Occurrence and Pathogenicity of Fusarium Species in Cereals in Finland1

Several Fusarium species occurred abundantly in spring cereal seed samples studied in 1966, 1968 and 1972 in Finland. A total of 17 species of Fusarium were isolated and identified. The most frequently isolated species were F. avenaceum (Fr.) Sacc., F. culmorum (W. G. Sm.) Sacc., F. poae (Peck) Wr. and F. tricinctum (Cda) Sacc. These species were found in more than 40% of the seed samples from 1972. The same 4 Fusarium species in addition to F. graminearum Schwabe were used in the pathogenicity test. Artificially inoculated spring wheat and barley seeds were grown in the field during the growing season of 1973. Of these 5 species F. culmorum was clearly the most pathogenic. This species significantly lowered the shooting percentage and the yield of both cereals. Foot rot assessment also revealed F. culmorum as the most damaging species. Natural soil-borne infection obviously increased the foot and root rot, and thus the soil-borne F. culmorum strongly affected all treatments. In the harvested seed F. poae dominated in all treatments, but F. culmorum still occurred most abundantly in the seed lot harvested from plots originally inoculated with this species.     

Molecular Diversity of Agriculturally Important Aspergillus Species

Although Aspergillus species are not usually considered as serious plant pathogens, Aspergilli are frequently encountered in plant products. The most important consequence of their presence is mycotoxin contamination. The main mycotoxins produced by Aspergilli are the aflatoxins, ochratoxin A and patulin, which are produced by a variety of Aspergillus species in different plant commodities. Phylogenetic analysis of sequences of the ribosomal RNA gene cluster is useful for clarifying taxonomic relationships among toxigenic Aspergilli causing pre- and postharvest contamination of agricultural products. Molecular data has enabled us to clarify the taxonomy of black Aspergilli, A. flavus and its relatives, and sections Circumdati and Clavati, which include ochratoxin and patulin-producing species. Phylogenetically unrelated species were found to produce the same mycotoxins, indicating that mycotoxin-producing abilities of the isolates have been lost (or gained) several times during the evolution of the genus. The data also indicate that biosynthetic gene-based probes are necessary for molecular detection of these mycotoxin-producing organisms. The organisation of the biosynthetic genes of patulin and ochratoxins is unknown, although experiments are in progress in several laboratories to clarify the genetic background of biosynthesis of these mycotoxins. Identification of biosynthetic genes responsible for mycotoxin production is essential for clarifying the evolution of mycotoxin biosynthesis in Aspergilli, and to develop specific gene probes for the detection of mycotoxin-producing Aspergilli in agricultural products.     

分子检测土壤中南美大豆猝死综合症病菌和北美大豆猝死综合症病菌

对南美大豆猝死综合症病菌（Fusarium tucumaniae）和北美大豆猝死综合症病菌（Fusarium virguliforme）rDNA基因间间隔区（IGS）进行分析,设计并筛选出3对特异性引物FT1/FT6、FT1/FT9和FV1/FV1A。分别利用FT1/FT6、FT1/FT9进行PCR反应,对F.tucumaniae分别扩增出250bp、656bp的特异性片段,而F.virguliforme、F.brasiliense、F.cuneirostrum和F.phaseoli等近似种均无特异性PCR产物出现。利用FV1/FV1A进行PCR反应,F.virguliforme出现228bp特异性PCR产物,而F.tucumaniae、F.brasiliense、F.cuneirostrum和F.phaseoli等近似种无特异性PCR产物。引物FT1/FT6、FV1/FV1A检测F.tucumaniae和F.virguliforme的最低DNA含量为1pg/μL,利用FT1/FT6和FT1/FT9对土壤中的病菌进行巢式PCR,能检测到接种量为每g土壤含100个大分生孢子的F.tucumaniae,FV1/FV1A能检测到接种量为每g土壤含1000个大分生孢子的F.virguliforme。     

Genetic Diversity of Fusarium Graminearum in Europe and Asia

The biodiversity and phylogeny of Fusarium graminearum isolates originating from different geographical areas was analyzed using isozyme variation, β-tubulin and IGS sequences. Geographically distinct groups of isolates originating from different areas of the Russian Federation, China, Germany and Finland were studied. The complex enzymes (α- and β-esterases, aspartate aminotransferase and superoxide dismutase) and the IGS sequences demonstrated a high level of genetic diversity in F. graminearum. Diversity in the Asian population was higher than in the European one. There was a correlation between genetic clusters of the IGS sequences and geographical origin in F. graminearum. Knowledge of biodiversity and identification of some phylogenetic lineages in F. graminearum will be useful in defining the risk of pathogen evolution as well as benefiting disease management strategies.     

