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1.
Restriction enzyme-mediated integration (REMI) mutagenesis was used to isolate mutants of Fusarium oxysporum f. sp. melonis impaired in pathogenicity. The race 2 strain Mel02010 was transformed with linearized pSH75, conferring resistance to hygromycin B, with or without the enzyme used to linearize the plasmid. Addition of restriction enzymes did not affect the transformation frequency. A total of 2929 REMI transformants were tested for pathogenicity to three melon cultivars, Amus, Ogon 9 and Ohi. The race 2 strains are pathogenic to Amus and Ogon 9, but not to Ohi. Of 43 transformants with reduced pathogenicity on susceptible melon cultivars, 12 mutants were examined in detail for pathogenicity, vegetative growth and integrative mode of pSH75. The levels of pathogenicity varied among these mutants. Two mutants (B48 and B137) almost completely lost pathogenicity to both susceptible cultivars, and the others had reduced pathogenicity. Mutants B48, B241, B886 and X36 were also impaired in vegetative growth. Mutant B809 was a biotin auxotroph. By DNA gel blot analysis, nine mutants were found to contain a single copy of the transformation vector. These mutants may thus be useful in isolating genes involved in pathogenicity. Received 22 December 2000/ Accepted in revised form 16 April 2001  相似文献   

2.
Pseudomonas syringae pv.pisi, pea bacterial blight agent, is seed-transmitted. Some aspects of its life cycle and its biology were investigated. The colonization of pea plants obtained from naturally infected seeds was studied in natural conditions while high populations of bacteria developed on plants showing no symptoms. Two streptomycin-resistant mutants were used to study the epiphytic life of the pathogen. Populations were monitored in different host-parasite compatibilities. When race 2 or race 6 of the pathogen was surface-inoculated on susceptible cultivars, a decrease of population size was observed during the following one to three days but was followed by an increase to levels 1000 times greater than the initial number detected, without symptoms for most of the plants. When race 2 was surface-inoculated on resistant genotypes or race 6 on non-host plants, bacteria did not multiply but population levels slightly decreased.Pseudomonas syringae pv.pisi shows a resident phase and its development is race-specific. Weeds collected in naturally contaminated pea fields, diseased or not, often harboured the pathogen but with levels smaller than those observed on peas. Pea crop debris and volunteers kept high levels of bacteria for at least eight months after the harvest of a diseased crop. As long as two pea crops are not grown one after the other in the same field, it is unlikely that debris and volunteers will act as an important inoculum source. The development of this pathogen during the growing season is considered as an important parameter to take into account for controlling the disease through seed health testing.  相似文献   

3.
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  相似文献   

4.
C. Zhang  H. Wu  X. Li  H. Shi  F. Wei  G. Zhu 《Plant pathology》2013,62(6):1378-1383
During 2009–2010, a total of 323 isolates of Xanthomonas oryzae pv. oryzae were obtained from rice with symptoms of bacterial leaf blight (BLB) in four provinces (Zhejiang, Jiangsu, Anhui and Hubei) in China. These isolates were tested for baseline sensitivity to zinc thiazole, a novel bactericide with strong antibacterial activity against Xanthomonas. The sampled pathogenic population had similar sensitivity to zinc thiazole (0·1–16·8 mg L?1) in all four regions and over the whole two‐year study period. The baseline sensitivity was distributed as a unimodal curve with a mean EC50 value of 6·79 ± 1·61 mg L?1. The risk of mutation to resistance of zinc thiazole in X. oryzae pv. oryzae was further evaluated in vitro and in vivo. Twelve zinc thiazole‐resistant mutants were obtained through ultraviolet (UV) irradiation, culturing on zinc thiazole‐amended nutrient agar (NA) plates, and culturing on zinc thiazole‐treated rice plants. These zinc thiazole‐resistant mutants had resistance factors (RF = EC50 value of a mutant / EC50 value of the wildtype parent of this mutant) of 12·4 to 186·1 with a mean RF value of 44·1. Mutants obtained via UV irradiation, culturing on NA plates and culturing on rice plants had mean RF values of 51·8, 24·5 and 14·4, respectively. All mutants showed decreases in resistance to zinc thiazole after 20 successive transfers on bactericide‐free media or 10 successive inoculation–reisolations on bactericide‐free rice plants. No significant difference was found in bacterial growth and sensitivity to bismerthiazol between zinc thiazole‐resistant mutants and their parents. However, a significant decrease was observed in the pathogenicity of zinc thiazole‐resistant mutants compared with their parents, especially for mutants obtained via UV irradiation.  相似文献   

