Characteristics of Phytophthora infestans Isolates from Uruguay

Isolates of Phytophthora infestans were obtained from late blighted plants from several potato-growing regions of Uruguay in 1998 and 1999. Of these, 25 representative isolates (4 from 1998, 21 from 1999) from the main potato-growing areas of the country, were characterised in terms of mating type, metalaxyl resistance, allozyme genotype, mitochondrial haplotype, RG57 fingerprint (1999 isolates only) and pathotype. All isolates proved to be A2 mating type, monomorphic and homozygous at the loci coding for glucose-6-phosphate isomerase and peptidase (Gpi 100/100, Pep 100/100) and to possess mitochondrial haplotype IIa. Metalaxyl-resistant isolates constituted 92% of the total. All the 1999 isolates possessed the same RG57 fingerprint, which was that previously reported as associated with the clonal lineage BR-1 from Brazil and Bolivia, which is also A2, Gpi 100/100, Pep 100/100. Most of the isolates displayed broad-spectrum virulence and five carried virulence to 10 of the 11 R genes tested despite the absence of R genes in commercially grown potato cultivars. It was concluded that the Uruguayan P. infestans isolates resembled isolates from neighbouring South American countries, notably Brazil, and belong to the new populations of the pathogen now predominant in many countries.     

Genetics of virulence in Ascochyta rabiei

In order to critically test the hypothesis that virulence variation in the Ascochyta rabiei/chickpea pathosystem is a discrete character under simple genetic control, a genetic cross was made between a highly virulent isolate of A. rabiei from Syria and a less virulent isolate from the USA. Two independent virulence assays conducted by inoculating susceptible and resistant chickpea cultivars under controlled conditions with 77 independent progeny isolates from this cross revealed a continuous distribution of disease phenotypes. Bimodality, as would be predicted for the segregation of virulence under simple genetic control, was not supported by statistical tests of the progeny phenotype distribution. anova revealed highly significant pathogen‐genotype × host‐genotype interactions demonstrating the segregation of genes controlling specialization on the two cultivars tested. These interactions could be localized to two isolates that changed virulence rank on the cultivars. It was concluded that variation in virulence to these two cultivars is under quantitative genetic control. If this conclusion applies to other cultivars, it can be speculated that the discrete categories of virulence variation identified in previous studies were probably the result of incomplete sampling of host resistance or pathogen virulence variation and/or of selection for increased virulence in contemporary A. rabiei populations.     

Population structure of Phytophthora infestans in Turkey reveals expansion and spread of dominant clonal lineages and virulence

Late blight, caused by the oomycete Phytophthora infestans, is considered the most important and destructive disease of potato in Turkey. In this study, characterization of 367 isolates of P. infestans obtained from the potato-growing areas of the country was carried out to evaluate the pathogen population structure over the 2017–2019 production seasons. The isolates were characterized by numerous features including mating type, in vitro mefenoxam sensitivity, simple-sequence repeat (SSR) markers, and virulence against a set of potato differential lines. Most isolates were A2 mating type (353 isolates). Also, 68% of isolates were resistant to mefenoxam; the remainder were intermediate in their sensitivity and there were no sensitive isolates. SSR-based genotypic analysis of P. infestans populations showed a low genetic diversity. The 13_A2 clonal lineage predominated with a frequency of 92.1%, followed by 34_A1 (3.3%) and 37_A2 (2.7%). Genotypes 34_A1 and 37_A2 were detected only in 2019. This is the first report of 34_A1 and 37_A2 clonal lineages causing late blight disease of potato in Turkey. The most abundant virulence type was one overcoming resistance genes R1, R2, R3, R4, R6, R7, R10, and R11. These results emphasized that the migration of individuals and the asexual generation of subclonal differences were the main factors driving the population structure of P. infestans in Turkey.     

