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1.
ABSTRACT One hundred two single zoospore isolates of Phytophthora infestans, derived asexually from four parental isolates of US-8 genotype and one isolate of US-1 genotype, were characterized for their virulence phenotypes to determine changes in virulence during asexual reproduction. Potato differentials, each containing a major gene for resistance to P. infestans (R1 to R11), were used to characterize the virulence patterns. Isolates were also characterized for mating type, glucose-6-phosphate isomerase (Gpi) banding pattern, and DNA fingerprints using probe RG57 to determine any genotypic changes in the single zoospore isolates. A subset of these single zoospore isolates was tested for response to mefenoxam to determine any shifts in sensitivity. Results showed that single zoospore isolates derived from parent PI-1 (US-8, 11 isolates) were identical to their parental virulence. Isolates derived from parent PI-191 (US-8, 29 isolates) showed some differences in virulence, mainly toward R8 and R9. Isolates derived from parent PI-126 (US-8, 14 isolates) demonstrated a higher level of virulence diversity. Isolates derived from parents PI-52 (US-1, 28 isolates) and PI-105 (US-8, 20 isolates) showed the highest level of virulence variability among the single zoospore isolates. Mating type, Gpi banding pattern, and DNA fingerprints for the single zoospore isolates were, in most cases, identical to the parental isolates. Single zoospore isolates showed different levels of sensitivity to mefenoxam. Virulence and other genetic changes during asexual reproduction are likely to play a major role in changing the race structure of P. infestans populations. This continuous change in the race structure is a serious problem and now poses a new challenge for utilization of race-specific resistance to manage late blight of potato. 相似文献
2.
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
3.
ABSTRACT Spot blotch, caused by the fungal pathogen Cochliobolus sativus, is an important disease of barley in many production areas of the world. To assess genetic diversity in this pathogen, a worldwide collection of C. sativus isolates was evaluated for virulence on barley and DNA polymorphism. Three pathotypes (0, 1, and 2) were identified among the 22 isolates tested in this study and the 36 isolates characterized previously on three barley differentials (ND5883, Bowman, and NDB112) that differ in their resistance to C. sativus. Pathotype 2, which exhibits high virulence on cv. Bowman, was only found in North Dakota, whereas the other two pathotypes occurred in many other regions of the world. Genetic diversity of the 58 C. sativus isolates, together with isolates of three related pathogenic Cochliobolus spp. (C. heterostrophus, C. carbonum, and C. victoriae) was analyzed using amplified fragment length polymorphism (AFLP) markers. A total of 577 polymorphic AFLP markers were recorded among the 70 isolates of the four Cochliobolus spp. using eight primer combinations. Cluster analysis revealed distinct groups corresponding to the four different species, except in one case where race 0 of C. carbonum was placed in an outgroup that may belong to a different species. In C. sativus, 95 polymorphic AFLP markers were detected with the eight primer pairs used, and each isolate exhibited a unique AFLP pattern. Allelic diversity in the pathotype 2 group was lower (0.10) than in the pathotype 0 (0.23) and pathotype 1 (0.15) groups, indicating that pathotype 2 may have arisen more recently. Cluster analysis did not reveal a close correlation between pathotypes and AFLP groups, although two AFLP markers unique to pathotype 2 isolates were identified. This low correlation suggests that genetic exchange may have occurred through parasexual recombination in the fungal population. Some isolates collected from different regions of the world were clustered into the same AFLP group, suggesting that migration of the fungal pathogen around these regions has occurred. 相似文献
4.
ABSTRACT Collections of Puccinia triticina, the wheat leaf rust fungus, were obtained from Great Britain, Slovakia, Israel, Germany, Australia, Italy, Spain, Hungary, South Africa, Uruguay, New Zealand, Brazil, Pakistan, Nepal, and eastern and western Canada. All single-uredinial isolates derived from the collections were tested for virulence polymorphism on 22 Thatcher wheat lines that are near-isogenic for leaf rust resistance genes. Based on virulence phenotype, selected isolates were also tested for randomly amplified polymorphic DNA (RAPD) using 11 primers. The national collections were placed into 11 groups based on previously established epidemiological zones. Among the 131 single-uredinial isolates, 105 virulence phenotypes and 82 RAPD phenotypes were described. In a modified analysis of variance, 26% of the virulence variation was due to differences in isolates between groups, with the remainder attributable to differences within groups. Of the RAPD variation, 36% was due to differences in isolates between groups. Clustering based on the average virulence distance (simple distance coefficient) within and between groups resulted in eight groups that differed significantly. Collections from Australia-New Zealand, Spain, Italy, and Britain did not differ significantly for virulence. Clustering of RAPD marker differences (1 - Dice coefficient) distinguished nine groups that differed significantly. Collections from Spain and Italy did not differ significantly for RAPD variation, neither did collections from western Canada and South America. Groups of isolates distinguished by avirulent/virulent infection types to wheat lines with resistance genes Lr1, Lr2a, Lr2c, and Lr3 also differed significantly for RAPD distance, showing a general relationship between virulence and RAPD phenotype. The results indicated that on a worldwide level collections of P. triticina differ for virulence and molecular backgrounds. 相似文献
5.