5.
One hundred and sixteen isolates of Fusarium oxysporum f. sp. lactucae obtained from 85 fields in three crisphead lettuce-producing areas in Nagano Prefecture, Japan were typed for races using differential cultivars Patriot, Banchu Red Fire and Costa Rica No. 4. They were also grouped into vegetative compatibility groups (VCGs) using complementation tests with nitrate non-utilizing (nit) mutants. Two California strains reported as F. oxysporum f. sp. lactucum, a type culture of F. oxysporum f. sp. lactucae, and 28 avirulent isolates of F. oxysporum obtained from crisphead lettuce were included for comparison. Among Nagano isolates, 66 isolates were identified as race 1, and 50 as race 2. Race 1 strains derived from Shiojiri and Komoro cities and race 2 from Kawakami village and Komoro city. All isolates of race 2 were biotin auxotrophs, and the race could be distinguished based on its requirement for biotin on minimal nitrate agar medium (MM). Pathogenic isolates were classified into two VCGs and three heterokaryon self-incompatible isolates. Strong correlations were found between race and VCG. All the race 1 strains were assigned to VCG 1 except self-incompatible isolates, and all the race 2 strains to VCG 2. The 28 avirulent isolates of F. oxysporum were incompatible with VCG 1 and VCG 2. California strains was vegetatively compatible with VCG 1, and they were assigned to race 1. Based on vegetative compatibility, these two races of F. oxysporum f. sp. lactucae may be genetically distinct, and F. oxysporum f. sp. lactucae race 1 is identical to F. oxysporum f. sp. lactucum. Received 7 May 2002/ Accepted in revised form 6 September 2002  相似文献   

6.
Mutants of Botrytis cinerea with moderate and high resistance to pyraclostrobin, a Qo inhibitor of mitochondrial electron transport at the cytochrome bc 1 complex, were isolated at a high mutation frequency, after nitrosoguanidine mutagenesis and selection on medium containing pyraclostrobin and salicylhydroxamate (SHAM), a specific inhibitor of cyanide-resistant (alternative) respiration. Oxygen uptake in whole cells was strongly inhibited in the wild-type strain by pyraclostrobin and SHAM, but not in the mutant isolates. Cross-resistance studies with other Qo and Qi inhibitors (QoIs and QiIs) of cytochrome bc 1 complex of mitochondrial respiration showed that the mutation(s) for resistance to pyraclostrobin also reduced the sensitivity of mutant strains to other QoIs as azoxystrobin, fluoxastrobin, trifloxystrobin and picoxystrobin, but not to famoxadone and to the QiIs cyazofamid and antimycin-A. An increased sensitivity of pyraclostrobin-resistant strains to the carboxamide boscalid, an inhibitor of complex II, and to the anilinopyrimidine cyprodinil, a methionine biosynthesis inhibitor, was observed. Moreover, no effect of pyraclostrobin resistance mutation(s) on fungitoxicity of the hydroxyanilide fenhexamid, the phenylpyrrole fludioxonil, the benzimidazole benomyl, and to the phenylpyridinamine fluazinam, which affect other cellular pathways, was observed. Study of fitness parameters in the wild-type and pyraclostrobin-resistant mutants of B. cinerea showed that most mutants had a significant reduction in the sporulation, conidial germination and sclerotia production. Experiments on the stability of the pyraclostrobin-resistant phenotype showed a reduction of resistance, mainly in moderate resistant strains, when the mutants were grown on inhibitor-free medium. However, a rapid recovery of the resistance level was observed after the mutants were returned to a selective medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of high resistant mutants was higher than the moderate ones. Pathogenicity tests on cucumber seedlings showed that all mutant strains tested exhibited an infection ability similar with the wild-type parent strain. Preventive applications of the commercial product of F-500 25EC (pyraclostrobin) were effective against lesion development on cotyledons by the wild-type, but ineffective, even at high concentrations, against disease caused by the pyraclostrobin-resistant isolates. Boscalid (F-510 50WG) was found equally effective against the disease caused by the wild-type or pyraclostrobin-resistant mutants. This is the first report indicating the appearance of B. cinerea strains resistant to QoI fungicides by the biochemical mechanism of site modification and the risk for field resistance.  相似文献   

