Genetic Variability and Structure of Canadian Populations of the Sapstain Fungus Ceratocystis resinifera

ABSTRACT Genomic DNA was extracted from 129 isolates of Ceratocystis resinifera, a species belonging to the C. coerulescens complex, and 19 polymorphic random amplified polymorphic DNA markers were used to study the population genetic structure of this fungus. The analysis suggested a moderate value for genetic diversity (H(S) = 0.209). However, when monomorphic markers and rare alleles, representing 89 markers, also were included in the calculation, the genetic diversity of Canadian populations of C. resinifera appeared to be much lower (H(S) = 0.045). This could be explained by two hypotheses: (i) recent introduction of this species into North America and (ii) clonal reproduction (by selfing). No specialization by C. resinifera for coniferous tree species was observed based on genetic differentiation index between isolates sampled from Pinus and Picea spp. and on phylogenetic analysis using Dice coefficient of association. In spite of a low genetic diversity, a very high genetic differentiation was observed among the nine geographical populations studied (F(ST) = 20.8%). The genetic differences were especially striking when populations from Eastern Canada were compared with populations from Western Canada (phiST = 0.27%; P < 0.001), suggesting that a geographic reproductive barrier occurs in Central Canada. This barrier may be the consequence of a weak migration of insect vectors of C. resinifera due to reduced presence of hosts in the Canadian Great Plains, where extensive agriculture occurs. However, results from pairwise F(ST) matrix and phylogeny of haplotypes suggest that the barrier is not totally impenetrable because some gene flow occurred from the west and from the east in the Big River (Saskatchewan) population located in the middle of the Great Plains.     

内蒙古地区天然臭柏种群遗传多样性的RAPD分析

应用随机扩增多态性DNA(RAPD)标记法研究了内蒙古地区臭柏种群的遗传分化,所用的18条引物对4个臭柏种群的55个样本扩增出124个位点,其中多态位点100个,多态率达80.7%。种内的平均Nei's多样性指数和Shannon's多样性指数分别为0.286和0.428,种群内分别为0.232和0.347,基因多样性变化趋势为毛乌素沙地(0.258,0.386)>阴山山脉西部(0.258,0.376)>浑善达克沙地(0.231,0.345)>内蒙古贺兰山(0.184,0.281)。臭柏种群总的基因多样性(Ht=0.288),大于种群间的基因多样性(Hs=0.233)。种群间的遗传分化系数(Gst)为0.183,种群内的遗传变异占总遗传变异的81.7%。臭柏种群间的平均遗传距离为0.086,平均相似性系数为0.918,4个种群之间有比较相似的遗传多样性,由于地理隔离,种群间也存在分化。     

Association of Fusarium Species in the Wheat Stem Rot Complex

Data from a national survey were analysed to investigate whether there was interdependence among the Fusarium species, which cause the stem rot complex of wheat. About 25 wheat stems were sampled from each of 260 sites over the main wheat growing areas in the UK. Occurrence of each Fusarium species on individual stems was determined. Fusarium culmorum, F. avenaceum and Microdochium nivale were the three dominant species, detected in 248, 185 and 239 out of the 260 sites. There were no interactions among species in the distribution of the three species over the 260 sites. Several statistical tests were used to determine whether there was interdependence among the three species on the same stem within each site. Of the three species, there was only limited evidence of competition between F. culmorum and F. avenaceum.     

Genetic Variation and Population Structure of the Grape Powdery Mildew Fungus, <Emphasis Type="Italic">Erysiphe necator</Emphasis>, in Southern France

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.     

