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1.
ABSTRACT Sudden death syndrome (SDS) of soybean is caused by the soilborne Fusarium solani f. sp. glycines (synonym F. virguliforme). In a sequential approach, two multifactor factorial-design microplot experiments were conducted to investigate the effects of fungal infestation levels and soil moisture on both root necrosis and foliar SDS severity, and the interaction between F. solani f. sp. glycines and Heterodera glycines in fumigated versus nonfumigated soil. In 2003, soybean cv. Spencer was grown in nonfumigated or methyl bromide-fumigated soil and infested with increasing levels of F. solani f. sp. glycines, either under rainfall or irrigated after growth stage V6/R1. In 2004, interactions between F. solani f. sp. glycines and H. glycines were explored in a factorial inoculation design in fumigated or nonfumigated soil, planted to Williams 82 or Cyst-X20-18. In both years, higher levels of foliar SDS severity and root necrosis were found in F. solani f. sp. glycines-infested soils with H. glycines than in soils without the nematode on the soybean cultivars susceptible to both pathogens. Both natural infestations of H. glycines in 2003 and artificially amended populations of H. glycines in 2004 contributed to higher foliar SDS severity. More severe foliar SDS symptoms always were associated with more root necrosis, but elevated levels of root necrosis did not predict severe leaf symptoms. In contrast to the critical role of H. glycines, increasing fungal infestation levels had no significant effects on increasing either foliar SDS symptoms or root necrosis. Effects of moisture regime and fungal infestation levels also were examined in factorial greenhouse and growth chamber experiments. High soil moisture resulted in higher levels of SDS root necrosis. In the greenhouse, root necrosis increased at a higher rate in low soil moisture than the rate in high soil moisture. The two pathogens acted as a complex and the disease development was strongly dependent on high soil moisture. 相似文献
2.
ABSTRACT The soybean cyst nematode, Heterodera glycines, and the fungus that causes sudden death syndrome (SDS) of soybean, Fusarium solani f. sp. glycines, frequently co-infest soybean (Glycine max) fields. The interactions between H. glycines and F. solani f. sp. glycines were investigated in factorial greenhouse experiments with different inoculum levels of both organisms on a soybean cultivar susceptible to both pathogens. Measured responses included root and shoot dry weights, H. glycines reproduction, area under the SDS disease progress curve, and fungal colonization of roots. Both H. glycines and F. solani f. sp. glycines reduced the growth of soybeans. Reproduction of H. glycines was suppressed by high inoculum levels but not by low levels of F. solani f. sp. glycines. The infection of soybean roots by H. glycines did not affect root colonization by the fungus, as determined by real-time polymerase chain reaction. Although both pathogens reduced the growth of soybeans, H. glycines did not increase SDS foliar symptoms, and statistical interactions between the two pathogens were seldom significant. 相似文献
3.
Fusarium solani is a soilborne plant pathogen that infects many different hosts. Within the species, there is some specialization, and a number of forma specialis have been described based on host affiliation. One of these, F. solani f. sp. glycines, infects soybean and causes sudden death syndrome. To differentiate between F. solani f. sp. glycines and other F. solani isolates, a partial sequence of the mitochondrial small subunit (mtSSU) rRNA gene was amplified by polymerase chain reaction and sequenced from 14 F. solani f. sp. glycines and 24 F. solani isolates from various plant hosts. All F. solani f. sp. glycines isolates had identical sequences. A single, unique insertion of cytosine occurred in all F. solaniisolates but not in any of the F. solani f. sp. glycines isolates. Two major lineages, distinguished by sequence divergence and the presence or absence of multiple insertions, occurred in F. solani isolates. Cladistic analysis produced a single most-parsimonious tree with three major clades. The first clade contained all F. solani f. sp. glycines isolates. A second clade grouped together all of the F. solani isolates that had only a single nucleotide insertion difference from the first clade. Genetic distance between these two clades was 0.016. A third clade was formed by five F. solaniisolates that had multiple insertions. Isolates in the third clade had a genetic distance of 0.040 from the first and second clades. Based on the sequence data, it is likely that F. solani f. sp. glycineshas a shorter evolutionary history than other F. solaniisolates that have either single or multiple nucleotide insertions. The differences in nucleotide insertions in part of the mtSSU rRNA gene between F. solani f. sp. glycinesand other F. solani isolates provide a direct and reliable way to distinguish isolates of F. solani. 相似文献
4.
