乙烯丰诱导橡胶树对条溃疡病抗性的超微结构

用透射电镜技术研究了柑桔褐腐疫霉(Phytophthora citrophthora)引起的橡胶树(Hevea brasiliensis Mull.Arg.)无性系 RRIM600条溃疡病,比较了正常树皮(即不施乙烯丰树皮)和施用乙烯丰(2—氯乙基磷酸,乙烯释放剂)树皮接种后菌丝的侵染和寄主细胞的反应。在正常树皮中,菌丝主要在寄主细胞间生长并引起细胞壁分解。菌丝也常常垂直穿透寄主细胞壁而在细胞内形成吸器和细胞内菌丝。当开始垂直穿透时,寄主细胞壁常常形成乳头状突起。吸器具有吸器外基质(extrahaustorial matrix),也常常具有领(collar)。施用乙烯丰的树皮病斑比正常树皮上的较早停止扩展而且病斑小得多,但是未发现菌丝侵染和寄主细胞反应的方式有明显差别。据用商品纤维素酶和果胶酶对树皮细胞壁试验,乙烯丰处理的树皮比正常树皮较能抵抗这些酶的分解。这些事实表明,乙烯丰诱导的寄主细胞的变化使菌丝侵染延迟,但并未杀伤病原,也没有完全阻止穿透。     

Amplification of the <Emphasis Type="Italic">Phytophthora infestans</Emphasis> RG57 loci Facilitates <Emphasis Type="Italic">in planta</Emphasis> T-RFLP Identification of Late Blight Genotypes

Late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary, is a devastating disease in potato and tomato and causes yield and quality losses worldwide. The disease first emerged in central America and has since spread in North America including the United States and Canada. Several new genotypes of P. infestans have recently emerged, including US-22, US-23 and US-24. Due to significant economic and environmental impacts, there has been an increasing interest in the rapid identification of P. infestans genotypes. In addition to providing details regarding the various phenotypic characteristics such as fungicide resistance, host preference, and pathogenicity associated with various P. infestans genotypes, information related to pathogen movement and potential recombination may also be determined from the genetic analyses. Restriction fragment length polymorphism (RFLP) analysis with the RG57 loci is one of the most reliable procedures used to genotype P. infestans. However, the RFLP procedure requires propagation and isolation of the pathogen and relatively large amounts of DNA. Isolation of the late blight pathogen is sometimes impossible due to the poor condition of the infected tissues or the presence of fungicide residues. In this study, we describe a procedure to identify P. infestans at the molecular level in planta using terminal restriction fragment length polymorphism (T-RFLP) of the RG57 loci. This T-RFLP assay is sufficiently sensitive to detect and differentiate P. infestans genotypes directly in planta without propagation and isolation of the pathogen, to facilitate the timely implementation of best management practices.     

Exploiting Knowledge of Pathogen Effectors to Enhance Late Blight Resistance in Potato

Late blight disease of potato, caused by Phytophthora infestans, is the most serious disease of this crop on a global scale and is thus a threat to food security. Use of resistant potato cultivars to prevent late blight does not have a very successful history, as P. infestans genotypes have overcome the deployed resistances. Thus, there is a need to identify more durable resistances, or identify and develop novel forms of resistance that exploit vulnerabilities in the biology of P. infestans. Application of molecular biology tools in P. infestans research has recently culminated in the identification of several avirulence effectors matching specific resistances in potato, the sequencing of the P. infestans genome and identification of hundreds of candidate translocated pathogen ‘RXLR’ effector proteins that may promote disease progression. Strategies for prioritising these effectors for further research are revealing those that are highly expressed during infection, difficult for the pathogen to alter rapidly, essential for P. infestans pathogenesis and recognized by resistant accessions of Solanum spp. These effector characteristics are being used to identify and characterise resistances from Solanum germplasm that may prove more durable. In addition to RXLR effectors, P. infestans also produces a broad spectrum of additional secreted proteins. These are exposed to plant cells and may potentially act to trigger resistance, either as broad spectrum pathogen-associated molecular patterns or as specific effectors of resistance. Alternatively, conserved secreted proteins may be attractive targets for novel agrichemical development. We have silenced a diverse selection of these candidate secreted proteins in P. infestans and demonstrated their effects on late blight disease development. Results from these studies are aiding a deeper understanding of P. infestans disease development and identifying potential pathogen weaknesses for exploitation in future control measures.     

