Gene and Genotypic Diversity of Phytophthora cinnamomi in South Africa and Australia Revealed by DNA Polymorphisms

Phytophthora cinnamomi isolates from South Africa and Australia were compared to assess genetic differentiation between the two populations. These two populations were analysed for levels of phenotypic diversity using random amplified polymorphic DNAs (RAPDs) and gene and genotypic diversity using restriction fragment length polymorphisms (RFLPs). Sixteen RAPD markers from four decanucleotide Operon primers and 34 RFLP alleles from 15 putative loci were used. A few isolates from Papua New Guinea known to posses alleles different from Australian isolates were also included for comparative purposes. South African and Australian P. cinnamomi populations were almost identical with an extremely low level of genetic distance between them (D_m=0.003). Common features for the two populations include shared alleles, low levels of phenotypic/genotypic diversity, high clonality, and low observed and expected levels of heterozygosity. Furthermore, relatively high levels of genetic differentiation between mating type populations (D_m South Africa=0.020 and D_m Australia=0.025 respectively), negative fixation indices, and significant deviations from Hardy–Weinberg equilibrium, all provided evidence for the lack of frequent sexual reproduction in both populations. The data strongly suggest that both the South African and Australian P. cinnamomi populations are introduced.     

Nuclear and Mitochondrial DNA Polymorphisms in Three Mitotic Parthenogenetic Meloidogyne Spp.

In order to expand our understanding of the genetics of root-knot nematodes, variation in nuclear DNA and mitochondrial DNA in Meloidogyne incognita, M. arenaria and M. javanica was investigated. Despite the obligate mitotic parthenogenetic mode of reproduction, a large number of AFLP polymorphisms were observed among all 16 populations studied. Both UPGMA and principle coordinate analyses revealed three distinct groups that corresponded with the respective species identities of the 16 populations. M. incognita was genetically most distinct. Amplification of 63-bp tandem repeats (TR) in mtDNA from single individuals enabled the calculation of diversity measures at three hierarchical levels: within individuals, among individuals of a single population and among populations. For all three species, the highest diversity was observed within individuals explaining 43–65% of the total diversity. Many individuals contained more than one mtDNA size variant. M. incognita harboured the most heteroplasmic individuals and was the most homogenous at the population level. Only 13% of the total diversity was observed among populations, while this figure was 35% for M. arenaria. Both TR and AFLP data showed that M. arenaria is the most heterogeneous species. The comparison of the genetic distances based on AFLPs and mtDNA size variants revealed a significant correlation for the six M. arenaria populations, whereas no consistent correlation was observed for the populations of the other two species.     

Population Genetic Structure of Phytophthora infestans in Ecuador

ABSTRACT The population genetic structure of Phytophthora infestans in Ecuador was assessed from 101 isolates collected from 1990 to 1992 and 111 isolates collected in 1993. All isolates were analyzed for mating type and allozyme genotype. Both samples were dominated (>95%) by a clonal lineage (EC-1) defined from neutral markers: 90/100 genotype for glucose-6-phosphate isomerase, 96/100 genotype for peptidase, A1 mating type, and a previously unreported nuclear DNA fingerprint. The remaining isolates belonged to the US-1 clonal lineage, which has a worldwide distribution. Isolates in the 1993 sample were analyzed for virulence and metalaxyl sensitivity. All representatives of EC-1 had complex patho-types, with three pathotypes representing >60% of the collection. There was variation for metalaxyl sensitivity. There was no evidence for geographical substructuring on the basis of neutral markers, but there was evidence for limited substructuring based on metalaxyl sensitivity and specific virulence. We hypothesize that EC-1 has been recently introduced to Ecuador.     

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.     

中国不同地区致病疫霉遗传多样性的RAPD分析

 本文应用RAPD技术检测了我国主要马铃薯产区致病疫霉的遗传分化情况及不同地区菌株间的亲缘关系。用筛选出的10个随机引物对1997-2001年间采自我国9省市的82株及3株来自日本的致病疫霉DNA进行了PCR扩增,获得了79条谱带,其中多态性标记75条,占95%。根据扩增结果,运用UPGMA分析,获得了表现菌株间亲缘关系的树状图。菌株间的最大遗传距离为0.5,以距离0.3为阈值,可将供试菌株划分为10个组(RG1-10)。结果发现:A1交配型菌株群体内的差异大于A1和A2菌株群体之间的;RAPD分组与菌株的地理来源、交配型及对甲霜灵的敏感性无明显相关性。研究结果显示,来自中国北方甘肃、内蒙、吉林、黑龙江地区的菌株与一些来自云南、四川等西南地区的菌株亲缘关系相近。病原菌随种薯的迁移可能是导致这种现象的原因之一。     