Population genetic analysis of Phytophthora infestans in northwestern China

Late blight caused by Phytophthora infestans is the most devastating disease of potato worldwide. To understand the P. infestans population structure and dynamics in northwestern China, 959 single‐lesion isolates were purified in three consecutive years (2009–2011) and were characterized for mating type, pathotype, mtDNA haplotype and molecular variation at eight SSR loci. The results showed that the distribution of mating types changed significantly over years, with self‐fertile isolates dominant in 2010 and 2011. SSR genotyping distinguished 959 isolates into 151 genotypes, and association analysis indicated that P. infestans populations in 2010 and 2011 were strictly asexual while in 2009 they showed signs of sexual reproduction. Population analysis showed that the majority of genetic variation was within P. infestans populations. Isolates sharing identical SSR genotypes were detected in distant regions, indicating that migration of P. infestans could have occurred between regions. Pathogenicity assays on a set of potato differential lines containing R1 to R11 resistance genes detected four pathotypes from 74 selected isolates, with the pathotype virulent against all 11 R genes being dominant. Three mtDNA haplotypes (Ia, IIa, IIb) were detected with Ia being dominant among 507 isolates examined. Phylogenetic analysis indicated that P. infestans populations in northwestern China are distant from European lineages including 13‐A2 (blue‐13) at the time of this survey. The results have implications for the trade of healthy seed tubers as a means of managing late blight.     

The role of aggressiveness and competition in the selection of Phytophthora infestans populations

Selection within populations of Phytophthora infestans was investigated by comparing the aggressiveness of single‐lesion isolates on detached leaflets of four potato cultivars with differing levels of race‐nonspecific resistance to P. infestans. The isolates included 23 representative of Northern Ireland genotypes from the early 2000s, used to inoculate previously reported field trials on competitive selection (2003–2005), plus 12 isolates recovered from the 2003 trial. The cultivars were those planted in the previous trials: Atlantic (blight‐susceptible) and Santé, Milagro and Stirling (partially resistant). Very highly significant variation for latent period, infection frequency and lesion area was found between genotypes and cultivars; differences between genotypes were more marked on the more resistant cultivars, but no one genotype was the most aggressive across all. Detached leaflets were also inoculated with mixtures of isolates from each genotype group at three sporangial concentrations: differences in aggressiveness between genotypes were more apparent at lower concentrations and on the more resistant cultivars. Genotype groups that were the most aggressive on the more resistant cultivars tended to be those selected by the same cultivars in the field. A mixture of all isolates of all genotypes was used to inoculate detached leaflets of the same cultivars. With one exception, single spore isolates recovered from any one leaflet belonged to a single genotype, but different genotypes were recovered from different cultivars. Phytophthora infestans isolates from Northern Ireland showed significant variation for foliar aggressiveness, and pathogen genotypes exhibited differential aggressiveness to partially resistant cultivars and interacted competitively in genotype selection.     

江西省稻瘟病菌毒性的群体结构分析

用江苏省的吴江、赣榆、通州、高邮和宜兴等5个代表地区的324个稻瘟病菌株接种13个日本已知抗病基因品种，结果表明，江苏省稻瘟病菌对携带有Pi-Ka,Pi-ta,Pi-ta2和Pi-sh抗病基因品种的可致病比例较高，毒力频率达64．2％－97．2％，对携带有Pi-iPi-z和Pi-b抗病基因的品种毒力频率较低，在0－11．9％之间；江苏省稻瘟病菌毒性的群体结构年度间有一定变化，表现为对某些抗病基因品种毒力频率的明显上升或下降。从江苏省10年主栽品种上分离的稻瘟病菌株对13个已知抗病基因品种的毒力频率测定结果表明，不同类型主栽品种上分离的菌株其毒性结构组成有一定差异,表明江苏省水稻主栽品种的抗性类型存在一定差异。     

Eucablight – collating and analysing pathogenicity and resistance data on a European scale*

