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1.
In DAS-ELISAs of 86 grapevine samples from northwestern Iran, Grapevine fanleaf virus (GFLV) was detected in 18 samples. RT-PCR with two primer pairs (M2/M4 or M0/M4) corresponding to GFLV movement protein (MP) amplified the expected 854- and/or 1,489-bp fragment(s), respectively, from all ELISA-positive samples. Four smaller and three larger PCR products were cloned and sequenced, which revealed that the MP region of the isolates was 1,044 nucleotides (nt) long, corresponding to the GFLV MP. There were 83–86% nucleotide and 93–94% amino acid identities deduced between the MPs of the sequenced isolates. Nucleotide sequence identities of 81–87 and 75–79% were found between the MP regions of these isolates and that of previously published GFLV and Arabis mosaic virus (ArMV) strains/isolates, respectively. On a consensus parsimony tree based on the nucleotide sequences, isolates La208 and X300 remained distinct from previously reported GFLVs. This is the first molecular characterization of GFLV MP isolates from Iran. The sequence data reported in this paper have been submitted to the DDBJ/EMBL/GenBank databases and have been assigned accession numbers DQ286901 to DQ286916.  相似文献   

2.
Fusarium graminearum and F. verticillioides are among the most important pathogens causing ear rot of maize in Central Europe. Our objectives were to (1) compare eight isolates of each species on two susceptible inbred lines for their variation in ear rot rating and mycotoxin production across 3 years, and (2) analyse two susceptible and three resistant inbred lines for potential isolate x line interactions across 2 years by silk-channel inoculation. Ear rot rating, zearalenone (ZEA) and deoxynivalenol (DON) concentrations were evaluated for all F. graminearum isolates. In addition, nivalenol (NIV) concentrations were analysed for two NIV producers. Fumonisin (FUM) concentrations were measured for all F. verticillioides isolates. Mean ear rot severity was highest for DON producers of F. graminearum (62.9% of the ear covered by mycelium), followed by NIV producers of the same species (24.2%) and lowest for F. verticillioides isolates (9.8%). For the latter species, ear rot severities differed highly among years (2006: 24%, 2007: 3%, 2008: 7%). Mycotoxin concentrations among isolates showed a broad range (DON: 100–284 mg kg−1, NIV: 15–38 mg kg−1, ZEA: 1.1–49.5 mg kg−1, FUM: 14.5–57.5 mg kg−1). Genotypic variances were significant for isolates and inbred lines in all traits and for both species. Isolate x line interactions were significant only for ear rot rating (P < 0.01) and DON concentration (P < 0.05) of the F. graminearum isolates, but no rank reversals occurred. Most isolates were capable of differentiating the susceptible from the resistant lines for ear rot severity. For resistance screening, a sufficiently aggressive isolate should be used to warrant maximal differentiation among inbred lines. With respect to F. verticillioides infections, high FUM concentrations were found in grains from ears with minimal disease symptoms.  相似文献   

3.
4.
为明确大豆胶孢炭疽菌Colletotrichum gloeosporioides对苯醚甲环唑的抗性风险及其抗性机制,采用药剂驯化法诱导获得其抗性突变体,并测定抗性突变体的适合度和交互抗性,同时分析抗性突变体和敏感菌株cyp51A和cyp51B基因的cDNA序列、启动区序列及相对表达量。结果表明:从大豆胶孢炭疽菌2株敏感菌株中筛选获得7株低抗苯醚甲环唑的突变体,突变频率为1.75×10-4,其抗药性可以稳定遗传。抗性突变体与亲本敏感菌株在对温度的敏感性、菌丝生长速率、产孢量和致病力方面均存在一定差异。苯醚甲环唑与丙环唑之间存在中等交互抗性(相关系数为0.80,P<0.05),苯醚甲环唑与咪鲜胺锰盐、多菌灵、氟啶胺和吡唑醚菌酯之间无交互抗性。抗性突变体cyp51A和cyp51B基因的cDNA序列未发生核苷酸突变,cyp51A和cyp51B基因的启动区无插入片段。经苯醚甲环唑处理后,4株抗性突变体lq27-7-1、lq27-7-2、lq30-2-1和lq30-2-2的cyp51A基因相对表达量被诱导上调倍数显著高于亲本敏感菌株;3株抗性突变体lq27-7-2、lq...  相似文献   