Susceptibility of five nightshade (Solanum) species tophytophthora infestans

The susceptibility of leaves and berries of five night-shade species to infection by the potato late blight pathogenPhytophthora infestans was tested. American black nightshade (Solanum americanum) and eastern black nightshade (Solanum ptycanthum) were resistant to infection. However, two accessions (entire-edged and dentate-edged leaf) of hairy nightshade (Solanum sarrachoides), cutleaf nightshade (Solanum triflorum), and bitter or climbing nightshade (Solanum dulcamara) were susceptible. Survival of the pathogen in susceptible species was tested under different environmental conditions. The pathogen survived for at least 2 wk in both hairy nightshade accessions when plant material was kept in soil at a temperature of 20 C. However,P. infestans survived 48 h at ?15 C only in the entire-leaf hairy nightshade accession. In the laboratory, oospores of the pathogen were formed in inoculated plants of the two hairy nightshade accessions, cutleaf nightshade, and ‘Russet Burbank’ potato, but not in American black nightshade, bitter nightshade, or eastern black nightshade.     

Characterisation of Phytophthora infestans Isolates Collected from Potato and Tomato Crops in Tunisia During 2006–2008

Severe late blight epidemics in Tunisia in recent years prompted population studies on the pathogen responsible for this disease, Phythophthora infestans. Characterisation of 165 Tunisian P. infestans isolates collected from 2006 to 2008 was performed for the mating type and mt haplotype, while subsets were analysed for metalaxyl sensitivity (n?=?65), virulence on differential set of 11 R genes of Solanum demissum (n?=?31), aggressiveness on cv. Bintje (n?=?36) and measurement of the radial growth on agar medium at three temperatures (n?=?38). Most isolates from potato and all isolates from tomato had the A1 mating type. The A2 mating type was detected in the north-east and northern areas, but not in the north-west. All the A2 mating type isolates were metalaxyl resistant and seem to be part of a new generation of the P. infestans isolates which are more aggressive, with more complex races, and tolerant to higher temperatures. The increased severity of epidemics during 2006 to 2008 can be attributed to favourable weather conditions during growing seasons, adaptation of new genotypes, widespread phenylamide resistance in potato production regions and most probably incorrect spray programmes. In contrast to the presence of complex pathotypes in two major potato crop regions (north-east and northern areas), the P. infestans population detected in the other regions and in tomato crops was still relatively simple. Compared with the situation in Europe and the American continent, or even compared with neighbouring countries such as Algeria, the genetic changes in Tunisia are still comforting and require strict management decision on late blight control to avoid the spread of new P. infestans populations from Europe or neighbouring countries.     

胶孢炭疽菌侵染芒果的过程及寄主的组织病理学研究

炭疽菌是一类地理分布广泛、寄主范围广泛的真菌,在作物成熟前和成熟后都能侵染植株和果实,特别对成熟的热带作物危害极大,造成果实腐烂,导致严重的经济损失并影响产品的出口质量.为探讨胶孢炭疽菌侵染芒果的过程及其与寄主的互作关系,揭示该病菌对果实和叶片的侵染方式和扩展途径,以元江本地品种'三年芒'病果上分离、筛选的强致病菌株C...     

Comparative studies on the mode of action of aluminium ethyl phosphite in four Phytophthora species

Aluminium ethyl phosphite (fosetyl-aluminium) at concentrations of 250, 500, 750 and 1000 ppm caused severe morphological changes in mycelia of Phytophthora cactorum and P. capsici; P. megasperma was susceptible only at the two highest concentrations, whereas P. infestans was not affected at the concentrations tested. Thickening of the mycelial tips and knot-like swellings along the hyphae were particularly characteristic changes seen in the sensitive Phytophthora species. The fungicide caused a reduction in ion efflux from mycelia of P. cactorum (reduced membrane permeability), whereas the ion efflux from mycelia of P. megasperma and P. infestans was increased (enhanced membrane permeability). The compound did not affect the metabolism of neutral lipids in the four fungi tested. It had marked effects on the metabolism of phospholipids in P. cactorum and P. capsici but only slightly altered these components in P. megasperma and P. infestans. The relatively insensitive P. megasperma and the insensitive P. infestans contain two additional phospholipid components in their membranes compared with those in the sensitive P. cactorum and P. capsici. The fungicide induced a significant increase in the incorporation of glucose into the cellulose/cell-wall fraction of P. cactorum and P. capsici, whereas the synthesis of cellulose in P. megasperma and P. infestans was not affected. The results suggest that toxicity of aluminium ethyl phosphite may be related to the disturbance of physiological processes involved directly or indirectly with membrane function and cell-wall synthesis.     