Genetic variability among isolates of Fusarium oxysporum from sugar beet

Fusarium yellows, caused by the soil‐borne fungus Fusarium oxysporum f. sp. betae (Fob), can lead to significant yield losses in sugar beet. This fungus is variable in pathogenicity, morphology, host range and symptom production, and is not a well characterized pathogen on sugar beet. From 1998 to 2003, 86 isolates of F. oxysporum and 20 other Fusarium species from sugar beet, along with four F. oxysporum isolates from dry bean and five from spinach, were obtained from diseased plants and characterized for pathogenicity to sugar beet. A group of sugar beet Fusarium isolates from different geographic areas (including nonpathogenic and pathogenic F. oxysporum, F. solani, F. proliferatum and F. avenaceum), F. oxysporum from dry bean and spinach, and Fusarium DNA from Europe were chosen for phylogenetic analysis. Sequence data from β‐ tubulin, EF1α and ITS DNA were used to examine whether Fusarium diversity is related to geographic origin and pathogenicity. Parsimony and Bayesian MCMC analyses of individual and combined datasets revealed no clades based on geographic origin and a single clade consisting exclusively of pathogens. The presence of FOB and nonpathogenic isolates in clades predominately made up of Fusarium species from sugar beet and other hosts indicates that F. oxysporum f. sp. betae is not monophyletic.     

浑善达克沙地榆遗传多样性的研究

浑善达克沙地榆 ( Ulmuspumila var. sabulosa)是白榆的一个变种 ,天然分布于浑善达克沙地 ,在维护草原生态系统的稳定方面起着重要作用。本文应用 RAPD技术对浑善达克沙地榆的遗传多样性和系统地位进行了研究。在浑善达克沙地榆六个居群和对照白榆两个居群中 ,1 6个有效引物共获得了 1 1 5个位点 ,其中居群水平多态性位点为 49个 ,占 42 .6 ,高于 H amrick等的统计得出的平均多态位点比率 ( 3 4 ) ,表明在长期的进化过程中 ,居群内发生了较高的遗传变异 ,个体水平多态性位点为 89个 ,占 77.4。根据遗传距离进行聚类得出 :居群水平上以 0 .1为阈值 ,浑善达克沙地榆和对照白榆聚为明显的两支 ,本实验结果支持浑善达克沙地榆作为白榆一个变种的结论 ;遗传多样性指数分析表明 ,个体水平上浑善达克沙地榆各居群之间遗传距离有一定的差异 ,并与地理分布范围相关。浑善达克沙地榆由于各居群都存在相对较大的遗传变异 ,反映了在居群内具有较大的变异潜力。本实验结果也表明 ,RAPD可应用于浑善达克沙地榆遗传多样性及系统学的研究。文中还对浑善达克沙地榆的遗传变异与繁育系统的关系进行了讨论。     

不同干扰生境中荒漠小灌木红砂种群遗传多样性研究

应用RAPD标记技术对荒漠小灌木红砂(Reaumuria soongorica)种群在不同扰动下的遗传多样性进行了分析。18条随机引物对红砂6个种群的120个个体进行扩增,共检测102个位点,其中多态位点99个。研究结果表明:红砂种群的多态位点比率(P)为96.86%,显示了不同生境中红砂种群内存在较高的遗传多样性。Shannon多样性指数(0.5007)、Nei基因多样性指数(0.3307)和基因分化系数(Gst=0.1952)揭示了红砂种群遗传变异多存在于种群内,种群间的遗传分化则较小。聚类分析表明:红砂种群遗传距离与地理距离之间存在一定相关性;遗传多样性水平与物种特性和不同干扰生境有关,与生态因子无相关性。     

Genetic diversity assessed by SSR markers and chemotyping of Fusarium culmorum causal agent of foot and root rot of wheat collected from two different fields in Tunisia

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.     

The Relationship Between Wheat Seed Weight, Infection by Fusarium culmorum or Microdochium nivale, Germination and Seedling Disease

The distribution of seeds by weight for three lots of winter wheat cv. Avalon infected by Fusarium culmorum and three lots of winter wheat cv. Riband infected by Microdochium nivale was determined. The distribution of infected seeds within each seed lot was then determined by isolating F. culmorum from seeds on moist filter paper and M. nivale from seeds on potato dextrose agar. The distribution of M. nivale infected seeds between seeds of different weight was similar to that of the seed lot as a whole, whereas the distribution of F. culmorum was greater in light seeds than heavy seeds. The percentage germination of infected seeds decreased with seed weight. A similar situation was found with respect to seedling emergence in compost for F. culmorum infected seeds. However, with M. nivale infection, similar numbers of seedlings emerged from both light and heavy infected seeds. Seed treatment with guazatine increased seedling emergence for both light and heavy seed infected by M. nivale. However, seedling emergence from F. culmorum infected seed was poor even following treatment with guazatine. Poor emergence was most evident from light seed.     