ABSTRACT Sudden death syndrome of soybean, caused by Fusarium solani f. sp. glycines, is a disease of increasing economic importance in the United States. Although the ecology of sudden death syndrome has been extensively studied in relation to crop management practices such as tillage, irrigation, and cultivar selection, there is no information on the effects of herbicides on this disease. Three herbicides (lactofen, glyphosate, and imazethapyr) commonly used in soybean were evaluated for their effects on the phenology of F. solani f. sp. glycines and the development of sudden death syndrome in four soybean cultivars varying in resistance to the disease and in tolerance to glyphosate. Conidial germination, mycelial growth, and sporulation in vitro were reduced by glyphosate and lactofen. In growth-chamber and greenhouse experiments, there was a significant increase in disease severity and frequency of isolation of F. solani f. sp. glycines from roots of all cultivars after application of imazethapyr or glyphosate compared with the control treatment (no herbicide applied). Conversely, disease severity and isolation frequency of F. solani f. sp. glycines decreased after application of lactofen. Across all herbicide treatments, severity of sudden death syndrome and isolation frequency were lower in disease-resistant than in susceptible cultivars. Results suggest that glyphosate-tolerant and -nontolerant cultivars respond similarly to infection by F. solani f. sp. glycines after herbicide application. 相似文献
5.
Morphological and molecular characterization of the sudden-death syndrome pathogen of soybean in Brazil 总被引：1，自引：0，他引：1
G. M. T. Arruda R. N. G. Miller † M. A. S. V. Ferreira A. C. Café-Filho 《Plant pathology》2005,54(1):53-65
Brazilian Fusarium isolates causing soybean sudden death syndrome (SDS) were characterized by comparing them with other Fusarium isolates associated with soybean root rot, as well as F. solani f.sp. glycines isolates associated with the disease in the USA, using molecular (mitochondrial and nuclear rDNA), morphological, cultural and pathogenic characteristics. On the basis of pathogenicity data, and restriction fragment length polymorphism and sequence analysis of the rDNA internal transcribed spacer (ITS) regions, isolates formed a group distinct from nonSDS F. solani isolates, as well as other Fusarium species. ITS sequence analysis also revealed that Brazilian isolates were distinct from the majority of SDS pathogens from the USA ( Fusarium virguliforme ) and conformed to Fusarium tucumaniae . 相似文献
6.
Fusarium oxysporum f.sp. gladioli (FOG) race 1 infects both large- and small-flowered Gladiolus cultivars. Race 2 isolates infect only small-flowered cultivars but can be present as epiphytes on large-flowered plants. When 160 arbitrary 10-mer oligonucleotide primers were tested on FOG by PCR to find RAPD markers specific for race 1, the RAPD primer G12 amplified two discriminating DNA fragments, AB (609 bp) and EF (1196 bp), in race 1 isolates only. Both fragments were cloned and sequenced. Two pairs of race 1-specific primers for multiplex PCR were designed. Tests of 112 F. oxysporum isolates by PCR showed that, in almost all cases, race 1 isolates of vegetative compatibility group 0340 could be distinguished with these primers. Seven putative race 1 isolates did not react in multiplex PCR; hybridization studies with labelled AB and EF DNA fragments showed that these isolates belong to separate groups. A bioassay was developed to detect corms that were latently infected with FOG race 1. Gladiolus corms were homogenized and incubated for 5 days at 28°C in a semiselective medium to induce growth of Fusarium . Cultivated mycelium was isolated and subjected to the developed multiplex PCR after standard DNA isolation or disruption by microwave treatment. 相似文献
7.
