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1.
The population structure of the grape powdery mildew fungus, Erysiphe necator (formerly Uncinula necator), has been hypothesized to vary from being clonal to highly diverse and recombining. We report here on the structure of an E. necator population sampled during a 4-year period from an isolated vineyard in northern Italy (Voghera, Pavia Province). We obtained 54 isolates of E. necator that overwintered asexually as mycelium in grapevine buds and caused severe symptoms on the emerging shoots, known as flag shoots. All isolates were genotyped for mating type, four multilocus polymerase chain reaction (PCR)-based markers (a total of 64 loci were scored), and two single-copy loci designed to identify genetic subgroups in E. necator. All isolates had the same mating type and single-locus alleles that correlate to isolates from flag shoots in other areas. Only 2 of the 64 loci scored from multilocus markers were polymorphic; 46 of the 54 isolates had the same multilocus haplotype. Seven isolates had a second haplotype that was recovered over 3 years, and only a single isolate was found with a third haplotype. Both variant haplotypes differed from the main clonal haplotype by single loci. Spatial autocorrelation analyses showed that vines with flag shoots were not aggregated within years, but they were aggregated between consecutive years. These results demonstrate that this subpopulation of E. necator on flag shoots is composed of a single clonal lineage that has persisted for at least 4 years. We speculate that the lack of diversity in the flag shoot subpopulation in this vineyard is the result of restricted immigration from surrounding areas and genetic drift operating through founder effects and periodic bottlenecks. We propose a model that integrates epidemiology and population genetics to explain the variation observed in genetic structure of E. necator flag shoot subpopulations from different vineyards or viticultural regions. 相似文献
2.
ABSTRACT Didymella bryoniae (anamorph Phoma cucurbitacearum) is the causal agent of gummy stem blight, although other Phoma species are often isolated from cucurbit plants exhibiting symptoms of the disease. The molecular and phylogenetic relationships between D. bryoniae and these Phoma species are unknown. Isolates of D. bryoniae and Phoma obtained from cucurbits grown at various geographical locations in the United States were subjected to random amplified polymorphic DNA (RAPD) analysis and internal transcribed spacer (ITS) sequence analysis (ITS-1 and ITS-2) to determine the molecular and phylogenetic relationships within and between these fungi. Using RAPD fingerprinting, 59 isolates were placed into four phylogenetic groups, designated RAPD group (RG) I, RG II, RG III, and RG IV. D. bryoniae isolates clustered in either RG I (33 isolates), RG II (12 isolates), or RG IV (one isolate), whereas all 13 Phoma isolates clustered to RG III. There was greater than 99% sequence identity in the ITS-1 and ITS-2 regions between isolates in RG I and RG II, whereas isolates in RG III, P. medicaginis ATCC 64481, and P. exigua ATCC 14728 clustered separately. On muskmelon seedlings, a subset of RG I isolates were highly virulent (mean disease severity was 71%), RG II and RG IV isolates were slightly virulent (mean disease severity was 4%), and RG III isolates were nonpathogenic (disease severity was 0% for all isolates). The ITS sequences indicate that RG I and RG II are both D. bryoniae, but RAPD fingerprints and pathogenicity indicate that they represent two different molecular and virulence subgroups. 相似文献
3.
玉米新月弯孢菌(Curvularia lunata)的RAPD分析 总被引：3，自引：1，他引：3
对分离自玉米弯孢菌叶斑病标样中的77株新月弯孢菌和1株来自水稻的新月弯孢菌进行RAPD分析表明,菌株间具有丰富的遗传多样性,在相似系数约0.60处,所有菌株被聚为3个组,但88.0%的菌株聚入第Ⅰ组内,其余菌株被聚入另外2个组内。第Ⅰ组内共有69个菌株,包含来自不同区域的致病性较强的菌株,是玉米弯孢菌叶斑病的优势类群,其余2个类群主要是一些致病能力较弱或不致病的菌株。结果表明,新月弯孢菌种内菌株的遗传多样性与致病性相关,但与菌株地理来源无明显的直接关系。 相似文献
4.
