Production, survival and infectivity of oospores of Phytophthora infestans

The formation of oospores of Phytophthora infestans was studied in tomato and potato crops and volunteer plants under field conditions, and in laboratory tests with leaf discs of potato cultivars differing in their level of race-nonspecific resistance. Oospores were readily detected in blight-affected tomato leaflets and fruits, and in leaflets of field crops and volunteer potato plants. Oospores extracted from blighted potato leaflets yielded 13 oospore-derived progeny. Oospores were also produced following inoculation of leaf discs of eight potato cultivars expressing different levels of race-nonspecific resistance with a mixture of sporangia of A1 and A2 isolates. The highest numbers of oospores were produced in cvs Bintje (susceptible) and Pimpernel (resistant), and the lowest in Nicola (intermediate resistance). The relationship between lesions per leaflet and oospore incidence, affected by varying A1 : A2 ratios, was explored using a simple mathematical model, and validated by comparing actual oospore production in leaflets with multiple lesions of the race-nonspecific-resistant potato clone Lan 22-21 with the predictions generated by the model. Survival of oospores was investigated after their incorporation in either a sandy or a light clay soil in buried clay pots exposed to the local weather conditions. Over 6 years these soils were regularly assessed for their infection potential using floating leaflets in a spore-baiting bioassay. Sandy and clay soils contaminated with oospores remained infectious for 48 and 34 months, respectively, when flooded. Infections of floating potato leaflets occurred within 84–92 h and ceased after 11 days. Soil samples remained infective if dried and re-flooded on two, but not more, occasions.     

Viability, germination and infection potential of oospores of Phytophthora infestans

Matings between five A1 and five A2 wild-type isolates of Phytophthora infestans from potato and tomato crops in the United Kingdom produced oospores in vitro in all cases examined. Oospores from the majority of crosses germinated, albeit at a low level (max 13-4%), after extraction from agar cultures by high-speed blending and treatment with novoZym 234. Viability of oospores from 20 crosses was tested by three methods. Two methods involved stains, either tetrazolium bromide (MTT) or phloxine B, and the third measured plasmolysis in 2 M NaCl. Both staining methods indicated a high percentage viability but gave false-positive results with heat-killed oospores. The plasmolysis method gave a lower percentage viability but no false positives. Oospores produced in vitro and stored either in sterile H₂O or in soil at temperatures between 0 and 20 C survived for between 5 and 7 months, the length of the experiments. Oospores buried in non-sterile field soil survived for up to 8 months (January-August). Inoculation of potato with zoospores of AI and A2 isolates produced oospores in stems but not in leaf tissue. In some, but not all cases, rapidly growing potato shoots (15-mm long) were successfully infected with oospores produced in vitro.     

Effect of metalaxyl on formation and germination of oospores of Phytophthora infestans

Oospores of Phytophthora infestans were produced in potato leaf discs floating on metalaxyl solution (100 μg mL⁻¹ a.i.) and inoculated with all combinations of two metalaxyl-sensitive and two -resistant parental isolates. Numbers of oospores produced varied between different matings, depending on parents, in the absence of the fungicide and when metalaxyl was added 0, 7, 14 and 21 days after inoculation. Oospores were not produced when metalaxyl was added at the time of inoculation (0 days) when either one or both parents were sensitive to metalaxyl. In two of three such matings further oospore formation was arrested when metalaxyl was added either 7 or 14 days after inoculation. Oospores extracted from leaf discs 14, 21 and 28 days after inoculation were assessed for germination on water agar after 21 days. Germination of oospores from water control treatments varied between 6 and 30% depending on the cross. Germination was significantly reduced in oospores of metalaxyl-sensitive parents extracted 28 days after inoculation of leaf discs treated with metalaxyl 0, 7 and 14 days after inoculation compared with the 21-day treatment. Minimal differences in germination were observed for oospores from the mating of resistant parents irrespective of metalaxyl treatment, although germination was generally low, not exceeding 8.5%.     

