A2 Mating Type Research Articles

Population studies on Phytophthora infestans on potatoes and tomatoes in southern Germany

Fifty-seven isolates of Phytophthora infestans from blighted potato foliage were collected in 1995 in southern Germany and analysed for mating type and sensitivity to metalaxyl. Fifty-six of them were characterised as A1 and one as A2 mating types. Resistance to metalaxyl was observed frequently: 53 isolates were resistant, three were partially sensitive, and one was sensitive. In a subsequent field study in 1999, 84 isolates collected from blighted potato and tomato foliage were analysed for mating type. Seventy-two were characterised as A1 and twelve as A2 mating types. The response of 76 isolates to metalaxyl and to propamocarb was tested. The majority (42) of the 76 isolates was classified as resistant to metalaxyl; 31 were partially sensitive and only three isolates were sensitive. The results with propamocarb were less discrete; 10 isolates were classified as resistant and three were clearly sensitive. AFLP fingerprinting was used to examine the genetic structure of the southern German P. infestans population collected in 1999 and indicated that the tested population can be sub-divided into a tomato group, a potato group and a mixed group containing isolates collected from both crops. The presence of Ia and IIa mitochondrial DNA haplotypes indicates that the German P. infestans isolates belong to the new pathogen population that has also been reported in neighbouring regions of Europe. The present study indicates that at the beginning of the season only a few genotypes were present, and the population became genetically more variable at the end of the growing season.

Phenotypic and Genotypic Characterization ofPhytophthora infestansIsolates from China

A total of 288 (202 from potato and 86 from tomato) isolates of Phytophthora infestans were collected from 1998 to 2007 in China. The isolates were characterized based on mating type, in vitro metalaxyl sensitivity, virulence on potato differentials, allozymes of glucose-6-phosphate isomerase (Gpi), peptidase (Pep), and mitochondrial DNA (mtDNA) haplotype and examined by DNA-based simple sequence repeat (SSR) and random amplified polymorphic DNA (RAPD) fingerprinting. The majority (283 of 288) of the isolates were of the A1 mating type, the other three were the A2 mating type and two were the A1A2 mating type. Resistance to metalaxyl was frequently observed, with 248 (86.1%) resistant, 21 (7.3%) intermediate and 19 (6.6%) sensitive isolates identified. Virulence was assessed for 125 isolates on a set of 11 potato differentials and 61 races were detected. Most isolates were virulent on the differential genotype with gene R3, and all known virulence genes were found, with race 3.4.7.11 being the most common. This pattern did not appear to be associated with geographic origin, sample type, mating type or metalaxyl sensitivity. The dominant banding patterns for Gpi were 100/100/111 (176 isolates) and 100/100 (109 isolates), but genotypes 86/100 and 100/111 were also identified. All isolates tested were homozygous (100/100) at the Pep locus. The majority (205 of 288) of isolates tested was of mtDNA haplotype IIb, 76 were haplotype IIa and seven were the rare Ib haplotype. The genetic diversity of 60 representative isolates from China was assayed by two types of molecular markers, RAPD and SSR. A high level of polymorphism was found. The results demonstrated the diverse phenotypic and genotypic structure of the current populations of P. infestans in China.

Phenotypic and Genotypic Changes in the Phytophthora infestans Population in Taiwan – 1991 to 2006

