Phenotypic and genotypic diversity of Phytophthora infestans populations in Scotland (1995–97)

In a survey of Scottish potato late blight ( Phytophthora infestans ) populations from 1995 to 1997, nearly 500 isolates were collected from over 80 disease outbreaks in commercial potato crops and gardens/allotments. The isolates were characterized by mating type, resistance to the fungicide metalaxyl and almost 300 were examined by DNA-based AFLP fingerprinting. These data were examined alongside cropping details to determine the population structure in the context of existing disease management strategies. A1 and A2 mating type isolates were present in both commercial potato crops and gardens or allotments although they coexisted more frequently in the latter sites. One-fifth of the isolates collected were of the A2 mating type and the frequency was similar over the 3 years and amongst sites. In 1995 the proportions of isolates that were sensitive and resistant to metalaxyl were equal (∼40%) but, over the following 2 years, the frequency of resistant isolates decreased and that of intermediate isolates increased. The mating type response to metalaxyl differed markedly, with 52% of A1 and only 5% of A2 isolates being resistant. Considerable molecular diversity was observed, with over half of the isolates having unique AFLP patterns. Analysis of the molecular and phenotypic data revealed a broad clustering of the population into three groups. Many factors point to an A2 population restricted by its sensitivity to phenylamides. The majority of the A2 isolates were found in a single AFLP group, but the presence of mixed mating type samples, an increasing frequency of isolates of intermediate metalaxyl resistance and the extent of the AFLP diversity suggest occasional sexual recombination, and thus gene flow, between groups.     

The Northern Ireland Phytophthora infestans population 1998–2002 characterized by genotypic and phenotypic markers

A total of 204 isolates of Phytophthora infestans from Northern Ireland, almost all from commercial potato crops, were collected over 5 years (1998–2002). Phenotypic diversity was assessed using mating type and metalaxyl resistance; genotypic diversity was assessed using two allozyme loci (glucose-6-phosphate isomerase, Gpi, and peptidase, Pep ), mitochondrial DNA haplotype and the multilocus RFLP probe RG57. All isolates were A1 mating type and Gpi 100/100 . The majority were Pep 100/100 , but four Pep 83/100 and six Pep 96/100 isolates were identified. Three mtDNA haplotypes were detected; haplotype IIa was the most common, but each year up to 2001 its frequency declined, with a concomitant increase in Ia isolates. Three isolates had the rare haplotype IIb, but this was only detected in 1998. Metalaxyl resistance and mtDNA haplotype were markedly associated: most haplotype Ia isolates were metalaxyl-resistant, whereas haplotype IIa was more commonly associated with metalaxyl sensitivity. Analysis of a subsample of 91 isolates revealed nine RG57 genotypes, three associated exclusively with haplotype IIa and six exclusively with haplotype Ia. The most common RG57 genotype (51% of isolates) comprised both metalaxyl-resistant and -sensitive haplotype IIa isolates. However, of haplotype Ia isolates, all metalaxyl-resistant phenotypes belonged to one of four RG57 types, one of which was the second most frequent overall (29% of isolates), while all metalaxyl-sensitive isolates belonged to one of two other types. The P. infestans population in Northern Ireland appears to consist of a limited number of clones related to, but differentiated from, the populations in mainland Britain and elsewhere in Europe.     

Phenotypic variation in Nordic populations of Phytophthora infestans in 2003

A total of 743 single-lesion isolates of Phytophthora infestans were collected in summer 2003 from Denmark, Finland, Norway and Sweden. Most of the isolates were tested for mating type, and subsets were tested for sensitivity to fungicides and virulence (host specific pathogenicity). Approximately 60% of the isolates were A1 mating type in each country. Both mating types were present in 40% of the fields where more than one isolate was tested, indicating strong potential for sexual reproduction. The proportion of metalaxyl-resistant isolates dropped to under 15% from the 60% observed in the early 1990s in Norway and Finland, possibly due to lower selection pressure because of decreased use of metalaxyl. Propamocarb-HCl sensitivity remained unchanged in the Nordic countries compared to the situation in 1997–2000 in Finland. Four isolates collected from Finland and Sweden were able to sporulate in the presence of this fungicide at a concentration of 1000 mg L⁻¹. In Norway and Finland the frequencies of virulence factors and pathotypes remained nearly unchanged since the 1990s, but the mean number of virulence factors per isolate increased from 5·6 to 6·3. In Denmark and Sweden virulence factors 2 and especially 6 were more common than in Norway and Finland. In addition, in the Swedish population the frequencies of pathotypes were quite even while in other countries pathotype 1,3,4,7,10,11 was most prevalent.     