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.     

大豆疫霉卵孢子致死温度的测定

大豆疫霉根腐病是由大豆疫霉菌(Phytophthora sojae)侵染引起的、危害极其严重的最具毁灭性的大豆病害之一.病原菌以卵孢子在土壤中越冬,在条件适宜时导致下一年大豆发生根茎腐烂.病原菌卵孢子也可以存在于患病种皮内,但其在病害循环中的作用还不清楚.卵孢子是大豆疫霉的主要传播方式.本研究中对大豆疫霉卵孢子的致死条件进行了筛选.结果表明,使用60℃湿热处理大豆疫霉卵孢子20min或80℃干热处理大豆疫霉卵孢子20min,其萌发率为0,且死亡率为100%.     

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.     

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.     

Phytophthora infestans: populations,pathogenicity and phenylamides

Isolates of Phytophthora infestans (Mont) de Bary (the potato and tomato late blight pathogen) resistant to phenylamides appeared in Europe and North America in the late 1970s and early 1990s respectively. Concurrent, but coincidentally, with both these events there were radical structural shifts in the pathogen populations as immigrant genotypes from Mexico displaced the indigenous populations. Both A1 and A2 mating type isolates are now present in blighted crops, permitting alternative inoculum via germinating sexually produced oospores to influence dynamics of late blight populations. Studies of inheritance of ploidy, host-specific pathogenicity, mating type and resistance to antibiotics and phenylamide fungicides have provided insight into mechanisms of variation in this potent pathogen.     

Survival of Phytophthora infestans in Surface Water

ABSTRACT Coverless petri dishes with water suspensions of sporangia and zoospores of Phytophthora infestans were embedded in sandy soil in eastern Washington in July and October 2001 and July 2002 to quantify longevity of spores in water under natural conditions. Effects of solar radiation intensity, presence of soil in petri dishes (15 g per dish), and a 2-h chill period on survival of isolates of clonal lineages US-8 and US-11 were investigated. Spores in water suspensions survived 0 to 16 days under nonshaded conditions and 2 to 20 days under shaded conditions. Mean spore survival significantly increased from 1.7 to 5.8 days when soil was added to the water. Maximum survival time of spores in water without soil exposed to direct sunlight was 2 to 3 days in July and 6 to 8 days in October. Mean duration of survival did not differ significantly between chilled and nonchilled sporangia, but significantly fewer chilled spores survived for extended periods than that of nonchilled spores. Spores of US-11 and US-8 isolates did not differ in mean duration of survival, but significantly greater numbers of sporangia of US-8 survived than did sporangia of US-11 in one of three trials.     

Sprinkling irrigation enhances production of oospores of phytophthora infestans in field-grown crops of potato

ABSTRACT Two field experiments were conducted to study the effect of overhead sprinkling irrigation on oospore formation by the late blight fungus Phytophthora infestans in potato. Total rain (natural + sprinkling) accumulated in treatments of experiment 1 (winter 1997 to 1998) were 765, 287, and 219 mm and treatments of experiment 2 (winter 1999 to 2000) were 641, 193, and 129 mm. Sporangia from 11 isolates of P. infestans were combined in eight pairs, seven of A1 and A2 and one of A2 and A2 mating type, and were sprayed on field-grown potato crops (42 plants per plot at 7 m(2) each) and examined for their ability to form oospores in the host tissues. In experiment 1, oospores were recorded in a total of 132 of 1,680 leaflets (7.9%), 24 of 105 stems, and 2 of 90 tubers. In experiment 2, oospores were recorded in 40 of 519 leaflets (7.7%), but not in any of the 90 stems or the 45 tubers examined. Both the proportion of leaflets containing oospores and the number of oospores per leaflet increased with time after inoculation and were dependent on the rain regime, the position of leaves on the plant, and the isolate pair combination. In both field trials, increasing the rainfall significantly enhanced oospore production in leaves. Leaf samples collected from the soil surface had significantly more oospores than those collected from the midcanopy. Two pairs in experiment 1 were more fertile than the others, whereas the pair used in experiment 2 was the least fertile. The total number of oospores per leaflet usually ranged from 10 to 100 in experiment 1, but only from 2 to 10 in experiment 2. Maximal oospore counts in the field were 200 and 50 in experiments 1 and 2, respectively, but ranged from approximately 2,000 to 12,000 oospores per leaflet in detached leaves in the laboratory. We concluded that P. infestans can produce oospores in the foliage of field-grown potato crops, especially when kept wet by regular overhead sprinkling irrigation, but production was far below that in the laboratory.     

