Distribution and frequency of occurrence of potato cyst nematode pathotypes in Northern Ireland

The distribution and frequency of occurrence of potato cyst nematodes, Globodera rostochiensis and G. pallida , were examined in Northern Ireland. Globodera rostochiensis (pathotype Rol) was the most abundant species, while Pal and Pa3 pathotype reactions could be discerned within G. pallida. Unlike other parts of the UK, species proportions appear not to have changed over the last 20 years, probably as a result of the legislative control currently practised combined with the limited use of Rol-resistant cultivars within the country. Differences in the geographical distributions of the three pathotypes were observed, and these are discussed in relation to original potato cyst nematode introductions and subsequent agricultural practices.     

Molecular characterization of a morphologically unusual potato cyst nematode*

During routine screening of soil samples for potato cyst nematodes, a morphologically variant cyst was detected. The internal transcribed spacer regions of the ribosomal DNA repeat unit from individual juveniles hatched from the cyst were amplified by PCR. The resulting products were digested with a range of restriction enzymes and their RFLP profiles compared with standard samples of Globodera rostochiensis and Globodera pallida . Most of the RFLP profiles yielded patterns similar to those of G. pallida although some restriction enzymes gave hybrid patterns showing elements of both G. rostochiensis and G. pallida . Clones of individual ITS regions from a single nematode from the variant cyst displayed either a G. pallida or G. rostochiensis pattern.     

Morphological and molecular characterization of Globodera populations from Oregon and Idaho

An unusual population of cyst nematode was found in soils collected from a Powell Butte, OR field with a cropping history including potato, wheat, other crops, and significant weed presence. These nematodes could not be placed with certainty into any known species and exhibited some unique morphological features in some specimens. Compared with Globodera pallida, the cyst body length was slightly longer and the second-stage juvenile stylet length was slightly shorter. In some individuals, the J2 stylet knob height was greater and the tail annules were more prominent than in G. pallida, and the tail abruptly narrowed, with a slight constriction near the posterior third of the hyaline terminus. Compared with G. rostochiensis, the hyaline tail terminus had a larger number of refractive bodies, and cysts of this population had a smaller Granek's ratio and fewer cuticular ridges between the anus and vulva. In some individuals, the tail termini of second-stage juveniles were more bluntly pointed, and the stylet knobs were more anteriorly directed with greater height. Unlike G. tabacum, the cyst wall often lacked a network-like pattern and, in some individuals, the juvenile tail terminus distinctly narrowed after a constriction. Molecularly, the population was distinct from G. pallida, G. rostochiensis, and G. tabacum. Multiplex polymerase chain reaction of the internal transcribed spacer (ITS) rDNA region gave results similar to G. tabacum; however, ITS restriction fragment length polymorphism patterns were observed to have individual bands in common with G. rostochiensis and G. pallida. Phylogenetic analysis based on ITS1 and -2 rDNA sequences showed greatest similarity to populations from Argentina and Chile; together, they form a moderately supported clade, distinct from G. rostochiensis, G. tabacum, G. "mexicana," European type G. pallida, and several G. pallida populations from South America.     

Occurrence of Globodera pallida in Hungary

Golden potato cyst nematode, Globodera rostochiensis , has been known to occur in Hungarian potato fields for more than 20 years. The Hungarian NPPO has been carrying out a country-wide Globodera monitoring programme since 1995 and has inspected 16 000 samples. Until 2000, all records were of G. rostochiensis . However, in 2001, three localities in Pest county were found to be infested with Globodera pallida . This was the first official record of G. pallida in Hungary. The two species have been found together in five fields. Unmixed populations of G. pallida have not been found. Identification was based on morphometric data, as well as on biochemical methods (ELISA technique).     

Decline of potato cyst nematodes, Globodera rostochiensis and G. pallida, in spring barley microplots

In microplots of nine soils freed from volunteer potatoes and sown for four consecutive years with spring barley, the eggs of five populations of Globodera pallida and four populations of G. rostochiensis declined at annual rates of 12-8-40-5%. One population of G. rostochiensis declined faster than other populations of either species, although average rates of decline did not differ significantly between the two species of Globodera. These results are compared with earlier research and their relevance to control of the nematodes by crop rotation is discussed.     

