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1.
Fusarium crown rot (FCR), caused predominantly by Fusarium pseudograminearum (Fp) in Australia, is an important fungal disease of wheat and barley. FCR causes significant yield losses and reduced grain quality worldwide. This study investigated the population dynamics of FCR-causing F. pseudograminearum isolates from Western Australia (WA), a major wheat-growing region. Wheat samples were collected from a total of seven different sites in 2008 and 2015. Two sites, Tammin and Karlgarin, with moderate to high FCR incidence, were intensively sampled in both years. The results revealed significant increase in Fp isolation frequency between 2008 and 2015. Over 86% of 1100 Fusarium isolates were Fp in 2015 compared with 59% of 639 isolates from 2008. Mating type idiomorphs, toxin chemotypes and population genetic structures were determined for a subset of 279 Fp isolates (132 isolates from 2008 and 165 from 2015). Mating type analysis revealed differences in MAT1-1 and MAT1-2 distributions between Tammin and Karlgarin for both years. Results also showed that 97.6% of Fp isolates analysed had the 3-ADON trichothecene chemotype. Additionally, for the first time in Australia, the 15-ADON chemotype was identified in 2.3% and 2.4% of Fp isolates from 2008 and 2015, respectively. The genetic structure of Fp population determined using 21 cleaved amplified polymorphic sequence (CAPS) markers revealed a high level of genetic variation within and between populations. In addition, 2015 isolates from Tammin and Karlgarin were significantly more aggressive (P < 0.0001) than 2008 isolates. This finding may have implications in managing this significant fungal disease.  相似文献   

2.
Fusarium crown rot of wheat has been spreading in the Huanghuai wheat-growing area in China since 2010, leading to a potential yield loss. To investigate the pathogens associated with this disease in Jiangsu and Shandong provinces in recent years, 617 Fusarium isolates were isolated from nine sites in these two provinces between 2014 and 2016. Of these isolates, 372 were identified as Fusarium pseudograminearum, and the remaining isolates were identified as F. asiaticum and F. graminearum, suggesting that F. pseudograminearum is becoming a predominant causative pathogen of crown rot of wheat in eastern China. Trichothecene gene detection and chemical analyses of trichothecenes indicated that the F. pseudograminearum isolates belonged to the 3-ADON or 15-ADON chemotype, and one isolate had the NIV genotype but produced no detectable NIV. 3-ADON isolates were predominant in Jiangsu, whereas 15-ADON isolates were prevalent in Shandong. The mating type of the F. pseudograminearum isolates were identified. MAT-1 and MAT-2 existed, but in most collections, particularly those in Jiangsu, the ratios of the two mating types deviated significantly from an expected 1:1 ratio. The reason for the occurrence of F. pseudograminearum is hypothesized, and the chemotype and mating type distribution of this species in these two provinces are analysed.  相似文献   

3.
Fusarium species involved in the Fusarium crown rot (FCR) complex affect wheat in every stage of development from seedling to grain fill. This study was designed to compare the aggressiveness of the FCR complex members including F. culmorum, F. pseudograminearum and F. graminearum in causing seedling blight, decreased plant vigour and crown rot. To assess their relative pathogenicity, two hard red spring wheat cultivars and two durum wheat cultivars were inoculated in the field with five isolates from each of the three species for two years. Significant differences in patterns of pathogenicity were identified. In particular, F. culmorum caused greater seedling blight while F. pseudograminearum and F. graminearum caused greater crown rot. Greatest yield reductions were caused by F. pseudograminearum. Cultivar differences were identified with respect to seedling disease and late season crown rot. No interactions were identified between cultivar performance and isolates or species with which they were challenged.  相似文献   

