Variation for virulence in Erysiphe fischeri from Senecio vulgaris

Variation for virulence was investigated in two populations of Erysiphe fischeri , the cause of powdery mildew of the common annual weed Senecio vulgaris (groundsel). Infection types were recorded on detached leaf segments from 50 inbred lines of S. vulgaris following inoculation with 24 single-conidial-chain isolates of E. fischeri (12 each from two UK sites; one, at Glasgow, located about 480 km north of the other, at Wellesbourne). Mean infection types for each isolate/line combination were categorized by applying several upper limits below which combinations were considered to be incompatible. Regardless of the limit applied, numerous specific virulence and resistance phenotypes could be discriminated. Virulence phenotypes were complex and all isolates were capable of colonizing and reproducing on the majority of groundsel lines. However, all isolates were completely avirulent (no reproduction) on at least one groundsel line, so no isolate was universally virulent. Plants of several groundsel lines exhibiting different resistance phenotypes were exposed to natural infection at Wellesbourne to act as mildew traps, to examine the virulence characteristics of the pathogen population. Common components of the fungus population colonized groundsel lines exhibiting resistance to only a few or none of the single-conidial-chain isolates. Rarer components of the pathogen population colonized groundsel lines with resistance to the majority or all of the isolates previously tested. These latter components of the fungus population also detected resistance in groundsel lines previously recorded as susceptible to all isolates.     

Effects of preinoculation with an avirulent isolate of Pyricularia grisea on infection and development of leaf blast lesions caused by virulent isolates on near-isogenic lines of Sasanishiki rice

Eight near-isogenic Sasanishiki rice lines with different genes for complete resistance to rice blast were inoculated with an avirulent isolate 72 h before inoculation with a virulent isolate of Pyricularia grisea to clarify the mechanisms of induced resistance in the leaf blades. Subsequent lesions on the leaf blades were classified as brown spots (b type), as observed on Sasanishiki BL no. 8 with resistance gene Pii, or no symptoms (HR type), as observed on Sasanishiki BL no. 4 with the gene Piz-t and on the six other lines. Lesion expansion was significantly reduced in Sasanishiki BL no. 8 compared with that in Sasanishiki BL no. 4 when the leaf blades were preinoculated with a high concentration of a conidial suspension of the avirulent isolate. Moreover, after preinoculation with the conidial suspension of the avirulent isolate in silicon rings on the leaf blades, induced resistance was expressed only in areas close to the inoculation sites. These resistant areas were larger in Sasanishiki BL no. 8 than in BL no. 4. Hyphal growth was markedly inhibited in the epidermal cells of Sasanishiki BL no. 8, whereas inhibition was weak in those of Sasanishiki BL no. 4. In the epidermal cells of leaf blades of Sasanishiki BL no. 8 preinoculated with the avirulent isolate, the frequency of hyphal penetration of the virulent isolate in the presence of host cell browning decreased, as did the frequency of invading hyphae after inoculation with virulent isolates. The results indicate that induced resistance may play a role in the suppression of lesion development in the Sasanishiki near-isogenic lines and that the lines differ in the extent of suppression.     

An integrated multivariate approach to net blotch of barley: Virulence quantification,pathotyping and a breeding strategy for disease resistance

Knowledge of pathotype diversity and virulence in local populations of Pyrenophora teres is a prerequisite to screening for durable resistance to net blotch. The current study aimed to quantify the virulence level of Moroccan isolates, identify and designate existing pathotypes, and select resistant genotypes. We developed a method for virulence quantification of P. teres isolates based on a conversion of infection responses into frequencies for use in correspondence analysis. Coordinates of the first axis of this analysis had a virulence spectrum and ranked isolates from virulent to avirulent. Mixed model analysis was also devised for virulence quantification. Coordinates of the first dimension of correspondence analysis were linearly correlated to BLUPs (Best Linear Unbiased Predictors) of the mixed model. A genotype by genotype by environment model (GGE) coupled with cluster analysis differentiated P. teres isolates into ten and nine pathotypes for net- and spot-forms respectively. Populations of these two forms were dissimilar in terms of classes of virulence. For P. teres f. maculata, avirulent, moderately virulent and highly virulent isolates represented one-third of the population, whereas 90% of P. teres f. teres population was composed of avirulent to moderately avirulent isolates. Barley differential sets were subsequently reduced to two new sets that simplified pathotyping through a key code based on resistant or susceptible reactions. Dendrograms of cluster analysis based on GGE analysis depicted the stability of a genotype’s reactions across all isolates, and using only resistant cultivars as sources of resistance to control net blotch disease would, based on this analysis, fail to control all pathotypes. Therefore, we propose an alternative breeding strategy to control net blotch effectively.     

