Genetic Characterization by RAPD Analysis of Isolates of Fusarium oxysporum f. sp. erythroxyli Associated with an Emerging Epidemic in Peru

ABSTRACT An epidemic of vascular wilt caused by Fusarium oxysporum f. sp. erythroxyli is currently occurring on Erythroxylum coca var. coca in the coca-growing regions of the Huallaga Valley in Peru. Random amplified polymorphic DNA (RAPD) analysis of isolates of the pathogen was undertaken to elucidate its genetic complexity, as well as to identify a specific DNA fingerprint for the pathogen. Two hundred isolates of Fusarium were collected from 10 coca-growing regions in Peru. Of these, 187 were confirmed to be F. oxysporum, and 143 of the F. oxysporum were shown to be pathogens of coca by a root-dip pathogenicity test. The pathogens could be grouped into two subpopulations based on RAPD analysis, and no polymorphism in RAPD pattern was observed among isolates of either subpopulation. Both subpopulations were present in the central Huallaga Valley, where earliest reports of the epidemic occurred. RAPD analysis could easily distinguish the isolates of F. oxysporum f. sp. erythroxyli from the nonpathogenic isolates of F. oxysporum from E. coca var. coca, indicating its utility in DNA fingerprinting.     

Genetic diversity among isolates of Fusariumoxysporum f.sp. canariensis

Fusarium oxysporum f.sp. canariensis causes vascular wilt disease of Phoenix canariensis , the Canary Island date palm. Seventy-two isolates of this fungus were obtained from diverse geographic locations including France, Japan, Italy, the Canary Islands, and California, Florida and Nevada, USA. The isolates were tested for vegetative compatibility and for similarities based on mitochondrial DNA (mtDNA), single-copy sequences and repetitive DNA (pEY10) polymorphisms. Seventy-one percent of the isolates belonged to a single vegetative compatibility group (VCG 0240), and four closely related mitochondrial RFLP patterns were found. A subset of the isolates was further tested for single-copy RFLPs and repetitive DNA fingerprints. Only four single-copy RFLP haplotypes were found among 25 representative isolates of F. oxysporum f.sp. canariensis tested, using nine polymorphic single-locus probe/enzyme combinations. Finally, 32 different pEY10 DNA fingerprints were found out of 57 isolates examined. Overall the results indicate that F. oxysporum f.sp. canariensis is a single lineage with a low to moderate level of genetic diversity.     

Rapid Detection of the Fusarium oxysporum Lineage Containing the Canary Island Date Palm Wilt Pathogen

ABSTRACT Fusarium oxysporum f. sp. canariensis causes Fusarium wilt disease on the Canary Island date palm (Phoenix canariensis). To facilitate disease management, a polymerase chain reaction diagnostic method has been developed to rapidly detect the pathogen. A partial genomic library of F. oxysporum f. sp. canariensis isolate 95-913 was used to identify a DNA sequence diagnostic for a lineage containing all tested isolates of F. oxysporum f. sp. canariensis. Two oligonucleotide primers were designed and used to amplify a 567-bp fragment with F. oxysporum f. sp. canariensis DNAs. DNA from 61 outgroup isolates did not amplify using these primers. Once the primers were shown to amplify a 0.567-kb fragment from DNA of all the F. oxysporum f. sp. canariensis isolates tested, a rapid DNA extraction procedure was developed that led to the correct identification of 98% of the tested F. oxysporum f. sp. canariensis isolates.     

Identification of Race 1 of Fusarium oxysporum f. sp. lactucae on Lettuce by Inter-Retrotransposon Sequence-Characterized Amplified Region Technique

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.     

Genetic Diversity of Fusarium oxysporum Isolates from Cucumber: Differentiation by Pathogenicity, Vegetative Compatibility, and RAPD Fingerprinting