AbstractLate blight, caused by Phytophthora infestans, is one of the most destructive diseases of tomato in Taiwan. A total of 655 isolates of P. infestans, including 29 isolates from potato, was collected from major tomato and potato production areas of Taiwan during 1991 to 2006. Isolates were characterized for their pathogenicity, mating type, in vitro metalaxyl sensitivity and molecular genotype (including allozyme pattern, mitochondrial genomic haplotype and DNA fingerprint) to monitor population changes in P. infestans. The population of P. infestans in Taiwan underwent a dramatic genetic shift in the 1997–1998 cool growing season. Isolates collected from tomato before 1997 were aggressive to tomato but not potato; most isolates obtained after 1998, were aggressive to both hosts. Metalaxyl sensitivity of isolates changed from sensitive/intermediate before 1997 to resistant since 1998. Similarly, the isolates obtained before 1997 were all US‐1 clonal lineage (including variants US‐1.1, US‐1.2, US‐1.3 and US‐1.4). During the 1997–1998 cool growing season, the US‐11 clonal lineage and the TW‐1 genotype appeared, possibly introduced on imported table potatoes. The US‐11 lineage spread rapidly and since 1999 has almost completely displaced the old population in Taiwan. Mating type determined by pairing with A1 and A2 reference isolages of P. parasitica, showed all isolates were of the A1 mating type, suggesting that the A2 mating type had not become established in Taiwan. The increasing percentage (up to 42.3% in 2006) of the US‐11 variants (including US‐11.l, US‐11.2, US‐11.3 and US‐11.4) implied that genomic diversity of the pathogen is changing quickly. Therefore, it is important to continuously monitor the population changes of P. infestans and develop an integrated management strategy for this disease.

Chemistry of microbial signal compounds

The ComX pheromone is a post-translationally modified oligopeptide that stimulates natural genetic competence controlled by quorum sensing in Bacillus subtilis. The structure of the ComXRO-E-2 pheromone produced by strain RO-E-2 was determined. In the pheromone molecule, a geranyl group is bound to the tryptophan residue, which results in the formation of a tricyclic ring structure. Molds in the genus Phytophthora are some of the most destructive phytopathogens known. An important biological event in Phytophthora is sexual reproduction. A factor known as hormone α1 is secreted by the A1 mating type, and this induces the formation of oospores in the A2 mating type. The structure of hormone α1 is a C-20 linear isoprenoid and the absolute stereochemistry was determined by chemical synthesis.

Characterization of Phytophthora infestans Populations in Colombia: First Report of the A2 Mating Type

Phytophthora infestans, the causal agent of late blight in crops of the Solanaceae family, is one of the most important plant pathogens in Colombia. Not only are Solanum lycopersicum, and S. tuberosum at risk, but also several other solanaceous hosts (Physalis peruviana, S. betaceum, S. phureja, and S. quitoense) that have recently gained importance as new crops in Colombia may be at risk. Because little is known about the population structure of Phytophthora infestans in Colombia, we report here the phenotypic and molecular characterization of 97 isolates collected from these six different solanaceous plants in Colombia. All the isolates were analyzed for mating type, mitochondrial haplotypes, genotype for several microsatellites, and sequence of the internal transcribed spacer (ITS) region. This characterization identified a single individual of A2 mating type (from Physalis peruviana) for the first time in Colombia. All isolates had an ITS sequence that was at least 97% identical to the consensus sequence. Of the 97 isolates, 96 were mitochondrial haplotype IIa, with the single A2 isolate being Ia. All isolates were invariant for the microsatellites. Additionally, isolates collected from S. tuberosum and P. peruviana (64 isolates) were tested for: aggressiveness on both hosts, genotype for the isozymes (glucose-6-phosphate isomerase and peptidase), and restriction fragment length polymorphism fingerprint pattern as detected by RG57. Isolates from S. tuberosum were preferentially pathogenic on S. tuberosum, and isolates from P. peruviana were preferentially pathogenic on P. peruviana. The population from these two hosts was dominated by a single clonal lineage (59 of 64 individuals assayed), previously identified from Ecuador and Peru as EC-1. This lineage was mating type A1, IIa for mitochondrial DNA, invariant for two microsatellites, and invariant for both isozymes. The remaining four A1 isolates were in lineages very closely related to EC-1 (named EC-1.1, CO-1, and CO-2). The remaining lineage (the A2 mating type) had characteristics of the US-8 lineage (previously identified in Mexico, the United States, and Canada). These results have important epidemiological implications for the production of these two crops in Colombia.