The Concerted Action on potato late blight ‘Eucablight’ ( http://www.eucablight.org ) was launched in 2003 to promote collaboration between researchers across Europe and to collate the previously fragmented data on host resistance to late blight and characterization of populations of its causal agent, Phytophthora infestans. Fundamental to the project was the development of standardized protocols, databases and data collection tools with which data from across Europe could be centralized. The ultimate aim was to capitalize on the vast resource of data available in order to allow a pan‐European analysis to be conducted. The P. infestans database is currently populated with information relating to over 15 000 isolates from 20 European countries. The host resistance database holds primary disease data, and derived statistics, from more than 50 field trials assessing mainly foliage blight resistance. The use of five standard cultivars in these trials facilitated the comparison of resistance information across years and regions. The host database is structured, and made accessible, in such a way that DSS builders can access the model parameters they need to construct locally adapted forecasting systems. In this presentation we describe the database and the data analysis tools that have been developed in this project and how they are directly applicable to other host‐pathogen systems.     

Diversity of Phytophthora infestans from Poland

A collection of 96 Polish isolates of Phytophthora infestans sampled in the years 2006, 2008 and 2009 were analysed using phenotypic and genotypic markers. Mating type, virulence, resistance to metalaxyl, mitochondrial haplotype and polymorphism at 12 simple sequence repeat (SSR) loci were determined. The majority of isolates were of the A1 mating type, mitochondrial haplotype Ia and sensitive to metalaxyl. Virulence factors against potato R genes R1, R3, R4, R7, R10 and R11 were present in most isolates. Genotyping using SSR markers revealed high genetic diversity within the Polish P. infestans population. Amongst the 96 isolates 66 unique genotypes were identified, 49 of which were observed only in single isolates. Eight isolates of the genotype 13_A2 lineage that has been reported in other parts of Europe were also found in Poland. The implications of these results are discussed.     

Changes in specific virulence in Polish populations ofPhytophthora infestans: 1985–1991

Ninety-five isolates ofPhytophthora infestans collected throughout Poland during 1985–1991 and characterized for multilocus genotypes based on mating type, allozymes and DNA fingerprint, were analyzed for specific virulence to differential potato cultivars carrying ten major resistance genes. The multilocus analysis led to three groupings. The first group contained 22 isolates of a clonal lineage (PO-1) that is postulated to have been present in Europe during most of the twentieth century, but PO-1 isolates were recovered in Poland only during 1985–1988. This group contained, on average, virulence to 5.5 specific resistance genes per isolate. The second group consisted of 30 isolates in a clonal lineage (PO-4) that had not been detected before 1988. PO-4 isolates had virulence to a mean of 6.5 resistance genes per isolate. The third group was composed of 43 isolates representing 38 multilocus genotypes also not detected before 1988. These diverse genotypes had virulence to an average of 6.7 specific resistance genes per isolate. More than half (53%) of the PO-4 isolates shared a single pathotype. The group of 43 isolates was dominated by two pathotypes: the most common one (47% of the isolates) was the same pathotype that dominated PO-4 isolates; the next most common one (21%) differed from the most common one by the absence of virulence to resistance gene R5. The recent immigrant isolates (not detected before 1988) generally had virulence to a greater number of specific resistance genes than did isolates in the previous population [detected before 1988 (PO-1)]. Recent immigrant populations were dominated by one or two pathotypes, so their pathotypic diversity values were somewhat less than that of the previous population.     

The structure of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> populations from organic and conventional crops

The characteristics of populations of Phytophthora infestans from organic farms, small conventional farms and large conventional farms were determined from isolates collected in northern Estonia in 2004 and 2005. For the population as a whole 41% were A2; all virulence factors to the 11 R genes from Solanum demissum were found; and more than 70% had high or intermediate resistance to metalaxyl. Isolates from organic farms tended to have more complex pathotypes than isolates from either large or small conventional farms, but there was a higher proportion of metalaxyl resistant isolates from large conventional farms than from small conventional farms or from organic farms.     