7.
The phytotoxicity of the Fusarium trichothecene and fumonisin mycotoxins has led to speculation that both toxins are involved in plant pathogenesis. This subject has been addressed by examining virulence of trichothecene and fumonisin-nonproducing mutants of Fusarium in field tests. Mutants were generated by transformation-mediated disruption of genes encoding enzymes that catalyze early steps in the biosynthesis of each toxin. Two economically important species of Fusarium were selected for these studies: the trichothecene-producing species Fusarium graminearum, which causes wheat head blight and maize ear rot, and the fumonisin-producing species F. verticillioides, which causes maize ear rot. Trichothecene-non-producing mutants of F. graminearum caused less disease than the wild-type strain from which they were derived on both wheat and maize, although differences in virulence on maize were not observed under hot and dry environmental conditions. Genetic analyses of the mutants demonstrated that the reduced virulence on wheat was caused by the loss of trichothecene production rather than by a non-target mutation induced by the gene disruption procedure. Although the analyses of virulence of fumonisin-non-producing mutants of F. verticillioides are not complete, to date, the mutants have been as virulent on maize ears as their wild-type progenitor strains. The finding that trichothecene production contributes to the virulence of F. graminearum suggests that it may be possible to generate plants that are resistant to this fungus by increasing their resistance to trichothecenes. As a result, several researchers are trying to identify trichothecene resistance genes and transfer them to crop species.  相似文献   

8.
Mutants of Phytophthora parasitica Dast. more or less resistant to dimethomorph or metalaxyl were obtained by treating mycelium with ultraviolet radiation. Some metalaxyl-resistant mutants exhibited levels of resistance (i.e. the ratio between the EC50 values for the mutant and the wild-type strain) greater than 100, but those observed in the dimethomorph-resistant mutants never exceeded 25. Most mutants retained their resistance after sub-culturing on unamended agar medium. Single zoospore clones derived from a metalaxyl-resistant and a dimethomorph-resistant mutant, selected as amongst the most resistant, had levels of resistance similar to those of the parental strains. The metalaxyl-resistant mutants were also resistant to the related phenylamide fungicides, furalaxyl and benalaxyl, but remained sensitive to dimethomorph. The dimethomorph-resistant mutants were not resistant to phenylamide fungicides. The pathogenicity of some metalaxyl- or dimethomorph-resistant mutants on tobacco leaves was similar to that of the wild-type strains.  相似文献   

9.
Complementary auxotrophic nitrate-nonutilizing (nit) mutants were used to investigate vegetative compatibility within 27 strains ofVerticillium dahliae isolated from several hosts originating from Africa, Asia, Europe and the United States. Using about 500nit mutants generated from these strains, three vegetative compatibility groups, 1, 2 and 4, were identified. Simultaneously, virulence of each strain was assessed on cultivars ofGossypium hirsutum, G. barbadense andG. arboreum, based upon Foliar Alteration Index (FAI) and Browning Index (BI) estimation. The strains in VCG1 were of both the cotton-defoliating pathotype and race 3 (on cotton) but were non pathogenic on tomato; those in VCG2 and VCG4 were of the nondefoliating pathotype and belonged to different races on cotton and on tomato. Hyaline mutants deriving from parental wild-type strain showed differences in pathogenicity but were always assigned to the parental VCG. A relationship was established between VCGs and the taxonomic position of host plants. Data fromnit pairings indicated that the sub-populations ofV. dahliae (VCGs) may not be completely isolated genetically.  相似文献   

