我国部分地区蔬菜镰孢菌的分离及鉴定

对采自我国6省18市（县）的蔬菜根茎样本进行了组织分离和形态学鉴定,共鉴定出镰孢菌10个种.其中尖镰孢（Fusarium oxysporum）为优势茵,占镰孢菌总量的73.42％,其他种类的镰孢菌为锐顶镰孢(F.acuminatum)、弯角镰孢（F.camptoceras）、蓝色镰孢（F.coeruleunm）、木贼镰...     

湖北省蔬菜根际土壤中的镰孢菌分布特征

为明确湖北省蔬菜产区镰孢菌(Fusarium spp.)的种类及分布特征,本研究从湖北省13个地级市蔬菜产区采集根际土壤样品55份,对土样中的镰孢菌进行分离、纯化和鉴定,获得271株镰孢菌;通过镰孢菌特异性引物扩增、测序等分子生物学技术,鉴定出尖孢镰孢菌(F. oxysporum)118株,占分离菌株43.6%;木贼孢菌(F. equiseti)46株,占17.0%;共享镰孢菌(F.commune)45株,占16.6%;茄病镰孢菌(F. solani)31株,占11.4%;藤仓镰孢菌(F. fujikuroi)11株,占4.1%;镰孢霉菌(F.delphinoides)6株,占2.2%;轮纹镰孢菌(F. concentricum)5株,占1.8%;三线镰孢菌(F. tricinctum)3株,占1.1%;镰孢菌属(Fusarium spp.)6株,占2.2%。通过与致病菌尖孢镰孢菌番茄专化型(F. oxysporum f. sp. lycopersici)菌株进行对峙培养,筛选出拮抗镰孢菌16株,包括尖孢镰孢菌(F. oxysporum)4株,茄病镰孢菌(F. solani)4株,轮纹镰孢菌(F. concentricum)3株,三线镰孢菌(F.tricinctum)3株,木贼孢菌(F. equiseti)和藤仓镰孢菌(F. fujikuroi)各1株。本研究明确了湖北省蔬菜产区的镰孢菌属真菌的种类和多样性特征,为蔬菜镰孢菌病害的防治提供科学依据。     

我国东北地区玉米穗腐镰孢菌的种类及其分离频率

从辽宁、吉林、黑龙江三省采集的43份玉米病穗上共分离获得327株镰孢菌,根据形态特征鉴定属于7个种,其分离频率分别为半裸镰孢菌(Fusarium semitecturrt)37.77%,尖孢镰孢菌(F.oxysporum)3.51%,拟轮枝镰孢菌(F.verticillioides)3.57%,克鲁克威尔镰孢菌(F.c...     

甘肃大葱贮藏期镰孢菌腐烂病病原鉴定

【目的】大葱在贮藏期频繁发生镰孢菌腐烂病,损失严重。明确该病害病原种类对病害防治具有重要意义。【方法】利用组织分离法对采集自甘肃省兰州市(区)蔬菜市场的16份大葱贮藏期镰孢菌腐烂病病样进行病原物的分离、纯化培养,经单孢分离后根据形态学特征,再结合r DNA-ITS、EF-1a(tef)基因序列分析的方法进行鉴定。【结果】共分离得到80株镰孢菌,经鉴定分属3个种,即层出镰孢菌(Fusarium proliferatum)、尖孢镰孢菌(F.oxysporum)和燕麦镰孢菌(F.avenaceum),其中层出镰孢菌为大葱镰孢菌腐烂病的优势致病菌,分离频率为52.50%。对兰州白葱不同部位进行致病性测定,结果表明层出镰孢菌对大葱鳞茎的致病力最强,而燕麦镰孢菌对大葱鳞茎的致病力最弱。【结论】3种镰孢菌作为该病害的病原,属国内首次报道。     

乌头根腐病病原菌的分离与鉴定

【背景】四川江油地区栽培乌头极易发生根腐病,植株染病后枯萎死亡,最终影响药材产量。因发病因素和致病菌多样,乌头根腐病的防治向来困难。【目的】鉴定四川江油地区栽培乌头根腐病病原菌,为病害诊断和防治提供理论基础。【方法】采用组织分离法进行病原菌分离;按照Koch’s法则对分离的病菌进行致病性验证;通过形态学观察和真菌r DNA-ITS序列分析,鉴定病原菌种类。【结果】从具有根腐病症状的乌头根部分离得到47株真菌,其中菌株GF3-3、GF3-6和GF6-1是乌头根腐病的致病菌;通过形态特征结合rDNA-ITS序列分析将菌株GF3-3、GF3-6和GF6-1分别鉴定为Fusariumsolani、F.oxysporum和F.proliferatum。【结论】茄腐镰刀菌(F.solani)、尖孢镰刀菌(F. oxysporum)和层出镰刀菌(F. proliferatum)是乌头根腐病的主要病原菌,其中层出镰刀菌引起乌头患根腐病为首次报道。     