ABSTRACT Specific primers and polymerase chain reaction (PCR) assays that identify Fusarium oxysporum f. sp. ciceris and each of the F. oxysporum f. sp. ciceris pathogenic races 0, 1A, 5, and 6 were developed. F. oxysporum f. sp. ciceris- and race-specific random amplified polymorphic DNA (RAPD) markers identified in a previous study were cloned and sequenced, and sequence characterized amplified region (SCAR) primers for specific PCR were developed. Each cloned RAPD marker was characterized by Southern hybridization analysis of Eco RI-digested genomic DNA of a subset of F. oxysporum f. sp. ciceris and nonpathogenic F. oxysporum isolates. All except two cloned RAPD markers consisted of DNA sequences that were found highly repetitive in the genome of all F. oxysporum f. sp. ciceris races. F. oxysporum f. sp. ciceris isolates representing eight reported races from a wide geographic range, nonpathogenic F. oxysporum isolates, isolates of F. oxysporum f. spp. lycopersici, melonis, niveum, phaseoli, and pisi, and isolates of 47 different Fusarium spp. were tested using the SCAR markers developed. The specific primer pairs amplified a single 1,503-bp product from all F. oxysporum f. sp. ciceris isolates; and single 900- and 1,000-bp products were selectively amplified from race 0 and race 6 isolates, respectively. The specificity of these amplifications was confirmed by hybridization analysis of the PCR products. A race 5-specific identification assay was developed using a touchdown-PCR procedure. A joint use of race 0- and race 6-specific SCAR primers in a single-PCR reaction together with a PCR assay using the race 6-specific primer pair correctly identified race 1A isolates for which no RAPD marker had been found previously. All the PCR assays described herein detected up to 0.1 ng of fungal genomic DNA. The specific SCAR primers and PCR assays developed in this study clearly identify and differentiate isolates of F. oxysporum f. sp. ciceris and of each of its pathogenic races 0, 1A, 5, and 6. 相似文献
8.
ABSTRACT The capacity of the arbuscular mycorrhizal fungus Glomus intraradices in reducing the presence of Fusarium solani f. sp. phaseoli in bean plants and the surrounding mycorrhizosphere soil was evaluated in a compartmentalized experimental system. Quantification of the pathogen and the symbiont in plant tissues, the soil regions of the mycorrhizosphere (rhizosphere and mycosphere), and the bulk soil was accomplished using specific polymerase chain reaction (PCR) primers in real-time PCR assays, culture-dependant methods, and microscopic determination techniques. Nonmycorrhizal bean plants infected with the pathogen had distinctive Fusarium root rot symptoms, while infected plants previously colonized by G. intraradices remained healthy. The amount of F. solani f. sp. phaseoli genomic DNA was significantly reduced in mycorrhizal bean plants and in each mycorrhizosphere soil compartment. The presence of G. intraradices in the mycorrhizosphere was not significantly modified, although the mycorrhizal colonization of roots was slightly increased in the presence of the pathogen. The results suggest that the reduced presence of Fusarium as well as root rot symptoms are caused by biotic and/or abiotic modifications of the mycorrhizosphere as a result of colonization with G. intraradices. 相似文献
9.