Induction of a defense response in strawberry mediated by an avirulent strain of <Emphasis Type="Italic">Colletotrichum</Emphasis>

In the strawberry crop area of Tucumán (north-west Argentina) the three species of Colletotrichum causing anthracnose disease (C. acutatum, C. fragariae and C. gloeosporioides) were detected. Among all isolates characterized, one of them identified as C. acutatum (M11) and another as C. fragariae (F7) were selected due to their conspicuous interaction with the strawberry cultivar Pájaro. Whereas isolate M11 produced a strong compatible interaction in cv. Pájaro with clear disease symptoms (DSR = 5.0), the isolate F7 brought about a typical incompatible interaction (DSR = 1.0). When plants of cv. Pájaro were inoculated with F7 prior to the inoculation with M11, the former avirulent strain prevented the growth of the latter virulent pathogen. Experimental evidence indicated that the time elapsed between the first inoculation with the avirulent pathogen and the second inoculation with the virulent one was crucial to inhibit the growth of the latter. The growth of F7 on the plant without provoking damage and the fact that there was no in vitro antagonistic effect between the pathogens, suggests that the avirulent strain triggers a plant defensive response against M11. The defense response was further confirmed by the detection of an early oxidative burst occurring within 4 h after the first inoculation and by the observation of anatomical changes associated with defense mechanisms that lasted 50 days after the inoculation with F7. Results obtained support the hypothesis that the plant resistance against the virulent strain M11 is elicited by one or more diffusible(s) compound(s) produced by the avirulent strain F7.     

The identification of a gene for race-specific resistance to Peronospora parasitica (downy mildew) in Brassica napus var. oleifera (oilseed rape)

The oilseed rape cultivar Cresor was resistant to 14 isolates of Peronospora parasitica derived from crops of Brassica napus in the UK. Segregation for resistance to one isolate among F₂ plants and F₃ progeny of crosses between Cresor and the susceptible cultivars Victor and Jet Neuf indicated that resistance was controlled by a single gene. There was evidence that genetic background and environment could influence the phenotypic expression of this resistance. Two sexual progeny isolates derived from a homothallic isolate of P. parasitica avirulent on Cresor were completely virulent on this cultivar. This suggested that the parental isolate was heterozygous at a matching locus or loci for avirulence and demonstrated the race-specific nature of the resistance.     

Genetic Analysis of Avirulence/Virulence of an Isolate of Magnaporthe grisea from a Rice Field in Texas

ABSTRACT An isolate of Magnaporthe grisea, Tm4, from a rice field in Texas was crossed with a fertile laboratory strain, 70-6. The progenies showed segregation of avirulence/virulence on rice cvs. Newbonnet, Lemont, Lebonnet, Leah, and Katy. The avirulent/virulent segregation ratios were 29:6 on Newbonnet, Lemont, and Lebonnet; 28:7 on Leah; and 33:2 on Katy. There was cosegregation on the first three cultivars. Several avirulent progenies were backcrossed to virulent parent 70-6. Three generations of backcrossing avirulent progenies to 70-6 led to segregation ratios that suggested certain strains had only one avirulence gene. Strains avirulent only on cv. Katy or only on cvs. Newbonnet, Lemont, and Lebonnet were test crossed with virulent siblings. Strains that gave progeny ratios approximating 1 avirulent:1 virulent when crossed with virulent siblings were selected for further test crossing. Intercrosses between strains with possible single avirulence genes were made to determine whether these strains had the same or different avirulence genes. Many lines still segregated two genes for avirulence after three generations of backcrossing. This is based on the recovery of virulent progenies from crossing two avirulent siblings.     