Phytophthora infestans in a single field in southwest Sweden early in spring: symptoms, spatial distribution and genotypic variation

Sixty-eight isolates of Phytophthora infestans were collected from an early planted, fleece-protected potato field in southwest Sweden in May 2001. No infection was observed in the neighbouring fields. In the field investigated, infected plants were found in six discrete foci, with symptoms almost exclusively observed in the lower part of the canopy and numerous infections found on stems and on leaves touching the ground. The structure of the population was analysed based on mating type, mitochondrial haplotype and microsatellites (SSRs) as markers. Both mating types were present and haplotypes Ia and IIa were detected. Among 61 isolates analysed with microsatellite markers, 14 multilocus genotypes were distinguished based on six polymorphic loci. Out of the six foci, three included 3–5 genotypes each. There were unique genotypes in all foci, except two closely situated to each other. These findings strongly suggest that soilborne oosporic inoculum contributed significantly to initiate the late-blight epidemic in the investigated field.     

Phenotypic and genetic variation in Finnish potato-late blight populations, 1997–2000

This study was conducted on potato late blight samples collected for monitoring early attacks in Finland from 1997 to 2000. Most of the 1726 Phytophthora infestans isolates were collected soon after the onset of the epidemics. Most of the isolates were tested for mating type as well as metalaxyl and propamocarb hydrochloride (HCl) sensitivity, while virulence on potato R-gene differentials and mitochondrial DNA (mtDNA) haplotype were determined for a subset of the isolates. In half of the fields from which more than one isolate was tested, both mating types were detected, indicating strong potential for sexual reproduction. The mating types coexisted more often in organic fields or gardens than in conventional fields. The proportion of A1 mating type decreased from approximately 80% (1997 and 1998) to 22% by the year 2000. Simultaneously, the proportion of isolates resistant to metalaxyl decreased from approximately 40% to 16%. Resistance to metalaxyl was confined to the IIa mtDNA haplotype and clearly associated with the A1 mating type, as resistance was 10 times more common among A1 isolates than among A2 isolates. Resistance to metalaxyl therefore probably derives from common descent from an isolate with the IIa haplotype. Most of the regional variation in metalaxyl sensitivity was also linked to mating type, as both metalaxyl resistance and the A1 mating type were most common in the north and south-west of Finland. In contrast to metalaxyl, only three propamocarb-HCl-insensitive isolates were found, but propamocarb-HCl sensitivity decreased significantly during the study.     

云南省德宏州冬作马铃薯晚疫病菌表型特征研究

 2003~2005年,从云南省德宏州新兴冬作马铃薯主产区采集的马铃薯晚疫病病样中分离纯化到86株致病疫霉菌,对其代表性菌株的表型特征进行了较系统的研究。交配型实验结果表明:所测定的62个菌株都属于A1交配型,未发现A2交配型或自育型菌株。毒性基因鉴定、监测及生理小种组成分析显示:德宏州存在所有已知可鉴定的毒性基因,而且生理小种组成非常复杂;所测定的66个菌株可区分出36个生理小种,其中,出现频率最高的小种为1,2,3,4,5,6,7,8,9,10,11,占被测菌株的18.18%,在主产区潞西县和盈江县均有分布。对60个菌株的甲霜灵抗性测定结果表明:56.67%的菌株表现为敏感,25.00%的菌株中抗,18.33%的菌株对甲霜灵有抗性。本文对此实验研究结果作了初步的分析和讨论。     