Characterisation of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> isolates collected from potato in Estonia during 2002–2003

A collection of 101 isolates of Phytophthora infestans, obtained from seven sampling sites representing central, east and south-east Estonia during 2002 and 2003 were assessed for several phenotypic and genotypic markers. All 101 isolates were assessed for virulence and resistance to metalaxyl. Virulence to each of the 11 classic resistance genes was found among the tested isolates. The mean number of virulences per isolate was 6.3, with a very low frequency of virulence against resistance genes R5 (5%) and R9 (14%). The most common pathotypes were 1.3.4.7.8.10.11 and 1.3.4.7.10.11, representing altogether 12% of the studied strains. In terms of metalaxyl resistance, 30 resistant, 52 intermediate and 19 sensitive isolates were found. A subgroup of 50 isolates was assessed for mating type, allozymes [glucose-6-phosphate isomerase (Gpi) and peptidase (Pep)], DNA fingerprints with probe RG57 and mtDNA haplotype. Of this subset, 30 were A1 and 20 were A2. Collections from three of the seven fields contained both mating types. Allozyme analysis did not reveal any polymorphism. However, 19 diverse RG57 fingerprints were detected, and two mitochondrial DNA haplotypes, Ia and IIa, were detected. By combining the mating type, mtDNA haplotype and RG57 fingerprint data, 26 multilocus genotypes were identified, of which 18 were detected only once. Genotypic diversity measured by the normalised Shannon diversity index was high (0.76). The large number of multilocus genotypes and the presence of both mating types in some fields indicate that sexual reproduction may take place in Estonian populations of P. infestans.     

Diversity of Avr‐vnt1 and AvrSmira1 effector genes in Polish and Norwegian populations of Phytophthora infestans

The oomycete Phytophthora infestans, the cause of late blight, is one of the most important potato pathogens. During infection, it secretes effector proteins that manipulate host cell function, thus contributing to pathogenicity. This study examines sequence differentiation of two P. infestans effectors from 91 isolates collected in Poland and Norway and five reference isolates. A gene encoding the Avr‐vnt1 effector, recognized by the potato Rpi‐phu1 resistance gene product, is conserved. In contrast, the second effector, AvrSmira1 recognized by Rpi‐Smira1, is highly diverse. Both effectors contain positively selected amino acids. A majority of the polymorphisms and all selected sites are located in the effector C‐terminal region, which is responsible for their function inside host cells. Hence it is concluded that they are associated with a response to diversified target protein or recognition avoidance. Diversification of the AvrSmira1 effector sequences, which existed prior to the large‐scale cultivation of plants containing the Rpi‐Smira1 gene, may reduce the predicted durability of resistance provided by this gene. Although no isolates virulent to plants with the Rpi‐phu1 gene were found, the corresponding Avr‐vnt1 effector has undergone selection, providing evidence for an ongoing ‘arms race’ between the host and pathogen. Both genes remain valuable components for resistance gene pyramiding.     

Dominance of a single clonal lineage in the Phytophthora infestans population from northern Shaanxi,China revealed by genetic and phenotypic diversity analysis

Late blight caused by Phytophthora infestans is the most serious disease of potato worldwide. To understand the P. infestans population structure in northern Shaanxi, an emerging potato production region in China, 125 single‐lesion isolates were randomly collected from farmers' fields in 2009 and characterized phenotypically and genotypically. A mating type assay showed that 94 isolates were A1 mating type. Virulence determination of selected isolates on a set of differential potato lines containing R1 to R11, respectively, showed the presence of two pathotypes, of which the pathotype lacking avirulence genes Avr3, Avr4 and Avr10 was dominant. Isolates lacking all avirulence factors Avr1 to Avr11 were detected but at lower frequency (13·6%). Analysis for mtDNA haplotype showed all 61 examined isolates were IIa. A total of seven multilocus genotypes were distinguished among 125 isolates, as determined with seven polymorphic microsatellite markers. The genotype SG‐1 was dominant in the population with a frequency of 75·2% and was present throughout the region. Analysis of the phenotypic and genotypic structures of P. infestans populations indicated strict clonal reproduction of the pathogen and suggested that sexual reproduction probably does not occur. Potential implications for disease management are discussed.     