The random amplified polymorphic DNA (RAPD) technique was used to determine the mating groups of several members of the Fusarium section Liseola recovered from maize, rice and sorghum collected from different locations in Ghana. Three mating groups were identified, A, D and F, of which all A and F isolates were confirmed by mating studies. Fertile crosses were also obtained in crosses involving two of the isolates identified as belonging to the D population. Variability within the A population isolated from seeds and stem-bases of maize was investigated to determine whether the sub-structuring of this population was related to the host tissue from which the isolates were obtained. The relative merits of the RAPD procedure, compared to the mating procedure, for determining the mating affiliations of isolates and for more detailed analyses of isolates within a population, as well as its possible advantages over established RFLP methodologies are discussed. 相似文献
5.
J. Feng R. Hwang K. F. Chang S. F. Hwang S. E. Strelkov B. D. Gossen R. L. Conner G. D. Turnbull 《Plant pathology》2010,59(5):845-852
Isolates of Fusarium spp. were recovered from the roots of field pea (Pisum sativum) collected from 15 commercial fields in Alberta, Canada. Most of the isolates (75 out of 96) were identified as F. avenaceum, based on morphology, phylogeny and species‐specific PCR amplification. Molecular differences in the F. avenaceum isolates were detected based on putative mating type, and on ITS and CPN60 sequences. MAT‐1 and MAT‐2 were equally distributed among the isolates. Phylogenetic analysis based on ITS and CPN60 sequences clustered most of the F. avenaceum isolates into a single group. In some cases, isolates with low aggressiveness clustered together in additional groups. There was no correlation between phylogenetic profile and either mating type or geographic origin. This population of F. avenaceum has a low level of genetic variation and consists of isolates derived from the two mating types. Isolates with low aggressiveness are also retained in the population. 相似文献
6.
Hajime KATO Masaaki YAMAMOTO Tomona YAMAGUCHI-OZAKI Hiroyuki KADOUCHI Yutaka IWAMOTO Hitoshi NAKAYASHIKI Yukio TOSA Shigeyuki MAYAMA Naoki MORI 《Journal of General Plant Pathology》2000,66(1):30-47
Eighty-five Pyricularia isolates were collected from 29 host species of Gramineae, Bambusideae and Zingiberaceae plants sampled in Brazil, Uganda,
Ivory Coast, India, Nepal, China, Indonesia and Japan. These isolates were compared on the basis of pathogenicity, mating
ability and restriction fragment length polymorphisms with single-copy DNA probes. Based on the pathogenicity to eight differential
gramineous plants, these isolates were classified into seven pathotypes: finger millet type, foxtail millet type, common millet
type, rice type, crabgrass type, Italian ryegrass/ weeping lovegrass type, and non-cereal/grass type. Genetic variation among
these isolates was assessed by RFLP analysis with two restriction enzymes and nine single-copy DNA probes isolated from a
finger millet strain. An UPGMA dendrogram based on the RFLPs revealed that the 85 isolates could be classified into seven
major groups. Isolates from cereal crops (finger millet, foxtail millet, common millet, wheat and rice) and a grass, Brachiaria plantaginea, were clustered into a single group. They were further divided into six subgroups corresponding to the pathotypes. Among
cereal crop isolates only an isolate from pearl millet was located into a different group. The remaining isolates were clustered
into five groups designated as the crabgrass group, the buffelgrass and jungle rice group, the rice cutgrass, knotroot bristlegrass
and Setaria tomentosa group, the bamboo and bamboo grass group and the Zingiber mioga group. The isolates from cereal crops were generally capable of mating with finger millet strains and constituted a closed
mating compatibility group. These results suggested that the isolates from cereal crops form a single group with a common
ancestor although they are pathogenic to taxonomically diverse plants. A combined analysis of the pathogenicity and genetic
similarity suggested that the transmission of M. grisea isolates occurs in natural agroecosystems between finger millet and Eleusine africana, goosegrass or Bambusa arundinacea, between foxtail millet and green bristlegrass, and between rice and tall fescue, Italian ryegrass, sweet vernalgrass, reed
canarygrass or Oryza longistaminata.