基于小麦白粉病菌rDNA ITS序列的PCR分子检测 总被引：6，自引：0，他引：6
Wheat powdery mildew(Blumeria graminis f.sp.tritici) is the one of main wheat diseases in China.Based on the internal transcribed spacer(ITS) sequences of ribosome of B.graminis f.sp.tritici,three molecular primer pairs(F1/R,F2/R and F3/R) were designed to detect the fungal pathogen of wheat powdery mildew.The species specificity of these primers was confirmed.F1/R was demonstrated a higher sensitivity than the other two primer pairs,and could detect as low as 1 pg DNA of B.graminis f.sp.tritici.Furthermore,F1/R primer pair was used to detect the pathogen DNA extracted from wheat leaves showing chlorosis and typical symptoms of powdery mildew caused by artificial inoculation with B.graminis f.sp.tritici.The preliminary results demonstrated the usefulness of this primer pair and its potential applications in efficient detection of wheat powdery mildew pathogen from leaves with latent infections at early growth stages of wheat. 相似文献
10.
菠菜枯萎病是由致病性镰刀菌(F.o.f.sp.spinaciae)引起的,是菠菜生产中的重要病害之一。利用常规方法鉴定菠菜枯萎病病原菌需耗费大量时间,并且很难得到正确的结论。随机扩增多态性DNA序列标签(Randomly amplified polymorphic DNA-sequence tagged sites,RAPD-STS)为病原菌鉴定提供了一种有效方法。本研究通过对供试菌株的RAPD分析,克隆出了1个菠菜枯萎病病原菌的特异片段(GenBank登录号:AY337463)。根据测序结果设计了1对菠菜枯萎病病原菌的特异引物,并利用常规PCR和实时定量PCR(real-time PCR)2种方法对病原菌进行了鉴定,并对2种方法的敏感性进行了比较。结果表明,2种PCR方法都可以鉴定菠菜枯萎病病原菌(F.o.f.sp.spinaciae),但二者对病原菌DNA敏感程度不同,常规PCR检测的最低DNA量100Pg,而实时定量PCR检测的最低DNA量是1pg。同时,实时定量PCR还可以对病原菌DNA进行定量分析,并依此估算病原菌的数量。该方法可用于菠菜枯萎病病原菌的快速鉴定和病因诊断。 相似文献
11.
12.
Detection of Fusarium oxysporum f.sp. vasinfectum in cotton tissue by polymerase chain reaction 总被引：2，自引：0，他引：2
A polymerase chain reaction assay was developed for the detection of Fusarium oxysporum f.sp. vasinfectum (FOV), a serious wilt pathogen of cotton in many parts of the world. Based on small nucleotide differences in internal transcribed spacer sequences between 18S, 5.8S and 28S ribosomal DNAs, primers Fov1 (5'-CCCCTGTGAACATACCTTACT-3') and Fov 2 (5'-ACCAGTAACGAGGGTTTTACT-3') were selected. These primers unambiguously amplified a 400-bp DNA fragment of all the FOV isolates tested (from Angola, Brazil, China and the USA) but did not amplify any other isolates of mycoflora associated with cotton, such as F. moniliforme , Verticillium albo-atrum , V. dahliae , Aspergillus sp., F. oxysporum , F. sambucinum or F. solani . A control PCR assay was developed employing the universal primer pair ITS1 and ITS2 which amplified a fragment of approximately 220 bp from all isolates tested. This control assay demonstrated that all fungal DNAs were readily amplifiable, thus confirming that the lack of amplification with Fov1 and Fov2 primers was a result of primer specificity and not of other possible causes, such as DNA degradation or the presence of PCR inhibitors. The assay was effective on samples from the stems, leaves, roots and calli, and from plant tissues both with and without symptoms. This detection system proved to be accurate and sensitive and could aid not only diagnosis but also disease monitoring and forecasting. 相似文献
13.
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。 相似文献
14.