Isozyme and RAPD-PCR analyses of Fusarium avenaceum strains from Finland

Differences in isozyme and RAPD-PCR polymorphisms amongst 33 isolates of Fusarium avenaceum were compared using native polyacrylamide gel electrophoresis and agarose gel electrophoresis. The isolates were collected from different regions of Finland. Amongst eight enzymes analysed clear isozyme polymorphism was detected in five enzymes which could be grouped into 20 different electrophoretic phenotypes and three main groups at the similarity level of 70% in unweighted pair group method with arithmetic average (UPGMA) analysis. RAPD-PCR analysis differentiated all F. avenaceum strains from each other. The phenotypes resulting from RAPD-PCR analysis were grouped into five main groups by UPGMA analysis at the similarity level of 55%. These main groups had several similarities with the main groups from isozyme analysis. RAPD-PCR patterns of 16 isolates of Fusarium graminearum F. culmorum F. equiseti F. oxysporum and F. redolens were also studied and strains from each Fusarium species formed individual groups in UPGMA and principal components analyses. Thus, the extent of isozyme and RAPD-PCR polymorphisms found in Fusarium strains potentially provides a method for identifying the fungi both at strain and species level.     

New Leaf Spot Disease of Cymbidium Species Caused by Fusarium subglutinans and Fusarium proliferatum

Fusarium species were consistently isolated from yellow, swollen spots with reddishbrown centers and small black spots on leaves of Cymbidium plants in the greenhouse. Fusarium subglutinans caused the yellow spots and Fusarium proliferatum caused either the yellow or the black spots. We propose the name “yellow spot” for the new disease. To denote differences in their pathogenicity to orchid plants, we designate the population causing yellow spot as race Y and that causing black spot as race B of F. proliferatum. Received 29 October 1999/ Accepted in revised form 10 March 2000     

Quantitative Detection of Fusarium Species in Wheat Using TaqMan

Fusarium head blight (FHB) of wheat and other small-grain cereals is a disease complex caused by several fungal species. To monitor and quantify the major species in the FHB complex during the growing season, real-time PCR was developed. TaqMan primers and probes were designed that showed high specificity for Fusarium avenaceum, F. culmorum, F. graminearum, F. poae and Microdochium nivale var. majus. Inclusion of an internal PCR control and serial dilutions of pure genomic DNAs allowed accurate determination of the concentration of fungal DNA for each of these species in leaves, ears as well as harvested grains of winter wheat. The DNA concentration of F. graminearum in grain samples correlated (r ²= 0.7917) with the incidence of this species on the grain as determined by isolation from individual kernels. Application of the TaqMan technology to field samples collected in 40 wheat crops in the Netherlands during the growing season of 2001 revealed that M. nivale var. majus predominated on leaves early in the season (GS 45-65). Ears and harvested grains from the same fields, however, showed F. graminearum as the major species. In 2002, grain samples from 40 Dutch fields showed a much wider range of species, whereas in ears from 29 wheat crops in France, F. graminearum was the predominant species. The concentration of DON correlated equally well with the incidence of the DON-producing species F. culmorum and F. graminearum in the grain samples (r ²= 0.8232) as well as with total DNA of both these species (r ²= 0.8259). The Fusarium TaqMan technology is an important tool to quantify and monitor the dynamics of individual species of the complex causing FHB in cereals during the growing season. This versatile tool has been applied in a comparison of different genotypes, but can also be applied to other disease management systems, e.g. fungicide treatments.     