4.
Fusarium verticillioides (sexual stage Gibberella moniliformis) is a common fungal pathogen of maize worldwide that also produces fumonisin mycotoxins. Populations of this fungus can be diverse with respect to neutral and selectable genetic markers. We used vegetative compatibility groups (VCGs) and amplified fragment length polymorphisms (AFLPs) to evaluate the genetic structure of three F. verticillioides populations from commercial maize fields in Argentina. Based on work with similar populations from outside South America, we expected individuals within the populations to be genetically diverse, that genotypic variation would be distributed in a manner consistent with random mating, and that populations from different locations would be genetically indistinguishable from one another. We analysed 62 AFLP loci for 133 fungal isolates. All three populations were genotypically diverse but genetically similar and potentially part of a larger, randomly mating population, with significant genetic exchange occurring between the three subpopulations. There was no evidence for linkage disequilibrium at P = 0.05. The low values of G ST , the lack of frequent private alleles, and the lack of a systemic pattern of linkage disequilibrium all suggest that sexual reproduction is sufficiently common in F. verticillioides and that the dispersal of strains is sufficiently efficient for the population of F. verticillioides in the main maize growing region to be a single randomly mating population with no detectable genetic subdivision. Thus differences in disease and/or toxin production observed in this region are best attributed to differences other than the genetic composition of the population.  相似文献   

5.
Fusarium head blight (FHB) is one of the most destructive diseases of wheat. Twelve small commercial wheat fields (size 1–3 hectares) were sampled in Germany for Fusarium populations at three spots per field with 10 heads each. PCR assays using generic primers confirmed 338 isolates as F.graminearum sensu stricto (s.s.) (64.9%) out of 521 Fusarium spp. that were further analyzed. Populations of F. graminearum s.s. in Germany contain three types of trichothecenes with a dominancy of 15-acetyldeoxynivalenol chemotype (92%) followed by 3-acetyldeoxynivalenol chemotype (6.8%) and a few isolates of nivalenol chemotype (1.2%). All these isolates were genotyped using 19 microsatellite loci. The 12 populations showed a high genetic diversity within the small scale sampling areas resulting in 300 different haplotypes. Genetic diversity within populations (71.2%) was considerably higher than among populations (28.8%) as shown by analysis of molecular variance. Gene flow (Nm) between populations ranged from 0.76–3.16. Composition of haplotypes of one population followed over 2 years changed considerably. No correlation between genetic and geographical distance was found. In conclusion, populations of F. graminearum s.s. in Germany display a tremendous genetic variation on a local scale with a restricted diversity among populations.  相似文献   

6.
Fusarium culmorum is a major pathogen able to cause foot and root rot and the incitant of Fusarium head blight in wheat in Tunisia. The aims of the present study were to evaluate by PCR the type of mycotoxins produced, to determine the mating type and to analyse the genetic diversity by microsatellite markers of 82?F. culmorum isolates recovered from two separated Tunisian fields. Specific sequences in the Tri6-Tri5 intergenic region, Tri7 and Tri13 were used to identify 3-AcDON- or 15-AcDON-. All studied F. culmorum isolates, were of the 3-AcDON- type. No 15-AcDON- and NIV types were detected in this research. Both mating types MAT1-1 and MAT1-2 were recovered from the two fields in approximately equal proportions. Five polymorphic microsatellite markers were applied to F. culmorum isolates, to determine the genetic variation in and among populations. Sixty-four haplotypes were identified; the analysis of the population structure did not reveal a strong variation between fields. Total gene diversity (H T ?=?0.505; H S ?=?0.497) and analysis of molecular variance confirmed that most of the genetic variability was within populations (Φ ST ?=?0.033; P?<?0.0039). Gene flow (N m ?=?31.05) indicated little differentiation among populations. Based on these results, the F. culmorum isolates collected from different fields might be part of one large panmictic population and in addition the low linkage disequilibrium values with high genetic variation within populations suggest that the population is recombining sexually.  相似文献   

7.
In this study, 18 microsatellite loci were used to examine the genetic structure and mode of reproduction of Phaeomoniella chlamydospora, the fungus that causes Petri disease of grapevine and is involved in another grapevine disease, esca. A total of 60 southeastern Australian isolates were tested and compared with 64 isolates from southern France. The French population possessed relatively high genotypic diversity (G = 29·2) whilst the Australian population showed low genotypic diversity (G = 11·1), consistent with a limited founder population. Haplotypes from four different grapevine cultivars were not significantly differentiated (Φpt values effectively zero). Likewise, genetic differentiation of haplotypes from different regions within each country was not significant, although small but significant genetic differentiation (9%) was identified between Australia and France. Based on bootstrapped cluster analysis, there did not appear to be any genetic groups within the overall sample of isolates. Significant linkage disequilibrium identified in both countries, together with overrepresented, widespread identical haplotypes, indicated limited genetic recombination and a largely clonal structure consistent with the absence of an observed sexual cycle in this species.  相似文献   