马铃薯晚疫病监测预警技术研究

结合我国马铃薯晚疫病发生为害规律和近年来测报调查工作经验,根据马铃薯晚疫病发生的主要影响因子,制定了科学的田间系统调查和大田普查监测方法,并总结概括了其他主要的预测预报方法。     

Characterization of some Asian isolates of Phytophthora infestans

A total of 401 isolates of Phytophthora infestans were collected from eight Asian regions (Korea, India, Taiwan, Indonesia, Thailand, Nepal, China and Japan) between 1992 and 2000 – 318 from potato and 83 from tomato. The isolates were analysed for mating type, metalaxyl resistance, RG57 fingerprinting, mitochondrial DNA (mtDNA) haplotype and the polymorphism of three allozyme loci, i.e. glucose-6-phosphate isomerase ( Gpi ), peptidase ( Pep ) and malic enzyme ( Me ). The isolates were multilocus-genotyped based on RFLP (RG57) fingerprint, dilocus allozyme genotype, mtDNA haplotype and mating type. Twenty multilocus genotypes were identified among 125 isolates. Of these genotypes, 14 had not been previously reported. Some of the multilocus genotypes were common to isolates from several geographical regions, suggesting migration. The metalaxyl-resistant isolates belonged to the multilocus genotypes JP-1, JP-2, and JP-3. Multilocus genotypes coexisting in a single field were found in following regions: Thailand (1994), central China (1996), Nepal (1997) and Japan (1998 and 2000). The possible origins of certain genotypes are discussed, including the possibility of sexual recombination within the P. infestans populations in Nepal and perhaps Thailand.     

Characterisation of Isolates of Phytophthora infestans From Hungary

A total of 36 single-lesion isolates were collected from 9 crops of potato and 13 of tomato in different regions of Hungary in the past decade, particularly in 1998. These were analysed for mating type, sensitivity to metalaxyl, allozyme genotype at glucose-6-phosphate isomerase and peptidase loci and genotype at 24 loci detected using the multilocus RFLP probe RG57. The ratios of the mating types A1 to A2 were 8:9 and 4:15 among isolates recovered from potato and tomato, respectively. Resistance to metalaxyl was found more frequently among isolates from potato and in the A1 mating type. The populations were not clearly differentiated on the basis of host origin. All isolates were homozygous (100/100) at the locus for glucose-6-phosphate isomerase. Unlike in other European countries, the most common peptidase allele was 96. Genotypes at the peptidase locus were 96/96 (50%), 96/100 (27.7%) and 100/100 (16.6%). In addition, one isolate from 1993 and another from 1998, were defined as 83/96, a genotype that had not been described elsewhere. The 18 RG57 fingerprints that were observed among 36 isolates, with one exception, seem to be unique to Europe. On the basis of combined genotypic traits, 20 multilocus genotypes were designated. These data, which reveal a remarkable variability with unique genotypes of the late blight pathogen, suggest that migration and sexual and/or asexual recombination have a role in the recent evolution of the pathogen in Hungary.     

