我国小麦条锈菌体细胞遗传重组的分子证据

 小麦条锈病是影响我国小麦生产的重要病害之一,条锈菌毒性变异是引致小麦品种抗病性丧失的主要原因。本文应用SSR分子标记技术,研究了陇南越夏区小麦条锈菌群体遗传结构,以期寻找条锈菌群体遗传重组的分子证据。研究结果表明,陇南地区小麦条锈菌群体遗传多样性比较丰富而遗传分化较小,遗传变异主要存在于群体内部,而在群体之间,遗传多样性有显著的差异。在11对SSR引物中,有4对能够检测到陇南地区小麦条锈菌群体存在遗传重组,而且重组体出现的频率不同,CPS15揭示的条锈菌重组体出现的频率为20.0%,CPS34揭示的条锈菌重组体出现的频率为18.5%,RJ20揭示的条锈菌重组体出现的频率为12.8%,RJ18揭示的条锈菌重组体出现的频率为15.0%,陇南地区小麦条锈菌群体重组体出现频率平均为16.6%。本文揭示的遗传重组现象表明陇南地区条锈菌体细胞结合十分普遍,由此推测我国小麦条锈菌在自然条件下通过遗传重组而导致毒性变异的可能性。     

Attempted somatic hybridization of Puccinia striiformis f. sp. tritici and P. striiformis f. sp. hordei

Attempts were made to produce somatic hybrids between isolates of Puccinia striiformis f. sp. tritici and f. sp. hordei. A mixed infection was produced on a common susceptible barley host, Fong Tien, using white-spored isolates of P. striiformis f. sp. tritici and yellow-spored isolates off. sp. hordei. Selection was made for non-parental spore colour on selective wheat and barley hosts, and variants thus isolated were analysed for virulence markers, and for isozyme and double-stranded RNA (dsRNA) markers, all of which clearly differentiated the parental isolates. Two white-spored (non-parental) isolates were found on the selective barley host which otherwise resembled the parental f. sp. hordei isolate in virulence, isozyme and dsRNA markers. The most likely explanation of the origin of these isolates is mutation to white spore colour in the f. sp. hordei isolate.     

The recent history of Puccinia striiformis f.sp. tritici in Denmark as revealed by disease incidence and AFLP markers

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.     

Genetic Diversity in Australian Populations of Puccinia graminis f. sp. avenae

ABSTRACT Sequence-tagged microsatellite profiling was used to develop 110 microsatellites for Puccinia graminis f. sp. tritici (causal agent of wheat stem rust). Low microsatellite polymorphism was exhibited among 10 pathogenically diverse P. graminis f. sp. tritici isolates collected from Australian cereal growing regions over a period of at least 70 years, with two polymorphic loci detected, each revealing two alleles. Limited cross-species amplification was observed for the wheat rust pathogens, P. triticina (leaf rust) and P. striiformis f. sp. tritici (stripe rust). However, very high transferability was revealed with P. graminis f. sp. avenae (causal agent of oat stem rust) isolates. A genetic diversity study of 47 P. graminis f. sp. avenae isolates collected from an Australia-wide survey in 1999, and a historical group of 16 isolates collected from Australian cereal growing regions from 1971 to 1996, revealed six polymorphic microsatellite loci with a total of 15 alleles. Genetic analysis revealed the presence of several clonal lineages and subpopulations in the pathogen population, and wide dispersal of identical races and genotypes throughout Australian cereal-growing regions. These findings demonstrated the dynamic population structure of this pathogen in Australia and concur with the patterns of diversity observed in pathogenicity studies.     

Support for a stepwise mutation model for pathogen evolution in Australasian Puccinia striiformis f.sp. tritici by use of molecular markers

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.     