Variation in populations of Phytophthora infestans in Finland and Norway: mating type, metalaxyl resistance and virulence phenotype

Phytophthora infestans was isolated from potato leaves and tubers collected from different parts of Finland in 1990–96 and Norway in 1993–96. Isolates were assessed for mating type, resistance to metalaxyl and virulence phenotype. In Finland 15% of 200 isolates tested and in Norway 25% of 642 tested were A2 mating type. Differences in the A1/A2 ratio were evident among regions and A2 was not found in northern areas. In Finland the frequency of metalaxyl-resistant isolates, among 1834 tested, decreased from 59% in 1990–95 to 2% in 1996. In Norway 59% of 491 isolates tested were resistant to metalaxyl in 1996. Among 269 Finnish and 105 Norwegian isolates, all known virulence genes were found in both countries and race 1.3.4.7.10.11 was the most common (resistance gene R9 was not included in the differential set). Oospores were observed in potato leaves from three locations in the southern part of Norway. The implications of the 'new' populations in the Nordic countries are discussed.     

甘肃省部分地区马铃薯晚疫病菌交配型及线粒体DNA单倍型分析

由致病疫霉Phytophthora infestans(Mont.) de Bary侵染引起的马铃薯晚疫病是一种世界范围内的毁灭性病害。为了明确甘肃省马铃薯晚疫病菌的遗传结构,对2018年甘肃省定西市、张掖市和陇南市采集的130株晚疫病菌进行了交配型和线粒体DNA (mtDNA)单倍型测定。结果表明：从130株晚疫病菌中检测到A1和A2两种交配型,A1交配型为14株(10.77%),A2交配型为116株(89.23%);检测到Ⅰa、Ⅱa和Ⅱb 3种线粒体DNA单倍型,发生频率分别为88.46%、3.85%和7.69%。Ⅰa单倍型菌株的交配型为A1和A2,Ⅱa单倍型菌株交配型为A1和A2,Ⅱb单倍型菌株交配型为A1。该结果表明甘肃省部分地区马铃薯晚疫病菌以A2交配型,Ⅰa mtDNA单倍型为主。甘肃省各采集地晚疫病菌群体结构或与当地种薯繁育、调运有关。     

French isolates of Phytophthora infestans from potato and tomato differ in phenotype and genotype

Prior to 1996, the A2 mating type of Phytophthora infestans was not detected on potato in France, but was found at one site on tomato in 1995. This finding lead to the question of the extent of differences and relationships existing between the populations of P. infestans present on each host. A collection of 76 isolates collected in France, mainly in 1996, from potato and tomato was characterised for mating type, allozyme genotype at the Gpi and Pep loci, and mitochondrial DNA haplotype; 74 of these isolates were also characterised for multilocus RFLP fingerprint, and 62 for virulence. All isolates except four showed allozyme genotypes (Gpi 90/100 or 100/100, Pep 83/100 or 100/100) and mtDNA haplotypes (Ia or IIa) characteristic of the populations introduced into Europe in the late 1970s. The four exceptions were isolates collected from tomato in Southern France in 1988-1991, which showed some characteristics of the former European populations (Gpi 86/100, Pep 92/100, mtDNA Ib). Both mating types were present among the collections from both hosts, but isolates with the A2 mating type were found on potato only in one garden crop, adjacent to tomato. Nine different RG57 fingerprints were observed, with a greater diversity among tomato isolates. Furthermore, tomato and potato collections differed markedly in the frequencies of genotypes present. Finally, tomato isolates generally had a lower virulence complexity than potato isolates. These data suggest that P. infestans populations on tomato and potato are largely separated, despite the occurrence of limited gene flow.     