5.
Fusarium verticillioides (teleomorph: Gibberella moniliformis = G. fujikuroi mating population A) is one of the most important fungal pathogens of corn worldwide. The pathogen produces fumonisins, mycotoxins that are potentially harmful to humans and animals. Thirty-five Fusarium isolates from Laguna and Isabela, Philippines were identified morphologically and molecularly as F. verticillioides and characterized by PCR for mating type (MAT). Twenty-six isolates were MAT1-2, while nine isolates were MAT1-1. The isolates from Isabela were tested for aggressiveness, rated according to a disease index (%) on ‘Super Sweet’ corn IPB variety 1 under field conditions across two trials using the toothpick inoculation method. Other aggressiveness traits such as inhibition of seedling emergence, decrease of seedling height, fresh and dry mass were also determined in two greenhouse trials. All isolates were pathogenic to corn seedlings and mature plants compared to the noninoculated control. Significant genotypic variation was observed (P = 0.01) in trial, isolate, and isolate × trial interaction for all traits across two greenhouse trials and that aggressiveness was highly influenced by the trial conditions. Similarly, significant genotypic variation was observed in trial, replication within trial and isolate × trial interaction. Heritability was high for the five traits in the greenhouse (h 2 = 0.80–0.90) but moderate for disease index in the field (h 2 = 0.49). In an analysis of fumonisin production in corn culture by high performance liquid chromatography, 30 of 35 strains produced a detectable level of fumonisins, varying from 0.44 to 742 μg FB1/g corn, 0.51–222 μg FB2/g and 0.12–37 μg FB3/g. Isabela isolates produced more fumonisins than the Laguna isolates did. In vitro fumonisin production had little correlation with the field disease index (r = 0.32) or with greenhouse seedling germination (0.25).  相似文献   

6.
Grapevine virus A (GVA) is considered one of the viruses associated with rugose wood (RW), one of the most economically important diseases of grapevine. Thirty-seven GVA isolates collected from grapevine cultivars from Marche (central-eastern Italy), Apulia and Campania (southern Italy), were subjected to molecular characterization. The genetic and population diversity was studied in the coat protein (CP) gene by RT-PCR-RFLP analysis with three restriction enzymes (MseI, AluI, and AciI), and nucleotide sequencing. A new primer pair (CP1F/R) allowing amplification of the whole CP gene (621 bp) was developed. RFLP with AciI yielded the highest number of variants in GVA isolates, showing seven different ‘simple’ profiles (A, B, C, D, E, F, and G). ‘Complex’ profiles were also found, and the most common variant combination was A + B in 39% of isolates. The analysis of GVA sequences confirmed the presence of plants infected with more than one GVA variant and suggested that RT-PCR-RFLP is suitable for evaluating population diversity of GVA enabling a screening of different haplotypes. The distribution of RFLP profiles and the phylogenetic analysis were not correlated with the location of infected plants, showing the presence of a GVA population with genetic diversity in the average with those of RNA viruses.  相似文献   

7.
为明确我国黄瓜花叶病毒株系分化及系统进化基本情况,从湖南、新疆、青海和海南4省区采集1 367个样品对其进行酶联免疫和RT-PCR检测,并对分离获得的15个黄瓜花叶病毒(Cucumber mosaic virus,CMV)纯化分离物CP、MP、2b核苷酸序列进行相似性和进化树分析及生物学性状比较。结果表明,辣椒、龙葵和黄瓜的CMV阳性检出率较高,分别为54.13%、29.19%和18.46%。进化树分析显示CMV-Q5与CMV亚组II的亲缘性较高;CMV-N7为新发现的重组株系,其CP、2b基因属于CMV亚组IB,MP基因却属于CMV亚组II;其余13个分离物均属于CMV亚组IB。CMV-N7和CMV-Q5在系统寄主心叶烟和枯斑寄主苋色藜上引发的症状相似,但比对照株系CMV-P3613(IB)的发病时间要晚1~2 d,系统花叶较温和,枯斑较小。表明在以上4省区常见农作物上广泛流行的CMV存在分子变异。  相似文献   