Variation at the b Mating Type Locus of Ustilago maydis

ABSTRACT Population level diversity at the Ustilago maydis b mating type locus was determined in samples from four Minnesota locations using a combination of plate mating techniques and a polymerase chain reaction (PCR)-based assay. The PCR method allows rapid identification of b types from samples of natural populations and utilizes the hypervariable regions of the b locus that determine mating type specificity. Results demonstrated high levels of b diversity within populations, with one population yielding 17 of the total 18 b types found in the study. Pairwise G(ST) values were in the range of 0.02 to 0.05, and common b mating types were found across broad geographic distances. These data demonstrated that very low levels of differentiation among U. maydis populations occur with respect to b locus variation. Consistent with frequency-dependent selection models, b types were represented at approximately equal frequencies within the entire Minnesota population. However, neutral evolutionary models for patterns of geographic distribution and variation at b cannot be entirely excluded. The importance to agricultural practices of understanding population genetic processes is discussed.     

Genetic variation among populations of Pythium irregulare in southern Australia

Isolates of Pythium irregulare were sampled from seven cereal crops throughout South Australia to determine the extent of genetic diversity within this pathogen and the scale of genetic differentiation among populations. Data derived from 29 individual restriction fragment length polymorphism (RFLP) loci differentiated 54 DNA fingerprints among the 92 isolates analysed. Some isolates had two alleles at several RFLP loci and were scored as heterozygous. One such isolate was selfed in vitro and segregation ratios in the progeny were not significantly different from those expected for allelic variation in a diploid. These data provided evidence that outcrossing occurs within P. irregulare and may contribute to the high level of genetic variation within the species ( D _T = 0·502). Allelic frequencies were significantly different among all seven populations and G _ST values showed significant genetic differentiation between populations. The average genetic identity among populations was low and hierarchical cluster analysis provided no clear evidence that populations formed geographically related groups. These analyses indicate low levels of interpopulation gene flow within P. irregulare and imply that population differentiation results from genetic drift.     

Comparison of natural populations of Mycosphaerella graminicola from single fields in Kansas and California

Kansas and California wheat-growing regions differ dramatically in soils, climate, wheat cultivars, crop rotation patterns, and cultural practices, which could select for different fungal populations of Mycosphaerella graminicola. Our objective in this study was to use amplified fragment length polymorphism (AFLP) loci to assess the genetic diversity of M. graminicola populations within single fields in two widely separated, and geographically isolated sites in Kansas and California. Three primer-pair combinations were used to resolve polymorphism at 177 loci in 67 and 63 isolates from Kansas and California, respectively. Genotypic variability was high, which is consistent with a genetically diverse initial inoculum. There was no evidence of genetic disequilibrium in either population, with only 4.6% of the locus pairs in Kansas, and 5.4% of the locus pairs in California in detectable disequilibrium. The migration rate calculated between the two sites was as low as 1.8 individuals per generation, and significant differences in allele frequencies were observed. Therefore, these two populations do not represent mere subsamples of a larger, randomly mating population. This is a rare report of isolation by distance occurring between two North American populations of M. graminicola, indicating that at least some of these populations may be differentiating. Although genetic isolation by distance may occur, we cannot exclude movement of new gene combinations such as fungicide resistance or virulence between these two locations.     

Global Hierarchical Gene Diversity Analysis Suggests the Fertile Crescent Is Not the Center of Origin of the Barley Scald Pathogen Rhynchosporium secalis

A total of 1,366 Rhynchosporium secalis isolates causing scald on barley, rye, and wild barley (Hordeum spontaneum) were assayed for restriction fragment length polymorphism loci, DNA fingerprints, and mating type, to characterize global genetic structure. The isolates originated from 31 field populations on five continents. Hierarchical analysis revealed that more than 70% of the total genetic variation within regions was distributed within a barley field. At the global level, only 58% of the total genetic variation was distributed within fields, while 11% was distributed among fields within regions, and 31% was distributed among regions. A significant correlation was found between genetic and geographic distance. These findings suggest that gene flow is common at the local level while it is low between regions on the same continent, and rare between continents. Analyses of multilocus associations, genotype diversity, and mating type frequencies indicate that sexual recombination is occurring in most of the populations. We found the highest allele richness in Scandinavia followed by Switzerland. This suggests that R. secalis may not have originated at the center of origin of barley, the Fertile Crescent, nor in a secondary center of diversity of barley, Ethiopia.     