Evidence of host‐range expansion from new powdery mildew (Blumeria graminis) infections of triticale (×Triticosecale) in France

This study aimed to determine whether powdery mildew caused by Blumeria graminis is an endemic pathogen of triticale (×Triticosecale: Triticum × Secale), emerging as a result of recent changes in its pathogenicity, or whether it is a new pathogen, possibly resulting from hybridization between ff. spp. tritici and secalis. A secondary aim was to consider breeding practices that may have favoured this emergence. Phylogenetic analyses based upon six genes revealed the close relatedness of the novel entity and the ff. spp. tritici and secalis, but the IGS marker finally grouped together the isolates collected on triticale and on wheat, supporting the scenario of a recent host‐range expansion from wheat to triticale. Pathotype analyses concluded that virulence spectra of B. graminis infecting triticale were new in comparison to those observed for other reference formae speciales, and lack of fungicide resistance in triticale isolates strengthens the hypothesis of no or little genetic exchange between wheat and triticale populations of powdery mildew. This adaptation may follow the breakdown of plant resistance genes, which are probably not very diverse in current triticale cultivars since this criterion was not considered as a major one until recent years. Moreover, the complex selection and genetics of this hybrid cereal makes it difficult to predict the transmission of powdery mildew resistance genes.     

Large subclonal variation in Phytophthora infestans populations associated with Ecuadorian potato landraces

The population of Phytophthora infestans on potato landraces in three provinces (Carchi, Chimborazo and Loja) of Ecuador was analysed. All isolates (n = 66) were of the A1 mating type. Simple sequence repeats (SSR) were used to assess the genetic diversity of the isolates. The P. infestans isolates from the potato landraces grouped in a single clade together with reference isolates belonging to the clonal lineage EC‐1. In the 66 SSR profiles obtained, 31 multilocus genotypes were identified. The 66 isolates constituted 49 different races according to the Solanum demissum differential set ( R1 to R11). The P. infestans population was complex and virulent on 4 to 11 R genes. Analysis showed that the subclonal variation in the Ecuadorian EC‐1 clone is increasing over time and is much larger than clonal variation in lineages in the Netherlands and Nicaragua, suggesting high mutation rates and little or no selection in Ecuador.     

Effect of rotation of canola (Brassica napus) cultivars with different complements of blackleg resistance genes on disease severity

Blackleg disease (phoma stem canker) caused by the fungus Leptosphaeria maculans is a major disease of canola (oilseed rape, Brassica napus) worldwide. Canola plants in pots were exposed to blackleg‐infested stubble of canola with different complements of resistance genes and then assessed for disease. Plant mortality was reduced when plants were exposed to stubble from a cultivar with a different complement of resistance genes compared to stubble of a cultivar with the same resistance gene. These findings were consistent with 7 years of field surveys, which showed that changes in selection pressure as a result of extensive sowing of cultivars with major‐gene resistance, termed ‘sylvestris resistance’, dramatically influenced the frequency of virulent isolates in the population towards particular resistance genes, and therefore disease severity. All these data were supported by PCR‐genotyping surveys of fungal populations whereby the frequency of virulence alleles of avirulence genes AvrLm1 and AvrLm4 changed significantly depending on the resistance gene present in the cultivar from which the isolates were cultured. This is the first example of a study showing that sowing of canola cultivars with different complements of resistance genes in subsequent years, i.e. rotation of resistance genes, minimizes disease pressure by manipulating fungal populations. This approach provides a valuable disease management strategy for canola growers and is likely to be applicable to other plant diseases.     