河北省苹果园根际土壤中疑似致病镰孢菌种类

为了解引起河北省苹果再植病害的病原菌,在河北省10个地区苹果园中采集土壤样品,在实验室进行病原菌的诱集分离培养,根据形态和分子特征对主要病原菌进行种类鉴定。结果表明,在分离得到的293株真菌中,有116株镰孢菌,为分离频率最高的真菌。在形态学鉴定的基础上,对供试镰孢菌进行了分子鉴定。在基于核糖体基因内转录间隔区（rDNA-ITS）序列与翻译延长因子1α（EF-1α）序列片段构建的系统发育树中,代表菌株分别与GenBank登记的所属菌株聚于同一群。研究结果明确了河北省苹果再植病害的疑似致病镰孢菌,包括：尖孢镰孢Fusarium oxysporum、木贼镰孢F. equiseti、锐顶镰孢F. acuminatum、层出镰孢F. proliferatum和茄腐镰孢F. solani。     

香蕉上的镰孢菌种类及其系统发育关系(英文)

镰孢菌属真菌是香蕉上的重要病原菌,主要引起香蕉枯萎病以及香蕉冠腐病,在我国已明确引起香蕉枯萎病的病原为尖孢镰孢古巴专化型 Fusarium oxysporum f. sp. cubense(FOC)1号和4号生理小种,但是引起香蕉冠腐病的镰孢菌种类还未明确。为了解香蕉上镰孢菌在种间及种内水平上的多样性,2008–2011 年间作者从华南地区不同的水果市场及香蕉果园采集香蕉样品90份,分离得到143株镰孢菌。通过形态学观察及基于 EF-1α基因的系统进化分析鉴定出10种镰孢菌,即F. oxysporum、F. solani、F. camptoceras、F. pallidoroseum、F. stiloides、F. chlamydosporum、F.verticillioides、F. proliferatum、F. concentricum、F. sacchari,以及藤仓赤霉复合种(Gibberella fujikuroi species complex,GFC)中 3 个未定名的类群。轮纹镰孢 F. concentricum 及甘蔗镰孢 F.sacchari 是香蕉果实中最常见种,前菌为我国首次报道,后菌是首次报道与香蕉有关。对从香蕉上分离的藤仓赤霉复合种(GFC)及尖孢镰孢复合种(FOSC)的EF-1α序列进行了系统发育分析,其GFC中的27个菌株组成的单系群可分为7个不同的亚群,分别为 F.verticillioides、F. proliferatum、F. concentricum、F. sacchari 以及3个没有描述过的菌系 Fusarium sp. 1、Fusarium sp.2和 Fusarium sp.3;FOSC中的50个菌株形成2大分枝共12个谱系,分离自我国华南地区的21株尖孢镰孢形成7个谱系,其中 13株已知的香蕉枯萎病病原菌分布在3个谱系中,我国大陆的香蕉枯萎病病原菌菌株与来源于台湾地区及东南亚的菌株亲缘关系较近,FOC1号生理小种的遗传分化大于4号生理小种,FOC 1号生理小种与分离自香蕉果实上的尖孢镰孢菌的亲缘关系比与FOC 4号生理小种的亲缘关系更近。研究结果表明,我国香蕉上存在着丰富的镰孢菌种类,而且种内遗传多样性丰富。     

长期连作对大豆根际土壤镰孢菌种群的影响

利用中国科学院海伦农业生态试验站大豆连作长期定位试验区,进行了大豆根际土壤镰孢菌分离鉴定及大豆根腐病致病力检测,结合核酸序列和限制性片段长度多态性(RFLP)的系统发育分析,研究了长期连作(20年)较短期连作(3年)对大豆根际土壤镰孢菌种群密度和结构、致病力及遗传多样性的影响.结果表明： 3年连作大豆根际土壤镰孢菌种群密度为6.0×10⁴ CFU·g^-1,且以强致病力的尖镰孢菌、禾谷镰孢菌、轮枝镰孢菌及中等致病力的腐皮镰孢菌为优势种;连作20年大豆根际土壤镰孢菌种群密度和优势菌的优势度均显著低于3年连作,其中尖镰孢菌、禾谷镰孢菌和腐皮镰孢菌的种群密度仅为3年连作的36%、32%和22%,没有分离到致病力最高的轮枝镰孢菌,而种群多样性和均匀度显著高于3年连作;仅分离自20年连作土壤的三线镰孢菌、砖红镰孢菌及燕麦镰孢菌均为非致病菌种,且与强致病力镰孢菌种在基于转录间隔区(ITS)和转录延长因子(EF-1α)序列的聚类分析中显示了系统进化亲缘关系的差异性.因此,大豆20年连作会导致根际土壤镰孢菌种群生长受抑制、使其种群结构和遗传多样性发生改变,同时降低大豆根腐病菌种群致病力.     