Molecular Characterization of Fusarium oxysporum and Fusarium commune Isolates from a Conifer Nursery

ABSTRACT Fusarium species can cause severe root disease and damping-off in conifer nurseries. Fusarium inoculum is commonly found in most container and bareroot nurseries on healthy and diseased seedlings, in nursery soils, and on conifer seeds. Isolates of Fusarium spp. can differ in virulence; however, virulence and colony morphology are not correlated. Forty-one isolates of Fusarium spp., morphologically indistinguishable from F. oxysporum, were collected from nursery samples (soils, healthy seedlings, and diseased seedlings). These isolates were characterized by amplified fragment length polymorphism (AFLP) and DNA sequencing of nuclear rDNA (internal transcribed spacer including 5.8S rDNA), mitochon-drial rDNA (small subunit [mtSSU]), and nuclear translation elongation factor 1-alpha. Each isolate had a unique AFLP phenotype. Out of 121 loci, 111 (92%) were polymorphic; 30 alleles were unique to only highly virulent isolates and 33 alleles were unique to only isolates nonpathogenic on conifers. Maximum parsimony and Bayesian analyses of DNA sequences from all three regions and the combined data set showed that all highly virulent isolates clearly separated into a common clade that contained F. commune, which was recently distinguished from its sister taxon, F. oxysporum. Interestingly, all but one of the nonpathogenic isolates grouped into a common clade and were genetically similar to F. oxysporum. The AFLP cladograms had similar topologies when compared with the DNA-based phylograms. Although all tested isolates were morphologically indistinguishable from F. oxysporum based on currently available monographs, some morphological traits can be plastic and unreliable for identification of Fusarium spp. We consider the highly virulent isolates to be F. commune based on strong genetic evidence. To our knowledge, this is the first reported evidence that shows F. commune is a cause of Fusarium disease (root rot and dampingoff) on Douglas-fir seedlings. Furthermore, several AFLP genetic markers and mtSSU sequences offer potential for development of molecular markers that could be used to detect and distinguish isolates of F. oxysporum nonpathogenic to conifers and highly virulent isolates of F. commune in forest nurseries.     

Virulence and Molecular Diversity in Cochliobolus sativus

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.     

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.     

物种多样性与病虫草害管理研究进展

农业生物多样性有许多重要的生态功能,控制农业有害生物的发生和流行是其中重要组成部分。生物多样性包括形态的多样性、功能多样性及互作的多样性,互作物质的多样性和互作方式的多样性决定了互作结果的多样性。充分利用品种之间,作物之间、作物与非作物之间的正向作用和负向作用来促进植物生长,减少病虫害的发生,是有效控制农林有害生物的重要措施。本文介绍该领域近年来的物种多样性研究的主要进展,同时提出以后研究的建议和方向,即应用宏基因组学、蛋白组学和代谢组学等后基因组学及植物生理生化等综合技术,系统分析农田生态系统中各种生物之间直接和间接互作机制的基础,根据不同的防控对象及环境条件确定多样性的结构和模式,通过大数据进行管控,才能实现病虫害的绿色防控和农业可持续发展。     

Effect of Trichothecenes Produced by Fusarium graminearum during Fusarium Head Blight Development in Six Cereal Species

Fusarium head blight (FHB) is a complex cereal disease associated with trichothecene production; these mycotoxins are factors of aggressiveness in wheat. Six species (bread and durum wheat, triticale, rye, barley and oats) were submitted to point inoculations with two isogenic strains of Fusarium graminearum; a wild strain (Tri5 +) produced trichothecenes and the mutated strain (Tri5 –) did not. The trichothecene-producing strain was generally more aggressive than the non-producing strain, but this varied according to crop species. The difference in aggressiveness was less pronounced in rye, a very resistant species. High resistance levels were observed in oats due to the large spacing between florets. In six-row barley, despite the existence of a moderate Type II resistance, the fungus was often observed to move externally from one floret to another within the dense spike, without penetrating the rachis. Bread wheat had low resistance to the trichothecene-producing strain and good resistance to the non-producing strain. Triticale responded to the strains in a similar way but was somewhat more resistant to both: symptoms on the spikelets and rachis of the triticales were restricted to below the point of inoculation. Durum wheat was susceptible to the trichothecene-producing strain and only moderately resistant to the non-producing strain, which was able to cause serious damage only to this species. Our study confirmed that the role of trichothecenes in FHB pathogenesis differs among species. The failure of the trichothecene non-producing F. graminearum strain to spread within the inflorescence of wheat, triticale, rye and barley, and the significant reduction of spread in the durum wheat spike strongly suggested that trichothecenes are a major determinant of fungal spread and disease development in Triticeae.     