ABSTRACT A total of 106 isolates of Fusarium oxysporum obtained from diseased cucumber plants showing typical root and stem rot or Fusarium wilt symptoms were characterized by pathogenicity, vegetative compatibility, and random amplified polymorphic DNA (RAPD). Twelve isolates of other formae speciales and races of F. oxysporum from cucurbit hosts, three avirulent isolates of F. oxysporum, and four isolates of Fusarium spp. obtained from cucumber were included for comparison. Of the 106 isolates of F. oxysporum from cucumber, 68 were identified by pathogenicity as F. oxysporum f. sp. radicis-cucumerinum, 32 as F. oxysporum f. sp. cucumerinum, and 6 were avirulent on cucumber. Isolates of F. oxysporum f. sp. radicis-cucumerinum were vegetatively incompatible with F. oxysporum f. sp. cucumerinum and the other Fusarium isolates tested. A total of 60 isolates of F. oxysporum f. sp. radicis-cucumerinum was assigned to vegetative compatibility group (VCG) 0260 and 5 to VCG 0261, while 3 were vegetatively compatible with isolates in both VCGs 0260 and 0261 (bridging isolates). All 68 isolates of F. oxysporum f. sp. radicis-cucumerinum belonged to a single RAPD group. A total of 32 isolates of F. oxysporum f. sp. cucumerinum was assigned to eight different VCGs and two different RAPD groups, while 2 isolates were vegetatively self-incompatible. Pathogenicity, vegetative compatibility, and RAPD were effective in distinguishing isolates of F. oxysporum f. sp. radicis-cucumerinum from those of F. oxysporum f. sp. cucumerinum. Parsimony and bootstrap analysis of the RAPD data placed each of the two formae speciales into a different phylogenetic branch.     

Mechanisms of date palm resistance to Bayoud disease: Current state of knowledge and research prospects

The Bayoud disease, caused by Fusarium oxysporum f. sp. albedinis (Foa), represents a major limiting factor of date palm culture in Morocco and constitutes a serious threat to the date palm plantations in Algeria and all other countries. Efficient disease prevention requires the development of resistant cultivars. In Morocco, among the cultivars listed, only six appear to be resistant to Bayoud disease, but they produce poor quality fruit. Thus, the Moroccan program of date palm genetic improvement is based on directed crossing between resistant cultivars and susceptible cultivars with good date quality traits to select resistant genotypes producing high quality fruits. In addition to the separation of the resistance to Bayoud disease and quality of the fruits characters, this breeding program is really complex due to the sex separation in the date palm, the duration of juvenile phase which is very long, and the lifespan of the date palm which requires a durable polygenic resistance. Then, the selected genotypes must be of female sex, of good date quality, and possess effective defense mechanisms against the pathogen. Moreover, the selection of the date palm resistance must necessarily take into account the mechanisms of pathogen aggressiveness. In this review, we will present and discuss studies developed on the Bayoud disease of the date palm, particularly on the disease control, the biochemical and molecular markers of resistance, the program of date palm genetic improvement of resistance, the Foa pathogenicity factors, and host defense mechanisms. It will also highlight the recent studies that showed that differential behaviour of the resistant and susceptible cultivars was not related to a difference of induction of the defense mechanisms, but to the suppression of their elicitation in the susceptible cultivars.     

Reliable detection of the fungal pathogenfusarium oxysporum f.sp.albedinis, causal agent of bayoud disease of date palm, using molecular techniques

Bayoud, caused by the soilborne fungusFusarium oxysporum f.sp.albedinis (FOA), is the most serious disease of date palm. Since the disease is located in the North African countries of Morocco and Algeria, and advancing steadily eastwards, the ultimate goal is to prevent spread of the pathogen to other date-growing areas in the region and farther afield. Molecular diagnostic techniques have been developed for detection of FOA. In view of the fact that the fungus does not exist in Israel, DNA of FOA was obtained to determine the reliability of these methods for diagnostic purposes. Random amplified polymorphic DNA was not reliable enough for differentiation between FOA and various pathogenic and saprophyticFusarium isolates. However, the polymerase chain reaction utilizing FOA-specific primers was accurate and enabled amplification of a unique band specific to FOA DNA alone, and not that of the other tested pathogenic and saprophyticFusaria. The availability of a rapid and reliable diagnostic tool for detection of FOA will enable the Plant Protection and Inspection Services of the Israel Ministry of Agriculture to test date palm tissue for the presence of the pathogen. Contribution no. 513/00 from the Inst. of Plant Protection, ARO, The Volcani Center, Bet Dagan, Israel.     

Development of a Specific Polymerase Chain Reaction-Based Assay for the Identification of Fusarium oxysporum f. sp. ciceris and Its Pathogenic Races 0, 1A, 5, and 6

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.     

