黄瓜枯萎病菌毒力、营养体亲和性及ISSR分析

 本研究对来自哈尔滨、长春、沈阳、北京、西宁5个城市的70个尖孢镰刀菌黄瓜专化型菌株进行了毒力、营养体亲和性及ISSR分析。毒力测定结果显示黄瓜枯萎病菌在东农803品种上存在明显的毒力分化。在营养体亲和群的测定中有8个菌株没有产生nit突变体,2个菌株经测定为异核体自身不亲和性菌株,不能进行营养体亲和群的测定;其余60个菌株可分为5个营养体亲和群。利用筛选的7个引物对70个菌株进行了ISSR分子标记,聚类分析可将70个菌株分为3个类群,其中IGⅠ的41个菌株均来自东北三省,IGⅡ的21个菌株均来自北京,IGⅢ的8个菌株全部来自西宁。VCGs和ISSRs与菌株的地理来源及毒力存在一定的相关性。     

Pathogenicity and vegetative compatibility grouping among Indian populations of Fusarium oxysporum f. sp. ciceris causing chickpea wilt

Thirty-nine isolates of Fusarium oxysporum f. sp. ciceri – the causal agent of chickpea (Cicer arietinum) wilt – collected from different parts of India and representing eight races of the pathogen, were analyzed for virulence and classified on the basis of vegetative compatibility grouping (VCG). The wilt incidence ranged from 24% to 100% on a highly susceptible cultivar JG 62. Six isolates, from Delhi, Gujarat, Karnataka, Punjab and Rajasthan and belonging to six different races of the pathogen, caused 100% wilt incidence. Five isolates belonging to four different races, namely, Foc 143 from Andhra Pradesh, Foc 161 from Chhattisgarh, Foc 146 from Karnataka, Foc 158 from Madhya Pradesh and Foc 50 from Rajasthan, caused low wilt incidence. For VCG analysis, nitrate non-utilizing mutants (nit) were obtained by culturing wild-type isolates on 2.5% potassium chlorate and selecting resistant sectors. Complementary nit mutants were paired in all possible combinations to determine varying degrees of heterokaryon formation within the isolates, which showed that most of the isolates were self-compatible. Pairing of all the mutants showed that the isolates included in the present study belonged to a single VCG (0280). Thus, in spite of variability in the virulence, the Indian populations of the pathogen have only one VCG.     

Physiological races and vegetative compatibility groups of butterhead lettuce isolates of Fusarium oxysporum f. sp. lactucae in Japan

Races were identified among butterhead lettuce isolates of Fusarium oxysporum f. sp. lactucae collected from three geographical areas of Hokkaido, Shizuoka, and Fukuoka in Japan by inoculation tests using Fujinagas race differential cultivars of lettuce (i.e., Patriot, Costa Rica No. 4, and Banchu Red Fire). Eighteen isolates from Shizuoka and Fukuoka were designated race 3, with two unknown vegetative compatibility groups (VCGs) that differed from Ogisos VCG 1 and 2. These two new VCGs were obtained from both Shizuoka and Fukuoka. On the other hand, three isolates from Hokkaido were classified as race 1 and identified as VCG 1, which represents a VCG of crisphead isolates from Nagano.     

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.     

我国小麦禾谷镰刀菌(Fusarium graminearum)营养体亲合性与致病性研究

 赤霉病是在世界范围内多种作物上反复发生的病害,小麦受害尤其严重。     

Variability amongst strains of Fusarium poae assessed by vegetative compatibility and RAPD polymorphism

Vegetative compatibility tests and random amplification of polymorphic DNA (RAPD) were used to assess genetic relationships amongst 54 strains of Fusarium poae obtained from various geographical regions. Twenty-seven strains were assigned to eight multiple member vegetative compatibility groups (VCGs), while the other 27 isolates were found to form single-member VCGs. There was a partial correlation between VCG and geographical origin, but the relationship was not always clear. However, no correlation was observed between the VCG and the host plant of origin. RAPD patterns were closely associated with VCGs in all cases. Members of VCGs that were interconnected by bridging isolates formed common branches in the phenogram constructed on the basis of the RAPD patterns, while strains that belonged to single-member VCGs were scattered throughout the phenogram. These data demonstrate that the combination of traditional and molecular methodologies allows reliable intraspecific subdivisions in an asexual fungus, which is a secondary invader of a wide range of host plants, and so has never been subject to the intense selection pressure of a single host species and lacks pathogenic subgroups.     