Ancient Trans-specific Polymorphism at Pheromone Receptor Genes in Basidiomycetes

In the majority of sexual organisms, reproduction occurs almost exclusively through the combination of distinct and alternate forms, called sexes or mating types. In some fungi, there can be dozens to hundreds of alternate alleles that determine compatible mating types. Such extensive polymorphism is expected to be maintained by balancing selection, and in extreme cases may give rise to trans-specific polymorphism. Here, we analyzed sequences of two pheromone receptors in the Microbotryum fungal species complex (Basidiomycota), which has only two alternate mating types. Several lines of evidence strongly suggest that the pheromone receptors are two allelic sequences acting to determine the alternate A1 and A2 mating types required for mating in Microbotryum. Phylogenetic trees of pheromone receptors in the Microbotryum species complex indicated a trans-specific polymorphism: the Microbotryum sequences from a given mating type were all more similar to the pheromone receptors of distantly related classes of fungi than to the alternate pheromone receptor in the Microbotryum species. A phylogenetic tree built using other known pheromone receptors from basidiomycetes showed that trans-specific polymorphism is widespread. The pheromone receptor alleles from Microbotryum appeared as the oldest, being at least 370 million years old. This represents the oldest known trans-specific polymorphism known in any organism so far, which may be due to the existence of sex chromosomes, obligate sexuality, mitochondrial inheritance linked to the mating type, and a highly selfing mating system in Microbotryum.

Phytophthora infestans isolates from Northern China show high virulence diversity but low genotypic diversity

Phenotypic and genotypic characteristics of 48 Phytophthora infestans isolates, collected in five provinces in Northern China between 1997 and 2003, were determined and compared with reference isolates. Characterisation included mating type, virulence, mitochondrial DNA (mtDNA) haplotype and DNA fingerprinting patterns based on simple sequence repeats (SSR) and amplified fragment length polymorphisms (AFLP). All isolates had the A1 mating type, mtDNA haplotype IIa and an identical SSR genotype (designated as SG-01-01) that differed from SSR genotypes found in the reference isolates, including those representing the 'old' US-1 lineage that dominated the P. infestans population worldwide prior to 1980. In contrast, the virulence spectra were highly variable and virulence to all resistance genes present in the standard differential set (R1 to R11) was found. AFLP analysis revealed some diversity; eight different AFLP genotypes were found that could be grouped into two major clusters. This study shows that there is very little genotypic diversity in the P. infestans population in Northern China. The occurrence of many different races within this rather uniform population is discussed in the framework of recent insights into the molecular determinants of avirulence in potato-P. infestans'gene-for-gene' interactions.

Occurrence of isolates of Phytophthora colocasiae in Taiwan with homothallic behavior and its significance

Leaf blight and corm rot caused by Phytophthora colocasiae are the most devastating diseases of taro. Oospores of P. colocasiae have not been considered important in the survival in natural soil because the pathogen is heterothallic and there is essentially no chance for the presence of both A1 and A2 mating types in the same host tissue. During our recent survey of the mating type distribution of P. colocasiae in Taiwan seven homothallic isolates of Phytophthora were obtained from diseased taro leaves at Tsu Chi in central Taiwan. These organisms were identified as P. colocasiae based on morphological characteristics, ITS sequence homology and pathogenicity to taro plants. The homothallic isolates of P. colocasiae segregated into A1 and A2 types in addition to the original A1A2 type during asexual reproduction and vegetative growth. The homothallic isolate and the mixture of its A1 and A2 segregants produced abundant oospores in live tissue of taro petioles on or away from soil, indicating the possibility of oospores as a survival structure and the source of genetic variation in certain areas in nature.