Changes in race structure of Canadian populations ofPhytophthora infestans based on specific virulence to selected potato clones

Summary A collection of 80 isolates ofPhytophthora infestans, representing multilocus genotypes of the pathogen found in Canada from 1994 to 1996, was screened on a series of nine differential potato host cultivars to determine the physiological race of the isolates. A total of 28 pathotypes were found among the 11 genotypes present in the collection. There was a significant increase in the complexity of pathotypes from 1994 to 1996 which reflected the displacement of the US-1 genotype (mean of 2.2 host differentials infected) by the US-8 genotype (mean of 8.1 host differentials infected). Eighteen of the US-8 isolates could overcome all nine of the resistance genes tested. Virulence genes which overcame host resistance conferred by R8 were the rarest in the populations studied. The predominance of complex populations ofP. infestans in Canada implies that the introduction of cultivars with specific R genes for resistance would be an ineffective control measure. AAFC Contribution No. 901     

Impact of Different US Genotypes of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on Potato Seed Tuber Rot and Plant Emergence in a Range of Cultivars and Advanced Breeding Lines

Seed pieces of different potato cultivars and advanced breeding lines (ABLs) from north central US breeding programmes were inoculated with different genotypes of Phytophthora infestans (US-1, US-1.7, US-8, US-11 and US-14). The effect of these genotypes of P. infestans on seed piece rot severity after re-storage was assessed using an image analysis technique. P. infestans genotypes demonstrated variable ability to cause seed piece rot and to reduce plant emergence measured as final plant stand (%) and the relative area under the plant emergence curve (RAUEPC). The US-8 genotype of P. infestans was the most aggressive genotype, as indicated by tuber rot severity across all cultivars/ABLs tested, followed by US-14 in both years. The US-1, US-1.7 and US-11 genotypes were the least aggressive, causing only moderate seed piece rotting across cultivars/ABLs tested. Similar trends were observed in two field experiments, where the US-8 and US-14 genotypes delayed or reduced emergence. Values of final plant stand (%) and RAUEPC demonstrated that the cultivars/ABLs Atlantic, MSJ453-4Y and Torridon were the least susceptible across all P. infestans genotypes. In both experiments cv. Pike was the most susceptible. Other cultivars/ABLs demonstrated variable responses to different genotypes of P. infestans. No symptoms of P. infestans were observed on emerged plants up to 60 days after planting. The variability of susceptibility of tubers to different genotypes of P. infestans has implications for plant breeding efforts in that the major emphasis in the past has been to breed for foliar resistance, with limited emphasis on the reaction of the tuber. Results from this study suggest that highly aggressive genotypes of P. infestans such as US-8 may lead to severe tuber rotting and deterioration of tubers before emergence, but despite this observation the US-8 genotype is still predominant in North America.     

Identification of late blight, Colorado potato beetle, and blackleg resistance in three Mexican and two South American wild 2x (1EBN)Solanum species