M. López D. Ruano-Rosa C. J. López-Herrera E. Monte R. Hermosa 《European journal of plant pathology / European Foundation for Plant Pathology》2008,121(2):201-205
Fifty-five isolates of Rosellinia necatrix, the cause of common avocado white root rot disease, were collected from south-east Spain and characterised according to
their virulence behaviour and their molecular patterns to assess broader levels of genetic diversity. Virulence properties
were revealed by in vitro inoculation on avocado plants. Differences in reaction types showed variability among these isolates.
No sequence differences were observed when the internal transcribed spacer 1 (ITS1) and ITS2 regions and DNA fragments of
the β-tubulin, adenosine triphosphatase and translation elongation factor 1 genes were explored in representive isolates from
five virulence groups. Random amplified polymorphic DNA (RAPD) amplifications were also performed for each isolate using 19
random primers. Four of these primers revealed polymorphism among isolates and repetitive and discriminative bands were used
to build an unweighted pair group with arithmetic mean tree. However, RAPD clustering showed low stability, and no correlation
between RAPD and virulence groups was observed, possibly indicating high levels of sexual recombination. 相似文献
6.
ABSTRACT The role of asexual reproduction in the production of pathogenic and genotypic variation in Aphanomyces euteiches was investigated. Variation was studied among three groups of 18 single-zoospore progeny of A. euteiches derived from each of three single-zoospore parental strains. Pathogenicity was assessed by evaluating disease severity (DS) on roots of five pea lines possessing different levels of resistance to Aphanomyces root rot and of a susceptible cultivar of snap bean and alfalfa. None of the single-zoospore progeny incited significantly higher DS levels than their parental strain on any of the seven hosts; however, 3 or 4 of the 18 progeny in each group incited significantly lower DS than their parental strains. The host range of the progeny either decreased or remained the same as compared with parental strains. Genotypic variation was assessed with randomly amplified polymorphic DNA (RAPD) analysis. Polymorphic RAPD markers that distinguished parental and progeny strains were detected within two of the three groups of strains with two of four RAPD primers used. Of 76 total RAPD markers that were detected among all strains in all groups, four (5%) were polymorphic. The polymorphic markers were not associated with the pathogenic variation. 相似文献
7.
ABSTRACT Isolates of Sclerotium rolfsii, the causal organism of stem rot or southern blight of groundnut, can be placed in mycelial compatibility groups (MCGs) based on hyphal interactions between isolates. The aim of this study was to determine whether amplified fragment length polymorphism (AFLP) analysis was a suitable technique to assess genetic variability between isolates and MCGs of S. rolfsii. For preliminary genetic analysis, 10 isolates were selected from each of two MCGs and compared with each other using the restriction enzymes EcoRI and MseI and 4 primer pairs. The number of polymorphisms ranged from 10 to 36 per primer combination, with an average of 22.5. AFLP analysis clearly showed genotypic differences (22%) among MCGs B and C, with a maximum variation of 6.41% between any two isolates per group using four primer pairs. Certain isolates could not be distinguished from each other. A more in-depth study of 10 isolates from MCG B, using 8 additional primer pairs, showed small genetic differences (maximum of 4.2% and minimum of 0.2%) between isolates. These results suggested that DNA could be pooled for comparison of MCGs. Pooled DNA from isolates within groups using 20 primer pairs confirmed differences between 9 MCGs. This technique effectively differentiated MCGs of S. rolfsii from each other and also detected differences between isolates within a single MCG. 相似文献
8.