Received 9 August 1999/ Accepted in revised form 12 November 1999 相似文献
7.
Denoyes-Rothan B Guérin G Délye C Smith B Minz D Maymon M Freeman S 《Phytopathology》2003,93(2):219-228
ABSTRACT Ninety-five isolates of Colletotrichum including 81 isolates of C. acutatum (62 from strawberry) and 14 isolates of C. gloeosporioides (13 from strawberry) were characterized by various molecular methods and pathogenicity tests. Results based on random amplified polymorphic DNA (RAPD) polymorphism and internal transcribed spacer (ITS) 2 sequence data provided clear genetic evidence of two subgroups in C. acutatum. The first subgroup, characterized as CA-clonal, included only isolates from strawberry and exhibited identical RAPD patterns and nearly identical ITS2 sequence analysis. A larger genetic group, CA-variable, included isolates from various hosts and exhibited variable RAPD patterns and divergent ITS2 sequence analysis. Within the C. acutatum population isolated from strawberry, the CA-clonal group is prevalent in Europe (54 isolates of 62). A subset of European C. acutatum isolates isolated from strawberry and representing the CA-clonal and CA-variable groups was assigned to two pathogenicity groups. No correlation could be drawn between genetic and pathogenicity groups. On the basis of molecular data, it is proposed that the CA-clonal subgroup contains closely related, highly virulent C. acutatum isolates that may have developed host specialization to strawberry. C. gloeosporioides isolates from Europe, which were rarely observed were either slightly or nonpathogenic on strawberry. The absence of correlation between genetic polymorphism and geographical origin in Colletotrichum spp. suggests a worldwide dissemination of isolates, probably through international plant exchanges. 相似文献
8.
A previous study of the diversity and population structure of the rice blast fungus, Magnaporthe oryzae, over a 20-year period in Korea, found novel fingerprint haplotypes each year, and the authors hypothesized that populations might experience annual bottlenecks. Based on this model, we predicted that M. oryzae populations would have little or no genetic differentiation among geographic regions because rice blast is commonly found throughout Korea each year and M. oryzae would have to disperse from small populations surviving annually between rice crops. To test this hypothesis, we sampled M. oryzae from rice fields in eight provinces in Korea in a single year (1999). In four provinces, we sampled from a set of rice cultivars commonly grown in commercial fields (group I); because of low disease incidence in four other provinces, we could not sample from commercial fields and instead sampled from scouting plots of different cultivars set up for detecting new pathotypes of M. oryzae (group II). All isolates were genotyped with DNA fingerprint probes MGR586 and MAGGY, a telomere-linked gene family member TLH1, the PWL2 host specificity gene and mating type. Fingerprint haplotypes clustered into two distinct lineages corresponding to the two sets of cultivars (groups I and II), with haplotype similarities of 71% between lineages and >76% within lineages. Isolates from the same cultivar within group I were genetically differentiated among locations, and isolates within the same location were differentiated among cultivars. Differentiation for TLH1 and PWL2 was significant (P < 0.03), but not as strong as for fingerprint markers. Similar analyses were not possible among group II isolates because too few isolates were available from any one cultivar. All isolates were in the same mating type, Mat1-1, ruling out sexual reproduction as a source of novel haplotypes. When the 1999 samples were compared with the historical samples from the previous study, haplotypes of group I formed a separate cluster, while those of group II clustered with haplotypes from the historical sample. Altogether, geographic subdivision, monomorphism of mating type, and correlation of haplotypes to sets of cultivars are not consistent with the hypothesis of repeated turnover of haplotypes. Instead, the previous correlations of haplotypes to year might have been caused by inadequate sampling of haplotypes each year, highlighting the need for studies of population genetics to be conducted with systematic samples collected to address specific questions. 相似文献
9.