Variation in Pathogenicity Associated with the Genetic Diversity of Fusarium graminearum

We screened 188 isolates of Fusarium graminearum, which originated from northwest Europe, the USA and Nepal, for genetic diversity using a sequence-characterised amplified region polymorphism (SCAR). On the basis of this analysis, 42 of the 118 isolates were selected for random amplified polymorphic DNA (RAPD) analysis. Three groups were identified, two of which, A and B, contained the isolates from Nepal, and a third, group C, contained the isolates from Europe and the USA. In pathogenicity tests on wheat and maize seedlings, group C isolates were more pathogenic than the group A and B isolates. The isolates were assigned chemotypes based on their ability to produce the trichothecene mycotoxins nivalenol (NIV) and deoxynivalenol (DON). Isolates from group A were equally likely to produce NIV or DON while group B isolates produced predominantly NIV, and group C isolates produced predominantly DON. Within group A, isolates of the two chemotypes were equally pathogenic to wheat but isolates with the NIV chemotype were significantly more pathogenic to maize. The results confirm that distinct genetic groups exist within F. graminearum and demonstrate that these groups have different biological properties, especially with respect to their pathogenicity to two of the most economically important hosts of this pathogen.     

Genetic variation and phylogenetic relationships of triazine-resistant and triazine-susceptible biotypes of Solanum nigrum– analysis using RAPD markers

The expansion of weed species is a major problem in agriculture, especially when the number of herbicide-resistant biotypes is rising continuously. The major ecological questions associated with the evolution of herbicide resistance involve an intricate understanding of the interplay between gene frequency, fitness, inheritance and gene flow. In this study, the RAPD (Random Amplified Polymorphic DNA) technique, which facilitates detection of variability at DNA level, was used to examine the spread of Solanum nigrum L. populations. Twenty-five populations, from Poland, France and the UK, were analysed. Six populations from Poland and one from France showed target site-based triazine resistance. The genetic relationship between individuals was studied using the RAPD technique. It was found that some resistant populations from the Gabin and Grojec areas show very high affinity levels compared with individuals from France. Three groups of populations in which resistance had developed independently were distinguished. The results of the present investigation suggest that migratory birds, such as Turdus pilaris L. and Sturnus vulgaris L., play an important in spreading S. nigrum seeds.     

Competition Effects Among Isolates of Fusarium culmorum Differing in Aggressiveness and Mycotoxin Production on Heads of Winter Rye

Fusarium culmorum is a phytopathogenic, toxigenic fungus causing seedling diseases, foot rot and head blight of cereals. For estimating competition effects in mixtures of two single-spore isolates, two winter rye single crosses were tested with either four isolates individually or four 1 : 1 mixtures of the same isolates in six field environments. Two isolates (FC46, FC64) were highly aggressive deoxynivalenol (DON) and 3-acetyl DON-producers, the other two (FC30, FC71) were medium aggressive nivalenol-producers. Rye heads were inoculated during flowering with conidia of pairs of isolates expressing similar (FC46 + FC64, FC30 + FC71) or contrary (FC46 + FC71, FC30 + FC64) levels of aggressiveness and similar or different concentrations and chemotypes of mycotoxins, respectively. Head blight rating and yield components relative to the non-inoculated plots were recorded as aggressiveness traits. Additionally, mycotoxin concentrations were measured in the rye grain. Random pathogen samples were re-isolated from heads at the onset of symptom development and analysed by molecular markers (RAPD–PCR) in one environment. Aggressiveness of the isolate mixtures was significantly lower than that of the isolates applied individually on both rye genotypes. Similarly, mycotoxin concentrations were significantly lower in the mixtures in seven out of eleven comparisons. Among the re-isolates, the component genotypes of a mixture significantly deviated from the inoculated 1 : 1 ratio when a particular isolate (FC46) was present in the mixture. This isolate displayed a superior competitive ability irrespective of the aggressiveness or mycotoxin profile of the mixing partner illustrating that pathogenic fitness is caused by additional factors that have not, as yet, been identified.     