10.
The type virus of the family Hypoviridae, Cryphonectria hypovirus 1 strain EP713 (CHV1-EP713), infects Cryphonectria parasitica, the filamentous causal fungus of chestnut blight, and reduces its virulence. This pathosystem serves as a model to study fungus-mycovirus interactions. We previously developed a genetic screening protocol for host factors associated with symptom induction by CHV1-EP713 and its mutants. In the procedure the standard field fungal isolate EP155 was transformed by cDNA from a mild hypovirus mutant Cys(72), launching virus infection, and mutagenized by random plasmid insertion with pHygR conferring hygromycin resistance. We now report an extension of the study to characterize different mutant strains, with different phenotypes than their parental strain TCys(72)-1. TCys(72)-1 is moderately reduced in pigmentation and sporulation compared to the uninfected wild-type strain EP155. Mutants sfb1, sfb2 and k202 were characterized biologically and molecularly in comparison to the previously isolated mutant (namA) and the parental strain. These mutants harbored one (sfb1) or more copies (sfb2 and k202) of the mutagenic plasmid, pHygR. The three mutants had similar biological attributes; that is, vegetative growth rate, conidiation and virulence (assay on apples) was reduced on potato dextrose agar media, relative to the parental strain and pigmentation was the same or slightly increased. Interestingly, viral dsRNA accumulation levels were apparently unaltered in these mutants. The screening method was efficient for mining fungal mutants with unusual hypovirus symptoms. Further, characterization of the mutants provides interesting insights into symptom induction by the hypovirus.  相似文献   

11.
Thirty-nine genotypes ofCucumis melo (plant introduction entries, open-pollinated cultivars and F1 hybrids) were evaluated for resistance to powdery mildew under either natural field conditions or artificial inoculation in growth chambers at the cotyledonary stage and the 2-true-leaf stage. Results confirmed that susceptibility in cotyledons was not necessarily associated with susceptibility in either true leaves in growth chambers or adult plants in the field. However, resistance at the 2-true-leaf stage in growth chambers was highly correlated with resistance of field-grown plants. Results also showed that 20 muskmelon genotypes resistant to race 1 at the cotyledonary stage were also resistant at the 2-leaf-stage and as adult plants in the field. The same was true for ten genotypes with race 2 inoculations. Because muskmelon genotypes expressing resistance in cotyledons were also resistant in true leaves in growth chambers or the field, the use of plants at the cotyledonary stage is recommended for screening for powdery mildew resistance caused by race 1 or race 2 ofS. fuliginea. When cotyledons are susceptible, screening should be done at the 2-true-leaf stage.  相似文献   

12.
Fusarium oxysporum f. sp. pisi (Fop) is an important pathogen of field pea (Pisum sativum) worldwide. The constant evolution of the pathogen drives the necessity to broaden the genetic basis of resistance to Fop. To achieve this, it is important to have a large germplasm collection available and an accurate and efficient method for disease assessment. Here, a detailed evaluation method coupling disease incidence, disease rating over time and its related area under the disease progression curve (AUDPC) was established and used to screen a Pisum spp. germplasm collection against one isolate of Fop race 2. A large variation in the disease response of specific pea accessions ranging from highly resistant to susceptible was observed within the collection, indicating the quantitative expression of the resistance. The repetition of the inoculation experiments on a subset of 19 accessions, including two susceptible accessions, indicated that the scoring method was robust and reproducible and confirmed the highly resistant phenotypes of 11 accessions. To initiate the characterization of resistance mechanisms within these accessions, the external and internal stem symptoms were compared between these selected pea accessions, together with the extent of fungal colonization within plants. All these tests indicated that, in all resistant accessions, the resistance mechanisms efficiently stopped pathogen progression at the crown. Incorporation of these sources of resistance to breeding programmes will contribute to improved Fop resistance in pea cultivars.  相似文献   