8.
Isolates of an unidentified Rhizoctonia sp. (UR isolates) were obtained from creeping bentgrass and Kentucky bluegrass with reddish brown sheath and foliar rots. Because the UR isolates anastomosed with isolates of three varieties of Waitea circinata (var. oryzae, var. zeae, and var. circinata), colony morphology, hyphal growth rate at different temperatures, pathogenicity, sequence analysis of the internal transcribed spacers (ITS) region of ribosomal RNA genes (rDNA) were compared. The colony color of mature UR isolates was distinct from isolates of the other three varieties of W. circinata. In pathogenicity tests on creeping bentgrass, the severity of the disease caused by UR isolates was significantly higher than that caused by the three varieties of W. circinata. Sequence similarities of the rDNA-ITS region between UR isolates and between isolates within each variety were high (97–100%), but they were lower among isolates from UR and the varieties of W. circinata (88–94%). In a phylogenetic tree based on the rDNA-ITS sequences, UR isolates formed a cluster separate from each of the clusters formed by the three varieties of W. circinata. These results indicate that the UR isolates clearly differ from the three varieties of W. circinata. We therefore propose that the UR isolates be classified as new Rhizoctonia sp. that are closely related to W. circinata and that the disease on creeping bentgrass should be called Waitea reddish-brown patch disease (Sekikasshoku-hagusare-byo in Japanese).  相似文献   

9.
Xanthomonas citri subsp. citri, the causal agent of citrus canker, relies extensively on a type III secretion system for infection by delivering type III effectors into host cells. In the genus Xanthomonas, two major regulators, HrpG and HrpX, are involved in the expression of genes encoding the type III secretion system. Twenty-three candidate type III effectors were identified as targets for analysis. The involvement in pathogenicity of 20 candidate effector genes in X. citri strain 306 (Xcc-306) was investigated using site-directed mutagenesis. Pathogenicity assays in grapefruit of 19 genes using site-directed mutagenesis revealed that none of the mutants demonstrated to have reduced ability to cause canker disease. A mutation in the TAL effector pthA4 resulted in loss of hypertrophy although no changes were observed in bacterial growth in leaves. Mutations in hrpG, hrpX, or hrpA genes displayed a complete loss of pathogenicity. Moreover, all mutants maintained the ability to trigger a hypersensitive response (HR) in non-host tomato. In contrast to previous studies, hrpG , hrpX and hrpA mutants also retained the ability to elicit an HR in tomato, indicating the presence of an Hrp independent elicitor in Xcc-306.  相似文献   

10.
Fusarium proliferatum can occur on a wide range of economically important vegetable plants but its role in disease is not always well established. In 2000 and 2001, from forty-one field samples of wilting onion and garlic plants in Serbia, F. proliferatum as the predominant fungal species was isolated from root and bulbs. Seventy isolates were firstly characterized for their sexual fertility and were shown to be mostly members of Gibberella intermedia (sixty-seven of seventy isolates, the remaining three isolates were unfertile), the sexual stage of F. proliferatum (syn. mating population D of G. fujikuroi complex). A selected set of eleven F. proliferatum isolates from both hosts were also tested for their pathogenicity and toxigenicity. Although onion and garlic plants were susceptible to all isolates, onion plants showed a significantly higher disease severity index. Six of the eleven isolates of F. proliferatum produced fumonisin B1 from 25 to 3000 μg g−1, and beauvericin from 400 to 550 μg g−1; ten isolates produced fusaric acid from 80 to 950 μg g−1 and moniliformin from 50 to 520 μg g−1. Finally, all isolates produced fusaproliferin up to 400 μg g−1. These results confirm F. proliferatum as an important pathogen of garlic and onion in Europe and that there is a potential mycotoxin accumulation risk in contaminated plants of both garlic and onion.  相似文献   