香蕉枯萎病菌RAPD分析及4号生理小种的快速检测

 用随机扩增多态性DNA(RAPD)技术,对采自广东、广西的香蕉和粉蕉上的30个香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)菌株和3个其它尖孢镰刀菌专化型的菌株进行比较及聚类分析。在遗传相似系数0.67时,可将供试菌株划分为3个RAPD群(RGs),其中香蕉枯萎病菌4号生理小种(FOC4)共15个菌株属于RGⅠ,1号生理小种(FOC1)共15个菌株属于RGⅡ,供试的其它尖孢镰刀菌专化型的3个菌株则属于RGⅢ。这说明香蕉枯萎病菌和供试3个其它专化型菌株与致病性间存在明显的相关性。1号生理小种内菌株间的遗传分化大于4号生理小种内菌株间的遗传分化。从90条RAPD随机引物中筛选出2条引物可产生4号生理小种的RAPD标记2个。将这2个RAPD标记电泳切胶回收、克隆及测序,并根据这2个特异片段序列设计SCAR上下游特异引物,通过对30个菌株的PCR扩增检验,其中一个RAPD标记成功地转化为SCAR标记,初步建立了以此为基础的4号生理小种快速检测技术,其检测灵敏度为2 ng新鲜菌丝。对采自不同地区的显症样品、吸芽、室内接种未显症的香蕉苗以及发病的香蕉植株不同部位进行检测,能够准确灵敏地鉴定出4号生理小种,从而为香蕉枯萎病菌的快速检测及防治奠定了基础。同时,快速检测结果发现,田间发病植株果柄的各部位及果实内并没有枯萎病菌的存在。     

Phylogeny of Fusarium oxysporum f. sp. lactucae Inferred from Mitochondrial Small Subunit, Elongation Factor 1-alpha, and Nuclear Ribosomal Intergenic Spacer Sequence Data

ABSTRACT Fusarium oxysporum f. sp. lactucae, causal agent of Fusarium wilt of lettuce, is a serious pathogen recently reported in Arizona. Sequence analysis of the mitochondrial small subunit (mtSSU), translation elongation factor 1-alpha (EF-1alpha) gene, and the nuclear ribosomal DNA intergenic spacer (IGS) region was conducted to resolve relationships among f. sp. lactucae isolates, F. oxysporum isolates from other hosts, and local non-pathogenic isolates. Analysis of mtSSU sequences provided limited phylogenetic resolution and did not differentiate the lactucae isolates from 13 other F. oxysporum isolates. Analysis of EF-1alpha sequences resulted in moderate resolution, grouping seven formae speciales with the lactucae isolates. Analysis of the IGS region revealed numerous sequence polymorphisms among F. oxysporum formae speciales consisting of insertions, deletions, and single nucleotide transitions and substitutions. Repeat sequence analysis revealed several duplicated subrepeat units that were distributed across much of the region. Based on analysis of the IGS sequence data, lactucae race 1 isolates resolved as a monophyletic group with three other formae speciales of F. oxysporum. In all analyses, lactucae race 2 isolates composed a separate lineage that was phylo-genetically distinct and distantly related to the lactucae race 1 isolates.     

A Review of Fusarium Wilt of Oil Palm Caused by Fusarium oxysporum f. sp. elaeidis

ABSTRACT Vascular wilt is the most destructive disease of oil palm in Africa and causes severe losses in some areas. Symptoms include initial wilting followed by desiccation of the fronds, which finally break and hang around the trunk. Internally, characteristic browning of the vascular elements is seen both in adult palms and in seedlings. Two disease syndromes are commonly seen in the field in adult palms-"acute wilt" where the palm dies within a few weeks and "chronic wilt" where the palm may remain alive for many months and even years but becomes progressively stunted. The pathogen (Fusarium oxysporum f. sp. elaeidis) is a soilborne fungus and the perennial nature of the crop ensured that, in the past, disease management was difficult. Over a period of 30 to 40 years, screening for resistance at the nursery stage was introduced in many plantations and research stations, and successful breeding programs in West Africa, notably in Ivory Coast, have resulted in more resistant oil palm material becoming available. The disease has not yet been detected in South East Asia (largest producer of palm oil) and rigorous quarantine measures have been imposed to prevent introduction of the pathogen into these highly productive areas.     

Molecular identification of two vegetative compatibility groups of Fusarium oxysporum f. sp. cepae

Fusarium oxysporum f. sp. cepae, which causes basal rot of onion, consists of seven vegetative compatibility groups (VCGs 0420 to 0426) and several single-member VCGs (SMVs). F. oxysporum f. sp. cepae populations in South Africa and Colorado each consist of one main VCG (namely, VCG 0425 and 0421, respectively). The aim of this study was to develop sequence-characterized amplified region (SCAR) markers for the identification of VCGs 0425 and 0421, using 79 previously characterized F. oxysporum isolates. A second aim was to investigate the prevalence of VCG 0425 among 88 uncharacterized South African onion F. oxysporum isolates using (i) the developed SCAR markers and (ii) inter-retrotransposon (IR)- and random amplified polymorphic DNA (RAPD) fingerprinting. Only two RAPD primers provided informative fingerprints for VCG 0425 isolates but these could not be developed into SCAR markers, although they provided diagnostic fragments for differentiation of VCG 0425 from VCG 0421. IR fingerprinting data were used to develop a multiplex IR-SCAR polymerase chain reaction method for the identification of VCG 0421, VCG 0425, and SMV 4 isolates as a group. Molecular identification of the uncharacterized collection of 88 F. oxysporum isolates (65 F. oxysporum f. sp. cepae and 23 F. oxysporum isolates nonpathogenic to onion) confirmed that VCG 0425 is the main VCG in South Africa, with all but 3 of the 65 F. oxysporum f. sp. cepae isolates having the molecular characteristics of this VCG. Genotyping and VCG testing showed that two of the three aforementioned isolates were new SMVs (SMV 6 and SMV 7), whereas the third (previously known as SMV 3) now belongs to VGC 0247.     