Wild potatoes are important sources of genes for resistance to disease and insect pests. A collection of wild Mexican and South AmericanSolarium species from the US potato Genebank was evaluated under laboratory and/or field conditions for their reaction to late blight (Phytophthora infestans), Colorado potato beetle (CPB,Leptinotarsa decemlineata Say), and blackleg (Erwinia carotovora subsp.atroseptica (van Hall) Dye) in order to identify individual genotypes with multiple resistance genes. Late blight inoculations using aggressive isolates (US-8/A2 and US-11/A1 mating types) of P.infestans revealed a wide range of variation for resistance between and within the accessions of the wild species tested. For late blight, susceptible as well as moderately to highly resistant genotypes were observed in all the species tested. However, at least one accession from the three Mexican and one South American wild diploid species tested showed a relatively uniform high level of resistance toP. infestans. These includedS. bulbocastanum, S. pinnatisectum, S. cardiophyllum, andS. circaeifolium. Two accessions from South American speciesS. commersonii were highly susceptible to late blight. For the Colorado potato beetle test, only one species,S. pinnatisectum appeared uniformly resistant to CPB under field conditions. Results of screening for blackleg resistance showed that there were major differences between genotypes in the wild species. Accessions ofS. circaeifolium PI 498119 andS. bulbocastanum PI 243504 were identified as having significantly higher blackleg resistance than cultivated potato and the other wild species tested. However, genotypes from these two accessions were more susceptible to late blight and CPB. Characterization of theP. infestans isolate P1801C.16 used for late blight evaluation and multi-locus isolate tests using US-8/A2 and US-11/A1 races revealed that the resistance inS. pinnatisectum genotypes tested corresponded to a race-non-specific genetic system, which was different from any existing R genes.Solanum pin-natisectum genotypes with both high levels of late blight and CPB resistance as well as blackleg resistance genotypes identified in the present study represent a diverse gene pool that may be useful for development of new potato cultivars with multiple disease and insect resistance. The potential utilization of these valuable sources for improvement of cultivated potato is discussed.     

Molecular Analysis of Phytophthora infestans (Mont.) de Bary from Chapingo,Mexico. Phylogeographic Referential

An Andean origin of Phytophthora infestans was originally based on written records, and recently inferred from mitochondrial and nuclear gene genealogies, which emphasize a common South American origin of the European isolates, and absence of the I-b haplotype in Mexico (recently identified in Tlaxcala, Mexico). In order to compare such results, we sequenced the nuclear genes β-tubulin, Ras, and Intron Ras, and rDNA ITS regions in 24 P. infestans genotypes isolated, identified, and selected, during three crop growing cycles in Chapingo, Mexico. The pathogen showed homology and 100 % sequence similarities to the ones reported for the Andes and Europe β-tubulin, Ras, and Intron Ras genes. Results for the ITS region were inconclusive. The molecular oomycete similarities from the three geographical regions (South America, Europe, and Mexico), and the Mexican genotypes characteristics reported in the species supports the hypothesis of the location of P. infestans diversity and a center of dispersal in the central highlands of Mexico.     

Comparative analysis of quantitative trait loci for foliage resistance toPhytophthora infestans in tuber-bearingSolanum species

Previously published data for potato foliage resistance to late blight were examined in a combined analysis to determine how the genetic backgrounds of the host and pathogen affect the occurrence of resistance QTLs. Data from 19 diploid populations and one tetraploid population that originated from at least 12 different tuber-bearingSolanum species were included. Comparative analysis across all populations revealed three highly active genomic regions on the distal parts of chromosomes 3,4, and 5. The region most consistently detected that conferred foliage resistance inSolanum was located on chromosome 5, near marker locus GP21. A previously identified cluster of three race-specific R-genes on chromosome 11 was not associated with polygenic resistance. Statistical examination of active QTLs indicates that congruence among QTLs is significantly affected by both genetic relatedness of mapping populations and race ofPhytophthora infestans used for resistance tests. The proportion of congruent QTLs for late blight resistance decreased from 0.52 detected in relatedSolanum populations tested with the same race ofPhytophthora infestans to 0.20 found in unrelated mapping populations tested with different races ofPhytophthora infestans. Analysis of resistance mapping tests provided statistical evidence for the occurrence of race-specific QTLs in tuber-bearingSolanum. The highly conserved genomic regions identified in the comparative analysis are likely to be good candidates for gene cloning or marker assisted selection in potato breeding programs.     

Enhanced defense responses of tomato plants against late blight pathogen Phytophthora infestans by pre-inoculation with rhizobacteria

The efficacy of resistance in tomato plants, induced by rhizobacteria isolated from Jeju Island in Korea, against late blight disease caused by Phytophthora infestans was tested. Among the bacterial isolates tested, pre-inoculation with four isolates induced an effective defense against late blight. The four isolates, TRH423-3, TRH427-2, KRJ502-1 and KRY505-3, were identified as Burkholderia gladioli, Miamiensis avidus, Acinetobacter quenomosp and Bacillus cereus, respectively, by sequencing the ribosomal intergenic spacer region. They also promoted the growth of tomato seedlings. To illustrate resistance mechanisms, the infection process by P. infestans was examined using a fluorescence microscope. There were no noticeable differences in the rate of germination and appressorium formation between the untreated and pre-inoculated plants. However, callose was more frequently formed at the penetration sites on the leaves of pre-inoculated plants than the untreated plants, suggesting that these rhizobacterial isolates induce defense responses against P. infestans.     