11.
Three isolates of Chrysanthemum stem necrosis virus (CSNV) were obtained from chrysanthemum plants in distinct regions of Japan in 2006 and 2007. All the original host plants showed severe necrotic symptoms on the leaves and stems. Amino acid sequence data of the nucleocapsid protein genes of the three isolates (CbCh07A, TcCh07A, and GnCh07S) showed high identities with those of two other CSNV isolates, HiCh06A L1 from Japan and Chry1 from Brazil. Furthermore, for the first time the complete nucleotide sequence of the S RNA was determined for CSNV (isolate HiCh06A). In phylogenetic analysis based on the non-structural protein genes from the genus Tospovirus, HiCh06A L1 was placed in the same genetic group as Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus. Host range examination for isolates HiCh06A L1 and CbCh07A showed that green pepper (cv. ‘Kyoyutaka’, ‘Saitamawase’, ‘Tosakatsura’, ‘L3 sarara’ and ‘L3 miogi’) and tomato (cv. ‘Sekaiichitomato’) were systemically susceptible hosts, whereas TSWV-resistant Solanaceae species, Capsicum chinense, Lycopersicon peruvianum and a TSWV-resistant cultivar of green pepper (cv. TSR miogi), were resistant.  相似文献   

12.
Biological and molecular characterisation of ten Peanut stunt virus (PSV) isolates from Robinia pseudoacacia was carried out. The host range of these isolates was similar to that of the previously described PSV strains in most cases, but on Pisum sativum and Lens culinaris latent infection was induced. Variability in systemic symptoms was observed only on Nicotiana glutinosa. The partial RNA3 sequences were determined, including the carboxyl terminal region of the movement protein gene, the intergenic region, the entire coat protein gene and the 3′ untranslated region. Nucleotide sequence comparison of the coat protein genes showed 77.6–84.2% identity with most of the known PSV strains and 96.3–98.0% identity with PSV-Rp the typical member of subgroup IV. Phylogenetic analysis indicated the presence of the ancient homologous recombination in all of the examined black locust isolates and all the isolates were members of the fourth PSV subgroup. These results showed that the isolates of the fourth subgroup are widely distributed in black locust in this region.  相似文献   

13.
A weed survey conducted in 2004 and 2005 in Aydin province of Turkey showed that Solanum nigrum, Xanthium strumarium, Amaranthus retroflexus, Portulaca oleracea, Sonchus oleraceus and Datura stramonium were the most prevalent weeds in the cotton fields exhibiting Verticillium wilt. Verticillium dahliae Kleb. was recovered from A. retroflexus and X. strumarium in those cotton fields. This is the first report of V. dahliae occurring naturally in A. retroflexus in Turkey. Pathogenicity tests on cotton and weeds showed that the virulence of V. dahliae isolates from weeds was higher on cotton plants than on weeds, with the disease severity ranging from 31.7% to 98.0%. Disease severity of V. dahliae isolates was 54.7–93.9% on eggplant, 23.7–51.6% on cucumber and 11.0–16.4% on tomato, whereas it did not cause any disease symptoms, or only low levels, on pepper and bell pepper. Two vegetative compatibility groups (VCGs) were identified among seven tested weed isolates: VCG2A (two isolates) and VCG2B (three isolates) using international reference strains.  相似文献   

14.
Tomato black ring virus (TBRV) is a worldwide-distributed RNA virus infecting a wide range of different host plants, including crop species, trees, shrubs, and weeds. Here, we investigated the molecular evolution of TBRV and its adaptability to different plant species. The TBRV-Pi isolate was used to generate five independent evolution lineages serially passaged in either quinoa, tobacco, or tomato plants. After 15 passages, the genetic variability present in all the lineages was characterized for the movement (MP) and coat (CP) coding cistrons. We addressed two main questions: to what extent does the amount of genetic variability in the TBRV genome depend on the host species, and are there host species-specific adaptive mutations? Overall, 201 different nucleotide substitutions emerged during the evolution experiment, with some of them appearing multiple times in different lineages; two of them (one in CP and one in MP) were unique for a particular host plant. We have shown that the degree of genetic differentiation depends on the host species in which the virus evolved, and that positive selection is operating upon certain residues, particularly in CP. Moreover, we have characterized new types of defective RNAs that arose during the TBRV-Pi evolution in tobacco. Furthermore, this is the first report of a defective RNA from the RNA2 of TBRV.  相似文献   