中国不同地区致病疫霉遗传多样性的RAPD分析 总被引：4，自引：0，他引：4
本文应用RAPD技术检测了我国主要马铃薯产区致病疫霉的遗传分化情况及不同地区菌株间的亲缘关系。用筛选出的10个随机引物对1997-2001年间采自我国9省市的82株及3株来自日本的致病疫霉DNA进行了PCR扩增,获得了79条谱带,其中多态性标记75条,占95%。根据扩增结果,运用UPGMA分析,获得了表现菌株间亲缘关系的树状图。菌株间的最大遗传距离为0.5,以距离0.3为阈值,可将供试菌株划分为10个组(RG1-10)。结果发现:A1交配型菌株群体内的差异大于A1和A2菌株群体之间的;RAPD分组与菌株的地理来源、交配型及对甲霜灵的敏感性无明显相关性。研究结果显示,来自中国北方甘肃、内蒙、吉林、黑龙江地区的菌株与一些来自云南、四川等西南地区的菌株亲缘关系相近。病原菌随种薯的迁移可能是导致这种现象的原因之一。 相似文献
10.
Erysiphe necator, the causative agent of powdery mildew in grapevine, was introduced into Europe from North America during the middle of the
19th century. Our objective was to analyze the genetic variation and the population structure of the fungus in southern France.
The sample comprised 101 isolates and was mainly of flag shoot origin, i.e., infection of sprouting shoots after overwintering
of mycelium in buds. RAPD analysis identified different haplotypes that clustered in two genetic groups (A and B). The most
frequent haplotypes of each group were found in several different locations in two areas separated by 100 km and throughout
the 3 year period. Several haplotypes of both groups originated from flag shoots and were recovered over successive years
indicating that there is no correlation between genetic group and overwintering mode. All isolates of group A were of mating
type +, but those in group B could be either + or −. Lower genotypic diversity was detected within group A than within group
B. These results were consistent with the hypothesis that group A reproduces only asexually. 相似文献
11.
Christophe Dlye Frdric Laigret Marie-France Corio-Costet 《Pest management science》1997,51(3):309-314
Using a Random Amplified Polymorphic DNA (RAPD) assay, we investigated the genetic polymorphism existing among 62 European isolates of the grape powdery mildew fungus (Uncinula necator [Schw.] Burr.). Isolates overwintering as mycelium in buds were genetically distinct from isolates overwintering as ascospores, suggesting the existence of two genetically isolated powdery mildew populations, and consequently of two independent sources of inoculum in the vineyard. Isolates resistant to fungicides inhibiting sterol 14α-demethylation (DMIs) were found in both populations, suggesting that resistance to DMIs may arise independently in the two powdery mildew populations. A PCR assay targeting the gene encoding U. necator 14α-demethylase has been developed which will permit an early, specific detection of U. necator infections, and may be useful for spraying programmes. ©1997 SCI 相似文献
12.
ABSTRACT Erysiphe necator overwinters as ascospores in cleistothecia and mycelium in dormant buds of grapevines. Shoots developing from infected buds early in the growing season are covered with dense mycelium and are known as "flag shoots". Combining epidemiological and genetic analyses, the objective of this study was to analyze the spatial and genetic structure of a flag shoot subpopulation of E. necator as a way to assess the contribution of flag shoots as primary inoculum, and to determine if flag shoot subpopulations are clonal with only one mating type. One vineyard in Tuscany, Italy was surveyed intensively for flag shoots for 8 years; isolations of E. necator were made from flag shoots for 5 years. We observed distinct disease foci developing around flag shoots early in epidemics, demonstrating a steep dispersal gradient of conidia and the importance of flag shoots as primary inoculum sources. Flag shoots were spatially aggregated within and between years, most likely as a result of short-distance dispersal of conidia from flags early in the season when dormant buds for the next year's shoots are formed and are susceptible to infection. The two mating types were found in 1:1 ratios in this flag shoot subpopulation. Genotypic diversity, based on inter-simple sequence repeat markers, was high in all years with only two haplotypes occurring twice, and subpopulations were genetically differentiated between years. Similarities between haplotypes were not spatially autocorrelated. One multilocus analysis of population structure is consistent with the hypothesis of random mating but another is not. These results are not consistent with expectations for a strictly clonal or strictly randomly mating flag shoot subpopulation. Instead, the hypothesis that the flag shoot subpopulation of E. necator may reproduce clonally and sexually needs further testing. 相似文献
13.
14.