13.
The objective of the current study was to characterize Fusarium oxysporum f. sp. radicis-cucumerinum isolates from cucumbers in Turkey in terms of pathogenicity, vegetative compatibility and amplified fragment length polymorphism (AFLP) variation. In the 2007 and 2008 greenhouse cucumber-growing seasons, surveys were conducted in Adana, Antalya, Hatay and Mersin provinces of the Mediterranean region of Turkey. Forty-seven fungal isolates of F. oxysporum were recovered from diseased cucumber plants. The pathogenicity of each isolate was tested on cucumber seedlings at the one-true-leaf stage. Forty of the 47 isolates of F. oxysporum were virulent on cucumber seedlings. Based on disease symptoms, the differential effect of temperatures of 17°C and 29°C on disease development, and the virulence on cucumber seedlings, these 40 isolates were identified as F. oxysporum f. sp. radicis-cucumerinum. Nitrate non-utilizing mutants were generated on minimal medium containing 1.5% KClO3 and their phenotypes were determined. Mutants in different phenotypic classes were paired on minimal medium; of 40 F. oxysporum f. sp. radicis-cucumerinum isolates, thirty-eight were placed into VCG 0260. Remaining two strains were assigned to VCG 0261. The AFLP primers produced a total of 180 fragments between 200 and 500 bp in length for the 30 isolates tested. At a genetic similarity of 0.71, the UPGMA analysis separated the isolates into two distinct clusters. The first cluster, AFLP I, included 28 isolates, of which all belonged to VCG 0260. Two strains in the second AFLP cluster both belonged to VCG 0261.  相似文献   

14.
Fourty-four strains of Fusarium oxysporum were isolated from plants of melon with Fusarium wilt symptoms. Among these strains, thirty-nine were characterized for their pathogenicity on melon. Thirty-seven strains belonged to known races of F. oxysporum f. sp. melonis, while two strains were non-pathogenic. Four strains belonged to race 0, seven to race 1, four to race 2, and twenty-two to race 1,2. Beauvericin was produced by thirty-six strains in a range from 1 to 310gg–1. Eight isolates of race 1,2 did not produce the toxin. In addition, of the two non-pathogenic strains, only one strain produced the toxin (290gg–1). The production of enniatin A1, enniatin B1, and enniatin B was also investigated. Enniatin B was the only enniatin detected, being produced by eleven strains belonging to all the races, with a range of production from traces to 60gg–1. Finally, melon fruits belonging to two different cultivars (Cantalupo and Amarillo) were artificially inoculated with one strain of F. oxysporum f. sp. melonis (ITEM 3464). Beauvericin was detected in the fruit tissues of both cultivars at a level of 11.2 and 73.8gg–1, respectively. These data suggest that the production of both the toxins is not related to the pathogenicity of F. oxysporum f. sp. melonis, nor to the differential specificity of the races. The results confirm that beauvericin is a common metabolite of phytopathogenic Fusarium species.  相似文献   

15.
Nitrate non-utilising (Nit) mutants of six field strains and three single-ascospore strains of Botryotinia fuckeliana were selected by plating mycelial plugs onto a nitrate-containing minimal medium amended with chlorate. Mutants were characterised by growth responses on minimal medium amended with various sole nitrogen sources. For each parental strain two mutants were produced: nit1, defective in nitrate reductase apoenzyme; and NitM, defective in the molybdenum cofactor pathway. Complementation on nitrate minimal medium was observed between nit1 and NitM mutants from the same parent in all instances. However, complementation was not observed between any such combinations when the mutants were derived from different parents. It is concluded that a vegetative incompatibility system operates in B. fuckeliana resulting in multiple vegetative compatibility groups. One of the single-ascospore strains was derived from a cross between two of the field strains, indicating that new compatibility groups can be generated during sexual reproduction. Mycelial interaction zones were formed between all parental strains when they were paired on NaCl-amended medium, indicating congruence between mycelial incompatibility and vegetative incompatibility.  相似文献   