Detection of Fusarium oxysporum f. sp. dianthi in Carnation Tissue by PCR Amplification of Transposon Insertions

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.     

A DNA-Based Procedure for In Planta Detection of Fusarium oxysporum f. sp. phaseoli

ABSTRACT We have characterized strains of Fusarium oxysporum from common bean fields in Spain that were nonpathogenic on common bean, as well as F. oxysporum strains (F. oxysporum f. sp. phaseoli) pathogenic to common bean by random amplified polymorphic DNA (RAPD) analysis. We identified a RAPD marker (RAPD 4.12) specific for the highly virulent pathogenic strains of the seven races of F. oxysporum f. sp. phaseoli. Sequence analysis of RAPD 4.12 allowed the design of oligonucleotides that amplify a 609-bp sequence characterized amplified region (SCAR) marker (SCAR-B310A280). Under controlled environmental and greenhouse conditions, detection of the pathogen by polymerase chain reaction was 100% successful in root samples of infected but still symptomless plants and in stem samples of plants with disease severity of >/=4 in the Centro Internacional de Agricultura Tropical (CIAT; Cali, Colombia) scale. The diagnostic procedure can be completed in 5 h and allows the detection of all known races of the pathogen in plant samples at early stages of the disease with no visible symptoms.     

新疆棉花枯萎病菌群体结构的研究

 采自新疆24个不同植棉县(市或团场)的37株棉花枯萎病菌代表菌株,经人工接种于国际通用鉴别寄主,致病性反应均表现为典型的7号生理小种特征。RAPD分析结果也显示出这37个供试菌株与7号小种各对照菌株间基因组DNA的指纹图谱高度相似,属同一遗传相似组,而与3号和8号小种的对照菌株间遗传差异较大,亲缘关系较远,即7号生理小种是组成目前新疆棉花枯萎病菌群体的优势小种,而原分布于新疆吐鲁番等地的3号小种在本研究中未被发现。结合部分自选辅助鉴别寄主对其中18个菌株进行的致病力分化研究表明,在7号小种内部还存在着侵染力的分化,显示出棉花枯萎病菌较强的变异性和适应性。     

利用RAPD-STS和实时定量PCR鉴别菠菜枯萎病病原菌(Fusarium oxysporum f.sp.spinaciae)

 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。     

Genetic Variation Among Vegetative Compatibility Groups of Fusarium oxysporum f. sp. cubense Analyzed by DNA Fingerprinting

ABSTRACT Genetic variation within a worldwide collection of 208 isolates of Fu-sarium oxysporum f. sp. cubense, representing physiological races 1, 2, 3, and 4 and the 20 reported vegetative compatibility groups (VCGs), was analyzed using modified DNA amplification fingerprinting. Also characterized were 133 isolates that did not belong to any of the reported VCGs of F. oxysporum f. sp. cubense including race 3 isolates from a Heliconia species and isolates from a symptomatic wild banana species growing in the jungle in peninsular Malaysia. The DNA fingerprint patterns were generally VCG specific, irrespective of geographic or host origin. A total of 33 different genotypes were identified within F. oxysporum f. sp. cu-bense; 19 genotypes were distinguished among the isolates that belonged to the 20 reported VCGs, and 14 new genotypes were identified among the isolates that did not belong to any of the existing VCGs. DNA fingerprinting analysis also allowed differentiation of nine clonal lineages within F. oxysporum f. sp. cubense. Five of these lineages each contained numerous closely related VCGs and genotypes, and the remaining four lineages each contained a single genotype. The genetic diversity and geographic distribution of several of these lineages of F. oxysporum f. sp. cubense suggests that they have coevolved with edible bananas and their wild diploid progenitors in Asia. DNA fingerprinting analysis of isolates from the wild pathosystem provides further evidence for the coevolution hypothesis. The genetic isolation and limited geographic distribution of four of the lineages of F. oxysporum f. sp. cubense suggests that the pathogen has also arisen independently, both within and outside of the center of origin of the host.     