8.
An extensive survey was carried out to collect Fusarium species colonizing the lower stems (crowns) of bread wheat (Triticum aestivum L.) and durum wheat (T. durum Desf.) from different wheat growing regions of Turkey in summer 2013. Samples were collected from 200 fields representing the major wheat cultivation areas in Turkey, and fungi were isolated from symptomatic crowns. The isolates were identified to species level by sequencing the translation elongation factor 1-alpha (TEF1-α) gene region using primers ef1 and ef2. A total of 339 isolates representing 17 Fusarium species were isolated. The isolates were identified as F. culmorum, F. pseudograminearum, F. graminearum, F. equiseti, F. acuminatum, F. brachygibbosum, F. hostae, F. redolens, F. avenaceum, F. oxysporum, F. torulosum, F. proliferatum, F. flocciferum, F. solani, F. incarnatum, F. tricinctum and F. reticulatum. Fusarium equiseti was the most commonly isolated species, accounting for 36% of the total Fusarium species isolated. Among the damaging species, F. culmorum was the predominant species being isolated from 13.6% of sites surveyed while F. pseudograminearum and F. graminearum were isolated only from 1% and 0.5% of surveyed sites, respectively. Six out of the 17 Fusarium species tested for pathogenicity caused crown rot with different levels of severity. Fusarium culmorum, F. pseudograminearum and F. graminearum caused severe crown rot disease on durum wheat. Fusarium avenaceum and F. hostae were weakly to moderately virulent. Fusarium redolens was weakly virulent. However, F. oxysporum, F. equiseti, F. solani, F. incarnatum, F. reticulatum, F. flocciferum, F. tricinctum, F. brachygibbosum, F. torulosum, F. acuminatum and F. proliferatum were non-pathogenic. The result of this study reveal the existence of a wide range of Fusarium species associated with crown rot of wheat in Turkey.  相似文献   

9.
Fusarium head blight (FHB), caused principally by Gibberella zeae (Fusarium graminearum), is a devastating disease of small grains such as wheat and barley worldwide. Grain infected with G. zeae may be contaminated with trichothecene mycotoxins such as deoxynivalenol (DON) and nivalenol (NIV). Strains of G. zeae that produce DON may also produce acetylated derivatives of DON: 3‐acetyl‐DON (3‐ADON) and 15‐acetyl‐DON (15‐ADON). Gradients (clines) of 3‐ADON genotypes in Canada have raised questions about the distribution of G. zeae trichothecene genotypes in wheat fields in the eastern USA. Tri3 and Tri12 genotypes were evaluated in 998 isolates of G. zeae collected from 39 winter wheat fields in New York (NY), Pennsylvania (PA), Maryland (MD), Virginia (VA), Kentucky (KY) and North Carolina (NC). Ninety‐two percent (919/998) of the isolates were 15‐ADON, 7% (69/998) were 3‐ADON, and 1% (10/998) was NIV. A phylogenetic analysis based on portions of three genes (PHO, RED and URA) from 23 isolates revealed two species of Fusarium (F. graminearum sensu stricto and one isolate of F. cerealis (synonym F. crookwellense)). An increasing trend of 3‐ADON genotypes was observed from NC (south) to NY (north). Punctuated episodes of atmospheric transport may favour a higher frequency of 3‐ADON genotypes in the northeastern USA, near Canada, compared with the mid‐Atlantic states. Discoveries of the NIV genotype in NY and NC indicate the need for more intensive sampling in the surrounding regions.  相似文献   

10.
The objective of this study was to determine whether genetically differentiated groups of Puccinia triticina are present in Europe. In total, 133 isolates of P. triticina collected from western Europe, central Europe and Turkey were tested for virulence on 20 lines of wheat with single leaf rust resistance genes, and for molecular genotypes with 23 simple sequence repeat (SSR) markers. After removal of isolates with identical virulence and SSR genotype within countries, 121 isolates were retained for further analysis. Isolates were grouped based on SSR genotypes using a Bayesian approach and a genetic distance method. Both methods optimally placed the isolates into eight European (EU) groups of P. triticina SSR genotypes. Seven of the groups had virulence characteristics of isolates collected from common hexaploid wheat, and one of the groups had virulence characteristics of isolates from tetraploid durum wheat. There was a significant correlation between the SSR genotypes and virulence phenotypes of the isolates. All EU groups had observed values of heterozygosity greater than expected and significant fixation values, which indicated the clonal reproduction of urediniospores in the overall population. Linkage disequilibria for SSR genotypes were high across the entire population and within countries. The overall values of RST and FST were lower when isolates were grouped by country, which indicated the migration of isolates within Europe. The European population of P. triticina had higher levels of genetic differentiation compared to other continental populations.  相似文献   