US-8 and US-11 genotypes ofPhytophthora infestans from potato and tomato respond differently to commercial fungicides

Isolates ofPhytophthora infestans collected in Canada in 1997 from both potatoes and tomatoes, were tested on potato leaf discs for their response to an equal active ingredient concentration (10 μg a.i./mL) of the following commercial fungicides: Acrobat MZ (Dimethomorph and Mancozeb), Ridomil Gold (metalaxyl-m), Dithane (Mancozeb), Curzate (Cymoxanil), Bravo (Chlorothalonil), and Tattoo C (Propamocarb and Chlorothalonil). Relative percent leaf infection values, estimated on fungicide-treated vs fungicide-free leaf discs, were compared among isolates from the US-8 and US-11 genotypes isolated from the two host plants. Based on an equal concentration of each fungicide’s active ingredients, variations in relative percent leaf infection were recorded between US-8 and US-11 genotypes, and between potato and tomato isolates within each genotype. Bravo and Tattoo C used with similar active ingredients concentrations were the most inhibitory to all groups of isolates. Dithane and Ridomil Gold provided uniform low inhibition againstP. infestans when tested on potato leaf discs. The different behavior ofP. infestans isolates from potato vs tomato suggests that management of late blight in these two important crops must take such differences into consideration. In particular, the nature and concentration of the fungicides to be applied must take into account any information available about genotypes present on each crop.     

Variation for 2n pollen production and male fertility in wildSolanum germplasm resistant toPhytophthora infestans (Mont.) de Bary (US-8)

Wild potato species possess genetic variability for valuable traits including resistance toPhytophthora infestans, the causal agent for potato late blight disease. Breeding schemes using 2n gametes are a powerful tool for introgressing these traits. Forty-five plant introductions (PIs) across 12Solanum species representing various Endosperm Balance Numbers (EBN) and having resistance toP. infestans (US-8, isolate ND 95-2) were screened for 2n pollen production and male fertility. Species evaluated were 2x(lEBN)S. bulbocastanum, S. cardiophyllum, S. pinnatisectum, andS. trifidum; 2x(2EBN)S. berthaultii, S. megistacrolobum, S. microdontum, andS. verrucosum; 4x(2EBN)S. fendleri andS. stoloniferum; 6x(4EBN)S. guerreroense; and 2x(unknown EBN)S. polyadenium. Acetocarmine staining and cytological analyses were used to determine 2n pollen production and pollen stainability from genotypes grown across three locations. Based on frequency, genotypes were assigned to 2n pollen production categories (0≤1%, Low=1%–10%, Medium=11%–60%, and High≥60%) and pollen stainability categories (0≤1%, 1=196–5%, 2=6%–10%, 3=11%–20%, 4=21%–50%, and 5≥50%). Eleven of the 12 species and 37 of the 45 PIs examined contained genotypes producing ≥1% 2n pollen with ranges of 0%–63.6% and 0%–81.8%, respectively. Dependence was found between location and pollen stainability, EBN and pollen stainability, but not for location and frequency of genotypes with 2n pollen. Stability for 2n pollen production was observed across environments. Among the 1EBN germplasm, dependence was found between 2n pollen and pollen stainability. Resistance toP. infestans was found to be independent of 2n pollen production; therefore, identifying genotypes combining 2n pollen production, fertility, and resistance toP. infestans was possible.     

Role of Solanum dulcamara L. in Potato Late Blight Epidemiology

Four sites with naturally growing Solanum dulcamara were surveyed during 2006 and 2007 for the presence of late blight. Despite 2 years of observations, no late blight was detected among natural populations of bittersweet. Nevertheless, repeated infections occurred on few S. dulcamara plants from a collection growing in a botanical garden in the same years. These plants were used to investigate the possibility of survival of the inoculum between seasons. In the respective years, a set of 21 and 52 S. dulcamara accessions inoculated with Phytophthora infestans under field conditions resulted in a wide range of responses to the disease. More susceptible reactions were found among genotypes collected at greater distance from commercial potato fields indicating the possibility of genetic selection caused by P. infestans. However, both scarceness of natural infections and no overwintering, suggest that bittersweet may not play a role in late blight epidemiology.     

