Stepwise Evolution of Races in Fusarium oxysporum f. sp. ciceris Inferred from Fingerprinting with Repetitive DNA Sequences

ABSTRACT Plant pathogens often exhibit variation in virulence, the ability to cause disease on host plants with specific resistance, evident from the diversity of races observed within pathogen species. The evolution of races in asexual fungal pathogens has been hypothesized to occur in a stepwise fashion, in which mutations to virulence accumulate sequentially in clonal lineages, resulting in races capable of overcoming multiple host plant resistance genes or multiple resistant cultivars. In this study, we demonstrate a simple stepwise pattern of race evolution in Fusarium oxysporum f. sp. ciceris, the fungus that causes Fusarium wilt of chickpeas. The inferred intraspecific phylogeny of races in this fungus, based on DNA fingerprinting with repetitive sequences, shows that each of the eight races forms a monophyletic lineage. By mapping virulence to each differential cultivar (used for defining races) onto the inferred phylogeny, we show that virulence has been acquired in a simple stepwise pattern, with few parallel gains or losses. Such a clear pattern of stepwise evolution of races, to our knowledge, has not been demonstrated previously for other pathogens based on analyses of field populations. We speculate that in other systems the stepwise pattern is obscured by parallel gains or losses of virulence caused by higher mutation rates and selection by widespread deployment of resistant cultivars. Although chickpea cultivars resistant to Fusarium wilt are available, their deployment has not been extensive and the stepwise acquisition of virulence is still clearly evident.     

Physiological Specialization of Fusarium oxysporum f. sp. lactucae, a Causal Organism of Fusarium Root Rot of Crisp Head Lettuce in Japan

In 1995, Fusarium root rot of crisp head lettuce, caused by Fusarium oxysporum f. sp. lactucae, was simultaneously found in the Shiojiri and Kawakami areas of Nagano Prefecture, Japan. The Shiojiri and Kawakami isolates differed in pathogenicity to lettuce cultivars. Because of this distinct physiological specialization, these Shiojiri and Kawakami isolates should be designated as race 1 and race 2, respectively, using lines VP1010 (highly resistant to race 1), VP1013 (highly resistant to race 2) and variety Patriot (highly susceptible to both races) as differential varieties. This is the first report of races of Fusarium oxysporum f. sp. lactucae, Received 21 September 2000/ Accepted in revised form 21 March 2001     

Physiological races of Fusarium oxysporum f.sp. melonis in Tunisia

Fusarium wilt of melon, caused by Fusarium oxysporum f.sp. melonis (Fom), is an important disease; races of the pathogen were identified by inoculating differential standard host cultivars. A total of ten isolates that were obtained from 23 fields located in four different geographical regions were identified as pathogenic. Results indicate that all four known Fom races, namely, 0, 1, 2 and 1.2, were found in north and middle Tunisia. Race 1.2 was the most prevalent.     

Interactions between Fusarium oxysporum f. sp. racheiphilum and Meloidogyne spp. in Vigna unguculata. 2. Specificity of different taxa

A California isolate of Meloidogyne javanica increased Fusarium wilt symptoms in cowpea cultivars California Blackeye No. 3 (CB3) (resistant to wilt) and Grant (tolerant) inoculated with each of the three races of Fusarium oxysporum f. sp. tracheiphilum. The same isolate of M. javancia did not similarly increase wilt in wilt-resistant cultivar CB7977 inoculated with two isolates of race 3 of F. o. tracheiphilum. Six of seven isolates of M. javanica caused similar increases in vascular discoloration in cultivar CB3. but one isolate of M. javanica and seven of M. incognita did not. Vascular discoloration rating was positively correlated with galling severity. However, increasing the initial inoculum density, and thus galling index, of one isolate of M. incognita did not increase vascular discoloration. The vascular discoloration ratings for the wilt-susceptible CB5 controls in each experiment were higher than those for the wilt-resistant cultivars infected with M. javanica. It is hypothesized that M. javanica but not M. incognita reduces, but does not eliminate, resistance to all races of F. o. tracheiphilum in cultivars CB3 and Grant.     