16.
Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification. All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation, and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA.  相似文献   

17.
Four carnation cultivars, Novada (resistant to races 1 and 2 ofFusarium oxysporum f.sp.dianthi), Elsy (susceptible to race 1), Lena (susceptible to race 2) and Sam's Pride (susceptible to both races), were selfed and crossed. When three months old, the seedlings were inoculated via the roots or via the stems, after which wilting was recorded weekly according to a 5-point ordinal scale.Analyses were carried out on the proportions of diseased plants. For race 1 variation between the progenies could be described by means of general combining abilities only; GCA values were not affected by the inoculation method used. Also for race 2 GCAs were most important but the GCA values appeared different for the two inoculation methods. It is concluded that resistance to both races is inherited in an additive way.Indications for independently inherited root-specific resistance components (extravascular resistance) were only found with race 2. With both races, the ability to confine the pathogen at the infection site appeared the most important resistance component. Resistant progenies were also characterized by longer latent periods and lower wilting rates.Both race 1 and race 2 induced the accumulation of the phytoalexins dianthalexin and methoxydianthramide S, but race 2 induced higher amounts than race 1. The accumulation of phytoalexins was positively correlated to the resistance level of the progenies against the respective races. The progenies of the double-resistant cultivar Novada appeared to produce particularly high levels of phytoalexins.  相似文献   

18.
BACKGROUND: QoI fungicides, inhibitors of mitochondrial respiration, are considered to be at high risk of resistance development. In several phytopathogenic fungi, resistance is caused by mutations (most frequently G143A) in the mitochondrial cytochrome b (cytb) gene. The genetic and molecular basis of QoI resistance were investigated in laboratory and field mutants of Botryotinia fuckeliana (de Bary) Whetz. exhibiting in vitro reduced sensitivity to trifloxystrobin. RESULTS: B. fuckeliana mutants highly resistant to trifloxystrobin were obtained in the laboratory by spontaneous mutations in wild‐type strains, or from naturally infected plants on a medium amended with 1–3 mg L?1 trifloxystrobin and 2 mM salicylhydroxamic acid, an inhibitor of alternative oxidase. No point mutations were detected, either in the complete nucleotide sequences of the cytb gene or in those of the aox and Rieske protein genes of laboratory mutants, whereas all field mutants carried the G143A mutation in the mitochondrial cytb gene. QoI resistance was always maternally inherited in ascospore progeny of sexual crosses of field mutants with sensitive reference strains. CONCLUSIONS: The G143A mutation in cytb gene is confirmed to be responsible for field resistance to QoIs in B. fuckeliana. Maternal inheritance of resistance to QoIs in progeny of sexual crosses confirmed that it is caused by extranuclear genetic determinants. In laboratory mutants the heteroplasmic state of mutated mitochondria could likely hamper the G143A detection, otherwise other gene(s) underlying different mechanisms of resistance could be involved. Copyright © 2012 Society of Chemical Industry  相似文献   