Population structure of Phytophthora infestans collected on potato and tomato in Italy

Late blight caused by the oomycete Phytophthora infestans is a disease of potato and tomato of worldwide relevance and is widespread throughout Europe and the Mediterranean region. While pathogen populations in northern Europe have been sampled and characterized for many years, the genetic structure of populations from southern Europe, including Italy, has been less studied. Between 2018 and 2019, we collected 91 samples of P. infestans from potato and tomato crops in Italy, Algeria, and Tunisia on FTA cards and genotyped them using 12-plex microsatellites. These samples were compared to genotypes of P. infestans previously collected within the framework of the EuroBlight network and from published sources. Four clonal lineages were identified: 13_A2 (Blue 13), 2_A1, 23_A1, and 36_A2. Two other isolates collected could not be matched to any currently known clonal lineage. The 13_A2 and 36_A2 lineages were found exclusively in southern Italy and Algeria, while 2_A1 was only found in Algeria. This is the first report of the 36_A2 lineage in Italy. Two isolates from Solanum nigrum were 13_A2, suggesting this weed host could be a reservoir of inoculum. The 23_A1 lineage was found widely on infected tomato crops in Italy and is the same as the lineage US-23 that is widespread in North America. Differences in genotypes across the country suggests that there may be different sources of introduction into Italy, possibly via infected seed tubers from other countries in Europe, tubers for consumption from North Africa, or tomatoes.     

Population Structure of Phytophthora infestans in the Toluca Valley Region of Central Mexico

ABSTRACT We tested the hypothesis that the population of Phytophthora infestans in the Toluca valley region is genetically differentiated according to habitat. Isolates were sampled in three habitats from (i) wild Solanum spp. (WILD), (ii) land-race varieties in low-input production systems (RURAL), and (iii) modern cultivars in high-input agriculture (VALLEY). Isolates were sampled in 1988-89 (n= 179) and in 1997-98 (n= 389). In both sampling periods, the greatest genetic diversity was observed in RURAL and VALLEY habitats. Based on the Glucose-6-phosphate isomerase and Peptidase allozymes, the subpopulations from the three habitats were significantly differentiated in both sampling periods. In contrast to allozyme data for 1997-98, no differences were found among the three subpopulations for sensitivity to metalaxyl. Two groups of isolates identical for allozyme and mating type were further investigated by restriction fragment length polymorphism fingerprinting; 65% of one group and 85% of another group were demonstrated to be unique. The genetic diversity data and the chronology of disease occurrence during the season are consistent with the hypothesis that populations of P. infestans on wild Solanum populations are derived from populations on cultivated potatoes in the central highlands of Mexico near Toluca.     

Oospore Formation by Phytophthora infestans in Potato Tubers

ABSTRACT The ability of Phytophthora infestans, the causal agent of potato and tomato late blight, to produce oospores in potato tuber tissue was studied in the field and under laboratory conditions. In 1998 and 2000 field experiments, the canopy of potato cvs. Alpha and Mondial, respectively, were coinoculated with A1 + A2 sporangia of the fungus, and the infected tubers collected at harvest were examined for the presence of oospores. In 1998, only 2 of 90 infected tubers had oospores, whereas none of the 90 tubers examined in 2000 had any oospores. In the latter experiment, infected tubers kept in storage up to 12 weeks after harvest had no oospores. Artificial co-inoculations of whole tubers with A1 + A2 sporangia resulted only rarely in the formation of oospores inside the tubers. Co-inoculations of potato tuber discs taken from dormant tubers 0 to 16 weeks after harvest failed to support any oospore production, whereas discs taken from sprouting tubers of >/=18 weeks after harvest allowed oospores to form. Tuber discs showed enhanced oospore formation when treated before inoculation with either sugars, amino acids, casein hydrolysate, beta-sitosterol, or chloroethylphosphonic acid. In contrast, reducing airflow into the petri dishes where potato tuber discs were incubated reduced the number of oospores produced. The number of oospores produced in tuber tissue was lower compared with that in leaf tissue regardless of the origin of isolates used. The data show that the ability of Phytophthora infestans to produce oospores in potato tuber tissue is very limited and increases with tuber aging.     