Distribution of race-specific resistance against <Emphasis Type="Italic">Bremia lactucae</Emphasis> in natural populations of <Emphasis Type="Italic">Lactuca serriola</Emphasis>

A metapopulation approach was applied to population studies of a common weed, Lactuca serriola (prickly lettuce). Seedlings grown from seed samples collected from 752 individual L. serriola plants in 50 populations occurring along an east-to-west transect across four European countries (Czech Republic, Germany, Netherlands and United Kingdom) were screened for resistance to 10 common races of Bremia lactucae. Based on the recorded reaction patterns, host individuals were characterized into specific resistance (R-) phenotypes. Diversity of R-phenotypes, their variation and distribution among and within European populations, was evaluated at different spatial scales, i.e. from a metapopulation involving the entire European study area to individual plants occurring in local populations. Generally, European populations of L. serriola have been shown to be highly susceptible to B. lactucae. However, large variation in L. serriola resistance was found both among and within individual countries. There was a clear gradient of increasing uniformity of race-specificity moving from central to western Europe, as well as a slight decrease in the diversity of R-phenotypes. Populations in the United Kingdom were the most divergent in terms of resistance structure from other geographic regions, and also were the most homogeneous, most likely a consequence of the relatively greater degree of spatial isolation from other regions. Metapopulation, inter- and intra-population variation in host resistance is discussed from the viewpoint of occurrence of race-specific interactions in this wild plant pathosystem.     

Gene Flow and Sexual Reproduction in the Wheat Glume Blotch Pathogen Phaeosphaeria nodorum (Anamorph Stagonospora nodorum)

ABSTRACT Restriction fragment length polymorphisms (RFLPs) were used to characterize the genetic structures of three field populations of Phaeosphaeria nodorum from Texas, Oregon, and Switzerland. Data from seven nuclear RFLP loci were used to estimate gene diversity and genetic distances and to make indirect measures of gene flow between populations. Three of the seven RFLP loci differed significantly in allele frequencies across populations. On average, 96% of the total gene diversity was found within populations. There was little evidence for population subdivision, suggesting that gene flow was not restricted among populations. Based on an average population differentiation of 0.04, we estimated that the exchange of 11 migrants among populations per generation would be needed to account for the present level of population subdivision. Genotype diversity based on DNA fingerprints was at a maximum for the Swiss population, whereas populations in Texas and Oregon had lower genotype diversities. Many multilocus haplotypes were found in each population. Ninety-five percent of RFLP allele pairs were in gametic equilibrium. The data were consistent with random mating within each population.     

警惕检疫性有害生物马铃薯孢囊线虫(Globodera rostochiensis和G. pallida)入侵我国

马铃薯孢囊线虫包括马铃薯金线虫Globodera rostochiensis和马铃薯白线虫G. pallida, 是马铃薯生产上危害最为严重的植物寄生线虫, 一般造成30%的产量损失, 在热带发病严重地区, 产量损失高达80%~90%, 甚至绝收?由于其危害严重性, 包括我国在内的100多个国家将其列为重要检疫性有害生物?我国目前尚无马铃薯金线虫和白线虫的发生报道, 但随着贸易全球化, 马铃薯孢囊线虫传入我国的风险日趋增高?本文主要对马铃薯孢囊线虫的发生分布?危害症状?经济损失?生物学特性?传入我国的潜在风险和预防控制措施进行综述, 旨在为防止马铃薯孢囊线虫入侵我国提供参考?     