11.
The heterothallic ascomycete Fusarium fujikuroi (teleomorph: Gibberella fujikuroi) is the causal agent of bakanae of rice, a disease of increasing economic importance in the major rice‐producing areas in the world and a serious threat for Italian rice cultivation. A few studies have characterized F. fujikuroi isolates in America and the Philippines but no data are available on the genetic structure of Italian pathogen populations. Microsatellite SSRs are useful tools to study the intraspecific diversity at population level. In this study, 19 polymorphic SSRs have been identified and applied to characterize the genetic variation of 334 isolates of F. fujikuroi coming from eight Italian rice‐growing areas. A high degree of diversity at haplotype level has emerged: in the eight populations, 107 unique haplotypes were scored. Analysis of molecular variance (amova ) showed that 98% of genetic variability occurred within F. fujikuroi Italian populations, as confirmed by the allelic Shannon index ranging from 0.56 to 1.06. The presence of a 1:1 ratio of mating type alleles in six out of eight of the Italian fungal populations suggests a potential for sexual reproduction in the field. However, the high fraction of clonality (43%), confirmed by neighbour‐joining clustering analysis, and the high level of linkage disequilibrium observed, indicates that reproduction of F. fujikuroi is mostly clonal in Italy. All data suggest that the observed genetic variability was probably mediated by human activity and transmission by rice seeds.  相似文献   

12.
The objective of this study was to assess the genetic diversity and to infer the mode of reproduction of Botrytis elliptica and B. tulipae in the Netherlands. First, three molecular typing methods were compared for their ability to differentiate isolates of B. tulipae, B. elliptica, and B. cinerea. The methods compared were multilocus sequencing, restriction analysis of the ribosomal intergenic spacer (IGS) region, and amplified fragment length polymorphism (AFLP) analysis. AFLP fingerprinting provided the most efficient method to differentiate isolates within each Botrytis species and therefore this method was used for population analyses of B. elliptica and B. tulipae. Isolates of both species were sampled during successive growing seasons in experimental field plots in Lisse and other locations in the Netherlands. Among 174 B. elliptica isolates, 105 genotypes could be discriminated and 87 genotypes were found only once, reflecting high genotypic variation. Clonal genotypes were found only within growing seasons and in one location. Linkage disequilibrium analyses indicated that between 9.4% and 19.3% of the loci in clone-corrected samples were linked. The multilocus association index provided no evidence for random mating. We conclude that sexual recombination occurs in the B. elliptica population. Among the 170 B. tulipae isolates, 25 genotypes could be discriminated and four genotypes were found only once, reflecting a low genotypic variation. Clonal genotypes were frequently found in different growing seasons and different locations. Linkage disequilibrium analyses indicated that between 25.2% and 48.6% of the loci in clone-corrected samples were linked. We conclude that the B. tulipae population is mainly clonal with some recombination.  相似文献   

13.
董杰  张金良  杨建国  张昊  冯洁 《植物保护》2016,42(6):116-121
本文分析了北京市与河北省小麦赤霉病菌群体遗传结构以及基础生物学特性。结果表明所有菌株均为禾谷镰刀菌(Fusarium graminearum),属于一个大的单一群体,群体内具有较高的遗传多样性。毒素化学型测定表明,北京与河北地区小麦真菌毒素污染的主要风险为DON与15ADON毒素。表型测定显示,与F.asiaticum群体相比,F.graminearum具有较高的产孢能力,而生长速率和产毒能力较低。该群体对主要杀菌剂多菌灵、戊唑醇和氰烯菌酯均无抗药性。  相似文献   