致病疫霉有性生殖在晚疫病流行中的意义

对致病疫霉有性生殖的发生、有性生殖生理、遗传变异及可能造成的危害进行概述 ,对致病疫霉有性生殖在晚疫病流行中的意义作了探讨 ,为制定致病疫霉可靠的防治策略提供科学依据。     

防治马铃薯晚疫病药剂的筛选试验

试验结果表明:在马铃薯晚疫病发病初期,采用68%精甲霜·锰锌WG 75g/667m2、72%代森锰锌·霜脲氰WP100g/667m2或68.75%[噁]唑菌酮·锰锌WG 65g/667m2,施用2次后,均可收到87%以上的防治效果.     

Variation in sensitivity of Phytophthora infestans to fosetyl-Al

Twenty-three field isolates of Phytophthora infestans from North America, Europe and Israel were tested for sensitivity to fosetyl-Al on rye-seed-agar medium and on leaf discs from 12 potato cultivars. Sensitivity varied greatly between fungal isolates; their sensitivity in vitro was not significantly correlated with sensitivity in vivo. In vivo , there was a significant effect of potato cultivars on sensitivity, which interacted with fungal isolates. The fungicidal effect of fosetyl-Al was greater on older than on younger leaves, and greater with a 12-h photoperiod than in continuous light. Sensitivity of isolates to fosetyl-Al was significantly correlated with sensitivity to phosphorous acid (tested in vitro ) but was not correlated with sensitivity to metalaxyl (tested in vitro and in vivo ). The results explain some of the controversial reports on the efficacy of fosetyl-Al in controlling potato late blight.     

Genetic Structure of Peruvian Populations of Phytophthora infestans

ABSTRACT Isolates of the late blight pathogen Phytophthora infestans (n = 327) from the central to southern Peruvian Andes were systematically collected in 1997 to 1999 and analyzed to determine the pathogen's population structure at its host's center of diversity. No isolates of the A2 mating type were detected. Cluster analysis of DNA fingerprinting data indicated that the collection consisted of five major groups that were interpreted to be clonal lineages. Two of the lineages (US-1 and EC-1) have been previously described, and three (PE-3, 5, and 6) are described here for the first time. Collections from three areas in the central Peruvian Andes, including two key sites used in an international potato breeding program, consisted of isolates of the EC-1 lineage, which has been reported to dominate the pathogen population in Andean countries to the north of Peru. The collections from Cusco and Puno were more diverse. More than one lineage was detected in 10 of the 20 fields sampled in Cusco. Data on virulence, metalaxyl sensitivity, and band data for allozymes, mitochondrial DNA, and ipiB1 suggested that PE-3 may have been produced through recombination events between US-1 and EC-1. Restriction fragment length polymorphism and amplified fragment length polymorphism marker data were not consistent with this hypothesis.     

3株放线菌对致病疫霉抑制作用的比较研究

采用对峙培养法、菌悬液混合法和块茎切片法分别比较了3株拮抗放线菌Sy11、NB8和A5295对致病疫霉(Phytophthora infestans)的抑制作用及其对马铃薯晚疫病的预防效果。结果显示,放线菌活体菌株以NB8对致病疫霉菌丝生长的抑制作用最强(87.18%),Sy11次之(76.92%),A5295无明显抑制作用。3株拮抗株菌对致病疫霉游动孢子释放和孢子囊直接萌发的抑制作用均随菌液浓度下降而减弱。其中,Sy11的抑制作用最强,A5295最弱;其原液对游动孢子释放的抑制率分别为92.3%、79.0%和63.7%,对孢子囊直接萌发的抑制率分别为75.8%、72.4%和55.8%。在块茎切片上对马铃薯晚疫病的预防效果也以Sy11最高(75.90%),显著优于NB8和A5295。受Sy11和NB8抑制致病疫霉部分菌丝体形态发生畸变,而A5295对菌丝形态未表现出明显的致畸作用,表明Sy11防治马铃薯晚疫病潜力最大。