青海马铃薯晚疫病菌线粒体DNA单倍型鉴定及分析

 为查明青海地区晚疫病菌单倍型分布情况,本研究对青海不同地区采自2006～2007年的70个晚疫病菌材料的线粒体单倍型进行了分析,结果表明：94.3%的菌株为Ⅱa型,5.7%的菌株为Ⅱb型,其中Ⅱb型均分离自番茄。这说明青海地区马铃薯晚疫病菌群体类型比较单一,主要为Ⅱa型,线粒体单倍型不存在多态性。     

Phenotypic and genotypic variation in Michigan populations of Phytophthora infestans from 2008 to 2010

Severe potato and tomato late blight epidemics in Michigan since 2008 prompted characterization of Phytophthora infestans isolates from the region. From eight counties in Michigan, 124 isolates were collected from infected potato and tomato plants from 2008 to 2010 and characterized using ‘classical’ markers and microsatellites. The classical markers included mating type, Gpi allozyme, mitochondrial DNA haplotype, sensitivity to metalaxyl‐M and tuber pathogenicity. All isolates from 2008 to 2010 were A2 mating type and Ia mtDNA haplotype (124 isolates), 105 isolates had Gpi profile 100/122, 17 isolates had the profile 100/100/111 and the remaining two isolates had 100/111/122. Sensitivity to metalaxyl‐M, expressed as EC₅₀ for mycelial growth in vitro, ranged from <0·1 to 91 μg mL^?1, where 95 and 96% of isolates were classified as either sensitive or intermediate in 2008–2009 and 2010 respectively. The metalaxyl‐M sensitivity and dominant Gpi profile were typical of clonal lineage US‐22, first isolated in 2008 in North America from tomato plants. Tuber pathogenicity, characterized as severity of tuber late blight, was also variable among isolates; however, isolates were less aggressive than previous genotypes present in Michigan, such as US‐8. Microsatellites (simple sequence repeats; SSRs) revealed a shift in the population, characterized by two clusters differentiated over time. These results suggested displacement of the US‐8 genotype by US‐22 from 2008 to 2010 in Michigan. Continuous tracking of changes within P. infestans populations provides evidence of genetic shifts due to migration, prompting modification of management strategies based on the phenotypic characteristics of causal genotypes.     

Phenotypic and Genotypic Characterisation of Northern Ireland Isolates Of Phytophthora Infestans

Genetic and phenotypic diversity of the population of the late blight pathogen Phytophthora infestans in Northern Ireland was assessed from 223 single-lesion isolates collected from 29 sites in 1995 and 1996. The proportion of metalaxyl-resistant isolates was approximately 14%. The A2 mating type was not detected. Allozyme analyses revealed that the Northern Ireland isolates were monomorphic and homozygous at the loci coding for allozymes of glucose-6-phosphate isomerase and peptidase (Gpi 100/100, Pep 100/100). The majority of isolates tested (156 out of 162) were mitochondrial DNA haplotype IIa. Among the remaining isolates, four were haplotype Ia, two were the rare IIb haplotype and none was of the old Ib type. RAPD-PCR analysis of selected isolates revealed relatively little diversity among the Northern Ireland isolates, which clustered separately from isolates from GB and the European mainland.     

云南省马铃薯晚疫病菌的交配型、抗药性及生理小种分布的研究

 从云南省陆良县、建水县和南涧县采集2000年秋播的马铃薯晚疫病标本,分离获得124个致病疫霉菌株,连同1999~2000年分离自昆明、曲靖等地的春播马铃薯晚疫病的10个菌株共134个,采用常规技术对这些菌株进行了交配型、抗药性及生理小种的研究。交配型测定结果为:陆良县的18个菌株中有3个为A₂交配型,其余15个菌株为A₁交配型,建水县和南涧县的106个菌株及昆明、曲靖1999~2000年的10个菌株都是A₁交配型。测定了83个菌株对瑞毒霉(metalaxyl)和烯酰吗啉(dimethomorph)的抗药性,结果有12.0%的菌株对瑞毒霉表现抗性,16.9%表现中抗,71.1%表现敏感,其中陆良、昆明和曲靖的抗瑞毒霉菌株的比例大于建水和南涧的抗药性菌株的比例;所测定的83个菌株都对烯酰吗啉表现敏感。生理小种鉴定结果表明,云南晚疫病菌为3.4、0、3和4生理小种,分别占鉴定菌株总数的48.0%、32.5%、15.6%和3.9%。但南涧仅发现3.4和0小种,而昆明有0、3、3.4和4小种,陆良有0、3和3.4小种,可见昆明和陆良地区的晚疫病菌生理小种比南涧地区的生理小种的组成更趋多样。     