Physiological races and vegetative compatibility groups within Fusarium oxysporum f.sp. gladioli

The pathogenicity and vegetative compatibility of mainly Dutch isolates ofFusarium oxysporum collected from diseased gladioli and other Iridaceae were investigated. Based on their pathogenicity to two differential gladiolus cultivars, the isolates could tentatively be divided into two races. All self-compatible isolates ofFusarium oxysporum f.sp.gladioli belonged to one of three distinct vegetative compatibility groups, VCG 0340, 0341 or 0342, and were incompatible with isolates that were not pathogenic to gladiolus. Isolates of one of the two races were restricted to one VCG while isolates of the other race were present in all three VCGs.     

Evolution of Fusarium oxysporum f. sp. vasinfectum Races Inferred from Multigene Genealogies

ABSTRACT Fusarium wilt of cotton is a serious fungal disease responsible for significant yield losses throughout the world. Evolution of the causal organism Fusarium oxysporum f. sp. vasinfectum, including the eight races described for this specialized form, was studied using multigene genealogies. Partial sequences of translation elongation factor (EF-1alpha), nitrate reductase (NIR), phosphate permase (PHO), and the mitochondrial small subunit (mtSSU) rDNA were sequenced in 28 isolates of F. oxysporum f. sp. vasinfectum selected to represent the global genetic diversity of this forma specialis. Results of a Wilcoxon Signed-Ranks Templeton test indicated that sequences of the four genes could be combined. In addition, using combined data from EF-1alpha and mtSSU rDNA, the phylogenetic origin of F. oxysporum f. sp. vasinfectum within the F. oxysporum complex was evaluated by the Kishino-Hasegawa likelihood test. Results of this test indicated the eight races of F. oxysporum f. sp. vasinfectum appeared to be nonmonophyletic, having at least two independent, or polyphyletic, evolutionary origins. Races 3 and 5 formed a strongly supported clade separate from the other six races. The combined EF-1alpha, NIR, PHO, and mtSSU rDNA sequence data from the 28 isolates of F. oxysporum f. sp. vasinfectum recovered four lineages that correlated with differences in virulence and geographic origin: lineage I contained race 3, mostly from Egypt, and race 5 from Sudan; lineage II contained races 1, 2, and 6 from North and South America and Africa; lineage III contained race 8 from China; and lineage IV contained isolates of races 4 and 7 from India and China, respectively.     

麦类禾谷镰刀菌致病性鉴定

江苏各地禾谷镰刀菌菌株间的致病力有强弱的不同,小麦品种对江苏禾谷镰刀菌菌株的抗病性也有强弱不同,但无一品种免疫。江苏各地禾谷镰刀菌菌株的致病力和品种的抗病性都没有特异性差异,所以,还不能认为江苏的禾谷镰刀菌有不同的生理小种存在。建议在江苏可以当地毒性较强的菌株作为麦类赤霉病抗病性鉴定的对象。     

尖孢镰刀菌的遗传多态性

 尖孢镰刀菌是重要的植物维管束病原真菌。近年来,有关该菌的遗传多态性研究报道很多,本文着重综述了利用营养体亲和群、DNA多态性技术研究尖孢镰刀菌专化型、小种及其相互关系等方面的进展,同时介绍了对棉花枯萎病菌、瓜类枯萎病菌及尖孢镰刀菌小种起源等的研究概况。     

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.     

Examination of the relationships between vegetative compatibility groups and races in Fusarium oxysporum f.sp. dianthi