黑龙江省水稻纹枯病菌的致病力分化与AFLP分析 总被引：3，自引：0，他引：3
为了明确黑龙江省水稻纹枯病菌遗传多样性,为水稻抗病育种和水稻纹枯病的综合防治提供依据。本文对采自13个水稻种植地区的29个水稻纹枯病菌菌株进行了致病力测定和AFLP分析。结果表明9对AFLP引物对供试菌株扩增出396条带,其中多态性带187条,占总扩增带数的47.22%。黑龙江省水稻纹枯病菌的遗传距离变化在0.50~0.92之间,平均为0.71,群体遗传多样性较为丰富。UPGMA法可以将供试菌株分成4个AFLP聚类组群(Ⅰ、Ⅱ、Ⅲ和Ⅳ),相同地理来源的菌株基本上聚集在同一组群内,表明AFLP类群划分与菌株的地理来源有较强的相关性。黑龙江省水稻纹枯病菌致病性分化较为明显,并且AFLP类群划分与菌株的致病性鉴定之间存在一定相关性。 相似文献
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10.
ABSTRACT Two isolates of the barley net blotch pathogen (Pyrenophora teres f. teres), one possessing high virulence (0-1) and the other possessing low virulence (15A) on the barley cultivar Harbin, were crossed and the progeny of the mating were isolated. Conidia from cultures of the parent and progeny isolates were used as inoculum to determine the inheritance of virulence in the pathogen. Of the 82 progeny tested, 42 exhibited high virulence and 40 exhibited low virulence on 'Harbin' barley. The data support a model in which a single, major gene controls virulence in P. teres f. teres on this barley cultivar (1:1 ratio; chi(2) = 0.05, P = 0.83). Preparations of DNA were made from parental and progeny isolates, and the DNA was subjected to the random amplified polymorphic DNA (RAPD) technique in a search for molecular genetic markers associated with the virulence phenotype. Five RAPD markers were obtained that were associated in coupling with low virulence. The data indicate that the RAPD technique can be used to tag genetic determinants for virulence in P. teres f. teres. 相似文献
11.
Teklu Andebrhan Antonio Figueira Milton M. Yamada Julio Cascardo Douglas B. Furtek 《European journal of plant pathology / European Foundation for Plant Pathology》1999,105(2):167-175
Crinipellis perniciosa (Stahel) Singer is the causal agent of witches' broom disease in the Sterculiaceae, Solanaceae, and Bixaceae families. The disease is endemic to the Brazilian Amazon, and was first reported infecting Theobroma cacao (cocoa) in the State of Bahia, Brazil, in 1989. Random amplified polymorphic DNA (RAPD) analyses were performed on 46 isolates of C. perniciosa from cocoa that were collected from 15 counties in Bahia and the Brazilian Amazon. A total of 258 RAPD loci from 20 primers and three mixed primers were analyzed. Of these loci, 108 (42%) were polymorphic, with an average of 4.7 polymorphic loci per primer produced. Genetic similarities were estimated using Nei and Li's index and UPGMA clustering. Bootstrap analysis divided the phenogram into four significantly different clusters: two groups contained isolates from Ariquemes and from Ouro Preto, Rondônia, and the other two separated the isolates from Bahia into two major groups of C. perniciosa, classified as Group 1 (G1) and Group 2 (G2). The two groups of isolates from Bahia differed for their genetic similarity with the isolates from the Brazilian Amazon. The geographic distribution of the groups in Bahia suggests two independent focal points of introduction. Ongoing programs to screen for resistant cocoa genotypes should consider both groups of isolates. 相似文献
12.
Differentiation and quantification of the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis using a PCR assay 总被引：3，自引：1，他引：2
Isolates of Tapesia yallundae and Tapesia acuformis were subjected to Random Amplified Polymorphic DNA (RAPD) assay. Amplification products common to isolates of either species were cloned and primers were generated from each sequence for use in conventional PCR. The primer pair derived from a T. yallundae specific RAPD marker amplified a product only from DNA of T. yallundae isolates and not from DNA of a range of other fungal species associated with the stem base disease complex of cereals. Similarly, the primer pair generated from a T. acuformis -specific RAPD marker amplifed a product only from DNA of T. acuformis isolates. Quantitative assays were developed for both species of Tapesia from these primer pairs, using competitive PCR . Competitive PCR was used to determine the level of colonization of seedlings by each species in glasshouse- and field-inoculated cereal hosts and results compared to those for conventional seedling disease assessment. 相似文献
13.