瓜类保护地土壤镰孢菌种群及UP-PCR多样性分析

对采自辽宁省部分地区瓜类保护地的36份土壤样品进行镰孢菌（Fusarium）分离培养,共获得112株镰孢菌,采用传统形态学分类和现代分子生物学方法,确定属于11个种.对其中25株镰孢菌及3株对照镰孢菌进行了通用引物PCR（UP-PCR）多样性分析.结果表明：6条引物扩增出73条带,其中多态性条带66条,占总条带数的90.4%.对供试菌株进行UP-PCR聚类图谱分析,当相似系数为0.736时,可将其划分为8个类群,其中14株尖孢镰孢菌聚为一类.UP-PCR分析体现了镰孢菌菌株间的亲缘关系及差异性,可以作为镰孢菌分类的辅助方法.     

两种基因型大豆根分泌物对大豆根腐病菌的化感作用

采用生物模拟试验、化学分析等方法,研究了两种大豆基因型(9536、吉林30)的根分泌物中的糖、氨基酸、有机酸组分对大豆根腐病菌的化感作用.结果表明,两种大豆基因型(9536、吉林30)根分泌物糖组分表现出低浓度显著促进、高浓度显著抑制半裸镰孢菌、尖镰孢菌生长的规律,对粉红粘帚菌的生长影响不明显;氨基酸组分对上述三种病原菌所表现出的促进、抑制规律不同,9536基因型根分泌物氨基酸组分的中、高浓度处理对半裸镰孢菌、粉红粘帚菌、尖镰孢菌的生长表现出显著的抑制作用,而吉林30多表现出显著的促进作用;有机酸组分对半裸镰孢菌、粉红粘帚菌、尖镰孢菌生长都有显著的抑制作用.两种基因型大豆根分泌物(糖、氨基酸、有机酸组分)与根腐病害发生密切相关,大豆基因型不同,根分泌物对根腐病菌的化感促进或抑制作用有差异.     

Effect of dsRNA-containing and dsRNA-free hypovirulent isolates of Fusarium oxysporum on severity of Fusarium seedling disease of soybean in naturally infested soil

In the sandy soils of eastern Virginia, soybean seedlings are colonized by hypovirulent and virulent isolates of Fusarium oxysporum and F. solani. Our objectives were to determine if prior inoculation of soybean seeds with hypovirulent F. oxysporum isolates reduced severity of seedling disease in naturally infested soil, and to determine if there was an association between the presence of dsRNA mycovirus and hypovirulence in isolates of F. oxysporum and F. solani from soybean plants. The presence of dsRNA was not associated with hypovirulence in F. oxysporum since some hypovirulent isolates contained dsRNA while other hypovirulent isolates did not. Furthermore, of six dsRNA-containing F. oxysporum isolates, three were hypovirulent and three were virulent. Four segments of dsRNA, with sizes of 4.0, 3.1, 2.7 and 2.2 kb were detected in extracts of all six F. oxysporum isolates. No hypovirulent or dsRNA-containing of F. solani isolates were found. Prior inoculation of cv. Essex soybean seeds with conidia of dsRNA-free hypovirulent F. oxysporum isolates significantly (P < 0.05) reduced disease severity on cotyledons and hypocotyls, and increased the rate of seedling emergence in field soil, compared to control plants. No significant (P > 0.05) differences were found between dsRNA-containing and dsRNA-free hypovirulent F. oxysporum isolates in their effects on reducing disease severity. Hypovirulent isolates that colonize soybean tissues may play a role in reducing Fusarium seedling disease of soybean in natural soils.     

Sudden death syndrome of soybean in South America is caused by four species of Fusarium: Fusarium brasiliense sp. nov., F. cuneirostrum sp. nov., F. tucumaniae, and F. virguliforme

Soybean sudden death syndrome (SDS) pathogens and dry bean root-rot pathogens were studied taxonomically, phylogenetically, and pathologically. Detailed phenotypic comparisons of macro- and microscopic features and phylogenetic analyses of multilocus DNA sequence data, including those on the nuclear ribosomal intergenic spacer region and the single copy nuclear gene translation elongation factor 1-a, indicated that they comprised five distinct species of Fusarium. Two new species causing soybean SDS in Brazil, F. brasiliense and F. cuneirostrum, are formally described. Fusarium cuneirostrum is responsible for soybean SDS in Brazil and dry bean or mung bean root-rot in the United States, Canada, and Japan. Strains of each species, including F. cuneirostrum isolates from dry bean and mung bean and F. phaseoli isolates from dry bean, were inoculated on soybean cultivar Pioneer 9492RR to determine their pathogenicity. Although intraspecific variation in pathogenicity was observed, all the species were able to induce typical SDS symptoms on soybean plants in the artificial inoculation tests. Comparisons of the key diagnostic morphological features reveal that all five species can be diagnosed using conidial morphology.     