14.
Four-hundred-sixty-eightFusarium andFusarium-like isolates were obtained from crowns and subcrown internodes of winter wheat grown in Erzurum, Turkey. Of these isolates, 34.8% wereFusarium acuminatum, 32.3% wereF. equiseti, 16.9% wereF. oxysporum, 15.0% wereMicrodochium nivale (formerlyFusarium nivale), 0.6% wereF. tabacinum and 0.4% wereF. solani. In pathogenicity tests on wheat, the highest disease severity was caused by isolates ofM. nivale, whereas isolates ofF. acuminatum, F. equiseti, F. oxysporum andF. solani were slightly virulent; isolates ofF. tabacinum were nonpathogenic. This is the first report ofM. nivale andF. tabacinum from wheat in Turkey. http://www.phytoparasitica.org posting Jan. 29, 2003.  相似文献   

15.
Simple sequence repeat (SSR) markers were used to classify 116 isolates of Phakopsora pachyrhizi, the cause of soyabean rust, collected from infected soyabean leaves in four agroecological zones in Nigeria. A high degree of genetic variation was observed within the sampled populations of P. pachyrhizi. Eighty‐four distinct genotypes were identified among three of the four agroecological zones. Nei’s average genetic diversity across geographical regions was 0·22. Hierarchical analysis of molecular variance showed low genetic differentiation among all populations of P. pachyrhizi. The majority (> 90%) of the genetic diversity was distributed within each soyabean field, while approximately 6% of the genetic diversity was distributed among fields within geographic regions. Low population differentiation was indicated by the low FST values among populations, suggesting a wide dispersal of identical genotypes on a regional scale. Phylogenetic analysis indicated a strictly clonal structure of the populations and five main groups were observed, with group II accounting for 30% of the entire population. Because of the asexual reproduction of P. pachyrhizi, single‐step mutations in SSR genotypes are likely to account for the genetic differences within each group.  相似文献   

16.
Pitch canker on plantation-grown Pinus species, caused by the fungal pathogen Fusarium circinatum, first appeared in the western and southern Cape regions of South Africa. However, outbreaks have subsequently been reported from the major plantation growing regions of KwaZulu-Natal and Limpopo in the eastern, summer rainfall regions of the country. It is more than 10 years since the last detailed population genetics studies on F. circinatum in the region were conducted. To shed light on the population biology of F. circinatum in this region of South Africa, we used microsatellite markers and mating-type assays to study a collection of 296 isolates from different nurseries and plantation sites. Our results showed that populations in the region are highly diverse, but strongly interconnected, with various genotypes shared across nursery and plantation collection sites. In contrast to nursery populations, those associated with pitch canker outbreaks were characterized by the presence of relatively small numbers of dominant genotypes that were generally widespread across the region. Opposite mating-type individuals occurred in most of the isolate collections, but multilocus linkage disequilibrium analyses pointed towards clonality being the main reproductive mode of F. circinatum in the region. Most of the pathogen's genetic variation could probably have resulted from multiple different introductions into the country and more specifically, into the summer rainfall region. Because the spread and establishment of invasive pathogens are typically driven by aggressive clones, the results of this study provide important considerations for current and future Pinus disease management strategies.  相似文献   

17.
Fusarium head blight (FHB) in small grain cereals is primarily caused by the members of the Fusarium graminearum species complex. These produce mycotoxins in infected grains, primarily deoxynivalenol (DON); acetylated derivatives of DON, 3‐acetyl‐DON (3‐ADON) and 15‐acetyl‐DON (15‐ADON); and nivalenol (NIV). This study reports the isolation of Fusarium cerealis in infected winter wheat heads for the first time in Canada. A phylogenetic analysis based on the TRI101 gene and F. graminearum species‐specific primers revealed two species of Fusarium: F. graminearum sensu stricto (127 isolates) and F. cerealis (five isolates). Chemotype determination based on the TRI3 gene revealed that 65% of the isolates were 3‐ADON, 31% were 15‐ADON and 4% were NIV producers. All the F. cerealis isolates were of NIV chemotype. Fusarium cerealis isolates can often be misidentified as F. graminearum as the morphological characteristics are similar. Although the cultural and macroconidial characteristics of F. graminearum and F. cerealis isolates were similar, the aggressiveness of these isolates on susceptible wheat cultivar Roblin and moderately resistant cultivar Carberry differed significantly. The F. graminearum 3‐ADON isolates were most aggressive, followed by F. graminearum 15‐ADON and F. cerealis NIV isolates. The findings from this study confirm the continuous shift of chemotypes from 15‐ADON to 3‐ADON in North America. In Canada, the presence of NIV is limited to barley samples and the discovery of NIV‐producing F. cerealis species in Canadian wheat fields may pose a serious concern to the Canadian wheat industry in the future.  相似文献   