19.
A 2‐year comprehensive field survey was conducted across major tomato‐growing areas of Iran. Two hundred and thirty‐four tomato fields and six tomato‐producing greenhouses were surveyed for the potential presence of bacterial spot disease. Five hundred and ninety‐six tomato samples with and without symptoms were analysed. While Xanthomonas spp. were found in association with tomato plants both with and without symptoms from five surveyed counties, the bacterial spot disease was observed only in plants from three of them. Only strains isolated from plants with symptoms induced disease symptoms on tomato, while those isolated from symptomless plants caused symptoms only on cabbage and common bean. None of the isolates caused disease symptoms on pepper and eggplant. Phylogenetic analysis showed that X. perforans is the causal agent of tomato bacterial spot in Iran, although X. campestris and X. axonopodis were also associated with symptomless tomato plants. All X. perforans isolates in this study were sensitive to streptomycin, copper sulphate and copper oxychloride at concentrations of 50 mg L?1, 200 mg L?1 and 0.8 g L?1, respectively. Unlike the type strain of X. perforans, isolates in this study did not produce bacteriocin against other Xanthomonas spp., nor were they detected using the usual species‐specific primer pair Bs‐XpF/Bs‐XpR. This suggests an atypical nature of X. perforans strains in Iran, which leads to the hypothesis that X. perforans strains in Iran may have a separate origin to those causing disease epidemics elsewhere. The aggregated dispersal pattern of the diseased tomato fields signifies the seedborne introduction of the pathogen into the country.  相似文献   

20.
Mutants of Botrytis cinerea and Ustilago maydis highly resistant to fludioxonil were isolated at a high frequency, after nitrosoguanidine or UV mutagenesis, respectively, and selection on media containing fludioxonil. Tests on the response of mutant strains to high osmotic pressure resulted in the identification of two fludioxonil-resistant phenotypes (FLDosm/s and FLDosm/r), regarding the sensitivity to high osmolarity. Approximately 95% of fludioxonil-resistant mutants were found to be more sensitive to high osmotic pressure than the wild-type parent strains. Genetic analysis of phenylpyrrole-resistance in the phytopathogenic basidiomycete U. maydis, showed that fludioxonil-resistance was coded by three unlinked chromosomal loci (U/fld-1, U/fld-2 and U/fld-3), from which only the U/fld-1 mutation coded an osmotic sensitivity similar to that of the wild-types. Cross-resistance studies with fungicides from other chemical groups showed that the mutations for resistance to phenylpyrroles affect the sensitivity of mutant strains to the aromatic hydrocarbon and dicarboximide fungicides, but not to the benzimidazoles, anilinopyrimidines, phenylpyridinamines, hydroxyanilides or the sterol biosynthesis inhibiting fungicides. A study of fitness parameters in the wild-type and fludioxonil-resistant mutants of B. cinerea, showed that all osmotic sensitive (B/FLDosm/s) isolates had significant reductions in the characteristics determining saprophytic fitness such as mycelial growth, sporulation, conidial germination and sclerotial production. Contrary to that, with the exception of mycelial growth, the fitness parameters were unaffected or only slightly affected in most of the osmotic resistant (B/FLDosm/r) isolates. Tests on cucumber seedlings showed that the osmotic-sensitive strains were significantly less pathogenic compared with the wild-type and B/FLDosm/r strains of B. cinerea. Preventative applications of the commercial products Saphire 50 WP (fludioxonil) and Rovral 50 WP (iprodione) were effective against lesion development on cotyledons by the wild-type and the mutant strains of B. cinerea that were resistant to the anilinopyrimidine cyprodinil (B/CPL-27) and to the hydroxyanilide fenhexamid (B/FNH-21), but ineffective, even at high concentrations, against disease caused by the fludioxonil-resistant isolates (B/FLD) and a mutant strain resistant to the dicarboximide iprodione (B/IPR-1). Experiments on the stability of the fludioxonil-resistant phenotype showed a reduction of resistance, mainly in osmotic-sensitive isolates, when the mutants were grown on inhibitor-free medium. A rapid recovery of the high resistance was observed after mutants were returned to the selection medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain of B. cinerea, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of osmotic-resistant mutants was higher than the osmotic-sensitive ones. Furthermore, competition tests, in planta, showed a significant reduction of the frequency of both phenylpyrrole-resistant phenotypes, with a respective increase in the population of the wild-type strain of the pathogen.  相似文献   

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