Inoculum Sources and Genotypic Diversity of Phytophthora infestans in Southern Flevoland,The Netherlands

Genotypic changes in populations of Phytophthora infestans in Southern Flevoland (150km²) were analysed by characterising isolates from potato refuse piles, conventional and organic potato fields, and potatoes and tomatoes in allotment gardens for mating type (1712 isolates) and DNA fingerprint pattern using probe RG57 (1048 isolates). The overall percentages of genotypes (and of isolates) that were A2 varied from 32 (4) in 1994 to 45 (56) in 1996. Among the 1048 isolates 170 different genotypes were identified, of which 138 (81%) were rare (i.e., detected in only one sampling site in the research area during 1993–1996). Many rare genotypes were encountered in organic potato fields and in allotment gardens. In 1994 and 1995, four genotypes were abundant. The highest percentages of isolates with these common genotypes were encountered in refuse piles and conventional potato fields. The common genotypes were nearly absent in 1996, suggesting that the population may have passed through a bottleneck at the transition from 1995 to 1996. The Shannon index of genotypic diversity was high in allotment gardens and in organic potato fields. For the total populations the normalised Shannon index of genotypic diversity increased from 0.34 in 1994, with weather favourable to late blight, to 0.61 in 1996, with unfavourable weather. The high numbers of rare genotypes detected every year indicate that oospores may act as an infection source in commercial potato fields. However, refuse piles were identified as the most important infection sources for commercial fields in 1994 and 1995. In 1996 disease in commercial organic fields was probably initiated by a few genotypes originating from seed tubers. In allotment gardens oospores were probably the most important infection source.     

2009年黑龙江和吉林省马铃薯晚疫病菌表型结构研究

为明确2009年黑龙江和吉林两省马铃薯主产区晚疫病菌的表型特征,本研究对所采晚疫病菌的交配型、甲霜灵抗性及生理小种分别进行了测定.结果表明:被测黑龙江(46株)和吉林(48株)两省的菌株均为 A1 交配型.黑龙江省被测菌株甲霜灵高抗、敏感菌株各占50％;吉林省被测菌株中高抗、敏感菌株分别占70.8％、29.2％;两省均未发现中抗菌株.其中采自吉林敦化菌株均为高抗菌株,采自吉林榆树的菌株均为敏感菌株,甲霜灵抗/感比例地区间差异明显.在被测33个黑龙江菌株中发现24个生理小种,而吉林49个菌株中发现19个生理小种,其中9个生理小种两省共有.另外,两省被测菌株中还发现了18株“超级毒力小种”.因此,建议加强种薯监测,防止 A2交配型传入;在生产上使用甲霜灵类药剂防治马铃薯晚疫病应该以菌株的抗性监测数据为依据;进一步加强生理小种监测,合理布局抗病品种.     

Random amplified polymorphic DNA (RAPD) analysis of Phytophthora infestans isolates collected in Canada during 1994 to 1996

Mating type, glucose-6-phosphate isomerase ( Gpi ) allozyme banding patterns, response to the fungicide metalaxyl and random amplified polymorphic DNA (RAPD) markers were used to characterize genetic variability among 141 Canadian isolates of Phytophthora infestans collected between 1994 and 1996. Multiple correspondence analysis of RAPD profiles separated isolates into 21 groups that were not correlated to groups defined by mating type, Gpi allozyme banding patterns or response to metalaxyl. Population subdivision analysis showed that 97% of the total genetic variation was found among individuals within populations, compared with 3% among populations. The average similarity coefficient among isolates was 80%. No significant differences in haplotypic diversity were observed among the years under study, but levels of genetic diversity among local populations of P. infestans were high (0.76). All classes of response to the fungicide metalaxyl were observed, with 55% of isolates displaying moderate levels of insensitivity. The high level of genetic diversity detected within populations indicates that migration and sexual recombination probably play important roles in the population biology of P. infestans in Canada.     