Durability of resistance in lily to basal rot: evaluation of virulence and aggressiveness among isolates ofFusarium oxysporum f. sp.lilii

Prospects of durability of resistance in lily to basal rot have been evaluated by testing the virulence and aggressiveness of 31 isolates ofFusarium oxysporum f. sp.lilii towards a number of different resistance sources inLilium spp. Isolates differed strongly in aggressiveness as did species and cultivars ofLilium spp. in resistance. Significant interactions were observed between isolates of the pathogen and genotypes ofLilium spp., but the magnitude was very small compared to the main effects. The interactions were mainly due to a small group of isolates with low aggressiveness. It is argued that the interactions might be based on minor genes. No major break down of the resistance was found. For practical purposes it will be sufficient to use highly aggressive isolates in screening tests.     

High diversity, low spatial structure and rapid pathotype evolution in Moroccan populations of Blumeria graminis f.sp. hordei

Limited information is available about the spatial distribution and evolution of Blumeria graminis f.sp. hordei populations in North African countries, such as Morocco. Frequencies of virulence alleles in B. graminis populations are mainly driven by selection exerted by host resistance genes in addition to neutral processes such as migration and genetic drift. In Morocco, in contrast to Europe, there has been no systematic deployment of resistant cultivars, although some R genes are present in the traditional varieties. This is expected to result in the evolution of pathotypes with virulence to different R genes, and higher diversity in Morocco compared to Europe. To test this, we used 24 differential cultivars to characterise 72 isolates from Morocco in 2009. We assessed diversity and spatial structure of pathotypes and compared them to past isolates from the same area (collected in 1992). There was a high diversity of pathotypes. Isolates from 2009 were distinct from isolates from 1992, due to loss of virulence to Mla12, increased virulence to Mla8, Mla3 and Mlk1, and decreased virulence to Mla6, Ml(Ru2), Mlg and MlLa. Many virulences were different from those observed in European and Asian populations of B. graminis. At the spatial scale investigated, airborne dispersal and a lack of strong selection in the host population likely prevented the formation of population structure and allowed the accumulation of high isolate diversity. The evolution of novel and distinct pathotypes since 1992 is likely attributable to gene flow from Europe and selection by the host population in Morocco.     

Inheritance of resistance in <Emphasis Type="Italic">Solanum nigrum</Emphasis> to <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Solanum nigrum, black nightshade, is a wild non-tuber bearing hexaploid species with a high level of resistance to Phytophthora infestans (Colon et al. 1993), the causal agent of potato late blight, the most devastating disease in potato production. However, the genetic mode of resistance in S. nigrum is still poorly understood. In the present study, two S. nigrum accessions, 984750019 (N19) and #13, resistant (R) and susceptible (S), respectively, to three different isolates of P. infestans, were sexually crossed. The various kinds of progeny including F1, F2, F3, and backcross populations (BC₁; F1 × S), as well as two populations produced by self-pollinating the R parent and S parent, were each screened for susceptibility to P. infestans isolate MP 324 using detached leaf assays. Fifty seedling plant individuals of the F1 progeny were each resistant to this specific isolate, similarly to the seedling plants resulting from self-pollination of the resistant R parent. Thirty seedling plants obtained from self-pollination of the S parent were susceptible. Among a total of 180 F2 plants, the segregation ratio between resistant and susceptible plants was approximately 3: 1. Among the 66 seedling plants of the BC₁ progeny originating from crossing an F1 plant with the susceptible S parent, there were 26 susceptible and 40 resistant plants to P. infestans. The segregation patterns obtained indicated monogenic dominant inheritance of resistance to P. infestans isolate MP 324 in S. nigrum acc. 984750019. This gene, conferring resistance to P. infestans, may be useful for the transformation of potato cultivars susceptible to late blight.