18.
The ascomycete fungus Cryphonectria parasitica, causal agent of chestnut blight, is probably one of the best known invasive fungal pathogens in forests of Europe and North America. Mycovirus that reduces virulence of C. parasitica can be used as a biocontrol agent of the chestnut blight. However, anastomosis‐mediated virus transmission is limited by a vegetative (in)compatibility (vc) system involving at least six known diallelic vic genetic loci. This study looked at vegetative compatibility (vc) diversity in two populations of C. parasitica in Croatia. For that purpose, a PCR assay was validated and implemented using already known/published and newly designed primers for amplification of six known vic loci. The vc genotypes determined by PCR for 158 C. parasitica isolates investigated in this study were in complete agreement with the vc genotypes determined by pairwise co‐culturing of the same isolates, revealing the specificity and accuracy of the PCR‐based molecular vic genotyping assay. Twenty‐six unique vc genotypes were found among 158 isolates, and 19 vc types per population, which makes Croatian C. parasitica populations among the most diverse in Europe regarding the number of vc types and genetic diversity. Low values of multilocus linkage disequilibrium suggest sexual reproduction as a major contributor to high C. parasitica genetic diversity in studied populations.  相似文献   

19.
The causal agents of mango malformation disease in Brazil are a new Fusarium lineage in the Gibberella fujikuroi species complex and Fusarium sterilihyphosum; however information on the genetic and geographical diversity of these pathogens in Brazil is missing. Vegetative compatibility group (VCG) and amplified fragment length polymorphism (AFLP) analyses were used to measure the genetic diversity within these populations. Both techniques identified the same genetic groups. Six VCG and AFLP groups were identified amongst isolates of the new lineage from Brazil. FB-VCG 1/AFLP I was the most widespread group, found in seven of the 13 sites sampled. The second most frequent group was recovered from three sites. The remaining four groups were recovered from single-sites. We think that this lineage represents a genetically and geographically diverse indigenous population that reproduces clonally. In F. sterilihyphosum, group FS-VCG 1/AFLP VII was found at three sites in the southeast region of Brazil. FS-VCG 2/AFLP VIII contained isolates from South Africa but not from Brazil. Fusarium mangiferae isolates from India and South Africa formed one group, while isolates from Egypt and the USA formed a second group. F. sterilihyphosum at present is represented by a small population that might have been introduced only once into a restricted area. The clonal nature of the observed populations suggests that these fungi either occur naturally on indigenous hosts and have jumped to the introduced mango host (introduced in Brazil) or that they originated with mango and went through a severe population bottleneck when they were introduced to Brazil from India or Southeast Asia.  相似文献   

20.
Members of the Fusarium graminearum species complex (FGSC) cause Fusarium head blight in small cereal grains all over the world. To determine the species and trichothecene chemotype composition and population structure of FGSC in Jiangsu and Anhui provinces, an area where epidemics occur regularly, 891 isolates were collected in two consecutive years (2011 and 2012) and characterized with species- and chemotype-specific polymerase chain reaction. Of the 891 isolates typed, 83 were F. graminearum sensu stricto (s. str.) and 808 were F. asiaticum. All 83 F. graminearum s. str. isolates were of a 3ADON (26.51 %) or 15ADON (73.49 %) type, while F. asiaticum isolates included 696 3ADON producers, 46 15ADON producers, and 66 NIV producers. Eight variable number tandem repeat (VNTR) markers were tested on a representative 384 F. asiaticum isolates from 55 sampling sites. VNTR analysis showed high gene diversity and genotypic diversity but low linkage disequilibrium in both populations Fg2011 and Fg2012 grouped based on the year of collection. Low genetic differentiation (F ST ? =?0.026) and high gene flow (N m ?=?15.13) was observed between the two populations and among subpopulations within the same population (N m ?=?3.53 to 48.37), indicating that few influence of temporal and spatial variations on population differentiation in this area. Similar result was obtained from 3ADON, 15ADON and NIV populations or carbendazim resistant and sensitive populations, indicating that chemotype of Fusarium isolates and carbendazim application had minor influence on population subdivision.  相似文献   

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