Genetic structure of Fusarium oxysporum f. sp. cubense in different regions from Brazil

Panama disease, caused by Fusarium oxysporum f. sp. cubense (Foc), is ranked among the most destructive diseases of banana. The use of resistant varieties is the most desirable and effective control measure. Information on the pathogen population structure is essential, as durability of the resistance and effective cultivar deployment are strongly linked to this structure. In this study, 214 Foc isolates from different banana producing states in three regions of Brazil (northeastern, southeastern and southern) were analysed. Initially, nine microsatellite markers (SSR) were tested, which revealed 52 distinct haplotypes distributed in the different geographical regions and cultivars. While amova analysis showed that 68·01% of the total variation occurred within states, correlation between genetic and geographical distances was only found in the southern region. Results indicated that isolates from different states comprise a single population, which is predominantly clonal. When isolates representing different haplotypes were inoculated in four banana cultivars, differences in severity were found, with the high severity values being caused by isolates from haplotypes H7, H31 and H41. The diversity found here points to the need for additional studies, as this characteristic may be related to Foc's evolutionary potential and possibly to its ability to overcome the resistance from breeding programme‐generated cultivars. This is the most comprehensive study on population biology of Foc in Brazil.     

Genetic Structure of Mycosphaerella graminicola Populations from Iran, Argentina and Australia

Restriction fragment length polymorphism (RFLP) markers were used to assess the genetic structure of populations of Mycosphaerella graminicola collected from wheat fields. A total of 585 isolates representing 10 field populations were sampled from Iran, Argentina and Australia. The genetic structure of M. graminicola populations from Iran and Argentina is described for the first time. Results were compared to previously investigated populations from Israel, Uruguay and Australia. Populations from Iran exhibited high clonality and low gene diversity, suggesting an inoculation event. Populations from uninoculated fields in Argentina had gene and genotype diversities similar to previously described European and North American populations. Genotype diversity was high for populations from Australia and tests for multilocus associations were consistent with sexual recombination in these populations. Gene diversity was low and fixed alleles were found for several loci. These findings are consistent with a relatively small founding population for Australia. These 10 new populations were integrated into a genetic distance comparison with 13 global populations that were characterized earlier.     

Population Dynamics of Fusarium Oxysporum f. Sp. Radicis-lycopersici in Relation to the Onset of Fusarium Crown and Root Rot of Tomato

Fusarium oxysporum f. sp. radicis-lycopersici the causal agent of crown and root rot in tomato comprises two overlapping separate phases: monocyclic and polycyclic. Oversummering inoculum is the source of primary infection (the monocyclic phase) and the spread from plant to plant via root-to-root contact is the source of the secondary infection (the polycyclic phase). In the present work, relationships between initial inoculum density, population dynamics of the pathogen in the root zone of diseased plants, and disease onset were studied. For the monocyclic phase, 55.1% of the variance of disease onset was attributed to the rate of pathogen proliferation in the root zone of plants, and only 12.8% of the variance was attributed to the amount of initial inoculum density. For the polycyclic phase, disease onset was not related to either initial inoculum density or the rate of pathogen proliferation in the root zone. At disease onset, the inoculum density of the pathogen in the root zone of plants infected from oversummering inoculum reached an average of 4.08 log cfu g soil^–1. The inoculum density of the pathogen in the root zone of plants infected by their diseased neighbors was 3.23 log cfu g soil^–1. A large variation in pathogen proliferation rate in the root zone was found among individual plants, suggesting that differences in the level of soil suppressiveness may occur not only between fields, but even in the same field over short distances.     

产纳他霉素生防菌A01和A02与已知产生菌的RAPD比较

为了明确自主分离的天然抗真菌活性产物——纳他霉素产生菌A01和A02与已知纳他霉素产生菌的遗传相似性,采用RAPD技术对这2株菌和3个产纳他霉素的标准菌株进行了比较分析。利用筛选出的7个随机引物对5个菌株的PCR扩增共得到154条清晰稳定的DNA条带,其中同源性条带86条,多态性条带68条,分别占总条带数的55.8%和44.2%,平均每个引物扩增出9.7个多态性条带,得到了丰富的DNA指纹图谱。所得数据经NTSYS pc 2.10e软件聚类分析,表明5个菌株间的遗传距离为0.0991~0.4738,其中A02与其它菌株间的遗传距离均较远。结合前期的分类研究结果,确证了菌株A02为新的纳他霉素产生菌。     