Population changes in Phytophthora infestans in Taiwan associated with the appearance of resistance to metalaxyl

In recent years, late blight, caused by Phytophthora infestans (Mont) De Bary, has increased in severity in many parts of the world, and this has been associated with migrations which have introduced new, arguably more aggressive, populations of the pathogen. In Taiwan, late blight has been endemic on outdoor tomato crops grown in the highlands since the early 1900s, but recent epidemics have been more damaging. To ascertain the present status of the Taiwanese population of P infestans, 139 isolates of the pathogen collected and maintained by the Asian Vegetable Research and Development Center (AVRDC) were characterized using mating type, metalaxyl sensitivity, allozyme genotype, mitochondrial haplotype and RFLP fingerprinting. Up to 1997, all isolates were found to belong to the old clonal lineage of P infestans (US-1 and variants), but in isolates from 1998 a new genotype appeared, and by 2000 this had apparently completely displaced the old population. This new genotype was an A1 mating type and has the dilocus allozyme genotype 100/100/111, 100/100 for the loci coding for glucose-6-phosphate isomerase and peptidase, respectively. These characters, together with RG57 fingerprinting, indicated that these isolates belonged to the US-11 clonal lineage, a minority (11%) being a previously unreported variant of US-11. Whereas metalaxyl-resistant isolates were not detected in the old population, 96% of the new genotypes proved resistant, with the remainder being intermediate in sensitivity. It may be inferred from this sudden, marked change in the characteristics of the Taiwanese P infestans that a new population of the pathogen was introduced around 1997-98 and that this may well have already been metalaxyl-resistant when it arrived, although a role for in situ selection cannot be excluded.     

Genetic Structure of the Population of Phytophthora infestans Attacking Solanum ochranthum in the Highlands of Ecuador

Thirty-nine isolates of Phytophthora infestans were collected from the wild host Solanum ochranthum in the highland tropics of Ecuador and characterized with a set of phenotypic and molecular markers (mating type, metalaxyl sensitivity, the allozyme loci Gpi, and Pep, mitochondrial DNA haplotype, RFLP, and SSR), as well as for pathogenicity on various hosts. Three groups of isolates (A, B, and C) were identified based on their multilocus genotypes and variable abilities to cause disease on different hosts. Group A had a combination of alleles for the Gpi (86/100), Pep (96/100) and mtDNA (Ia) loci, as well as an RFLP fingerprint, that have not been reported for P. infestans in Ecuador, or elsewhere. Group B shares many marker characteristics with the US-1 lineage described in Ecuador on tomato, pear melon (S. muricatum), and S. caripense, but has SSR alleles not present in typical US-1 isolates. Group C for all markers tested is identical to the EC-1 lineage described on cultivated and wild potatoes in Ecuador. All isolates from S. ochranthum were able to re-infect their host of origin in the detached leaf assay; however, we did not draw clear conclusions as to the relative aggressiveness of the three groups on this host. Isolates of group A were the most specialized and were generally non-pathogenic or weakly pathogenic on all hosts other than S. ochranthum. Groups B and C infected tuber-bearing hosts, including the cultivated potato but were generally non-pathogenic on other non-tuber bearing hosts. Solanum ochranthum was infected by isolates coming from tuber-bearing Solanum hosts (i.e., the EC-1 lineage of P. infestans) and some US-1 isolates from non-tuber bearing hosts. Thus, in nature this species might be a potential reservoir of inoculum of different pathogen populations able to infect the cultivated hosts potato, tomato and pear melon (S.␣muricatum). Unlike potato and tomato in Ecuador, each of which is primarily attacked by a highly specialized pathogen population, S. ochranthum appears to harbour at least three pathogen groups of␣different genetic make-up. The unresolved issue of potential host specificity in isolates found on S.␣ochranthum could complicate efforts to use this species in tomato improvement.     