Restriction fragment length polymorphisms in Fusarium oxysporum f.sp. dianthi and other fusaria from Dianthus species

DNA restriction fragment length polymorphisms (RFLPs) among 46 isolates of Fusarium oxysporum from Dianthus spp., representing the known range of pathogenicity in carnation, were determined using total DNA digested with the restriction enzyme Hind III and a previously described probe, D4. Distinct multiple band RFLP patterns were found, which delineated RFLP groups as follows: (i) F. oxysporum f.sp. dianthi races I and 8; (ii) F. oxysporum f.sp. dianthi races 2, 5 and 6; (iii) F. oxysporum f.sp. dianthi race 4; (iv) a recently described race of F. oxysporum f.sp. dianthi (wilt-causing isolates from D. caryophyllus formerly classified as F. redolens); (v) wilt-causing isolates from D. barbatus formerly classified as F. redolens and (vi), (vii) and (viii), three further recently described races of F. oxysporum f.sp. dianthi. Isolate groups derived from analysis of RFLPs were consistent with existing and recently described vegetative compatibility groups (VCGs) in F. oxysporum f.sp. dianthi , but not in all cases with races. Isolates of F. oxysporum and F. proliferatum not associated with wilt disease had simpler RFLP patterns (with one exception) that were not associated with VCGs.     

Biological and Molecular Characterization of Fusarium oxysporum f. sp. lycopersici Divides Race 1 Isolates into Separate Virulence Groups

ABSTRACT A collection of race 1 and race 2 isolates of Fusarium oxysporum f. sp. lycopersici was screened for vegetative compatibility and characterized by random amplified polymorphic DNA (RAPD) analysis to establish the identity and genetic diversity of the isolates. Comparison of RAPD profiles revealed two main groups that coincide with vegetative compatibility groups (VCGs). In addition, several single-member VCGs were identified that could not be grouped in one of the two main RAPD clusters. This suggests that F. oxysporum f. sp. lycopersici is a polyphyletic taxon. To assign avirulence genotypes to race 1 isolates, they were tested for their virulence on a small set of tomato lines (Lycopersicon esculentum), including line OT364. This line was selected because it shows resistance to race 2 isolates but, unlike most other race 2-resistant lines, susceptibility to race 1 isolates. To exclude the influence of other components than those related to the race-specific resistance response, we tested the virulence of race 1 isolates on a susceptible tomato that has become race 2 resistant by introduction of an I-2 transgene. The results show that both line OT364 and the transgenic line were significantly affected by four race 1 isolates, but not by seven other race 1 isolates nor by any race 2 isolates. This allowed a subdivision of race 1 isolates based on the presence or absence of an avirulence gene corresponding to the I-2 resistance gene. The data presented here support a gene-for-gene relationship for the interaction between F. oxysporum f. sp. lycopersici and its host tomato.     

Characterization of Fusarium oxysporum f. sp. radicis-cucumerinum attacking melon under natural conditions in Greece

A severe root and stem rot disease of melon was observed during the 2001 growing season on four glasshouse crops in Heraklio, Greece. A total of 43 isolates of F. oxysporum , obtained in Crete from glasshouse-grown melon and showing fusarium wilt or root and stem rot symptoms, were characterized by pathogenicity and vegetative compatibility. The majority of these isolates was also fingerprinted via amplified fragment length polymorphic (AFLP) analysis. Of the total number of isolates, 22 were identified by pathogenicity tests as F. oxysporum f. sp. melonis , 20 as F. oxysporum f. sp. radicis-cucumerinum , while one isolate was nonpathogenic on cucumber, melon, sponge gourd and pumpkin. All 22 isolates of F. oxysporum f. sp. melonis were assigned to vegetative compatibility group (VCG) 0134, and all 20 isolates of F. oxysporum f. sp. radicis-cucumerinum to VCG 0260. Isolates of F. oxysporum f. sp. radicis-cucumerinum were incompatible with isolates of F. oxysporum f. sp. melonis. AFLP fingerprinting allowed for the clustering of the isolates of the two formae speciales of F. oxysporum along two separate phenetic groups: f. sp. melonis to AFLP major haplotype I, and f. sp. radicis-cucumerinum to AFLP major haplotype II. Overall, pathogenicity, vegetative compatibility grouping and AFLP analysis were correlated and effectively distinguished isolates of F. oxysporum from melon. This appears to be the first report of natural infection of melon by F. oxysporum f. sp. radicis-cucumerinum worldwide.