Resistance to Erysiphe fischeri in two populations of Senecio vulgaris

The frequency and distribution of different specific phenotypes for resistance to Erysiphe fischeri was studied in two populations of the annual weed Senecio vulgaris (groundsel) one located in Glasgow, Scotland the other located about 480 km south at Wellesbourne, England. Progeny of individual plants from the two host populations were tested for their response to up to 10 different isolates of E. fischeri , five from each location; each isolate had a different specific virulence phenotype. Most plants in each sample were susceptible to all 10 isolates. The proportion of plants whose progeny were resistant to a particular isolate ranged from 1% to 10% with the exception of resistance to one isolate that occurred with a frequency of 37% at Wellesbourne. Overall, resistance to one or more of the 10 isolates appeared to be more common in the plant population sampled at Wellesbourne than at Glasgow. Of the total number of groundsel line/isolate combinations tested, 10% involving Wellesbourne plants and 2% involving Glasgow plants were incompatible, i.e. resistant/avirulent. Both groundsel populations tended to be dominated by one or two resistance phenotypes but they were nevertheless highly heterogeneous when less frequent resistance phenotypes were considered. This was particularly evident at Wellesbourne where 10 different resistance phenotypes were recorded amongst a total of 75 plants growing within an area of 1 m².     

Host-mediated interactions between pathogen genotypes

Barley c ltivars with different resistance genes were inoculated with various sequences of virulent and non-virulent isolates of Erysiphe graminis f.sp. hordei. The resistance induced in incompatible reactions decreased infection by virulent isolates while, conversely, the degree of susceptibility induced in compatible reactions was sufficient to increase the ability of normally non-virulent isolates to infect a resistant host. The net effect of interactions was always overall reduction of conidial production when compared with inoculation of a virulent isolate alone.
Each interaction between isolates was dependent on the host cultivar, the pathogen isolates used and on the inoculation sequence. Often, induced resistance was most obvious when a nonvirulent isolate was inoculated prior to a virulent isolate. Induced susceptibility was usually more apparent when the virulent isolate was inoculated first. However, on the cultivar Wing, induced susceptibility was more apparent following prior inoculation of a non-virulent isolate. On the cultivar Hassan, prior inoculation with a virulent isolate sometimes induced susceptibility, but in some cases more susceptibility was induced by prior inoculation with a non-virulent isolate. In the latter instances, although relatively little infection developed, the majority of the spores produced were those of the normally non-virulent isolate.
There was some evidence of non-virulent isolates acquiring the ability to infect normally incompatible hosts. This effect disappeared after several generations in the absence of the virulent isolate.     

Race-specific elicitors from the Peronospora parasitica / Arabidopsis thaliana pathosystem

Intercellular washing fluids (IWFs) were isolated from compatible interactions ofPeronospora parasitica with Arabidopsis. IWFs were filter-sterilized then infiltrated into leaves of resistant and susceptible Arabidopsis genotypes. P. parasitica isolates WELA (virulent on Landsberg erecta (La-er) and Weiningen (Wei-0) but avirulent on Columbia (Col-0) and RLD) and NOCO (virulent on Col-0 and RLD but avirulent on La-er) were used. A “reciprocal check” pattern of necrosis-induction was observed in which IWFs from a compatible interaction caused necrosis in leaves of the Arabidopsis genotypes resistant to the “parent” fungal isolate but not in those genotypes susceptible to it. Analysis of variance showed that the interaction between fungal isolate and necrosis in host genotypes was highly significant. The necrosis-inducing activity was significantly correlated with a high degree of asexual sporulation in the compatible interaction from which the IWF was isolated. However, the necrosis-inducing activity was not correlated with the protein content of the IWF.     