Population dynamics of metalaxyl-sensitive and metalaxyl-resistant isolates of Phytophthora infestans in untreated crops of potato

Each of six pairs of metalaxyl-sensitive (MS) and metalaxyl-resistant (MR) field isolates of Phytophthora infestans was inoculated as a mixture of 90% MS sporangia with 10% MR sporangia onto fungicide-free potato plants (cv. Alpha) in six walk-in plastic tunnels in each of 2 years. In both experiments, late blight killed the crops in 26-31 days. In all tunnels, the frequency of MR sporangia increased during the logarithmic phase of the epidemic to 70-85% of the population. Later, however, the frequency of M R sporangia decreased to 7-38% in six tunnels but continued to increase to 94-100% in the other six. The increased frequency of MR during the logarithmic phase was attributed to the shorter latent period and larger lesion size of MR compared to MS isolates and to the unlimited availability of uninfected host tissue. The later decrease in the frequency of MR in six tunnels was attributed to the lower sporulation capacity and the shorter infectious period of MR compared to MS isolates, and to the limitation of uninfected host tissue. The data showed that the population dynamics of P. infestans was dependent not only on fitness components of isolates, but also on the availability of uninfected host tissue.     

Characterization of the Phytophthora infestans Population in the Columbia Basin of Oregon and Washington from 1992 to 1995

ABSTRACT Isolates of Phytophthora infestans collected from 1992 to 1995 from potato fields in the Columbia Basin of Oregon and Washington were analyzed for compatibility type, metalaxyl sensitivity, and glucose-6-phosphate isomerase (Gpi) genotype. In 1992, 30 of 31 isolates were of the US-1 multilocus genotype. A single metalaxyl-resistant isolate of the US-6 (A1 Gpi 86/100) genotype was found near the end of the growing season. In 1993, only 2 of the 59 isolates collected were A1 isolates with Gpi 86/100. Ten isolates were of the A2 compatibility type, seven with Gpi 100/111, two with Gpi 100/100, and one was undetermined. The remaining isolates were metalaxyl-resistant A1 compatibility types with either Gpi 100/100 or 100/111. The first A2 isolates in the Columbia Basin were found in 1993. In 1994, 10 of 18 isolates were of the US-1 genotype. The remaining isolates were US-6 and US-8 genotypes. In 1995, 97% of 268 isolates tested were of the US-8 genotype. Five isolates were A2 compatibility type with Gpi 100/122. One A2, metalaxyl-resistant isolate was Gpi 100/100/111, and two A1 isolates were Gpi 100/111/122. The population of P. infestans quickly changed between 1992 and 1995, from a population comprised almost exclusively of the US-1 genotype to a population represented by new or recombinant genotypes.     

Genetic Analysis of Metalaxyl Insensitivity Loci in Phytophthora infestans Using Linked DNA Markers

ABSTRACT Previous studies indicated that incompletely dominant loci determine insensitivity by oomycetes to phenylamide fungicides such as metalaxyl. To compare the bases of insensitivity in different strains of the late blight pathogen, Phytophthora infestans, crosses were performed between sensitive isolates and isolates from Mexico, the Netherlands, and the United Kingdom that displayed varying levels of insensitivity. Segregation analyses indicated that metalaxyl insensitivity was determined primarily by one locus in each isolate, and that two of the isolates were heterozygous and the other homozygous for the insensitive allele. Metalaxyl insensitivity was also affected by the segregation of additional loci of minor effect. DNA markers linked to insensitivity were obtained by bulked segregant analysis using random amplified polymorphic DNA (RAPD) markers and the Dutch and Mexican crosses. By studying the linkage relationships between these markers and the insensitivity in each cross by RAPD or restriction fragment length polymorphism analysis, it appeared that the same chromosomal locus conferred insensitivity in the Mexican and Dutch isolates. However, a gene at a different chromosomal position was responsible for insensitivity in the British isolate.     

Population structure and host range of the potato late blight pathogen Phytophthora infestans in Peru spanning two decades

The genetic diversity of the late blight pathogen Phytophthora infestans infecting cultivated potato and alternative hosts growing in the vicinity of fields in the main potato-growing areas of the Peruvian Andes was characterized using collections from 1997–2013 as reference. The Peruvian P. infestans population, including previously collected and current isolates, consists of four clonal lineages (EC-1, US-1, PE-3 and PE-7) that belong to the A1 mating type and have been present in the country for decades. The first report of US-1 was in isolates collected between 1982 and 1986; meanwhile, EC-1 and PE-3 appeared for the first time in isolates from 1992 and PE-7 was found in 1997. The pathogen has a very broad host range among the solanaceous plants infecting cultivated potato, tomato, pear melon and several wild species. The solanaceous species growing in the vicinity of the potato fields sampled were identified and surveyed for late blight-like symptoms. Phytophthora infestans was isolated from nine wild species, including three new host species: Solanum zahlbruckneri, Solanum grandidentatum and Iochroma grandiflorum. There was no clear host specialization, but geographical substructuring was found as well as changes in the pathogen populations at the regional level. The clonal lineage EC-1, which is mostly resistant to metalaxyl, has complex virulence and contains a high level of subclonal variation, continues to dominate the population. Some multilocus genotypes of the EC-1 lineage were sampled in high frequencies and were found among the previously collected and new samples.     