Population structure and mating system of Ascochyta rabiei in Tunisia: evidence for the recent introduction of mating type 2

The population structure of Ascochyta rabiei (teleomorph: Didymella rabiei ) in Tunisia was estimated among five populations sampled from the main chickpea growing regions using simple sequence repeat markers (SSR) and a mating type ( MAT ) marker. Mating type 2 isolates ( MAT1-2 ) had reduced genetic and genotypic diversity relative to mating type 1 isolates ( MAT1-1 ). This result, coupled with previous observations of lower overall frequency and restricted geographical distribution of MAT1-2 in Tunisia, and recent (2001) observation of the sexual stage, support the hypothesis of a recent introduction of MAT1-2 . Despite the presence of both mating types in Nabeul, Kef and Jendouba, the hypothesis of random mating was rejected in these locations with multilocus gametic disequilibrium tests. Highly significant genetic differentiation ( θ  = 0·32, G _ST = 0·28, P  < 0·001) was detected among populations and genetic distance and cluster analyses based on pooled allele frequencies revealed that populations from Nabeul and Kef were distinct from those in Beja, Bizerte and Jendouba. More than 70% of total gene diversity ( H _T = 0·55) detected was attributable to variation within populations compared to 28% among populations. This result, coupled with the occurrence of private alleles in each population, suggests that gene flow is currently limited among populations, even those separated by short geographic distances. The presence of two main genetic clusters was confirmed using Bayesian model-based population structure analyses of multilocus genotypes (MLGs) without regard to geographic origin of samples. The presence of MAT1-2 isolates in both clusters suggests at least two independent introductions of MAT1-2 into Tunisia that are likely to be the result of importation and planting of infected chickpea seeds.     

Cluster analysis of 36Globodera pallida field populations using two sets of molecular markers

Thirty-six populations of the potato cyst nematodeGlobodera pallida, all collected in the Netherlands, were analysed twice: by two-dimensional gel electrophoresis of proteins (2-DGE) and by random amplified polymorphic DNA fingerprinting (RAPD). Two-DGE revealed frequencies of 21 alleles at eight putative loci in each population. The same populations were subjected to RAPD analysis. This qualitative technique revealed 38 polymorphic DNA fragments. Both datasets were independently processed to determine the intraspecific variation. UPGMA analysis resulted in a 2-DGE- and a RAPD-based dendrogram with cophenetic correlation coefficients of 0.755 and 0.838 respectively. The correlation between the genetic similarity values for the populations was 0.572. Comparison between the 2-DGE- and the RAPD-based dendrogram revealed that only thirteen of the 36 populations analysed were clustered identically. It is concluded that the gene pool similarity concept is only in some instances applicable to Dutch populations ofG. pallida. For populations that could not be differentiated unequivocally on the basis of molecular markers, markers closely linked to avirulence genes should be identified. Approaches that will lead to the identification of such markers are discussed.     

黄淮麦区小麦全蚀病菌群体的遗传多样性分析

为了解小麦全蚀病病菌的群体组成和遗传多样性,采用8对多态性较好的微卫星标记,对我国黄淮麦区的116个小麦全蚀病菌株进行了分析。8个SSR标记的平均等位基因数为4.25个,多态性信息含量的平均值为0.66。供试菌株的平均有效等位基因数和基因遗传多样性指数分别为1.46和0.27,漯河群体的遗传多样性水平最高,周口群体最低。不同群体间遗传距离均较小,为0.0199~0.1153,其中徐州和周口群体间的遗传距离相对较大,而周口和驻马店群体间的遗传距离相对较小。分子方差分析结果显示,小麦全蚀病菌群体间和群体内均存在着一定的遗传分化,群体间的遗传变异占总变异的10%,群体内遗传变异占90%。6个群体间的基因流为3.5。根据SSR多态性,对来源不同菌株的UPGMA聚类分析结果显示,小麦全蚀病菌群体结构与地理来源有一定的关系。     

The use of isoelectric focusing as a tool in the identification and management of potato cyst nematode populations1

Isoelectric focusing on thin layers of polyacrylamide or agarose gels may be used to separate proteins from the potato cyst nematodes Globodera rostochiensis and G. pallida. General protein patterns may be used to identify the two species, even from single cysts. Densitometry of species-specific protein bands may be used to assess the proportions of G. rostochiensis and G. pallida in potato cyst nematode samples. Staining for the enzymes phosphoglucomutase and phosphoglucose isomerase also revealed species-specific patterns, and variation among G. pallida populations was observed. Thus calculations of coefficients of similarity based on six enzymes, phosphoglucose isomerase, phosphoglucomutase, glycerol-3-phosphate dehydrogenase, enolase, hexokinase and malate dehydrogenase and the construction of a dendrogram for several Northern Ireland populations indicated clear separations between G. pallida (Pa1), G. pallida (Pa3) and G. rostochiensis (Ro1). The use of the technique of isoelectric focusing as a routine research and advisory tool in nematology, and as a means of further understanding the genetic basis of pathotype schemes, is discussed.     