The recent history of Puccinia striiformis f.sp. tritici in Denmark as revealed by disease incidence and AFLP markers 总被引：2，自引：0，他引：2
Disease observations and amplified fragment length polymorphism (AFLP) markers were used to study recent developments in the Puccinia striiformis f.sp. tritici population in Denmark. The fungus appeared spontaneously at 10 locations in Denmark in 1997 after it was not observed under natural conditions in 1996. The pattern of disease development and prevailing winds suggested that the fungus reappeared by airborne spores from the south or west. In 1998, disease incidence was more evenly distributed throughout the country. Forty-eight single lesion isolates were collected from most crops where the disease was observed in these years; all except one from 1997 belonged to two pathotypes that were not previously detected in the country, and both possessed the newly discovered Yr17 virulence. The isolates were characterized with AFLP markers together with 28 isolates representing eight of 13 pathotypes observed prior to 1996. Initial screening of 240 Pst I/ Mse I AFLP primer combinations on four isolates showed that a primer combination, on average, revealed 0·4 polymorphisms between any isolate pair. A selection of 21 primer combinations resulted in 28 AFLP markers, which revealed 16 AFLP phenotypes among all 76 isolates. The two Yr17- virulent pathotypes consisted of three AFLP phenotypes, which were observed in both 1997 and 1998; the two most frequent AFLP phenotypes occurred at most sampling locations and often within the same crop. AFLP diversity was larger among samples collected prior to 1996, and also in this period most AFLP phenotypes were observed at different sampling locations. These results are consistent with the features of an entirely asexually reproducing pathogen dispersed by aerial spores across large areas. 相似文献
14.
Random amplified polymorphic DNA (RAPD) analysis of Phytophthora infestans isolates collected in Canada during 1994 to 1996 总被引：6，自引：0，他引：6
Mating type, glucose-6-phosphate isomerase ( Gpi ) allozyme banding patterns, response to the fungicide metalaxyl and random amplified polymorphic DNA (RAPD) markers were used to characterize genetic variability among 141 Canadian isolates of Phytophthora infestans collected between 1994 and 1996. Multiple correspondence analysis of RAPD profiles separated isolates into 21 groups that were not correlated to groups defined by mating type, Gpi allozyme banding patterns or response to metalaxyl. Population subdivision analysis showed that 97% of the total genetic variation was found among individuals within populations, compared with 3% among populations. The average similarity coefficient among isolates was 80%. No significant differences in haplotypic diversity were observed among the years under study, but levels of genetic diversity among local populations of P. infestans were high (0.76). All classes of response to the fungicide metalaxyl were observed, with 55% of isolates displaying moderate levels of insensitivity. The high level of genetic diversity detected within populations indicates that migration and sexual recombination probably play important roles in the population biology of P. infestans in Canada. 相似文献
15.
山东小麦纹枯病菌致病性与遗传分化关系的研究 总被引：4，自引：0，他引：4
对山东麦区被鉴定为双核丝核菌的35个菌株进行致病性测定的基础上,选用15个随机引物对上述菌株进行了RAPD分析,共标记出171条DNA片段,其中多态性片段161个,多态率为94.15%。用UPGMA法构建系统树,以遗传距离0.33为阈值,被鉴定为AG-D融合群的33个菌株隶属于同一个RAPD组,而3个未定融合群菌株(WK-6、WK-37和WK-13)均为独立的RAPD组。以遗传距离0.25为阈值,将属于同一个融合群的33个AG-D群菌株划分为7个亚组,说明受试小麦纹枯病菌株间存在丰富的遗传多样性。综合分析受试菌株的致病力测定结果与RAPD分析发现,供试菌株的RAPD组与菌株致病力的强弱无明显的相关性,但少数菌株的致病力强弱与亚RAPD组有一定的相关性。 相似文献
16.
Support for a stepwise mutation model for pathogen evolution in Australasian Puccinia striiformis f.sp. tritici by use of molecular markers 总被引：7，自引：0，他引：7
Since its initial detection in Australia in 1979, wheat yellow (stripe) rust ( Puccinia striiformis f.sp. tritici ) has evolved in Australia and New Zealand into more than 20 pathotypes with assorted virulence characteristics. This evolution is believed to have occurred in a stepwise fashion from an original single pathotype, with no subsequent new introductions. A combination of random amplified polymorphic DNA (RAPDs) and amplified fragment length polymorphisms (AFLPs) was used to examine the level of molecular variation in Australian and New Zealand isolates, and to compare this with variation amongst other isolates of P. striiformis . Using 60 RAPD primers on seven Australian isolates representing seven different pathotypes collected between 1979 and 1991, more than 300 potentially polymorphic loci were analysed and no polymorphisms were detected. Using the same primers on two UK isolates, 3% of loci showed a polymorphism. A similar level of polymorphism was found between UK isolates using AFLP primers, and between 5 and 15% of fragments were polymorphic between an isolate from the UK, an isolate from Denmark, and one from Colombia. However, no AFLP polymorphisms were found amongst 14 Australian and New Zealand isolates tested, at over 100 potentially polymorphic loci. The lack of molecular variation in the Australian and New Zealand collection is consistent with the stepwise mutation theory of pathotype evolution from a single introduction. 相似文献
17.