Genetic diversity in Phytophthora capsici from eastern China

Phytophthora capsici is a pathogen of several economically important crops, including pepper (Capsicum annuum). The pathogen attacks the roots, leaves, stems, and fruits of the plant. The variability among 37 strains of P. capsici from individual fields in Shandong, Anhui, and Fujian provinces in eastern China and 17 strains of P. capsici from other countries was assessed by determining virulence, mating type, and genetic profile using random amplified polymorphic DNA (RAPD) analysis. Five strains of Phytophthora nicotianae and two of Phytophthora infestans were also included in the RAPD analysis. No strains with mating type A2 were detected in P. capsici. Variation was found in the virulence assays based on pathogenicity to a set of nine pepper cultivar differentials, where the strains from eastern China grouped into seven pathotypes. RAPD analysis of all 37 strains from eastern China using 12 primers resulted in 129 fragments, of which 74.4% were polymorphic, revealing the existence of different patterns among strains. Cluster analysis using the unweighted pair-group method with arithmetic means separated the eastern China strains into two RAPD groups, and two non-Chinese strains clustered into group II. Among P. capsici strains derived from eastern China, no correlation was found between groups identified by RAPD analysis and pathotype groups defined by virulence. Phytophthora capsici was 36% similar to P. infestans and only 20% similar to P. nicotianae. Key words: pepper disease, phytophthora blight, RAPD, pathotypes. Phytophthora capsici est un pathogène de plusieurs récoltes économiquement importantes, y compris le poivre (Capsicum annuum). Le mycète attaque les racines, les feuilles, les tiges, et les fruits de la plante. La variabilité de 37 isole de P. capsici des champs individuels dans Shandong, les provinces d’Anhui et Fujian dans Chine d’est et 17 isolent de P. capsici des sources internationales a été évalué en déterminant la virulence, accouplant le type, et le profil génétique utilizant RAPD (random amplified polymorphic DNA) l’analyse. Cinq isolent de Phytophthora nicotianae et deux de Phytophthora infestans a été aussi inclus dans l’analyse RAPD. Non isole avec accoupler de type A2 a été détecté. Variation a été trouvé dans les essais de virulence a basé pathogenicity à une série de neuf arts de cultivar de poivre, où l’isole de Chine d’est a groupé dans 7 pathotypes. L’analyse RAPD de tout 37 isolent de l’utilization de Chine d’est 12 apprêts ont produit 129 fragments, dont 74,4 % était polymorphe, révélant l’existence de modèles différents parmi isole. L’analyse de groupe utilizant l’unweighted méthode de paire-groupe avec les moyennes arithmétiques a séparé la Chine d’est isole dans deux groupes RAPD, et deux international isole s’est rassemblé dans le groupe deux. Aucune corrélation a été trouvée entre les groupes révélés par l’analyse et pathotypes RAPD a défini par la virulence parmi P. capsici isole dérivé de Chine d’est. P. capsici était 36 % similaire à P. infestans et seulement 20 % similaire à P. nicotianae. Mots-clés : poivron, flétrissure phytophthoréenne, RAPD, pathovars.

Phytophthora nicotianae에 의한 아이비줄기썩음병

2007년 7월 전북 전주의 식물원에서 판매중인 아이비에서 줄기와 잎이 갈색으로 썩으며 급격히 말라죽는 증상으로부터 병원균을 분리 동정한 결과 Phytophthora nicotianae로 동정되었다. 병원균의 형태적 특성은 유두돌기가 있고 탈락성이 없는 난형의 유주자낭을 형성하였고, 크기는 평균 <TEX>$35.4{\times}25.2{\mu}m$</TEX>었고, 구형의 후막포자를 다량 형성하였으며, 크기는 평균 직경이 28.5 <TEX>${\mu}m$</TEX>였다. 유성생식형은 자웅이주로 A1 mating type이 관찰되었고 난포자의 크기는 평균 직경이 23.3 <TEX>${\mu}m$</TEX>였다. 균사의 생육적온은 <TEX>$25{\sim}30^{\circ}C$</TEX>였다. P. nicotianae에 의한 아이비줄기썩음병의 발생은 국내에서는 처음이다. A stem and leaf rot disease of English ivy(Hedera helix) was found in the flower market, Jeonju, Chonbuk province, Korea in July 2007. A Phytophthora species was isolated from the diseased part of the plant. Based on the mycological properties and pathogenesis the isolate was identified as Phytophthora nicotianae. The fungus produce ovoid sporangium which was ovoid to spherical which is noncaducous, papillate and averaged <TEX>$35.4{\times}25.2{\mu}m$</TEX> in dimension. Chlamydospores were abundantly produced on agar media and sized about 28.5 <TEX>${\mu}m$</TEX> in diameter. The fungus was heterothallic and A1 mating type. Oospores were measured 23.3 <TEX>${\mu}m$</TEX> in size. Optimum temperature for growth of the fungus was 25 to <TEX>$30^{\circ}C$</TEX>. Up to our knowledge, this is the first report demonstrating the stem rot on English ivy caused by P. nicotianae in Korea.