Durability of Resistance to Globodera pallida I. Changes in Pathogenicity, Virulence, and Aggressiveness During Reproduction on Partially Resistant Potato Cultivars

ABSTRACT We studied changes in pathogenicity, virulence, and aggressiveness of Globodera pallida populations over time. As a measure for pathogenicity, the reproduction factor on a partially resistant host was used; for aggressiveness, the reproduction factor on a susceptible reference host was used; and, for virulence, the ratio pathogenicity/aggressiveness was used. The G. pallida populations were reared in a glasshouse for four generations on potato cultivars with different levels of resistance. The cultivar Elkana did not increase pathogenicity significantly, but the more resistant cultivars Karakter and Darwina did. This increase in pathogenicity was caused by an increase in virulence, whereas aggressiveness generally was not altered significantly. The increase in virulence appeared to be caused by an enhanced ability of eggs to develop into cysts, and not by an increase in egg production per new cyst. The observed changes in virulence could be predicted reasonably well by a simple numerical model. The rate of selection depended strongly on the nematode population. Rearing a mix of two different populations on a susceptible host decreased the virulence strongly, as predicted by the Hardy-Weinberg equilibrium, but increased the aggressiveness because of heterosis.     

Problems of potato cyst nematode pathotypes in the Czech Republic

During 1983–1989 about 1,200 infested plots in the Czech republic were investigated for the occurrence of new pathotypes of the potato cyst nematodes (PCN) Globoderu rostochiensis and G. pallida. Only eight populations of the pest were suspected of containing another pathotype than Ro1. Glasshouse pot trials were performed with those eight natural populations, two populations of Ro1, 10 potato cultivars resistant to Ro1 and 10 cultivars resistant to more pathotypes, G. pallida included. The multiplication rates (Pf/Pi) showed that four of the PCN populations were probably mixtures, with pathotype Ro1 prevailing. Three populations were suspected of containing G. pallida , but their morphological characters corresponded only with G. rostochiensis. One population corresponded with pathotype Ro5. The extent of the occurrence of pathotypes or virulence groups different from Ro1 can be estimated at 0.3–0.4% of infested plots.     

Genetic diversity in populations of the fungi Phaeomoniella chlamydospora and Phaeoacremonium aleophilum on grapevine in France

Isolates of Phaeomoniella chlamydospora ( Phc ) and Phaeoacremonium aleophilum ( Pha ), two haploid, deuteromycetous fungi, were obtained from vines showing symptoms of esca disease in different localities in two French regions, and within a single vineyard in one of these regions. The population genetic structure was determined in both fungi using random amplified polymorphic DNA (RAPD) analysis. Populations of Phc showed similar levels of diversity at local and regional levels. The most frequent Phc haplotypes were found in every population, and the frequencies of positive alleles of markers were similar across populations. The hypothesis that recombination had occurred was rejected for the full set of samples, but not for the samples reduced to haplotypes, indicating that Phc may be a recombining species. Different features were identified in Pha populations. First, the southern population of Pha appeared more diverse than the south-western populations. Second, genetic differentiation was identified between Pha populations from southern and south-western regions for several RAPDs. Finally, in the southern population of Pha no evidence for recombination was obtained, even by reducing the sample to haplotypes. Within the single vineyard surveyed, several haplotypes of both fungi were recovered and randomly distributed. Thus different infection events appeared to have occurred on a low spatial scale. Data from this study showed that haplotypes of both fungi were distributed over long distances geographically, and that most of the vineyards surveyed were infested by more than one haplotype of Phc and Pha .