An analysis of the population structure of Erysiphe necator from Spain was carried out to determine if population structure of grapevine powdery mildew is partitioned by origin of primary inoculum, i.e. overwintering ascomata on grapevine bark and mycelia in dormant buds. Ten of the 31 isolates collected were from flag shoots, i.e. shoots covered with sporulating mycelia that arise from dormant infected buds. Genetic variation was assessed using 61 AFLP markers. Cluster analysis revealed two distinct groups with 63% genetic similarity. Principal coordinates analysis ( pccorda ) produced a similar distribution of isolates in two groups. Isolates were not well identified within their groups by the origin of primary inoculum. Analysis of molecular variance ( amova ) determined that the origin of primary inoculum was not a significant component in the genetic variation in the population of E. necator . It was concluded that these two subpopulations are not separated by the type of primary source of inoculum. 相似文献
15.
ABSTRACT Randomly amplified polymorphic DNA (RAPD) markers and mating type were used to examine regional population structure of Setosphaeria turcica in the eastern United States. Of 251 maize-infecting isolates studied, 155 multilocus haplotypes were identified using 21 RAPD markers. Twelve isolates of the most common haplotype were identified from seven states and represented 5.2% of the sample. Although variation in genetic diversity was greatest within states rather than between either regions or states within regions, multidimensional scaling based on average taxonomic distances among state samples showed a close association of samples from IL, OH, IN, IA, MN, MI/WI, and NC. Isolates from GA/SC, VA/TN, PA/NY, and FL were distant from this core group that included midwestern states and NC and were distinct from one another. The high genotypic diversity, near equal mating type frequencies, and gametic phase equilibrium in samples from several states are inconsistent with a strictly clonal population. The population genetic structure of S. turcica is likely the result of both asexual and sexual reproduction. It is not clear whether sexual recombination actually occurs in the eastern United States or occurs elsewhere in tropical America and recombinant genotypes migrate to North America. 相似文献
16.
The taxonomic placement of Spanish isolates of Gremmeniella abietina var. abietina (Gaa) (race and biotype) was determined, for the first time, using RAPD markers and comparison of RAPD patterns for Spanish isolates and those originating from different regions of Europe and North America. Fifteen Spanish isolates of Gaa were isolated from recently dead shoots of 10 Pinus halepensis trees. Additionally, 17 isolates of Gaa from Canada, Finland, Switzerland and the USA, previously characterized according to race and biotype, were used. Five primers, previously reported to yield readable, reproducible and polymorphic fragments, were used in PCR reactions. The results showed that the Spanish isolates represent the European race and not the Alpine or northern biotypes. Spanish isolates appear to be clearly separated from all other biotypes within the EU race and preliminary data suggest that Spanish isolates have low genetic variability. 相似文献
17.
ABSTRACT Setosphaeria turcica causes northern leaf blight, an economically important disease of maize throughout the world. Survey collections of S. turcica isolates from 1974 to 1994 provided a unique opportunity to examine temporal diversity in the eastern United States. Two hundred forty-two isolates of S. turcica from maize were studied with random amplified polymorphic DNA (RAPD) markers, mating type, and virulence on maize differential inbred lines with known Ht resistance genes to examine changes over time. One hundred forty-nine RAPD haplotypes were identified. Nearly 20% of haplotypes recurred in more than one year. Race 0 isolates declined in frequency from 83% in 1974 to near 50% in the 1990s, most likely in response to the widespread deployment of Ht1 in commercial maize hybrids. Races 23 and 23N were present in the collection at low levels throughout the study period and were also found among isolates from Virginia in 1957. The frequency of MAT1-2 isolates increased sharply after 1979 and was associated with the emergence of race 1 during the same period. RAPD markers were used to investigate the genetic diversity among a subset of isolates collected in the United States from 1976 to 1982, the period in which this dramatic shift in race frequency occurred. Multilocus haplotypes were not exclusively associated with known races of S. turcica. Based on shared haplotypes and cluster analysis, race 1 isolates share greater similarity with race 0 than with 23 or 23N isolates, indicating race 1 probably evolved from multiple lineages of race 0. Sorghum spp.-infecting isolates share greater similarity with one another than with maize-infecting isolates and represent a distinct subgroup. 相似文献
18.