Genetic Relatedness of African and United States Populations of Cercospora zeae-maydis

Two taxonomically identical but genetically distinct sibling species, designated groups I and II, of Cercospora zeae-maydis cause gray leaf spot of maize in the United States. Isolates of the gray leaf spot pathogen from Africa were compared with isolates from the United States by amplified fragment length polymorphism (AFLP) analysis and restriction digests of internal transcribed spacer (ITS) regions and 5.8S ribosomal DNA (rDNA), as well as by morphological and cultural characteristics. The isolates from Africa were morphologically indistinguishable from the U.S. isolates in both groups, but like isolates of group II, they grew more slowly and failed to produce detectable amounts of cercosporin in culture. Analysis of restriction fragments from the ITS and rDNA regions digested with five endonucleases indicated that all of the African isolates shared the profile of the C. zeae-maydis group II population from the eastern United States and, thus, are distinct from the group I population, which is more prevalent in the United States and other parts of the world. Cluster analysis of 85 AFLP loci confirmed that the African and U.S. group II populations were conspecific (greater than 97% average similarity) with limited variability. Among all group II isolates, only 8 of 57 AFLP loci were polymorphic, and none was specific to either population. Thus, although gray leaf spot was reported in the United States several decades prior to the first record in Africa, the relative age of the two populations on their respective continents could not be ascertained with confidence. The absence of C. zeae-maydis group I in our samples from four countries in the major maize-producing region of Africa as well as the greater AFLP haplotype diversity found in the African group II population, however, suggest that Africa was the source of C. zeae-maydis group II in the United States. The overall paucity of AFLP variation in this sibling species further suggests that its origin is recent or that the ancestral population experienced a severe bottleneck prior to secondary migration.     

2010年西藏小麦条锈菌生理小种群体结构与分析

西藏地区是中国相对独立的小麦种植区,小麦条锈病是西藏冬小麦上最重要的病害.长期以来,对西藏小麦条锈菌生理小种群体结构缺乏全面系统的了解.为了弄清西藏小麦条锈菌生理小种群体结构,本研究从西藏地区小麦条锈病发生的关键地区采集并鉴定了小麦条锈菌标样261份.西藏地区小麦条锈菌群体结构复杂,小种类型数多,主要优势小种以CYR32和CYR33为主,水源11类群为优势类群,Hybrid46类群结构简单,未发现CYR32以外的类型;CYR32之前的小种数较多、其中CYR17、CYR20、CYR31出现频率较高;西藏小麦条锈菌群体结构与内地有着较大的相似性,同时也有其自身的独特性,表现西藏小麦条锈菌优势小种组成与四川、云南两省相似,与青海省差异较大.推测西藏地区小麦条锈菌与四川和云南省存在较密切的菌源交流,与青海省交流较少.     

Inheritance and molecular mapping of barley genes conferring resistance to wheat stripe rust

ABSTRACT Most barley cultivars are resistant to stripe rust of wheat that is caused by Puccinia striiformis f. sp. tritici. The barley cv. Steptoe is susceptible to all identified races of P. striiformis f. sp. hordei (PSH), the barley stripe rust pathogen, but is resistant to most P. striiformis f. sp. tritici races. To determine inheritance of the Steptoe resistance to P. striiformis f. sp. tritici, a cross was made between Steptoe and Russell, a barley cultivar susceptible to some P. striiformis f. sp. tritici races and all tested P. striiformis f. sp. hordei races. Seedlings of parents and F(1), BC(1), F(2), and F(3) progeny from the barley cross were tested with P. striiformis f. sp. tritici races PST-41 and PST-45 under controlled greenhouse conditions. Genetic analyses of infection type data showed that Steptoe had one dominant gene and one recessive gene (provisionally designated as RpstS1 and rpstS2, respectively) for resistance to races PST-41 and PST-45. Genomic DNA was extracted from the parents and 150 F(2) plants that were tested for rust reaction and grown for seed of F(3) lines. The infection type data and polymorphic markers identified using the resistance gene analog polymorphism (RGAP) technique were analyzed with the Mapmaker computer program to map the resistance genes. The dominant resistance gene in Steptoe for resistance to P. striiformis f. sp. tritici races was mapped on barley chromosome 4H using a linked microsatellite marker, HVM68. A linkage group for the dominant gene was constructed with 12 RGAP markers and the microsatellite marker. The results show that resistance in barley to the wheat stripe rust pathogen is qualitatively inherited. These genes might provide useful resistance against wheat stripe rust when introgressed into wheat from barley.     