Phytophthora Species Causing Tanoak Stem Cankers in Southwestern Oregon.

Tanoak (Lithocarpus densiflorus) is a principal host of Phytophthora ramorum, cause of sudden oak death (SOD), in the western United States (1). In the course of SOD surveys in southwestern Oregon, other Phytophthora species were encountered to be causing stem cankers on tanoak that were indistinguishable from those caused by P. ramorum. In Oregon, SOD is subject to quarantine and eradication. Aerial surveys are flown two or more times a year to locate symptomatic tanoaks, which are then examined from the ground to determine the cause of death. Isolations on selective media were attempted from all trees with stem cankers typical of Phytophthora. Phytophthora species were identified by morphological features and DNA sequencing of either internal transcribed spacer (ITS) or the mitochondrial COX spacer region. ITS sequences were compared with validated GenBank records, and COX spacer sequences were compared with known reference isolates in the OSU collection. From 2001 through 2006, Phytophthora spp. were isolated from 482 of 1,057 tanoak stem cankers sampled. P. ramorum was isolated from 359 cankers, P. nemorosa was isolated from 102 cankers, P. gonapodyides was isolated from six cankers, P. cambivora was isolated from four cankers (all A1 mating type), P. siskiyouensis was isolated from four cankers, P. pseudosyringae was isolated from two cankers, P. cinnamomi was isolated from one canker (mating type A2), and P. taxon "Pgchlamydo" was isolated from one canker. Three cankers yielded isolates that were not identified but were closely related to P. pseudosyringae based on ITS sequence. No Phytophthora spp. were cultured from the remaining cankers. One isolate from each species identified (except P. ramorum and P. pseudosyringae) was tested for pathogenicity on tanoak stems (11.4 to 16.0 cm DBH) in the field. A 5-mm-diameter plug from the margin of a V8 agar culture was placed in a hole in the bark, covered with wet cheesecloth, and sealed with aluminum foil and duct tape. Each isolate was inoculated into five different stems. Each stem received three different isolates and an agar control. After 4 weeks, bark was removed to reveal lesion development. Lesions were measured (length by width), and pieces from four points on the lesion margin were plated in selective media to reisolate. P. cambivora, P. cinnamomi, P. gonapodyides, P. nemorosa, P. siskiyouensis and P. taxon "Pgchlamydo" all caused substantial lesions in inoculated tanoak trees (average area 11.5 to 18.6 cm2). In all cases, the species used for inoculation was recovered on reisolation from lesion margins. Control inoculations caused necrotic areas averaging 0.2 cm2. Isolations from these areas were clean. Prior to the recent SOD epidemic, no species of Phytophthora were known as pathogens of tanoak. The discovery of P. ramorum as a pathogen of tanoak in California was quickly followed by the discovery that P. nemorosa and P. pseudosyringae were also associated with tanoak cankers (2). Six years of diagnostic support for survey and detection of P. ramorum in tanoak forests of southwest Oregon has revealed the occurrence, at very low frequency, of at least five additional species of Phytophthora causing stem cankers in tanoak.

First Report of Crown and Root Rot Caused by Phytophthora capsici on Hydroponically Grown Cucumbers in Norway.