The feasibility of identifying races of Fusarium oxysporum f.sp. dianthi by tests for vegetative compatibility type was investigated. Nitrate non-utilizing nitl and NitM mutants were generated from 51 isolates of F. oxysporum f.sp. dianthi , 18 isolates of f. oxysporum from Dianthus spp. not belonging to f.sp. dianthi and, for comparison, 11 isolates of F. proliferatum from Dianthus spp. Vegetative compatibility groups (VCGs) among the isolates were identified by pairing all nitl with all NitM mutants.
Vegetative compatibility was found between isolates of F. oxysporum f.sp. dianthi races 1 and 8 (VCG 0022), races 2, 5 and 6 (VCG 0021) and race 4 (VCG 0020), and wilt-causing isolates previously classified as F. redolens from D. caryophyllus (VCG 0023) and D. barbatus (VCG 0024), Three self-compatible wilt-causing isolates were vegetatively incompatible with all other isolates (VCGs 0025,0026 and 0027), Two VCGs were found among isolates of F. oxysporum from D. caryophyllus not belonging to f.sp. dianthi ; six non-pathogenic isolates were self-compatible but vegetatively incompatible with all other isolates. The foot-rot-associated isolates of F. proliferatum from D. caryophyllus constituted a separate VCG.
Virulence analyses revealed at least four new races among VCGs 0023 to 0027, New Isolates could be categorized as races as a result of VCG analysis and VCG classification correctly indicated that the race identities previously ascribed to two old isolates had been incorrect. Vegetative compatibility tests offer the prospect for rapid identification of races, although inoculation tests continue to be necessary to differentiate races that belong to a single VCG.     

Genetic diversity in Fusarium oxysporum f.sp. dianthi and Fusarium redolens f.sp. dianthi

Pathogenic isolates were selected representing all known vegetative compatibility groups (VCGs) and races of Fusarium oxysporum sensu lato from Dianthus spp. On basis of differences in the internal transcribed spacer region of the ribosomal DNA, six VCGs were classified as F. oxysporum f.sp. dianthi and four as F. redolens f.sp. dianthi. All VCGs of F. oxysporum f.sp. dianthi were characterized by unique restriction fragment length polymorphisms (RFLPs), unique overall esterase profiles, and unique virulence spectra, supporting a clonal lineage concept. Two VCGs of F. oxysporum f.sp. dianthi nevertheless comprised more than one race, but races within the same VCG shared the same distinct overall virulence spectrum. VCGs belonging to F. redolens f.sp. dianthi also had unique RFLPs and unique virulence spectra, but had grossly identical esterase profiles. Three new races (9, 10 and 11) are described for F. oxysporum f.sp. dianthi, and four for F. redolens f.sp. dianthi. Two races previously considered lost were recovered; race 7 was identified as a member of VCG 0021 of F. oxysporum f.sp. dianthi while race 3 was identified as a distinct VCG and race of F. redolens f.sp. dianthi. A summary of races and VCGs in F. oxysporum f.sp. dianthi and F. redolens f.sp. dianthi is presented.     

Occurrence of Fusarium oxysporum f. sp. melonis Race 1 in Japan

In 1994, Fusarium wilt of melon cultivars which are resistant to races 0 and 2 of Fusarium oxysporum f. sp. melonis was observed in southern area of the Lake Biwa region, Shiga prefecture. In commercial fields, mature plants of cv. Amus which were grafted onto cv. Enken Daigi 2, and of cv. FR Amus showed yellowing, wilting and finally death before harvesting of fruits. Diseased plants had vascular and root discolorations, and their stem sections yielded typical colonies of F. oxysporum. When the Shiga strains were tested for their pathogenicity to 12 species of cucurbits, they caused wilts only on melon. Using race differential cultivars of melon, the Shiga strains were classified as race 1 of F. oxysporum f. sp. melonis, which has not been reported in Japan. To further characterize their pathogenicity, the strains were used to inoculate 46 additional cultivars of melon, oriental melon and oriental pickling melon. All the race 1 strains were pathogenic to the cultivars tested, and their host range was apparently different from those of strains belonging to other races (races 0, 2 and 1,2y). DNA fingerprinting with a repetitive DNA sequence, FOLR3, differentiated race 1 strains from strains of races 0 and 2, but not from race 1,2y strains. Received 2 July 1999/ Accepted in revised form 30 September 1999     

我国棉花枯萎菌生理小种特异性DNA扩增片段的筛选及序列测定

 利用RAPD技术,以随机引物对我国棉花枯萎菌的3个生理小种(3、7、8号小种)共26个菌株进行PCR扩增,从产生的140个RAPD分子标记中寻找到了不同小种的特征性条带,O PF-10₅₁₃(3号小种)、OPF-08₃₇₁(7号小种)及OPF-12₇₀₃(8号小种)。将其纯化后克隆到pGEM-TEasy质粒载体上,并获得了DNA特异性片段的核酸序列。     

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.     