瓜黑星病菌、枯萎病菌和蔓枯病菌的三重PCR检测 总被引：2，自引：0，他引：2
通过测定黄瓜黑星病菌(Cladosporium cucumerinum)rDNA的ITS序列,比对近缘种及瓜类几种重要病原菌的ITS序列,设计出特异性引物HX-1/HX-2,经过对引物HX-1/HX-2PCR条件的优化,可以扩增出1条190bp的黄瓜黑星病菌特异性DNA条带,灵敏度达到1pg/μL。进一步将引物HX-1/HX-2和瓜类枯萎病菌、瓜类蔓枯病菌特异检测引物Fn-1/Fn-2、Mn-1/Mn-2组合,建立三重PCR体系,可一次检测出瓜类黑星病菌、瓜类枯萎病菌、瓜类蔓枯病菌3种瓜类植物重要的病原菌。建立了可以应用于田间瓜类黑星病菌PCR检测技术和瓜类主要病害三重PCR检测技术,对瓜类病害的诊断和防治具有重要的指导作用。 相似文献
15.
山东省大豆根腐病病原菌及其生物学研究 总被引：10，自引：2，他引：10
1988-1989年采集山东大豆根腐病标样553个,经分离接种证明,致病菌有茄病镰刀菌[Fusarium solani(Mart.)Sacc.]、尖孢镰刀菌(F.oxysporum Schl.)和木贼镰刀菌[F.equiseti(Corda)Sacc.]三种,以茄病镰刀菌的分离频率最高,致病力最强,为主要致病菌。并对其培养特性、形态特征、寄主范围和光照、温度及pH对生长的影响进行了研究,根据这些研究结果,将大豆根腐病主要致病菌定名为茄病镰刀菌大豆专化型[Fusarium solani(Mart.)Sacc.f.sp.glycines Li et Luo],本专化型过去没有报告。 相似文献
16.
马铃薯粉痂菌(Spongospora subterranea f. sp. subterranea)是引起马铃薯粉痂病的病原。本研究根据粉痂菌内部转录间隔区和线粒体DNA的保守区域,分别设计了2对适用于普通PCR的引物A5/A9、C3/C8和1对适用于荧光定量PCR的引物QF/QR,用于检测块茎和土壤样品中的粉痂菌。特异性检测结果表明:引物对A5/A9和C3/C8,以马铃薯粉痂菌DNA为模板,能分别扩增出264和367 bp大小的单一条带,而对其他非靶标DNA无扩增;引物对QF/QR对马铃薯粉痂菌有单一的熔解峰,说明三对引物特异性良好。灵敏性检测结果表明:荧光定量PCR灵敏度为13.8 fg·μL-1,是普通PCR灵敏度的1 000倍。进一步建立循环域值(Ct)与质粒DNA含量的曲线关系,获得标准曲线y=-3.893 9 x+35.228,R2 = 0.9966,呈良好线性关系。通过对不同地区采集的18份带菌种薯和18份带菌土壤进行普通PCR和荧光定量PCR检测,引物A5/A9、C3/C8和QF/QR对带菌种薯检测率均为100%,对带菌土壤的检测率分别为44.44%、66.67%和100%。本研究建立的马铃薯粉痂病菌快速检测方法,能及时、准确地检测带菌种薯和土壤,为马铃薯粉痂病的早期诊断和防治提供依据。 相似文献
17.
Chiocchetti A Bernardo I Daboussi MJ Garibaldi A Gullino ML Langin T Migheli Q 《Phytopathology》1999,89(12):1169-1175
ABSTRACT Strains of the carnation wilt pathogen, Fusarium oxysporum f. sp. dianthi, can be distinguished by DNA fingerprint patterns, using the fungal transposable elements Fot1 and impala as probes for Southern hybridization. The DNA fingerprints correspond to three groups of F. oxysporum f. sp. dianthi strains: the first group includes isolates of races 1 and 8; the second group includes isolates of races 2, 5 and 6; and the third group includes isolates of race 4. Genomic DNAs flanking race-associated insertion sites of Fot1 (from races 1, 2, and 8) or impala (from race 4) were amplified by the inverse polymerase chain reaction (PCR) technique. These regions were cloned and sequenced, and three sets of primers overlapping the 3' or 5' end of the transposon and its genomic insertion were designed. Using fungal genomic DNA as template in PCR experiments, primer pairs generated amplification products of 295, 564 and 1,315 bp, corresponding to races 1 and 8; races 2, 5, and 6; and race 4, respectively. When multiplex PCR was performed with genomic DNA belonging to races 1 and 8, 2, or 4, single amplimers were generated, allowing clear race determination of the isolate tested. PCR was successfully performed on DNA extracted from susceptible carnation cv. Indios infected with isolates representative of races 1, 2, 4, and 8. 相似文献
18.