PSR(Puccinia Striiformis Repeat)序列的基因组特异性和指纹遗传稳定性研究

 以PSR331S₃(Puccinia striiformis repeat)为探针,对感染小麦条锈菌P. striiformis f. sp. tritici模式菌系的叶片和常用繁殖寄主健康叶片总DNA进行Southern分析,结果显示PSR331S₃具有良好的指纹分辨力和基因组特异性。对系列单孢系的指纹分析表明,PSR位点在有丝分裂中是稳定遗传的,可以用于条锈菌的遗传分析。     

西藏林芝与内地小麦条锈菌分子群体遗传结构及菌源关系

 2009-2010年春季,先后从甘肃、四川、陕西和西藏四省的29个县(市)采集到1 391份小麦条锈病标样,繁殖获得961个菌株,利用SSR分子标记进行群体遗传分析结果表明:甘肃天水、平凉和陇南,陕西宝鸡及汉中,四川阿坝和广元等地条锈菌的遗传多样性比较丰富,而四川宜宾及凉山、西藏林芝的遗传多样性水平相对较低。利用Arlequin软件中的AMOVA方法分析结果表明,小麦条锈菌的遗传变异主要存在于群体内部。内地各种群之间菌源交流频繁(Nm＞4),西藏与内地菌源交流很少(Nm＜1)。采用Structure及聚类分析表明,陕西宝鸡与甘肃平凉间,陕西汉中、甘肃陇南、甘肃天水及四川广元间,存在着频繁的菌源交流关系,四川宜宾和凉山与四川阿坝、陕西汉中和甘肃陇南间存在着菌源交流关系。而西藏与内地间几乎没有菌源交流。初步认为西藏林芝小麦条锈菌可能是一个相对独立的遗传群体。     

Genetic analysis and molecular mapping of wheat genes conferring resistance to the wheat stripe rust and barley stripe rust pathogens

ABSTRACT Stripe rust is one of the most important diseases of wheat and barley worldwide. On wheat it is caused by Puccinia striiformis f. sp. tritici and on barley by P. striiformis f. sp. hordei Most wheat genotypes are resistant to P. striiformis f. sp. hordei and most barley genotypes are resistant to P. striiformis f. sp. tritici. To determine the genetics of resistance in wheat to P. striiformis f. sp. hordei, crosses were made between wheat genotypes Lemhi (resistant to P. striiformis f. sp. hordei) and PI 478214 (susceptible to P. striiformis f. sp. hordei). The greenhouse seedling test of 150 F(2) progeny from the Lemhi x PI 478214 cross, inoculated with race PSH-14 of P. striiformis f. sp. hordei, indicated that Lemhi has a dominant resistance gene. The single dominant gene was confirmed by testing seedlings of the F(1), BC(1) to the two parents, and 150 F(3) lines from the F(2) plants with the same race. The tests of the F(1), BC(1), and F(3) progeny with race PSH-48 of P. striiformis f. sp. hordei and PST-21 of P. striiformis f. sp. tritici also showed a dominant gene for resistance to these races. Cosegregation analyses of the F(3) data from the tests with the two races of P. striiformis f. sp. hordei and one race of P. striiformis f. sp. tritici suggested that the same gene conferred the resistance to both races of P. striiformis f. sp. hordei, and this gene was different but closely linked to Yr21, a previously reported gene in Lemhi conferring resistance to race PST-21 of P. striiformis f. sp. tritici. A linkage group consisting of 11 resistance gene analog polymorphism (RGAP) markers was established for the genes. The gene was confirmed to be on chromosome 1B by amplification of a set of nullitetrasomic Chinese Spring lines with an RGAP marker linked in repulsion with the resistance allele. The genetic information obtained from this study is useful in understanding interactions between inappropriate hosts and pathogens. The gene identified in Lemhi for resistance to P. striiformis f. sp. hordei should provide resistance to barley stripe rust when introgressed into barley cultivars.     