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.     

香荚兰尖孢镰刀菌营养体亲和性测定的初步研究

 Eight hundred twenty four nit mutants were induced from 73 strains of Fusarium oxysporum f. sp. vanillae, and classified into four phenotypes by their abilities to utilize different nitrogen sources. Among these mutants, 64.9% were characterized as nit 1, 24.3% as nit 3, 9.8% as nit M, 1.0% as nit X. Based on complementary pairing tests of different nit mutants on the medium MM, 44 isolates belonged to 8 different VCGs, 29 isolates were classified into single and different VCGs. These results indicated that there was significant VCG diversity in Fusarium oxysporum f. sp. vanillae population. VCGs might be correlated with geographic origin of strains, but no close correlation was found between VCGs and pathogenicity.     

Pathogenicity and race characterization of Fusarium oxysporum f. sp. phaseoli isolates from Spain and Greece

Virulence (≡ severity of disease) and physiological specialization of nine isolates of Fusarium oxysporum f. sp. phaseoli recovered in El Barco de Avila (Castilla y León, west-central Spain) and of two isolates from Chryssoupolis (Greece) were determined. The susceptibility/resistance response showed by a differential set of common bean cultivars ( Phaseolus vulgaris ) selected at the Centro Internacional de Agricultura Tropical (CIAT) delineated the isolates into two new races: races 6 and 7. The results of pathogenicity tests did not show any significant differences in virulence among the isolates. However, the reactions of several Spanish common bean cultivars indicated the presence of two groups of isolates, highly virulent and weakly virulent, among the Spanish isolates analysed. These results indicate that isolates classified in the same race are not homogeneous with respect to virulence, and suggests that race analysis using the CIAT differential cultivars is insufficient to describe the physiological specialization of F. oxysporum f. sp. phaseoli .     

Population structure of Fusarium oxysporum f. sp. radicis-lycopersici in Florida inferred from vegetative compatibility groups and microsatellites

Fusarium crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL) is a serious soilborne disease reducing tomato yields in Florida, a leading state in fresh market tomato production in the United States. One hundred and twenty five isolates of FORL were collected from the three main tomato-growing counties in Florida between 2006 and 2008. Vegetative compatibility groups (VCGs) and 10 microsatellite loci were used to infer the population structure of FORL. Up to 69.8 % of the isolates could be assigned to one of three VCGs, 0094, 0098 or 0099, with frequencies of 38.6, 24.4, and 6.8 % respectively. A medium level of population differentiation (Φst?=?0.159) was detected among the three counties, and three optimal clusters (populations) were supported by discriminant analysis of principal components. In addition, each population had some individuals that likely migrated from other populations. Migration probably played an important role in shaping the population structure of FORL since repeated VCGs and multilocus genotypes were observed in the three counties. Considerable migrants (> 1.33 migrants per generation) were also detected between the three counties, resulting in an increase in the effective population size and genetic diversity of F. oxysporum f. sp. radicis-lycopersici. Although migration is an important evolutionary force, mutation and parasexual recombination could not be completely ruled out as contributing causes to the genetic diversity of FORL. Management strategies that limit the movement of F. oxysporum f. sp. radicis-lycopersici are necessary to reduce the evolutionary potential of the pathogen.     