Real-time PCR for Quantification of Toxigenic Fusarium Species in Barley and Malt

A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler® system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4–5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys.     

新疆棘豆属物种多样性分析

新疆棘豆属植物有98种2变种,占中国棘豆属总种数的67.1%,其中新疆特有种7种,中国新疆仅分布种58种,特有种与新疆仅分布种共占65%,所占比例较高.棘豆属在新疆的11个植被区域中均有分布,最丰富的地区为准噶尔和天山地区,尤以天山山系种类最多,它们是棘豆属现代分布与起源分化中心之一;绝大部分种类(约99%)集中分布在海拔1 001～5 000 m的山区,尤其在1 001～4 000 m最丰富,在2 001～3 000 m分布最多;植物生活型以地面芽植物为主,高达96%,地上芽植物仅占4%.植物区系与其邻近的青海省和西藏关系最为密切,相似性系数分别为28.8%和28.6%,与内蒙古和甘肃联系相对较弱;与邻国哈萨克斯坦关系最密切,相似性系数高达65.2%,其次是西西伯利亚和吉尔吉斯斯坦,相似性系数分别为41.8%和41.0%,与蒙古、塔吉克斯坦和巴基斯坦的相似性系数较低,分别为29.2%,22.8%和11.6%,联系相对较弱.     

中国花蝽科昆虫多样性及分布格局

在对中国花蝽科昆虫多样性分布统计的基础上,构建数据库,采用特有性简约分析方法,对中国花蝽科昆虫的地理分布格局和物种多样性进行探讨。结果表明,风蝶在中国8个生物地理大区均有分布,且表现出南部比北部丰富、北部比南部原始的特征,华南区和西南区的凤蝶多样性最为丰富。     

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.     

安徽大别山区绿僵菌属物种多样性

绿僵菌是一类常见且极具应用价值的生物防治材料。目前绿僵菌分类系统取得了突破性的进展,然而新分类系统在国内尚未普遍采用。本文用该系统研究了近年来分离自大别山区僵虫的21株绿僵菌的物种多样性。在对大别山绿僵菌菌株的形态学研究基础上;对其转录延长因子（EF-1a）进行了进一步的PCR扩增、测序和同源性分析,并选用MEGA软件依据简约法构建EF-1a系统发育树。结果表明,21株菌株在新系统中分别属于5个种,即贵州绿僵菌、大孢绿僵菌、蚱蜢绿僵菌、黄绿绿僵菌瘿绵蚜变种和棕色绿僵菌,并对中国新纪录棕色绿僵菌进行了形态学的描述。该研究基本明确了大别山区的绿僵菌物种多样性,为生物防治菌株提供了科学鉴定方法和丰富种质资源。     

Toxigenic Fusarium Species and Mycotoxins Associated with Maize Ear Rot in Europe

Several Fusarium species occurring worldwide on maize as causal agents of ear rot, are capable of producing mycotoxins in infected kernels, some of which have a notable impact on human and animal health. The main groups of Fusarium toxins commonly found are: trichothecenes, zearalenones, fumonisins, and moniliformin. In addition, beauvericin and fusaproliferin have been found in Fusarium-infected maize ears. Zearalenone and deoxynivalenol are commonly found in maize red ear rot, which is essentially caused by species of the Discolour section, particularly F. graminearum. Moreover, nivalenol and fusarenone-X were often found associated with the occasional occurrence of F. cerealis, and diacetoxyscirpenol and T-2 toxin with the occurrence of F. poae and F. sporotrichioides, respectively. In addition, the occurrence of F. avenaceum and F. subglutinans usually led to the accumulation of moniliformin. In maize pink ear rot, which is mainly caused by F. verticillioides, there is increasing evidence of the wide occurrence of fumonisin B₁. This carcinogenic toxin is usually found in association with moniliformin, beauvericin, and fusaproliferin, both in central Europe due to the co-occurrence of F. subglutinans, and in southern Europe where the spread of F. verticillioides is reinforced by the widespread presence of F. proliferatum capable of producing fumonisin B₁, moniliformin, beauvericin, and fusaproliferin.