ABSTRACT Coevolution of the angular leaf spot pathogen, Phaeoisariopsis griseola, with its common bean host has been demonstrated, and P. griseola isolates have been divided into Andean and Mesoamerican groups that correspond to defined bean gene pools. Recent characterization of P. griseola isolates from Africa has identified a group of isolates classified as Andean using random amplified polymorphic DNA (RAPD), but which are able to infect some Mesoamerican differential varieties. These isolates, designated Afro-Andean, have been identified only in Africa. Random amplified microsatellites, RAPD, and restriction digestion of amplified ribosomal intergenic spacer region were used to elucidate the relationships among the Afro-Andean, Andean, and Mesoamerican groups of P. griseola. Cluster and multiple correspondence analysis of molecular data separated isolates into Andean and Meso-american groups, and the Afro-Andean isolates clustered with Andean isolates. Analysis of molecular variance ascribed 2.8% of the total genetic variation to differences between Afro-Andean and Andean isolates from Africa. Gene diversity analysis revealed no genetic differentiation (G(ST) = 0.004) between Afro-Andean and Andean isolates from Africa. However, significant levels of genetic differentiation (G(ST) = 0.39) were observed between Afro-Andean or Andean isolates from Africa and Andean isolates from Latin America, revealing significant geographical differentiation within the Andean lineage. Results from this study showed that Afro-Andean isolates do not constitute a new P. griseola group and do not represent long-term evolution of the pathogen genome, but rather are likely the consequents of point mutations in genes for virulence. This finding has significant implications in the deployment of resistant bean genotypes. 相似文献
19.
ABSTRACT Profiles of fatty acids from 70 isolates of Rhizoctonia solani anastomosis group (AG)-4 clustered into three groups, corresponding to homogeneous group (HG)-I, HG-II, and a newly described HG-III. Isolates from Georgia peanuts exhibiting limb rot were characterized as gas chromatography (GC) subgroup 1 (GC-1) and contained HG-I isolates. Isolates from diseased soybean hypocotyls grown in North Dakota and sugar beet seedlings, taproots, and tare soil in Minnesota and North Dakota were characterized as GC subgroup 2 (GC-2) and contained predominantly HG-II isolates but also included three distinct isolates based on fatty acid methyl ester (FAME) analysis and morphological features. Selected isolates from North Carolina cucumbers clustered into three distinct groups that corresponded to HG-I, HG-II, and the newly described HG-III. Distinct isolates from the soybean and sugar beet populations clustered with HG-III. Fatty acid profiles of AG-4 were compared with FAME library profiles of AG-1, AG-2 type 2, and AG-3, which were developed in previous studies and were sufficiently different that they could be used to support speciation of this group from R. solani. It is suggested that binomial R. practicola may be appropriate for the portion of AG-4 identified as HG-II. 相似文献
20.
The biotrophic fungus Erysiphe necator (formerly Uncinula necator ), for which two genetic groups have been described in European vineyards, is the causal agent of grapevine powdery mildew. By analysing the pathogen population with respect to polymorphism in the sequence of the β-tubulin gene, which distinguishes two groups of isolates, a new tool was developed for epidemiological and population studies and tested in the vineyard. As in many ascomycetes, the β-tubulin gene of E. necator ( Entub ) includes six introns and seven exons and encodes a 447-amino-acid protein. A single nucleotide polymorphism (SNP) in the intron-3 region of the Entub gene distinguished two genetic groups (A and B). This method was used to examine differences in the ratio of the two groups from a total of 289 grape powdery mildew samples collected at the beginning of the growing season from either flag shoots or leaves with sparse-spot symptoms in four different vineyards. The SNP in the intron-3 region of the β-tubulin gene, similar to SNPs in the CYP51 gene, was associated with genotypes A and B of E. necator and confirmed the existence of two sympatric populations of the pathogen in the French vineyards. Differences in the relative proportions of each group varied with the presence or absence of flag-shoot symptoms and with the region in which isolates had been collected. 相似文献