O'Neill NR van Berkum P Lin JJ Kuo J Ude GN Kenworthy W Saunders JA 《Phytopathology》1997,87(7):745-750
ABSTRACT Amplified restriction fragment length polymorphism (AFLP) was used to assess the levels of genomic variations among species and isolates of the genus Colletotrichum. Our objective was to characterize at the molecular level two alfalfa pathogens, isolates Arl-NW and 57RR, which are unusually aggressive to anthracnose-resistant alfalfa cultivars and whose taxa has been uncertain based on morphological criteria. The fingerprint patterns obtained were complex but did enable us to place these two isolates within the species C. trifolii and C. gloeosporioides, respectively. The diversity detected with AFLP among and within Colletotrichum species from alfalfa and other crops corroborated their published taxonomy based on morphology, ribosomal DNA sequence, and random amplified polymorphic DNA analyses. Similarity matrices generated with three primer pairs were highly correlated and, thus, were combined to determine the similarity among the fungal species and isolates that were analyzed. Analysis of the data generated with each of the primer pairs individually and application of either distance or parsimony methods supported the placement of these two isolates. The parsimony method of data analysis was more confirmatory in the placement of Phoma medicaginis as an out-group than the distance method, using either simple matching or Jaccard's coefficients to generate the similarity matrices. Our conclusion is that the AFLP technique will be useful for identification of individual isolates within complex genera such as Colletotrichum because of its ability to generate large numbers of polymorphisms and the consistency of polymerase chain reaction amplification. 相似文献
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19.
双重PCR检测马铃薯晚疫病菌和青枯病菌方法的建立及应用 总被引：3，自引：0，他引：3
利用真菌通用引物ITS1和ITS4扩增马铃薯晚疫病菌转录间隔区并进行序列测定,通过序列比较,设计了1对马铃薯晚疫病菌的特异引物INF1/INF2,并对15种不同真菌、细菌和7种疫霉属和腐霉属卵菌基因组DNA进行PCR扩增,结果只有不同来源的马铃薯晚疫病菌株可获得324 bp的特异带。将引物INF1/INF2与卵菌通用引物进行巢式PCR扩增后,其检测灵敏度在DNA水平上可达30 fg。运用设计的引物与马铃薯青枯病菌特异引物结合建立了双重PCR体系,能从马铃薯晚疫病菌和马铃薯青枯病菌总基因组DNA以及人工接种和自然发病的马铃薯植株中分别或同时扩增到324 bp和281 bp的特异片段。实现了同时对马铃薯晚疫病菌和马铃薯青枯病菌的快速可靠检测。 相似文献
20.
Fusarium oxysporum f.sp. gladioli (FOG) race 1 infects both large- and small-flowered Gladiolus cultivars. Race 2 isolates infect only small-flowered cultivars but can be present as epiphytes on large-flowered plants. When 160 arbitrary 10-mer oligonucleotide primers were tested on FOG by PCR to find RAPD markers specific for race 1, the RAPD primer G12 amplified two discriminating DNA fragments, AB (609 bp) and EF (1196 bp), in race 1 isolates only. Both fragments were cloned and sequenced. Two pairs of race 1-specific primers for multiplex PCR were designed. Tests of 112 F. oxysporum isolates by PCR showed that, in almost all cases, race 1 isolates of vegetative compatibility group 0340 could be distinguished with these primers. Seven putative race 1 isolates did not react in multiplex PCR; hybridization studies with labelled AB and EF DNA fragments showed that these isolates belong to separate groups. A bioassay was developed to detect corms that were latently infected with FOG race 1. Gladiolus corms were homogenized and incubated for 5 days at 28°C in a semiselective medium to induce growth of Fusarium . Cultivated mycelium was isolated and subjected to the developed multiplex PCR after standard DNA isolation or disruption by microwave treatment. 相似文献