感染“中四”小麦条锈菌T4新菌系的致病范围测定

 为研究T4新菌系对我国条锈病流行区后备及生产品种、中四后代品种（系）的致病范围,预测T4新菌系对我国未来条锈菌群体结构的影响。在苗期和成株期,用T4菌系对供试品种进行了抗病性鉴定。结果表明：T4新菌系对我国条锈病流行区后备品种和陕西省生产品种致病范围较窄,毒性较弱;而对含有“中四”血缘的小麦品种（系）有较强的毒性,几乎能感染所有供试的“中四”衍生系品种。在现有生产品种大面积种植下,T4新菌系暂时不会构成对现有品种的威胁;但随着“中四”后代品种大面积种植,感染“中四”条锈菌新菌系将对我国条锈病持续控制构成潜在威胁,必须加强对新菌系的监测。     

Removal of urediniospores of brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rusts of wheat from infected leaves submitted to a mechanical stress

The passive spore removal from colonies due to mechanical stress was compared in the brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rusts of wheat. Mechanical stress was applied using either a miniaturized wind tunnel or a centrifuge. In wind-tunnel experiments, a wind of minimum velocity of 1.3 and 1.8 m s^-1 for P. recondita f.sp. tritici and P. striiformis, respectively, applied for at least 10 seconds, was necessary to remove spores. The interaction between wind velocity and cumulated duration was significant for both rusts. At low wind velocity, a longer duration was required to remove the spores than at high wind velocity, and vice versa. In centrifugation experiments, the maximum spore removal occurred for angular velocities of 10³ and 2 10³ rotations min^-1, for P. recondita f.sp. tritici and P. striiformis, respectively, applied for 5 min. Calculation of the aerodynamic and centrifugal forces showed that the forces necessary to remove spores are greater for P. striiformis than for P. recondita f.sp. tritici. This difference can be related to the size of the dispersal unit, which is larger in P. striiformis than in P. recondita f.sp. tritici due to spore clustering. These observations are consistent with the differences in the mean spore dispersal distance, which is usually smaller in P. striiformis than in P. recondita f.sp. tritici.     

Variation for isozymes and double-stranded RNA among isolates of Puccinia striiformis and two other cereal rusts

Gel electrophoresis was used to examine the variation in isozymes and dsRNA within and between the rust species Puccinia striiformis, P. recondita and P. hordei. No differences in isozyme phenotype were found among 29 diverse isolates of the wheat-attacking form of P. striiformis (WYR). Smaller numbers of isolates of the barley-attacking form (BYR) of P. recondita and of P. hordei showed similar intra-group uniformity. There were major differences in isozyme phenotypes between the three species, while WYR and BYR differed for two of 12 enzymes. Double-stranded RNA was detected in each species and in all 26 isolates examined. For WYR and BYR, all isolates within each group had the same or a similar phenotype. In contrast, each isolate of P. recondita and P. hordei had a unique phenotype. There were differences in dsRNA phenotypes both between the three species and between WYR and BYR.
The uniformity of these rust populations and species for isozyme phenotype is contrasted with their variability in pathogenicity and with the variability in isozymes encountered in higher organisms. Uniformity may result from a feature of the biology of the rust species and populations examined, or from the relative homogeneity of the environment of biotrophic pathogens. Consistent differences in isozyme and dsRNA phenotype between the WYR and BYR isolates of P. striiformis indicate that these are two discrete populations, supporting the view that they should be recognized as f.sp. tritici and f.sp. hordei , respectively.     