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.     

Resistance to metalaxyl in Phytophthora infestans in the Netherlands

Two hundred and twenty two isolates ofPhytophthora infestans from infected potato foliage or tubers, obtained in 1981 from different potato fields in various regions of the Netherlands, were tested for their ability to infect potato leaf discs floating on solutions of metalaxyl of various concentrations. Isolates were designated as resistant to metalaxyl when sporulation occurred at a concentration of 1 g ml^–1 or higher.Forty-one isolates appeared to be resistant. Thirty-five of the resistant isolates and 29 sensitive ones originated from three areas involving circa 15% of the total Dutch potato acreage. In these areas 28% of the fields sampled were metalaxyl-treated and 78% of these fields yielded resistant isolates. Only one of the three areas had a history of metalaxyl resistance in 1980. Six metalaxyl-resistant and 152 sensitive isolates were found in potato fields in other areas including those with major problems of resistance in 1980. All of four old potato cull piles sampled in the latter areas yielded metalaxyl-resistant isolates and two of these sites had possibly been inoculum sources for nearby potato fields. Only 9% of the fields sampled in these areas were metalaxyl-treated and none of these yielded a metalaxyl-resistant isolate.The data are compatible with the idea that infected seed potatoes were the major source of the incoculum that initiated the early and severe late blight epidemic in 1981 in the Netherlands. The severity has been enhanced by late-blight-conducive weather conditions during the emergence of the crop and its early stages of development, when fungicides had not yet been applied. Old potato cull piles are assumed to have been less important in starting the 1981 epidemic.Apparently, infected seed potatoes carried predominantly metalaxyl-sensitive strains, which explains the prevalence of metalaxyl-sensitive strains in most of the fields. In 1980, the seed potato crop had been harvested before the problem of metalaxyl-resistance surfaced. Hence, it might be assumed that infection of seed potatoes, favoured by late-blight-conducive weather in 1980, had been caused by metalaxyl-sensitive strains. Evidence that seed potatoes could also carry metalaxyl-resistant strains was obtained in two cases. It might explain their early occurrence in fields located in areas without a history of metalaxyl resistance in 1980 and the rapid build-up of a resistant population in a number of metalaxyl-treated fields.Development of resistance in potato fields, which were under heavy disease pressure after spraying with a mixture of metalaxyl and mancozeb, could not be proven definitely. In one experimental field the seed tubers very likely carried metalaxyl-resistant strains, which initiated an epidemic, and in a second field the influx of resistant strains from elsewhere could not be completely ruled out. In the latter field spraying the mixture at a two-week interval could not effectively control the disease. In this field a considerable degree of tuber infection by metalaxyl-resistant strains was noticed.No correlation existed between the behaviour of a particular strain towards metalaxyl and the genes for virulence present. Among a total of 79 isolates, the race of which has been identified, 23 different races were found. Race 1.3.4.7.10.11 was most frequently found, followed by race 1.3.4.7.11. Among 37 resistant isolates 10 different races could be identified.Samenvatting Gedurende de zomer en najaar van 1981 werden 222 veldisolaten vanPhytophthora infestans met behulp van een drijftoets getoetst op resistentie tegen metalaxyl. Per perceel werd meestal één isolaat, hetzij afkomstig van een door de aardappelziekte aangetast blad, hetzij van een zieke knol, getoetst.Eén en veertig isolaten bleken resistent te zijn. Van deze 41 waren er 35 afkomstig