In December 2004, symptoms of root and crown rot were observed on cucumbers (Cucumis sativus L.) in a greenhouse in Norway. Cucumbers were the only crop of the greenhouse that used rockwool as a growing substrate in a hydroponical system. The first symptoms were detected in propagation material. One week after planting, symptoms of root and crown rot were observed and approximately 10% of the plants died. Later, losses of 50% in some greenhouses were observed. A yield reduction as much as 65% was estimated in the winter period (January and February). The two main cucumber cultivars planted were Armada and Lopez. In February 2005, Phytophthora capsici (Leonian) (1) was isolated on potato dextrose agar from a sample of cv. Lopez. The isolate produced deciduous, papillate sporangia (occasionally with two or three papilla) and pedicels that were sometimes longer than the sporangia. Sequencing of amplicons of the internal transcribed spacer region (ITS) rDNA and of the mitochondrial cytochrome c oxidase subunit 1 (Cox1) gene (2) confirmed the identification. Three isolates collected through 2005 from the same greenhouse were crossed with tester strains of P. cryptogea. Formation of oogonia and amphigynous antheridia was always observed in crosses with mating type A2; thus, all isolates were the A1 mating type. All three isolates grew well at 35°C and did not produce chlamydospores. A pathogenicity test was performed with one isolate of P. capsici. Four plants of cucumber cvs. Indira and Jessica were grown in a growth chamber at 24°C. Plants at the two-leaf stage were drenched with 20 ml of a zoospore suspension of 106 zoospores per ml per plant. After 18 days, all plants of both cultivars developed symptoms of crown rot or wilted and died. P. capsici was reisolated from inoculated plants of both cultivars. The pathogenicity test was repeated in the same way, but in a greenhouse with temperatures that ranged between 18 and 29°C. In addition, four plants of both cultivars at the four-leaf stage were inoculated with a suspension of 105 zoospores per ml. After 1 week, all plants developed crown rot or were irreversibly wilted, independently of the plant age or the zoospore concentration. To our knowledge, this is the first report of P. capsici in Norway.

Biologically active Phytophthora mating hormone prepared by catalytic asymmetric total synthesis

A Phytophthora mating hormone with an array of 1,5-stereogenic centers has been synthesized by using our recently developed methodology of catalytic enantioselective conjugate addition of Grignard reagents. We applied this methodology in a diastereo- and enantioselective iterative route and obtained two of the 16 possible stereoisomers of Phytophthora hormone alpha1. These synthetic stereoisomers induced the formation of sexual spores (oospores) in A2 mating type strains of three heterothallic Phytophthora species, P. infestans, P. capsici, and P. nicotianae but not in A1 mating type strains. The response was concentration-dependent, and the oospores were viable. These results demonstrate that the biological activity of the synthetic hormone resembles that of the natural hormone alpha1. Mating hormones are essential components in the sexual life cycle of a variety of organisms. For plant pathogens like Phytophthora, sexual reproduction is important as a source of genetic variation. Moreover, the thick-walled oospores are the most durable propagules that can survive harsh environmental conditions. Sexual reproduction can thus greatly affect disease epidemics. The availability of synthetic compounds mimicking the activity of Phytophthora mating hormone will be instrumental for further unravelling sexual reproduction in this important group of plant pathogens.

First Report of the A2 Mating Type of Phytophthora infestans on Tomato Crops in Taiwan, Republic of China

In a study of the Phytophthora infestans population in Taiwan, samples with symptoms typical of late blight were collected from field crops in an important potato- (Solanum tuberosum) and tomato-(Lycopersicon esculentum) production area in the central highlands region. Isolates were obtained by surface disinfecting leaf sections and plating them onto antibiotic-amended rye A agar (1). After subculturing, the pathogen was confirmed as P. infestans on the basis of morphological characters (2). Mating type was determined by co-inoculating unamended rye agar plates with mycelial plugs of the test isolate and a reference P. infestans isolate of either the A1 or A2 mating type (four plates per test isolate, two with different A1, and two with different A2 reference isolates). After incubation (15°C darkness, 7 to 14 days), plates were examined microscopically for the presence of oospores where the colonies interacted. In 2004, one isolate of 200 tested, and in 2006, one isolate of 102 tested, produced oospores only with A1 reference isolates and were concluded to be A2 mating type. In vitro testing showed the two A2 isolates were metalaxyl-resistant (ED50 values >100 mg of metalaxyl per liter on rye grain agar), which is typical of recent P. infestans isolates from potato and tomato in this area (2). Twenty-one single-sporangial isolates from each of the two A2 strains were tested for mating type against two different A1 isolates of P. infestans and confirmed as A2. These isolates were characterized using the techniques described by Deahl et al. (1) and had the allozyme genotype 100/100/111, 100/100 at the loci coding for glucose-6-phosphate isomerase and peptidase, respectively, and were mitochondrial haplotype IIb. This multi-locus genotype is characteristic of recent P. infestans isolates from tomato and potato in Taiwan, but all previous such isolates were A1 mating type and attributed to the US-11 clonal lineage (1). When evaluated on differential hosts, both A2 isolates were tomato race PH-1 and complex potato race R 0,1,2,3,4,7,9,11. RG57 fingerprinting showed that the A2 isolates had fingerprints identical to each other and to A1 P. infestans isolates of the US-11 clonal lineage from tomato in Taiwan (101 011 100 100 110 101 011 001 1). Koch's postulates were completed and the two A2 isolates were found to be highly aggressive on cultivars of potato and tomato. To our knowledge, this is the first report of A2 mating type strains of P. infestans in the field in Taiwan, but currently, their incidence is very low (<1%). One crop from which an A2 isolate was obtained also yielded an A1 isolate, while A1 isolates were obtained from crops in the vicinity of the other. The concurrent presence of the two mating types of P. infestans poses a risk of sexual reproduction and oospore formation in tomato or potato in Taiwan.