Vegetative compatibility of Fusarium oxysporum from sweet basil in Israel

Fusarium wilt and crown rot of sweet basil, caused by Fusarium oxysporum f.sp. basilici (F.o.ba.), is widespread in Israel. Affected plants show a variety of symptoms, including vascular wilt as well as crown rot, and masses of macroconidia on stem surfaces. We used vegetative compatibility to determine whether F.o.ba. isolates associated with various symptoms and sources are genetically related. All 119 isolates previously described as F.o.ba., and 42 additional F. oxysporum isolates which had not been tested for pathogenicity, belonged to a single vegetative compatibility group (VCG). The various symptoms are therefore induced by a single pathogenic form which appears to be a specific clone of F. oxysporum. The isolates of F.o.ba. from Israel were vegetatively compatible with eight isolates of F.o.ba. from Italy and the USA, but not with nonpathogenic isolates of F. oxysporum from basil, or with F.o. lycopersici or F.o. radicis-lycopersici from tomato. We conclude that the population of F.o.ba. in Israel belongs to the common VCG of this pathogen described in the USA, and which includes American and Italian isolates.     

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.     

Genetic variation among Fusarium oxysporum isolates from cucumber

Isolates of Fusarium oxysporum obtained from cucumber worldwide were classified into 3 groups by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). All isolates of f. sp. radicis-cucumerinum fall into one group. Isolates of races 1 and 2 of f. sp. cucumerinum fall into a second group related to isolates of f. sp. melonis and niveum. Isolates of race 3 fall into a third group, related to f. sp. momordicae. Because f. sp. radicis-cucumerinum has relatively recently been introduced into Greece, where it is actively spreading and very damaging, RAPD-PCR may be valuable in monitoring populations of F. oxysporum.     

瓜类尖孢镰刀菌的营养体亲和群研究

 应用营养亲和性方法研究了尖孢镰刀菌菌株抗氯酸盐突变体和nit突变体的诱发规律及分布特性,以及菌株营养体亲和群(VCG)的划分。研究表明,不同寄主(黄瓜、甜瓜和西瓜)分离的尖孢镰刀菌菌株形成的抗氯酸盐突变体数目差异不显著,平均为每个接种点产生0.89~0.98个;但寄主不同部位(根部、茎基部和茎中部)分离的菌株间差异显著,形成的数目分别为1.27、0.75及0.76个。菌株产生的nit1突变体比例(75.40%)显著高于nitM突变体比例(13.17%);nit1突变体数目会因菌株的寄主及菌株寄主部位的不同而有差异,寄主为黄瓜、甜瓜和西瓜的菌株产生的比例依次为67.73%、83.71%和77.50%,根部、茎基部及茎中部分离菌株产生的比例依次为81.82%、78.48%和68.64%,而在致病菌株与非致病菌株间无显著差异,分别为74.43%和79.63%;nitM突变体数目受菌株寄主影响较小,所占比例在11.17%~13.92%之间;而在寄主不同部位分离的菌株及致病菌株与非致病菌株间差异显著,分离自茎基部的菌株所占比例最高为15.97%,茎中部菌株所占比例最低为9.87%,致病菌株与非致病菌株所占比例分别为14.08%和9.26%。供试菌株分为7个VCGs,其特点为来源于不同寄主的尖孢镰刀菌菌株互不亲和,同一寄主的致病菌株与非致病菌株均不亲和,同一寄主不同部位分离的菌株可亲和。     

Current status of vegetative compatibility groups in fusarium oxysporum

Thirty-eightformae speciales (ff.sp.) ofFusarium oxysporum which have been subjected to vegetative compatibility grouping (VCG) analysis are listed, along with their updated 3-digit numerical codes. The number of VCGs identified within a forma specialis ranges from one (in 11 cases) to 24. Between two and six VCGs were identified in each of 20 ff.spp., whereas seven VCGs or more were identified in the remaining ff.spp. VCGs to which 4-digit numerical codes have been given are listed for 30 ff.spp.     