Fusarium azukicola sp. nov., an exotic azuki bean root-rot pathogen in Hokkaido, Japan

We report on the phenotypic, molecular phylogenetic and pathogenic characterization of a novel azuki bean (Vigna angularis) root-rot (BRR) pathogen from Hokkaido, Japan, which formally is described herein as Fusarium azukicola. This species can be distinguished phenotypically from the other Phaseolus/Vigna BRR and soybean sudden-death syndrome (SDS) pathogens by the production of wider and longer four-septate conidia cultured on SNA. Molecular phylogenetic analyses of four anonymous intergenic loci, a portion of the translation elongation factor (EF-1α) gene and the nuclear ribosomal intergenic spacer region (IGS rDNA) strongly support the genealogical exclusivity of F. azukicola with respect to the other soybean SDS and BRR pathogens within Clade 2 of the F. solani species complex (FSSC). Evolutionary relationships of F. azukicola to other members of the SDS-BRR clade, however, are unresolved by phylogenetic analyses of the individual and combined datasets, with the exception of the IGS rDNA partition, which strongly supports it as a sister of the soybean SDS pathogen F. brasiliense. A multilocus genotyping assay is updated to include primer probes that successfully distinguish F. azukicola from the other soybean SDS and BRR pathogens. Results of a pathogenicity experiment reveal that the F. azukicola isolates are able to induce root-rot symptoms on azuki bean, mung bean (Vigna radiata), kidney bean (Phaseolus vulgaris) and soybean (Glycine max), as well as typical SDS foliar symptoms on soybean. Our hypothesis is that F. azukicola evolved in South America and was introduced to Hokkaido, Japan, on azuki bean but its possible route of introduction remains unknown.     

大豆内生细菌的分离及根腐病拮抗菌的筛选鉴定

内生细菌存在于健康植物体内,一些内生细菌具有促生长、抗病和固氮等生物学功能.本项研究采用化学药剂表面灭菌方法从黑龙江省大豆品种合丰25的根、茎、叶和种子中分离到大量内生细菌,其种群数量在根部最多,为3.4×103CFU/g,在叶部次之,为2.8×103CFU/g,在茎部和种子中最少,为2.9×102 CFU/g和1.4×102CFU/g.从121株内生细菌中筛选到31株对大豆根腐病菌Fusarium oxysporum f. sp.soybean具有较强抑制作用的拮抗内生细菌,其中菌株TF28抑菌谱广,抑菌率高,对不同植物的病原菌F. oxysporum的抑菌率为80.2%～96.7%.经形态、生理生化和16S rRNA鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens).     

Incidence and distribution of Fusarium species associated with feeds and seeds from Spain

Samples of seeds and feeds (corn-based and mixed) were collected during surveys in 1991-92 and 1992-93 from two regions of Spain, one in northern Spain where the annual rainfall is over 900 mm, and the other in southeastern Spain where the annual rainfall is about 400 mm. The level of Fusarium contamination was determined in the 657 samples analysed, and results were analysed statistically to assess the effects of type of sample and meteorological conditions on Fusarium proliferation. The predominant Fusarium species was Fusarium moniliforme, which represented 92.2% of the total Fusarium strains isolated. Other species isolated were Fusarium oxysporum (5.9%), F. oxysporum var. redolens (0.6%), Fusarium poae (0.6%) and Fusarium sporotrichioides (0.6%).     

Ribosomal variation in six species of shape Fusarium

Eighteen isolates representing six Fusarium species from diverse hosts and geographical origins were evaluated to determine ribosomal DNA variation using polymerase chain reaction and restriction fragment length polymorphisms. No length variation was observed for amplified 18S and 28S regions. However, amplification of the ITS region showed one isolate, a F. oxysporum, to be about 120 bp larger than the remaining 17. Restriction digestions in the 18S region revealed polymorphisms within species of F. oxysporum and F. solani. An amplified variable stretch of the 28S gene showed restriction site differences between F. avenecum, F. sambucinum and F. sporotrichioides. A large degree of polymorphism was observed both between and within species in the ITS region. Therefore, entire sequences of the ITS and the 5.8S subunit were obtained for 17 of the 18 isolates. These sequences, along with those from eight additional isolates, were analysed using PAUP to assess the occurrence of DNA sequence divergence within the ITS region. The lack of correlation between molecular-based relationships and species affinities inferred from morphology for some isolates indicates that species designation can be unreliable using morphological data alone. Possible reasons for the discordance of the sequence and morphological data are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.