Induced resistance in host mixtures and its effect on disease control in computer-simulated epidemics

The epidemic simulator EPIMUL was modified and used to study how induced resistance affected the development of epidemics in host mixtures. In the model, induced resistance resulted from the interaction of host tissue with avirulent spores and caused a reduction in the efficacy of virulent spores deposited afterwards. We denned three parameters to describe induced resistance: the level of protection, defined as the magnitude of reduction in the virulent spore efficacy for infecting host tissue; the host surface area protected by an interaction with one avirulent spore; and the duration of protection of the host tissue, in days. In our simulations, induced resistance slowed the epidemics and gave better disease control in the mixtures, even if protection lasted for only 2 days. The disease reduction in the mixture attributable to induced resistance was approximately proportional to the level of protection. The effect of induced resistance increased as the protected area increased. Epidemics were virtually unaffected by induced resistance restricted to the infection site, but the effect of induced resistance initially increased rapidly as larger areas were protected. There was little further gain as the protected area increased from 2·6% to 26%. The influence of induced resistance was reduced when the interactions between virulent and avirulent pathogens were reduced.     

Genetic Variation and Virulence on Lr26 in Puccinia triticina

ABSTRACT The genetic relationships between isolates of Puccinia triticina virulent on wheat with the Lr26 resistance gene were studied. The diversity within and between isolates of P. triticina from Israel, Europe, and the United States was determined by virulence on near-isogenic Thatcher lines and by random amplified polymorphic DNA. According to the molecular markers, isolates that were virulent on Lr26 had diversity levels similar to those of Lr26 nonpathogenic isolates. Distances between subpopulations of isolates virulent and avirulent on Lr26 varied and were unrelated to the Lr26 virulence phenotype. Cluster analysis suggested four groups, three of which were closely associated with the geographical origin of the isolates-Israel, the United States, and Europe. All four groups included both Lr26 virulent and avirulent pathotypes. The results showed that Lr26 virulent rust pathotypes are as genetically dissimilar as the rest of the population. The cluster analysis showed that the rust population in Israel includes at least two different subpopulations, both of which contain Lr26 virulent and Lr26 avirulent isolates.     

Efficacy of the R2 resistance gene as a component for the durable management of potato late blight in France

Many race-specific resistance genes to potato late blight are overcome in France, but the disease appears later on genotypes carrying the R2 gene. This study examined whether R2 could contribute to durable late-blight control in France, and analysed the conditions of its performance. Plants grown from tubers of different physiological ages showed no difference in R2 expression in field and climate-chamber experiments, demonstrating that the delay in epidemic onset provided by R2 was not the result of gene inactivation in old plants. Among isolates collected at one site, those virulent on R2 were classified into three AFLP profiles. AFLP-VII comprised exclusively isolates virulent to R2 , whereas AFLP-IV and AFLP-V included both virulent and avirulent isolates. No significant aggressiveness differences were observed between virulent and avirulent isolates from AFLP-V; however, isolates from AFLP-VII were significantly less aggressive than virulent isolates from AFLP-V. These results indicate that: (i) the delayed onset of epidemics on R2 cultivars is the result of the breakdown of R2 by virulent isolates; (ii) aggressiveness of isolates virulent to R2 depends primarily on the genetic background of the pathogen where the mutation to virulence occurs; and (iii) this mutation does not lead per se to lower pathogenic fitness. It is suggested that R2 is unlikely to make a lasting contribution to late-blight control in France, and that diversification strategies such as cultivar mixtures might not considerably increase its durability.     

小麦对白粉菌的抗性和感病性的诱导

 用小麦白粉菌(Erysiphe graminis f.sp.tritici)的无毒和有毒菌株,以3种接种顺序:1.先接无毒菌株,间隔一定时间后,再接有毒菌株;2.无毒和有毒菌株同时接;3.和"1"相反,接种具有不同抗性基因的小麦品种的幼苗或叶段,以研究小麦对白粉菌的诱导抗性和诱导感病性。结果表明:先接无毒菌株诱导的抗性最强;后接无毒菌株或无毒、有毒菌株同时接,当诱导菌(inducer)的密度很高时,也可以诱导出抗性,但较微弱;诱导的抗性随诱导菌的密度增高而增强;一经诱导,抗性可在被诱导部位维持到第6天;所使用的3种接种顺序都可以诱导小麦对白粉菌产生感病性,但较之诱导的抗性弱得多;在抗病性的诱导中,存在着小麦品种与诱导菌,或诱导菌与挑战菌(challenger)之间的相互作用;一个无毒菌株可以通过诱导而获得在抗病品种上生长的毒性,但当缺少有毒菌株存在时,获得毒性即消失;没有发现所诱导的抗性具有系统传导性。     