死亡植株对新疆野苹果种群遗传多样性的影响

新疆野苹果（Malus sieversii）为国家二级保护植物和重要的野生果树种质资源。选取遭受小吉丁虫等病虫害破坏最为严重的新源县种群为对象,利用微卫星（SSR）标记死亡植株和存活植株的基因型,对比两者间的遗传差异,探究树木死亡对新疆野苹果种群遗传多样性的影响。结果表明：（1）死亡植株与存活植株间没有显著的遗传分化,这说明新疆野苹果植株死亡不存在遗传相关性,为随机死亡过程;（2）树木死亡对新源县种群的遗传结构未产生重大影响,遗传多样性水平仅轻微减小,但丢失了许多低频度等位基因;（3）未来的监测和评价中,等位基因数（Na）比遗传多样性指数能更有效地反映新疆野苹果遗传资源变化。     

Parasitism of macroconidia, chlamydospores and hyphae of Fusarium culmorum by mycoparasitic Pythium species

Parasitism of macroconidia and endoconidial chlamydospores of Fusarium culmorum by Pythium oligandrum was studied on water agar (WA), corn-meal agar (CMA) and glass slides. Loss of cytoplasmic content in F. culmorum spores was followed by complete degradation, and P. oligandrum produced an abundance of oogonia on the parasitized macroconidia. A simple method for assessing the relative aggressiveness of isolates is presented, based on the percentage of macroconidial cells devoid of cytoplasm. Parasitism of macroconidia by P. acanthophoron , P. oligandrum and P. periplocum , but not by the plant pathogenic species, P. tracheiphilum , was demonstrated by this method. Interactions between hyphae of P. oligandrum and F. culmorum on WA resulted in an increase in the number of oogonia of P. oligandrum and a decrease in the sporulation of F. culmorum . The ability of isolates of P. oligandrum , P. periplocum , P. acanthophoron and P. mycoparasiticum to suppress disease symptoms caused by F. culmorum on barley seedlings was demonstrated in a greenhouse test.     

玫烟色棒束孢的遗传变异性及其芽孢子生产

本文评述了近年来在杀虫真菌玫烟色棒束孢Isaria fumosorosea Wize研究中的一个热点问题:遗传变异性与菌株的毒力、产品质量监控以及田间应用效果等方面的密切关系.同时介绍了一种有实用性的现场或家庭作坊式生产芽孢子的方法和相关制剂载体的研究.     

Evaluation and characterization of resistance to fusarium head blight caused by Fusarium culmorum in UK winter wheat cultivars

Fusarium head blight (FHB) resistance of 50 cultivars from the National List of winter wheat cultivars approved for sale (or were undergoing trails for approval in 2003) in the UK was compared with 21 reference cultivars from continental Europe which had previously been characterized for resistance. Only three UK National List cultivars (Soissons, Spark and Vector) had stable resistance over trial sites that was significantly greater than that of the FHB susceptible cultivar Wizard. In addition, under moderate disease pressure, 21 of the National List cultivars had levels of the trichothecene mycotoxin deoxynivalenol (DON) above the proposed European Union limit of 1·25 ppm in grain. Surveys show that levels of FHB and DON in the UK crop are currently very low, however, should disease pressure increase for any reason, then an improvement in the overall levels of FHB resistance of UK winter wheat germplasm will be required. In order to infer the origin of resistance and to identify potentially novel resistance, allele sizes of microsatellite (simple sequence repeat, SSR) markers linked to quantitative trait loci (QTL) for FHB resistance were compared between the test cultivars and known, characterized resistance sources. The major FHB resistance QTL from the Chinese cv. Sumai-3 (3BS, 5A and 6B), the Romanian cv. Fundulea F201R (1B and 5A) and the French cv. Renan (5AL) were screened with 17 SSRs. No National List cultivar had haplotypes similar to any of these QTL. However, the highly resistant German reference cultivar Petrus had an identical haplotype to cv. Fundulea F201R on 1B indicating that this cultivar has an allelic FHB resistance QTL at that location.