甘蓝枯萎病菌生理小种传统鉴定方法费时费力,不能满足生产的要求,因此需要建立一种快速、可靠的分子检测技术。本研究在甘蓝枯萎病菌1号和2号生理小种基因组测序的基础上,通过比较基因组学方法筛选1、2号生理小种各自的特异基因片段并设计引物,并分别以10个甘蓝枯萎病菌1号生理小种菌株、2个2号生理小种菌株、7个尖孢镰刀菌其他专化型菌株及4个外围菌株DNA为模板进行常规PCR扩增,筛选出甘蓝枯萎病菌1号和2号生理小种特异性引物,同时引入尖孢镰刀菌通用引物W106R/W106S,建立起一步三重PCR检测甘蓝枯萎病菌1、2号生理小种的分子检测技术。结果表明,该分子检测技术实现了在一次PCR反应中快速、准确地同步检测出甘蓝枯萎病菌DNA、罹病甘蓝组织和土壤中的甘蓝枯萎病菌1号和2号生理小种,对检测甘蓝植株是否感染枯萎菌及甘蓝种植区土壤是否受到枯萎菌的污染有实用价值。 相似文献
19.
ABSTRACT Fusarium wilt of lettuce, caused worldwide by Fusarium oxysporum f. sp. lactucae, is an emerging seed-transmitted disease on Lactuca sativa. In order to develop a molecular diagnostic tool for identifying race 1 (VCG0300) of the pathogen on vegetable samples, an effective technique is presented. Inter-retrotransposon amplified polymorphism polymerase chain reaction (PCR), a technique based on the amplification of genomic regions between long terminal repeats, was applied. It was shown to be useful for grouping F. oxysporum f. sp. lactucae race 1 isolates. Inter-retrotransposon sequence-characterized amplified regions (IR-SCAR) was used to develop a specific set of PCR primers to be utilized for differentiating F. oxysporum f. sp. lactucae isolates from other F. oxysporum isolates. The specific primers were able to uniquely amplify fungal genomic DNA from race 1 isolates obtained in Italy, Portugal, the United States, Japan, and Taiwan. The primers also were specific to pathogen DNA obtained from artificially infected lettuce seed and naturally and artificially infected plants. 相似文献
20.
为快速、准确地对番茄枯萎病菌Fusarium oxysporum f. sp. lycopersici(FOL)和番茄颈腐根腐病菌F. oxysporum f. sp. radicis-lycopersici(FORL)进行检测,基于尖孢镰刀菌F. oxysporum多聚半乳糖醛酸外切酶基因pgx4的单核苷酸多态性（single nucleotide polymorphism,SNP）位点,设计FORL、FOL生理小种1(FOL-R1)、2(FOL-R2)和3(FOL-R3)的竞争性等位基因特异性PCR-SNP(kompetitive allele specific PCR-SNP,KASP-SNP)引物,建立番茄颈腐根腐病菌和番茄枯萎病菌KASP-SNP检测技术,并通过与常规PCR比对及ITS与pgx4序列分析对该检测技术的可靠性进行验证。结果显示,在FORL、FOL-R1、FOL-R2和FOL-R3中存在35个变异SNP位点,设计出18对KASP-SNP引物,筛选出FORL＿KASP、FOLrace1＿KASP、FOLrace2＿KASP和FOLrace3＿KASP共4对分型清晰的... 相似文献