Virulent and avirulent isolates of Pseudomonas syringae subsp. savastanoi as colonizers of olive leaves: evaluation of possible biological control of the olive knot pathogen1

The epiphytic survival of virulent and avirulent (indoleacetic acid-deficient mutants) isolates of Pseudomonas syringae subsp. savastanoi, and their ability to colonize the olive phylloplane, was monitored for up to 30 days after artificial inoculation of several olive cultivars. After an initial decrease in numbers, the virulent bacteria multiplied and established a potential inoculum on olive leaf surfaces. In contrast, the numbers of avirulent bacteria rapidly diminished, so that they were eventually no longer detectable. Under the conditions used in this study, none of the avirulent mutants (producers or non-producers of bacteriocins) were able to compete with or successfully to inhibit the virulent isolate in vivo. This was probably due to the lack of epiphytic multiplication of the avirulent mutants.     

Field efficiency of Brassica napus specific resistance correlates with Leptosphaeria maculans population structure

Field experiments were conducted in Versailles, France, to assess blackleg resistance of Brassica napus cultivars Quinta and Glacier under natural infection conditions. Blackleg disease severity was assessed twice during growth of B. napus. Quinta resistance was highly expressed as only 13% to 18% of the plants exhibited leaf symptoms in December, whereas Glacier and other cultivars displayed more than 80% of infected plants. In June (harvest), 70% (first year) to 41.5% (second year) of Quinta plants were canker-free. In contrast, Glacier was as infected as the susceptible control cultivars, with more than 88% of plants displaying canker. The Leptosphaeria maculans population structure was examined in parallel. Based on soluble protein patterns, 9% of the 299 fungal isolates collected were characterized as Tox⁰ species, and belonged to the NA1 sub-group. All but two of Tox⁰ isolates were isolated from atypical dark necrotic leaf lesions, mainly occurring on Quinta. In contrast, the Tox⁺ isolates were recovered from typical leaf lesions. Following a cotyledon inoculation test on the differential set Westar, Quinta and Glacier, 92 to 95% of Tox⁺ isolates collected on susceptible cultivars were characterized as PG3 isolates, i.e. avirulent on Quinta. The remaining Tox⁺ isolates belong to PG4, i.e. virulent on the three cultivars. No PG2 isolate, i.e. avirulent on both Quinta and Glacier, was identified in the sampling. The present study suggests that specific resistance expressed at the cotyledon level can be efficient under field conditions where the corresponding avirulent races of the pathogen are prevalent.     

Quantitative trait loci associated with isolate specific and isolate nonspecific partial resistance to Phoma macdonaldii in sunflower

Black stem, caused by Phoma macdonaldii , is one of the most important diseases of sunflower in the world. Quantitative trait loci (QTLs) implicated in partial resistance to two single pycnidiospore isolates of P. macdonaldii (MP8 and MP10) were investigated using 99 recombinant inbred lines (RILs) from the cross between sunflower parental lines PAC2 and RHA266. The experimental design was a randomized complete block with three replications. High genetic variability and transgressive segregation were observed among RILs for partial resistance to P. macdonaldii isolates. QTL-mapping was performed using a recently developed high-density SSR/AFLP sunflower linkage map. A total of 10 QTLs were detected for black stem resistance. The phenotypic variance explained by each QTL (R²) was moderate, ranging from 6 to 20%. Four QTLs were common between two isolates on linkage group 5 and 15 whereas the others were specific for each isolate. Regarding isolate-specific and isolate-nonspecific QTLs detected for partial resistance, it is evident that both genetic effects control partial resistance to the disease isolates. This confirms the need to consider different isolates in the black stem resistance breeding programmes. The four SSR markers HA3700, SSU25, ORS1097 and ORS523_1 encompassing the QTLs for partial resistance to black stem isolates could be good candidates for marker assisted selection.     