15.
Pepper yellow mosaic virus (PepYMV) is the most important potyvirus infecting sweet pepper in Brazil. In this study, twenty isolates of PepYMV were obtained from commercial sweet pepper crops. To confirm virus identity, the coat protein gene was completely sequenced for eleven of these isolates, and partially sequenced for the other nine isolates. The amino acid identities obtained were above 93% when compared with the sequence of a characterized PepYMV isolate (AF348610). Extracts of Nicotiana tabacum cv. TNN plants infected with the different isolates were used to inoculate the differential series of Capsicum spp cultivars containing the genes pvr2 1 , pvr2 2 , pvr2 3 , pvr2 4 , and Pvr4. Using the same criteria established for Potato virus Y (PVY), fourteen isolates of PepYMV could be classified as known pathotypes described for PVY, that is: 1.2 (2 isolates), 1.3 (6) and 1.2.3 (6). The remaining six isolates, 1.3 (2) and 1.2.3 (4) could not be classified into the typical pathotypes of PVY because they were also virulent on Serrano Criollo de Morellos—334 (C.M 334) which carries the pvr2 3 and Pvr4 genes. To classify the PepYMV into pathotypes and counter the biological diversity found in this species we propose the utilization of 2x for the ability to overcome the correspondent allele of the pvr2 locus and 4 for the capacity to break down the Pvr4 gene. Using this criterion we could classify the PepYMV into five pathotypes: 21.22; 21.23; 21.22.23; 21.23. 4 and 21.22.23. 4.  相似文献   

16.
Wild type (WT) field isolates of Bremia lactucae failed to germinate in vitro or infect lettuce leaves in the presence of CAA (carboxylic acid amide) fungicides. Minimal inhibitory concentrations (MIC) for mandipropamid, dimethomorph, benthiavalicarb and iprovalicarb were 0.005, 0.5, 0.5 and 5 μg ml−1, respectively. Mutagenesis experiments showed that spores exposed to EMS (ethyl methane sulphonate) or UV irradiation (254 nm) could infect lettuce leaves in the presence of up to 100 μg ml−1 CAA. The proportion of infected leaves relative to the number of spores inoculated (infection frequency) was inversely related to the concentration of CAA used, ranging between 0 and 160 per 1 × 106 spores. Resistant mutants (RM) lost their resistance within 1–14 reproduction cycles on CAA-treated plants. Crosses were made between RMxWT isolates and RMxRM isolates with an attempt to obtain stable homozygous resistant off-springs. Such crosses yielded few resistant but unstable progeny isolates. Mutagenic treatments given to hybrid isolates also failed to produce stable resistance. Previous gene sequencing data showed that stable resistance to CAAs is based on a single SNP in the cellulose synthase 3 (CesA3) gene of Plasmopara viticola. Therefore, we sequenced a 582 bp DNA fragment of Ces3A of WT, RM and hybrid isolates of B.lactucae. No mutation in this gene fragment was found. We conclude that mutagenic agents like EMS or UV may induce resistance to CAA in Bremia lactucae but this resistance is not stable and not linked to mutations in CesA3 gene.  相似文献   

17.
Four hundred and sixty-one isolates of Blumeria graminis f.sp. hordei were obtained from eight populations occurring on cultivated barley (Hordeum vulgare) at four geographically distant locations in China during 2003 and 2004. Their virulence frequency was determined on 30 differential lines. No isolate was virulent on differential lines possessing the resistance genes Mla1, Mla3, Mla6, Mla7, Mla9, Mla12, Mla13, Mlat, Mlg, Mla10, Mla22, Mla23, Mlp1, Ml(N81) and Mlmw. Virulences to the first nine resistance genes are prevalent in Europe and constitute the main part of genetic distance between Chinese and European populations. Conversely, no isolate was avirulent on the differential lines possessing the genes Mla8 and Ml(Ch). The frequencies of isolates overcoming the genes Mla2, Mla11, Mlk1 and Mlk2 were .4–9.3%, and frequencies of isolates overcoming the genes Mlh, MlLa, Ml(Bw), Mlra, Ml(Ru2), mlw, MlGa, MlWo and Mlnn ranged from 18.2% to 98.7%. Based on reactions of differential lines possessing the genes Mlk1, Mlh, MlLa, Ml(Bw), Mlra and Ml(Ru2), pathotypes were identified and diversity parameters calculated. Eleven of 22 detected pathotypes were found in both years and comprised 94.6% of isolates. Generally, the populations from different locations in 1 year were more closely related than populations collected from the same locations in different years. Complete effectiveness of the resistance genes, for which no corresponding virulences were found, will allow Chinese breeders to access many modern European barley cultivars that are fully resistant to powdery mildew in China, including those possessing the non-host resistance gene mlo.  相似文献   