2010-2011年小麦条锈菌群体的温度敏感性与毒性及遗传多样性的关系

为了明确小麦条锈菌毒性、遗传多样性以及温度敏感性三者之间的关系,本研究利用21个已知抗条锈病基因近等基因品系对2010-2011年生长季采自6个省市78株已知温度敏感性(ET50)的小麦条锈菌群体进行毒性鉴定,并利用AFLP技术对其进行遗传多样性分析。苗期毒性鉴定结果表明,不同省市小麦条锈菌群体的毒性基因多样性存在一定差异,甘肃菌株群体毒性多样性指数H值最高,为0.269 3,云南菌株群体最低,为0.150 4。遗传多样性结果显示,6省市小麦条锈菌群体遗传多样性指数H值范围在0.125 5~0.165 3之间,遗传一致度GI为0.964 7~0.987 2,遗传距离GD为0.012 9~0.036 0。三者的相关性分析表明,条锈菌群体毒性多样性与平均ET50显著负相关,与ET50变异系数正相关,而与遗传多样性没有显著的相关关系。     

Genetic Diversity and the Presence of Two Distinct Groups in Ophiostoma clavigerum Associated with Dendroctonus ponderosae in British Columbia and the Northern Rocky Mountains

ABSTRACT The sapstaining fungal pathogen Ophiostoma clavigerum is associated with the mountain pine beetle (Dendroctonus ponderosae), which is currently the most destructive forest pest in North America. The genetic diversity of O. clavigerum populations collected from five sites in Canada and two sites in the United States was estimated with amplified fragment length polymorphism (AFLP) analysis. Genomic DNA from 170 O. clavigerum isolates was digested with EcoRI and PstI and amplified with six primer sets. A total of 469 AFLP markers consisting of 243 monomorphic and 226 polymorphic loci were scored. The overall genetic diversity of the O. clavigerum population was low (Hs = 0.0531) and the differentiation of the seven O. clavigerum populations was moderate (Phi = 0.143). Genetic distances among the populations were not significantly correlated with geographic distance (r = 0.3235, P = 0.074). Two genetically distinct groups in the O. clavigerum populations were shown by unique AFLP profiles and the unweighted pair group method with arithmetic averages. Further work to characterize biological differences between the two groups will be needed to confirm whether cryptic species are present in the O. clavigerum population.     

我国大麦条锈病菌的生理分化

 我国大麦条锈病主要由条形柄锈菌大麦专化型引起,病菌有明显的生理分化。对西藏大麦条锈菌鉴别寄主进行扩充改进,初步建立了一套在我国较为适用的大麦条锈菌鉴别寄主,由7个品种组成:喜马拉6号、矮秆齐、早熟3号、永1394、永802、科品2号、辉县红(小麦)。这套鉴别寄主可清楚地将大麦专化型和小麦专化型区分开;并将采自西藏、青海、甘肃、陕西和河南5省区的40个大麦专化型菌系区分为4个毒性类型。以毒性中等的大麦型1号和毒性较弱的大麦型2号分布较广,大麦型3号和大麦型4号则只见于西藏的标样中。     

Comparison of Virulence and Isozyme Phenotypes of Pgt-QCCJ and Great Plains Races of Puccinia graminis f. sp. tritici

ABSTRACT Stem rust race Pgt-QCCJ was first found in the Great Plains of the United States in 1989, collected primarily from barley. This race became a major part of the Puccinia graminis f. sp. tritici population, even though it is virulent to only a few hard red winter wheat cultivars in the central Great Plains and to barley in the northern Great Plains. It threatens barley production in the northern Great Plains of the United States and Canada due to virulence to Rpg-1. Six differences in virulence and two in isozyme banding patterns from the most similar stem rust races make it unlikely that QCCJ arose as a mutant. Thus, QCCJ likely arose through sexual or parasexual recombination. Sexual recombination in the Great Plains is unlikely, as it has not been detected in many years. Avirulence to 'McNair 70l' is only known from the Pacific Northwest of the United States and adjacent Canada. The rust population in this area is of sexual origin, and the pattern of virulence/avirulence and isozyme banding for QCCJ occurs there. Pgt-QCCJ likely originated in the Pacific Northwest during or before 1989 and was wind-transported into the Great Plains.     