Phylogenetic relationships between the lettuce root rot pathogen Fusarium oxysporum f. sp. lactucae races 1, 2, and 3 based on the sequence of the intergenic spacer region of its ribosomal DNA

The genetic relationship between the vegetative compatibility groups (VCGs) and between physiological races of Fusarium oxysporum f. sp. lactucae (FOL), the causal pathogen of lettuce root rot, was determined by analyzing the intergenic spacer (IGS) region of its ribosomal DNA. A total of 29 isolates containing a type strain were tested: 24 Japanese isolates, 2 Californian isolates, and 3 Italian isolates. Three races (races 1, 2, and 3) were found in Japan, and race 1 was also distributed in California and Italy. Races 1, 2, and 3 each belonged to a distinct VCG: VCG-1, VCG-2, and VCG-3 (VCG-3-1, VCG-3-3), respectively. Phylogenetic (neighbor-joining) analysis of the IGS sequences revealed that races 1, 2, and 3 coincided with three phylogenetic groups (PG): PG-1, PG-2, and PG-3, respectively. These results indicate that the three races are genetically quite different and have a strong correlation with VCGs and phylogenetic groupings. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession no. AB195218     

Real-time PCR for differential determination of the tomato wilt fungus, <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>, and its races

Five primer/probe sets to identify the tomato wilt pathogen, Fusarium oxysporum f. sp. lycopersici (FOL), and its three races selectively were designed based on the rDNA-intergenic spacer and avirulence genes. Real-time PCR using genomic DNA from mycelia and soil DNA with the primer/probe sets allowed the successful identification of FOL and its races.     

枯萎菌诱导棉花细胞壁富含羟脯氨酸糖蛋白积累与枯萎病抗性间的关系

 棉花枯萎病病菌(Fusarium oxysporum f.sp.vasinfectum)的侵染能激发起棉花对枯萎病的系统诱导抗性。不同品种间细胞壁富含羟脯氨酸糖蛋白(HRGP)的积累量存在差异,抗病品种86-1细胞壁内HRGP含量明显高于感病品种邯14。HRGP积累与细胞壁木质化有一定相关性。不同生理小种枯萎菌对棉花细胞壁HRGP的诱导能力不同。作者认为,HRGP在寄主与病原相互作用的专化性识别机制中起重要作用。     

Effectiveness of antagonistic bacteria,commercial fungicides,and fourth generation quaternary ammonium salts,against Fusarium oxysporum f. sp. cubense race “1 or 2”

European Journal of Plant Pathology - There are three races of Fusarium oxysporum f.sp. cubense (Foc) that significantly affect commercial banana plantations. Most banana cultivars are very...     

Gene genealogies support Fusarium oxysporum f. sp. ciceris as a monophyletic group

Fusarium oxysporum f. sp. ciceris (Foc), the causal agent of fusarium wilt of chickpea, consists of two pathotypes (yellowing and wilting) and eight races (races 0, 1B/C, 1A and 2–6) of diverse geographical distribution. Six Foc isolates, one each of races 0, 1B/C, 1A, 4, 5 and 6, representing the two pathotypes and the geographical range of the pathogen, showed identical sequences in introns of the genes for translation elongation factor 1α ( EF1 α), β-tubulin, histone 3, actin and calmodulin. Eleven additional Foc isolates representative of all races, pathotypes and geographical range, and three isolates of F. oxysporum (Fo) nonpathogenic to chickpea were further analysed for sequence variation in the EF1 α gene. All isolates pathogenic to chickpeas shared an identical EF1 α gene sequence, which differed from that shared by the three Fo isolates nonpathogenic to chickpea. EF1 α gene sequences from the 17 Foc isolates and the three Fo isolates were compared with 24 EF1 α gene sequences in GenBank from isolates of 11 formae speciales of F. oxysporum by parsimony analysis. Foc isolates formed a grouping distinct from other formae speciales and nonpathogenic isolates. These results indicate that F. oxysporum f. sp. ciceris is monophyletic.