Characterization ofPhytophthora infestans from Maine during 1999 and 2000

A total of 77 isolates ofPhytophthora infestans was collected in 1999 and 93 isolates were collected in 2000 from growers’ fields, home gardens, and research plots in Maine. Genotype diversity based onglucose-6-phosphate isomerase (Gpi) allozyme analysis was greater in the collections from 2000, with six genotypes identified vs two genotypes identified in 1999. The US-8 (Gpi 100/111/122) genotype constituted 88% of the isolates collected in 1999 and 75% of those collected in 2000. The US-14 (Gpi 100/122) genotype constituted 12% and 7% of the isolates collected in 1999 and 2000, respectively. Three newGpi genotypes constituted 7% of the isolates collected from potato in northern Maine in 2000. A new genotype (A1 mating type,Gpi 100/100) that was aggressive on both tomato and potato was identified from home gardens in western Maine and constituted 11% of the isolates collected in 2000. A1, A2, and A1A2 mating types as well as self-fertile isolates were collected in both years. Self-fertility in artificial culture appeared to be relatively common in field isolatesof P. infestans. Potential origins of the new genotypes are discussed.     

Phenotypic and Genomic Modifications Associated with <Emphasis Type="Italic">Globodera pallida</Emphasis> Adaptation to Potato Resistances

Studying phenotypic and genomic modifications associated with pathogen adaptation to resistance is a crucial step to better understand and anticipate resistance breakdown. This short review summarizes recent results obtained using experimentally evolved populations of the potato cyst nematode Globodera pallida. In a first step, the variability of resistance durability was explored in four different potato genotypes carrying the resistance quantitative trait loci (QTL) GpaV_vrn originating from Solanum vernei but differing by their genetic background. The consequences of the adaptation to resistance in terms of local adaptation, cross-virulence and virulence cost were then investigated. Finally, a genome scan approach was performed in order to identify the genomic regions involved in this adaptation. Results showed that nematode populations were able to adapt to the QTL GpaV_vrn, and that the plant genetic background has a strong impact on resistance durability. A trade-off between the adaptations to different resistant potato genotypes was detected, and we also showed that adaptation to the resistance QTL GpaV_vrn from S. vernei did not allow adaptation to the colinear locus from S. sparsipilum (GpaV_spl). Unexpectedly, the adaptation to resistance led to an increase of virulent individual’s fitness on a susceptible host. Moreover, the genome scan approach allowed the highlighting of candidate genomic regions involved in adaptation to host plant resistance. This review shows that experimental evolution is an interesting tool to anticipate the adaptation of pathogen populations and could be very useful for identifying durable strategies for resistance deployment.     

Novel genotypes ofPhytophthora infestans in Canada during 1994 and 1995

Populations ofPhytophthora infestans, causing late blight of potato, changed significantly in Canada in the mid-1990s. The US-1 genotype (A1, metalaxylsensitive [MS]) was displaced by novel genotypes (mainly US-8) in much of the country in 1994 and 1995. Many of the new genotypes were insensitive to metalaxyl and were of the A2 mating type, although metalaxyl-insensitive (MI) Al isolates of the gll (or US-11) genotype were found in British Columbia. A total of 14 unique multilocus genotypes were described over the two years of the study based on mating type (Al or A2), metalaxyl sensitivity (MS or MI), allozyme banding patterns (Gpi andPep), and random amplified polymorphic DNA (RAPD) markers. Six of these genotypes could be distinguished by allozyme banding patterns at theGpi locus alone; RAPD analysis revealed additional variation within allozyme groupings and defined additional genotypes. Among 82 isolates collected from various Canadian provinces in 1994, US-1 and US-8 constituted 30% and 49% of isolates, respectively; other genotypes occurred in minor frequency. Among 50 isolates collected in 1995, US-1 and US-8 constituted 10% and 74%, respectively. Isolates of gll (US-11) were found in British Columbia in 1995. Diversity of genotypes was greater in 1994 (11 genotypes) than in 1995 (six genotypes). We hypothesize that sexual reproduction may have been responsible for generating some of the genetic variability inP. infestans populations, during a time period in which A2 isolates first migrated into many provinces in Canada.