Survival and Spread of Phytophthora capsici in Coastal Peru

Phytophthora capsici is a soilborne pathogen that causes significant losses to pepper production in Peru. Our objective was to investigate the mechanisms by which P. capsici is able to survive and spread. During 2005 to 2007, 227 isolates of P. capsici were collected from four species of pepper (Capsicum annum, C. baccatum, C. chinense, and C. pubescens) and tomato (Solanum lycopersicum) at 33 field sites in 13 provinces across coastal Peru. All 227 isolates were of the A2 mating type and amplified fragment length polymorphism (AFLP) analysis indicates that 221 of the isolates had the same genotype. Analyses of six polymorphic single nucleotide polymorphism (SNP) loci showed fixed heterozygosity suggesting a single clonal lineage is widely dispersed. Members of the same clonal lineage were recovered during 2005 to 2007 from geographically separate locations from each of the host types sampled. Our results indicate that clonal reproduction drives the population structure of P. capsici in Peru. The impact of continuous cropping and irrigation from common river sources on the population structure in Barranca Valley are discussed.

A Foliar Blight and Tuber Rot of Potato Caused by Phytophthora nicotianae: New Occurrences and Characterization of Isolates.

Phytophthora spp. are pathogenic to many plant species worldwide, and late blight, caused by Phytophthora infestans, and pink rot, caused by P. erythroseptica, are two important diseases of potato. Another Phytophthora sp., P. nicotianae, was recovered from pink-rot-symptomatic tubers collected from commercial fields in Nebraska, Florida, and Missouri in 2005, 2006, and 2007, respectively. P. nicotianae also was recovered from foliage obtained from commercial potato fields in Nebraska and Texas exhibiting symptoms very similar to those of late blight. Isolates of P. cactorum also were recovered from foliar infections in a commercial potato field in Minnesota in 2005. Natural infection of potato foliage by P. cactorum and infection of wounded potato tuber tissue via inoculation with zoospores of P. capsici are reported here for the first time. Isolates of P. nicotianae, regardless of origin, were primarily of the A1 mating type. All isolates of P. nicotianae and P. cactorum were sensitive to the fungicide mefenoxam. Optimum growth of P. nicotianae, P. erythroseptica, and P. cactorum in vitro occurred at 25°C; however, only P. nicotianae sustained growth at 35°C. Regardless of the tissue of origin, all isolates of P. nicotianae and P. cactorum were capable of infecting potato tubers and leaves. However, isolates of P. nicotianae were less aggressive than P. erythroseptica isolates only when tubers were not wounded prior to inoculation. Pink rot incidence varied significantly among potato cultivars following inoculation of nonwounded tubers with zoospores of P. nicotianae, ranging from 51% in Red Norland to 19% in Atlantic. Phytophthora spp. also differed significantly in their ability to infect potato leaves. Highest infection frequencies were obtained with P. infestans and levels of infection varied significantly among P. nicotianae isolates. The rate of foliar lesion expansion was similar among isolates of P. nicotianae and P. infestans. Whereas P. infestans infections yielded profuse sporulation, no sporulation was observed with foliar infections of P. nicotianae.