Characterisation ofFusarium oxysporum isolated from carnation in Australia based on pathogenicity,vegetative compatibility and random amplified polymorphic DNA (RAPD) assay

Isolates ofF. oxysporum collected from symptomless carnation cuttings from Australian carnation growers properties, together with isolates from national collections, were screened for pathogenicity and grouped according to vegetative compatibility and random amplified polymorphic DNA (RAPD) patterns. The collection of 82 Australian isolates sorted into 23 different vegetative compatibility groups (VCGs). Of 69 isolates tested for pathogenicity, 24 were pathogenic to carnations, while the remaining 45 were non-pathogenic. All pathogenic isolates were within two VCGs, one of which was also compatible with an isolate obtained from an international culture collection, and which is known to represent VCG 0021 and race 2. Race status of the two pathogenic VCGs remains unknown. The RAPD assay revealed distinct DNA banding patterns which could distinguish pathogenic from non-pathogenic isolates as well as differentiate between isolates from the two pathogenic VCGs.     

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.     

福建省香蕉枯萎病菌硝酸盐营养突变体的营养体亲和性测定

 Fourteen strains of Fusarium oxysporum f. sp. cubense were induced to produce 146 nitrate-nonutilizing(nit) mutants on a chlorate-containing medium. Among them, there were 117 nit1 mutants(80.14%), 17 nit3 mutants(11.64%) and 12 nitM mutants(8.22%). These strains were divided into two vegetative compatibility groups(VCGs) by the vegetative compatibility tests. Twelve strains of F. oxysporum f. sp. cubense from Musa AAA belonged to VCG1, two trains from Musa ABB belonged to VCG2.     

Genetic variability among isolates of Fusarium oxysporum from sugar beet

Fusarium yellows, caused by the soil‐borne fungus Fusarium oxysporum f. sp. betae (Fob), can lead to significant yield losses in sugar beet. This fungus is variable in pathogenicity, morphology, host range and symptom production, and is not a well characterized pathogen on sugar beet. From 1998 to 2003, 86 isolates of F. oxysporum and 20 other Fusarium species from sugar beet, along with four F. oxysporum isolates from dry bean and five from spinach, were obtained from diseased plants and characterized for pathogenicity to sugar beet. A group of sugar beet Fusarium isolates from different geographic areas (including nonpathogenic and pathogenic F. oxysporum, F. solani, F. proliferatum and F. avenaceum), F. oxysporum from dry bean and spinach, and Fusarium DNA from Europe were chosen for phylogenetic analysis. Sequence data from β‐ tubulin, EF1α and ITS DNA were used to examine whether Fusarium diversity is related to geographic origin and pathogenicity. Parsimony and Bayesian MCMC analyses of individual and combined datasets revealed no clades based on geographic origin and a single clade consisting exclusively of pathogens. The presence of FOB and nonpathogenic isolates in clades predominately made up of Fusarium species from sugar beet and other hosts indicates that F. oxysporum f. sp. betae is not monophyletic.     

Restriction fragment length polymorphisms, races and vegetative compatibility groups within a worldwide collection of Fusarium oxysporum f.sp. gladioli

Isolates of Fusarium oxysporum f.sp. gladioli were collected from widely different geographic areas. These isolates were characterized by pathogenicity to two differential gladiolus cultivars, vegetative compatibility, and total genomic DNA restriction fragment length polymorphisms (RFLPs). RFLPs were used to estimate the genetic divergence and relationship among isolates of F. oxysporum. RFLPs were detected by Southern blot hybridization of total genomic DNA with a 3-4 kb DNA probe generated from total DNA off. oxysporum f.sp. dianthi. Cluster analysis allowed the division of pathogenic strains into three main RFLP groups, each group containing strains with similarity coefficients ranging from 78 to 100%. RFLP groups correlated with vegetative compatibility groups, not with races. Two single pathogenic isolates which could not be assigned to any of the three main vegetative compatibility groups also had distinctive RFLP patterns. Little genetic polymorphism was observed within vegetative compatibility groups, whereas the majority of RFLPs occurred between vegetative compatibility groups, suggesting a common ancestry for strains within a specific vegetative compatibility group and a polyphyletic origin for the present special form gladioli.