Single Cystosorus Isolate Production and Restriction Fragment Length Polymorphism Characterization of the Obligate Biotroph Spongospora subterranea f. sp. subterranea

ABSTRACT Spongospora subterranea f. sp. subterranea causes powdery scab in potatoes and is distributed worldwide. Genetic studies of this pathogen have been hampered due, in part, to its obligate parasitism and the lack of molecular markers for this pathogen. In this investigation, a single cystosorus inoculation technique was developed to produce large amounts of S. subterranea f. sp. subterranea plasmodia or zoosporangia in eastern black nightshade (Solanum ptycanthum) roots from which DNA was extracted. Cryopreservation of zoosporangia was used for long-term storage of the isolates. S. subterranea f. sp. subterranea-specific restriction fragment length polymorphism (RFLP) markers were developed from randomly amplified polymorphic DNA (RAPD) fragments. Cystosori of S. subterranea f. sp. subterranea were used for RAPD assays and putative pathogen-specific RAPD fragments were cloned and sequenced. The fragments were screened for specificity by Southern hybridization and subsequent DNA sequence BLAST search. Four polymorphic S. subterranea f. sp. subterranea-specific probes containing repetitive elements, and one containing single copy DNA were identified. These RFLP probes were then used to analyze 24 single cystosorus isolates derived from eight geographic locations in the United States and Canada. Genetic variation was recorded among, but not within, geographic locations. Cluster analysis separated the isolates into two major groups: group I included isolates originating from western North America, with the exception of those from Colorado, and group II included isolates originating from eastern North America and from Colorado. The techniques developed in this study, i.e., production of single cystosorus isolates of S. subterranea f. sp. subterranea and development of RFLP markers for this pathogen, provide methods to further study the genetic structure of S. subterranea f. sp. subterranea.     

Trichothecene profiling and population genetic analysis of Gibberella zeae from barley in North Dakota and Minnesota

Gibberella zeae, the principal cause of Fusarium head blight (FHB) of barley, contaminates grains with several mycotoxins, which creates a serious problem for the malting barley industry in the United States, China, and Europe. However, limited studies have been conducted on the trichothecene profiles and population genetic structure of G. zeae isolates collected from barley in the United States. Trichothecene biosynthesis gene (TRI)-based polymerase chain reaction (PCR) assays and 10 variable number tandem repeat (VNTR) markers were used to determine the genetic diversity and compare the trichothecene profiles of an older population (n = 115 isolates) of G. zeae collected in 1997 to 2000 with a newer population (n = 147 isolates) collected in 2008. Samples were from across the major barley-growing regions in North Dakota and Minnesota. The results of TRI-based PCR assays were further validated using a subset of 32 and 28 isolates of G. zeae by sequence analysis and gas chromatography, respectively. TRI-based PCR assays revealed that all the G. zeae isolates in both populations had markers for deoxynivalenol (DON), and the frequencies of isolates with a 3-acetyldeoxynivalenol (3-ADON) marker in the newer population were ≈11-fold higher than those among isolates in the older population. G. zeae populations from barley in the Midwest of the United States showed no spatial structure, and all the isolates were solidly in clade 7 of G. zeae, which is quite different from other barley-growing areas of world, where multiple species of G. zeae are commonly found in close proximity and display spatial structure. VNTR analysis showed high gene diversity (H = 0.82 to 0.83) and genotypic diversity but low linkage disequilibrium (LD = 0.02 to 0.07) in both populations. Low genetic differentiation (F(ST) = 0.013) and high gene flow (Nm = 36.84) was observed between the two populations and among subpopulations within the same population (Nm = 12.77 to 29.97), suggesting that temporal and spatial variations had little influence on population differentiation in the Upper Midwest. Similarly, low F(ST) (0.02) was observed between 3-ADON and 15-acetyldeoxynivalenol populations, indicating minor influence of the chemotype of G. zeae isolates on population subdivision, although there was a rapid increase in the frequencies of isolates with the 3-ADON marker in the Upper Midwest between the older collection made in 1997 to 2000 and the newer collection made in 2008. This study provides information to barley-breeding programs for their selection of isolates of G. zeae for evaluating barley genotypes for resistance to FHB and DON accumulation.