uit West Brabant, de Hoekse Waard en Oost Brabant/Noord Limburg, waar ongeveer 15% van het totale aardappelareaal in Nederland is gelegen. Daarnaast werden uit deze gebieden 29 gevoelige isolaten verkregen. Van de bemonsterde percelen uit de drie genoemde gebieden was 29% één- of tweemaal met metalaxyl behandeld, meestal in combinatie met andere middelen. Uit 78% van de aldus behandelde percelen werd een resistant isolaat verkregen. In de genoemde gebieden werden in 1980 alleen in Oost-Brabant/Noord Limburg problemen ondervonden van metalaxylresistentie.In percelen in overige gedeelten van Nederland, waar in 1980 algemeen metalaxylresistentie werd waargenomen, werden in totaal slechts zes metalaxyl-resistente en 152 gevoelige isolaten aangetroffen. Geen enkel van de met metalaxyl bespoten percelen (9% van het aantal bemonsterde percelen) leverde hier een resistent isolaat op. Uit een viertal aardappelafvalhopen in deze gebieden waarop de aardappelziekte voorkwam, werden metalaxyl-resistente stammen verkregen en twee ervan hadden waarschijnlijk een naburig perceel besmet.De resultaten van het onderzoek en de waarnemingen in de percelen wijzen erop dat de vroege en hevige epidemie van de aardappelziekte in 1981 is veroorzaakt door besmet pootgoed waaruit secundair zieke planten zijn ontstaan. Daarnaast werd het uitbreken van de epidemie vooral begunstig door de voorPhytophthora gunstige weersomstandigheden gedurende de periode eind mei tot half juni en het feit dat de eerste bespuiting te laat werd uitgevoerd, hetzij door overmacht, hetzij door het te traag reageren op de waarschuwingen via de radio.In 1980 hebben voornamelijk metalaxyl-gevoelige stammen het pootgoed kunnen besmetten, hetgeen verklaarbaar is uit het feit dat in dat jaar de problemen met metalaxylresistentie pas werden waargenomen nadat het loof van het pootgoed was vernietigd. In een tweetal gevallen zijn evenwel aanwijzingen verkregen dat pootgoed ook besmet kon zijn met resistente stammen. Een dergelijke besmetting is waarschijnlijk ook de verklaring voor het op grote schaal voorkomen van resistente stammen in West Brabant en de Hoekse Waard na een eenmalige toepassing van metalaxyl.In een tweetal proefvelden, waarvan er één was gelegen in Zuid-Flevoland en het andere te Heelsum, is het ontstaan van een resistante populatie bestudeerd bij een twee-wekelijkse toepassing van een mengsel van metalaxyl en mancozeb. Pootgoed, gebruikt in het op praktijkschaal uitgevoerde experiment in Zuid-Flevoland, bleek waarschijnlijk besmet te zijn met metalaxyl-resistente stammen, zoals op 1 juli werd vastgesteld in een onbehandeld gedeelte van het perceel. Op het behandelde gedeelte bleek het mengsel de ontwikkeling van de ziekte aanzienlijk te vertragen; evenwel werd na 16 juli een wekelijkse bespuiting met een maneb/fentin acetaat-bevattend produkt uitgevoerd voor een zo goed mogelijk ziektebestrijding.Op het in Heelsum gelegen proefveld werd op 5 augustus de ziekte het eerst waargenomen in het behandelde gedeelte, nadat de planten in het onbehandelde gedeelte, ten gevolgen van een doelbewuste inoculatie met gevoelige stammen op een naast gelegen aardappel-selectieveld, reeds voor meer dan 50% waren aangetast. Uit aangetaste bladeren van het behandelde gedeelte konden metalaxyl-resistente stammen worden geïsoleerd. In de loop van augustus bleek het metalaxyl/mancozeb-mengsel niet in staat verdere uitbreiding van de ziekte tegen te gaan en trad knolinfectie op. Begin september werden uitsluitend resistente stammen aangetroffen.Geen enkel verband kon worden vastgesteld tussen het optreden van metalaxyl-resistentie en het voorkomen van een bepaald fysio van de schimmel. Op een totaal van 79 isolaten konden 23 verschillende fysio's worden geïdentificeerd. Het fysio 1.3.4.7.10.11 werd de meeste keren gevonden, gevolgd door fysio 1.3.4.7.11. Tien verschillende fysio's werden aangetroffen onder de 37 getoetste metalaxyl-resistente isolaten.     