18.
The greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) is occasionally found in beehives and is a major pest of stored wax. Entomopathogenic fungi have recently received attention as possible biocontrol elements for certain insect pests. In this study, 90 isolates of Beauveria bassiana and 15 isolates of Metarhizium anisopliae were screened for proteases and lipases production. The results showed significant variations in the enzymatic action between the isolates. In the bioassay, the selected isolates evinced high virulence against the 4th instar of the G. mellonella larvae. The isolates BbaAUMC3076, BbaAUMC3263 and ManAUMC3085 realized 100% mortality at concentrations of 5.5 × 106 conidia ml−1, 5.86 × 105 conidia ml−1, and 4.8 × 106 conidia ml−1, respectively. Strong enzymatic activities in vitro did not necessarily indicate high virulence against the tested insect pest. The cuticle of the infected larvae became dark and black-spotted, indicating direct attack of fungus on the defense system of the insects. The LC50 values were 1.43 × 103, 1.04 × 105 and 5.06 × 104 for Bba3263AUMC, Bba3076AUMC and Man3085AUMC, respectively, and their slopes were determined by computerized probit analysis program as 0.738 ± 0.008, 0.635 ± 0.007 and 1.120 ± 0.024, respectively.  相似文献   

19.
Pythium and Phytophthora species were isolated from kalanchoe plants with root and stem rots. Phytophthora isolates were identified as Phytophthora nicotianae on the basis of morphological characteristics and restriction fragment length polymorphism (RFLP) analysis of the rDNA-internal transcribed spacer regions. Similarly, the Pythium isolates were identified as Pythium myriotylum and Pythium helicoides. In pathogenicity tests, isolates of the three species caused root and stem rots. Disease severity caused by the Pythium spp. and Ph. nicotianae was the greatest at 35°–40°C and 30°–40°C, respectively. Ph. nicotianae induced stem rot at two different relative humidities (60% and >95%) at 30°C. P. myriotylum and P. helicoides caused root and stem rots at high humidity (>95%), but only root rot at low humidity (60%).  相似文献   

20.
The genetics of host-pathogen interactions in the Hordeum vulgare – P. teres f. teres pathosystem was studied in twelve resistant barley accessions, i.e. CI 9825, CI 9819, Diamond, CI 4922, CI 5401, Harbin, c-8755, c-21849, c-8721 c-23874, c-19979, c-15811. F2 analyses of crosses with susceptible genotypes employing various isolates (from Europe, USA, Canada, and Australia) revealed that resistance is mostly isolate-specific and controlled by one or two genes. Segregation in ascospore progeny from two crosses between isolates of different origin revealed that avirulence in P. teres is also determined by one or two genes. An epistatic effect of suppressor genes on avirulence genes is proposed for the genetics of virulence to Diamond, Harbin, CI 5401 and c-8721 in the fungal crosses D (181-6 × A80) and F (H-22 × 92-178/9). Segregation in F2 of crosses of three new sources of resistance (c-23874, c-19979, c-15811) to the susceptible cv. Pirkka was studied in laboratory and greenhouse tests by using seven P. teres isolates, i.e. 181-6, d8-3, d8-4, d9-1, d9-4, F4 and F74. In addition, virulence to these barley accessions of ascospore progeny from crosses of the same isolates was studied. Based on these studies it was concluded that depending on the isolate used, resistance of c-23874 is determined at least by two genes and in c-19979 and c-15811 by three genes. The results of this parallel analyses of genetics of resistance and genetics of virulence allows the postulation of a gene–for–gene interaction in the P. teres – H. vulgare pathosystem.  相似文献   

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