Sporidiobolus johnsonii and Sporidiobolus salmonicolor revisited

The relationship between Sporidiobolus johnsonii and S. salmonicolor was investigated using rDNA sequence data. Two statistically well-supported clades were obtained. One clade included the type strain of S. johnsonii and the other included the type strain of S. salmonicolor. However, some mating strains of S. salmonicolor were found in the S. johnsonii group. These strains belonged to mating type A2 and were sexually compatible with mating type A1 strains from the S. salmonicolor group. DNA–DNA reassociation values were high within each clade and moderate between the two clades. In the re-investigation of teliospore germination, we observed that the basidia of S. salmonicolor were two-celled. In S. johnsonii, basidia were not formed and teliospore germination resulted in direct formation of yeast cells. We hypothesize that the S. johnsonii clade is becoming genetically isolated from the S. salmonicolor group and that a speciation process is presently going on. We suspect that the observed sexual compatibility between strains of the S. johnsonii and S. salmonicolor groups and the possible genetic flow between the two species has little biological relevance because distinct phenotypes have been fixed in the two taxa and intermediate (hybrid) sequences for LSU and ITS rDNAs have not been detected.

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.

Synthesis and absolute configuration of hormone α1

An important biological event in phytopathogens of the genus Phytophthora is sexual reproduction, which is conducted by two mating types, A1 and A2. A factor known as hormone alpha1 is secreted by the A1 mating type and induces the formation of sexual spores (oospores) in the A2 mating type. Here we describe the asymmetric synthesis and assignment of the absolute configuration of hormone alpha1 by oospore-inducing assays of the synthesized isomers.

First Report of the European Lineage of Phytophthora ramorum on Viburnum and Osmanthus spp. in a California Nursery.

Phytophthora ramorum S. Werres & A.W.A.M. de Cock is the causal agent of sudden oak death in California and Oregon forests and ramorum blight on a broad range of host species in wildlands and nurseries. It is thought to be an introduced pathogen and only three clonal lineages are known (3). The North American lineage (lineage NA1, mating type A2) is responsible for infections in California and Oregon forests. The European lineage (lineage EU1, predominantly A1) is responsible for infections in Europe, but has also been found in nurseries in Oregon and Washington. A third lineage (NA2) has only been isolated in a few instances from nurseries in Washington and California. In June 2006, P. ramorum was isolated from diseased Viburnum tinus, Osmanthus heterophyllus, and O. fragrans cultivars from a Humboldt County retail nursery in northern California. We genotyped isolates and placed them into clonal lineages using microsatellite markers developed for P. ramorum (3,4). Genomic DNA was extracted from mycelia with the FastDNA SPIN kit (Q-Biogene, Morgan, Irvine, CA). Primers used were PrMS6, Pr9C3, PrMS39, PrMS43a, PrMS43b, and PrMS45 (3) and 18, 64, and 82 (4). We sized fluorescently labeled amplicons using capillary electrophoresis (3100 Avant Genetic Analyzer, Applied Biosystems, Foster City, CA). Isolate genotypes were compared with control isolates of known clonal lineage, including BBA9/95 (EU1), Pr102 (NA1), and WSDA3765 (NA2). Three of four isolates belonged to genotype EU1. The fourth isolate, obtained from O. fragrans, belonged to genotype NA1. We repeated genotyping on independent genomic DNA extractions and obtained identical results. Two EU1 isolates and the single NA1 isolate were tested for mating type (1) and found to be of A1, A1, and A2 mating type, respectively. The coexistence of A1 and A2 mating types in the same retail nursery suggests the potential for sexual reproduction, as is the case in P. infestans where clonal and sexual populations exist (2), although to date, sexual reproduction in nature has not been documented in P. ramorum. The California retail nursery infestation highlights the risks associated with the unintentional transport of host nursery stock infested with P. ramorum.