基于无毒基因的晚疫病菌指纹类型与致病型关系研究

由致病疫霉Phytophthora infestans(Mont)de Bary引起的晚疫病是马铃薯生产上可导致绝收的世界性病害,在我国各地均有发生和流行[1].传统的致病型鉴别只局限于表型上的分析,无法对复杂多变的晚疫病菌致病型变化进行快速准确的预测,因此有必要探索新的方法来弥补常规手段的不足,以便更加准确有效地监测其变化趋势.晚疫病菌与寄主马铃薯这一病理体系中的单一抗性基因和相应的无毒基因符合典型"基因对基因"学说,因此基于抗病基因与无毒基因的关系对晚疫病菌分类将更为科学和实用.到目前为止,已知的晚疫病菌无毒基因序列有Avr1、Avr2、Avr3a和Avr4[2,3].本研究试图分析晚疫病菌基于无毒基因的指纹类型与马铃薯鉴别品种划分的致病型之间是否存在对应关系,以期进一步了解病原菌群体结构特征及遗传变异本质,从而更好地指导抗晚疫病育种和品种的合理布局.     

Studies of crop loss in potato blight caused by Phytophthora infestans

Relationships between healthy haulm area (HHA) and yield, and between losses of HHA and yield in the potato late blight system, were evaluated in several seasons and fields variously affected by the disease in Israel. The results showed that because of the different yield potential of potatoes cultivated in different conditions, evaluation of crop losses cannot be separated from evaluation of yield potentials of the crop as influenced by environmental and cultural factors.
In one calculation the yield and the HHA in the healthiest plot in each field were equated to 100% and losses in other plots were derived from comparisons with this plot. In such a calculation the relationships of yield loss to HHA loss in all fields and seasons fitted one regression equation, and a high correlation was obtained between both parameters. However, the actual haulm areas and yield weights differed widely from one field to another, even in the healthiest plots. Also, a similar percentage of destroyed haulms sometimes corresponded to different areas of the remaining haulms and to different yields. So no correlation was found between HHA and yields over all fields and seasons. Such a correlation was found only for several plots, variously affected by late blight, in the same field and season and in different fields and seasons yields responded in a different way to increases in HHA.     

Environmental and Genetic Factors Influencing Self-Fertility in Phytophthora infestans

ABSTRACT Phytophthora infestans is generally regarded as heterothallic-requiring physical proximity of two individuals of different mating type (A1 and A2) for oosporogenesis. Recent reports of limited selfing in young cultures of this oomycete stimulated us to investigate factors contributing to the phenomenon. The ability to produce oospores rapidly (within 2 weeks) in pure, single individual cultures (self-fertility) was tested in 116 individual isolates. The 116 isolates were from geographically diverse locations (16 countries) and were genetically diverse. Mating type and growth medium were the most prominent factors in determining if an isolate would be self-fertile. The majority of A2 isolates (45 of 47 tested) produced oospores when grown on a 50:50 mixture of V8 and rye B medium. In contrast, the majority of A1 isolates (65 of 69 tested) did not produce oospores on this medium. None of the 116 isolates produced oospores when grown on rye B medium (with no V8 juice). Further tests on representative A1 and A2 isolates revealed that oatmeal agar, tomato juice agar, and V8-juice agar all induced the A2 mating type isolate to produce oospores but did not induce the A1 mating type isolate to produce oospores. Calcium carbonate and pH did not alter the self-fertile oospore production in either A1 or A2 mating type isolates. For in vivo tests, the application of fungicide to potato or tomato leaf tissue either before or after inoculation did not stimulate any individual isolate (one A2 and one A1 isolate) to produce oospores in infected tissue. However, in all of the controls for all experiments (in vivo and in vitro), many oospores were produced rapidly if both strains grew in physical proximity.     

Activation of defence responses to Phytophthora infestans in potato by BABA

Late blight caused by Phytophthora infestans is one of the most devastating diseases of the potato crop. Resistance breeding and current fungicides are unable to control the rapidly evolving P. infestans and new control strategies are urgently needed. This study examined mechanisms of dl ‐β‐aminobutyric acid (BABA)‐induced resistance (IR) in the potato–P. infestans system. Leaves from two cultivars that differ in their degree of resistance, Bintje and Ovatio, were analysed after foliar treatment with BABA. Rapid activation of various defence responses and a significant reduction in P. infestans growth were observed in leaves treated with BABA. In the more resistant cultivar, Ovatio, the activation was both faster and stronger than in Bintje. Microscopic analysis of leaves treated with BABA revealed induction of small hypersensitive response (HR)‐like lesions surrounded by callose, as well as production of hydrogen peroxide (H₂O₂). Molecular and chemical analyses revealed soluble phenols such as arbutin and chlorogenic acid and activation of PR‐1. These results show a direct activation of defence responses in potato, rather than priming as reported for other plant species. They also show that the efficiency of BABA‐IR differs between cultivars, which highlights the importance of taking all aspects into consideration when establishing new methods for disease management.