A Molecular Marker Correlated with Selected Virulence Against the Tomato Resistance Gene Mi in Meloidogyne incognita, M. javanica, and M. arenaria

ABSTRACT Root-knot nematodes of the genus Meloidogyne are economically important pathogens of a wide range of crops. The tomato resistance gene Mi typically confers resistance to the three major species, M. incognita, M. javanica, and M. arenaria. However, virulent populations completely overcoming the Mi resistance still occur. In an attempt to develop molecular markers for virulence against Mi and gain insights into the genetic relationships among virulent populations of different species and origins, random amplified polymorphic DNA (RAPD) analyses of laboratory-selected virulent, field virulent, and avirulent populations of M. incognita, M. javanica, and M. arenaria were carried out. A RAPD marker, specific for selected virulent populations, was identified, and subsequently, converted to a sequence characterized amplified region (SCAR). Sequence characterization of the SCAR locus showed that alleles from laboratory- and field-selected virulent populations were highly similar to each other and clearly different from alleles from natural virulent and avirulent populations. This result suggests that the genetic mechanism for virulence against Mi may be similar among selected virulent populations of the three Meloidogyne spp., but different between selected and natural virulent populations. Based on the nucleotide polymorphisms at the SCAR locus, codominant and dominant polymerase chain reaction-based markers were developed enabling rapid diagnosis of selected virulent genotypes in M. incognita, M. javanica, and M. arenaria.     

Identification of isolate‐specific resistance QTLs to phytophthora root rot using an intraspecific recombinant inbred line population of pepper (Capsicum annuum)

Quantitative trait loci (QTL) for resistance to phytophthora root rot caused by Phytophthora capsici were investigated using two Korean P. capsici isolates and 126 F₈ recombinant inbred lines derived from a cross of Capsicum annuum line YCM334 (resistant parent) and local cv. Tean (susceptible parent). The experimental design was a split plot with two replications. Highly significant effects of pathogen isolate, plant genotype, and genotype × isolate were detected. QTL mapping was performed using a genetic linkage map covering 1486·6 cM of the pepper genome, and consisted of 249 markers including 136 AFLPs (Amplified Fragment Length Polymorphisms), 112 SSRs (Simple Sequence Repeats) and one CAPS (Cleaved Amplified Polymorphic Sequence). Fifteen QTLs were detected on chromosomes 5 (P5), 10 (P10), 11 (P11), Pb and Pc using two data processing methods: percentage of wilted plants (PWP) and relative area under the disease progress curves (RAUDPC). The phenotypic variation explained by each QTL (R²) ranged from 6·0% to 48·2%. Seven QTLs were common to resistance for the two isolates on chromosome 5 (P5); six were isolate‐specific for isolate 09‐051 on chromosomes 10 (P10) and Pc, and two for isolate 07‐127 on chromosomes 11 (P11) and Pb. The QTLs in common with the major effect on the resistance for two isolates explained 20·0–48·2% of phenotypic variation. The isolate‐specific QTLs explained 6·0–17·4% of phenotypic variation. The result confirms a gene‐for‐gene relationship between C. annuum and P. capsici for root rot resistance.     

Induction of Resistance in Mustard (Brassica juncea) against Alternaria Black Spot with an Avirulent Alternaria brassicae Isolate-D

Resistance in susceptible mustard cv. PR-15 against the highly virulent A. brassicae isolate A (AbA) and moderately virulent isolate C (AbC) was induced using an avirulent Alternaria brassicae isolate D (AbD). The induction of resistance due to AbD against AbA or AbC resulted in significant reduction in disease severity. The A. alternata (Aa) failed to induce resistance against AbA and AbC, on the contrary it induced susceptibility against them.