Aggressiveness and other factors relating to displacement of populations of Phytophthora infestans in England and Wales

Mating type, in vitro sensitivity to the phenylamide fungicide metalaxyl, and mitochondrial (mtDNA) haplotype were determined in some or all of 618 isolates of Phytophthora infestans from the years between 1978 and 1995. A2 mating type occurred infrequently in most but not all years and insensitivity to metalaxyl increased over time. After 1982, the mtDNA Ib haplotype was largely replaced (except for one isolate in 1986 and one in 1995) by two new haplotypes, Ia and IIa. Type Ia was much more common than type IIa.Approximately one quarter of these isolates (165) were compared using two components of fitness associated with aggressiveness (infection frequency × number of sporangia per lesion) on detached leaves of cultivars Maris Piper, Cara and Stirling, which were chosen as exhibiting increasing levels of race non-specific resistance. Isolates were compared with three standard isolates of low, intermediate or high aggressiveness, and the data standardised for comparison between experiments. On cvs. Cara and Stirling, but not on Maris Piper, mtDNA Ia and IIa haplotypes were more aggressive than type Ib in several separate experiments. Similarly, metalaxyl sensitive phenotypes were more aggressive than insensitive phenotypes on Cara and Stirling but not on Maris Piper. The displacement of mtDNA type Ib by types Ia and IIa over this period may have been a result of the lower aggressiveness and lack of complete insensitivity to metalaxyl in type Ib isolates.     

Production and Germination of Oospores of Phytophthora infestans (Mont.) de Bary in Morocco

One hundred and eight isolates of Phytophthora infestans were collected from infected potato and tomato crops in the middle-north of Morocco during 1997–2000. Pairings of these isolates with tester isolates of mating type A1 and A2 revealed that 60% of the isolates were mating type A2 (65/108) and 40% were mating type A1. After 10 days incubation at 20 °C and a 16-h photoperiod, approximately 25% and 18% of the oospores produced in-vitro germinated in potato soil extract and potato root extract, respectively. Oospores were observed in potato leaf tissues in pairings that were fertile in-vitro. Maximum production of oospores was obtained in potato leaves of cultivars that were moderately susceptible (Desirée, Nicola) after 10 days of incubation at 15 °C and a 16-h photoperiod. These results confirm the presence of P. infestans strains that are sexually compatible under Moroccan climatic conditions. Production of oospores constitutes a threat for these crops because of the occurrence of recombinants with new virulences which may be difficult to control and as a consequence survival of oospores in absence of the host plant in the soil.     

Late-blight epidemics on potato in Finland, 1933–2002; increased and earlier occurrence of epidemics associated with climate change and lack of rotation

Changes in the incidence and onset of potato late-blight epidemics in Finland were investigated and compared with possible changes in climate, presence of soil-borne inoculum, and aggressiveness of Phytophthora infestans populations. Datasets were constructed from leaf blight assessments in cultivar trials or fungicide tests carried out at eight experimental sites during the periods 1933–1962 and 1983–2002. Additional data were obtained from late-blight monitoring projects carried out from 1991 to 2002. From 1998 to 2002, the risk of blight outbreak was 17-fold higher compared with the periods 1933–62 and 1983–1997. Simultaneously, the outbreaks of the epidemics began 2–4 weeks earlier. The changes observed were associated with a climate more conducive to blight in the late 1990s. Lack of rotation also advanced blight epidemics by an average of 9 days in 1998–2002, but it did not have this effect in 1992–1997, suggesting that soil borne inoculum may not have been a significant threat to potato until the late 1990s. The aggressiveness of the P. infestans isolates seemed to have only minor effect on the onset of the epidemics after 1991, as the apparent infection rate remained unchanged despite weather conditions more favourable to late blight in the late 1990s. As a consequence of the more frequent and earlier epidemics, the sales of fungicides used against late blight in Finland increased 4-fold from the 1980s to 2002.