiment, demonstrating specificity in the wheat-M. graminicola pathosystem in the adult plant stage under field conditions. The reproducibility of the adult plant data was high. Genetic differences among the isolates were additionally demonstrated by randomly amplified polymorphic DNA (RAPD) patterns, which also showed that no significant contamination of the inoculated plots with the natural M. graminicola population had occurred. Rank correlations between seedling and adult plant data were significant for M. graminicola isolates IPO323 and IPO290, but not for isolate IPO001. Hence, evaluation of resistance and virulence may require seedling, as well as adult plant, tests.     

The incidence of barley yellow dwarf virus isolates in perennial ryegrass (Lolium perenne) in south-western Victoria

Barley yellow dwarf virus (BYDV) was detected in perennial ryegrass cultivars examined from three sites in south-western Victoria (Balmoral, Mininera and Hamilton); the frequency of BYDV infection varied with cultivar. PAV-related isolates were prevalent at all sites, but the incidence of MAV- and RPV-related isolates varied with the site and cultivar. The PAV-related isolate was less frequent in cv. Ellett than in cv. Victorian at all sites. The implications for the epidemiology of the disease and its agronomic importance in Victoria are discussed.     

Aggressiveness of Mycosphaerella graminicola Isolates from Susceptible and Partially Resistant Wheat Cultivars

ABSTRACT The selective effect of quantitative host resistance on pathogen aggressiveness is poorly understood. Because two previous experiments with a small number of bread wheat cultivars and isolates of Mycosphaerella graminicola had indicated that more susceptible hosts selected for more aggressive isolates, we conducted a larger experiment to test that hypothesis. In each of 2 years, six cultivars differing in their levels of partial resistance were planted in field plots, and isolates were collected from each cultivar early and late in the growing season. The isolates were inoculated as populations bulked by cultivar of origin, field replicate, and collection date on seedlings of the same six cultivars in the greenhouse. The selective impact of a cultivar on aggressiveness was measured as the difference in aggressiveness between early and late isolates from that cultivar. Regression of those differences on disease severity in the field yielded significance values of 0.0531 and 0.0037 for the 2 years, with moderately resistant cultivars selecting for more aggressive isolates. In a related experiment, the protectant fungicide chlorothalonil was applied to plots of two susceptible cultivars to retard epidemic development. When tested in the greenhouse, isolates of M. graminicola from those plots were significantly more aggressive than isolates from the same cultivars unprotected by fungicide.     

Dual control of avirulence in Leptosphaeria maculans towards a Brassica napus cultivar with 'sylvestris-derived' resistance suggests involvement of two resistance genes

Blackleg disease (phoma stem canker) of Brassica napus (canola, oilseed rape) is caused by the fungus Leptosphaeria maculans . In some regions of Australia, resistance in oilseed rape cultivars derived from B. rapa subs . sylvestris (e.g. cv. Surpass 400) became ineffective within three years of commercial release. The genetic control of avirulence in L. maculans towards cv. Surpass 400 is described. When Australian field isolates were screened on this cultivar, three phenotypic classes were observed; virulent, intermediate and avirulent. Analysis of crosses between fungal isolates varying in their ability to infect cv. Surpass 400 demonstrated the presence of two unlinked avirulence genes, AvrLm1 and AvrLmS . Complementation of isolates (genotype avrLm1 ) with a functional copy of AvrLm1 , and genotyping of field isolates using a molecular marker for AvrLm1 showed that virulence towards Rlm1 is necessary, but not sufficient, for expression of a virulent phenotype on cv. Surpass 400. Taken together, these data strongly suggest that cv. Surpass 400, with ' sylvestris -derived' resistance, contains at least two resistance genes, one of which is Rlm1 .