Systemic fungicides in the control of strawberry wilt (Verticillium dahliae)

In a series of experiments between 1970 and 1973 the application of benomyl or thiophanate methyl to field-grown strawberries, planted on Verticillium-mtested land, gave control of wilt for up to 5 months, the duration of control being related to the amount of fungicide applied in the spring. Treatment of inoculated plants grown on in chloropicrin-fumigated soil was effective for at least two seasons. An autumn-planted multi-factorial experiment in heavily-infested soil showed that, to achieve maximum wilt control, it was advantageous to grow cv. Gorella rather than cv. Cambridge Vigour; to drench the runners at planting rather than to dip them in the fungicide suspension; and to use a high concentration (0–075 % a-i-) and large volume (600 ml per plant) for a supplementary treatment in May rather than a lower concentration (0.025 %) ^or smaller volume (400 ml). There were small but significant advantages in applying benomyl rather than thiophanate methyl, and in using 0–2% a.i. suspension at planting rather than 0–05%. No advantage was gained by dividing the spring application into two equal doses applied 2 wk apart. Extrapolation from the logarithmic relation between wilt index and total dose of fungicide applied in the spring suggested that I.I g/plant would have given almost complete control until October; such control had been achieved in an earlier experiment in which 1–2 g/plant was applied. Crop yield in the second year was determined by the treatment applied in the first year, but although these treatments had given significantly better control of wilt in Gorella than in Cambridge Vigour, the greater growth and yield potentials of the latter cultivar had an over-riding effect on crop production; Cambridge Vigour yielded more than Gorella under all chemical treatments, but in the absence of treatment Gorella gave a larger crop than Cambridge Vigour. A proposed regime, entailing spring and autumn applications, is aimed at minimizing the colonization of the plant throughout the year, thus reducing the production of new inoculum and, by limiting the quantity of the pathogen in contact with the systemic chemicals, minimizing the probability of selecting fungicide-resistant variants of V. dahliae.     

Control of Verticillium wilt transmission in strawberry runners with systemic benzimidazole-type fungicides

Pre-planting root dips and soil drenches with benomyl, carbendazim (MBC), thiabendazole and thiophanate-methyl were used to reduce disease severity in mother plants. Transmission of the pathogen Verticillium dahliae through stolons and into young runners was prevented by two applications of benomyl or thiophanate-methyl at o-i% active ingredient applied at stolon appearance and 1 month later. Sufficient concentrations of fungistatic residues were detected in all plant tissues to suppress the internal spread of the pathogen.     

Effects of Pratylenchus penetrans on development of strawberry wilt caused by Verticillium dahliae

The nematode Pratylenchus penetrans or the closely related P. fallax occurred in three out of four strawberry plantations infested with Verticillium dahliae surveyed in Kent. When plants of cv. Cambridge Vigour were inoculated with P. penetrans and V. dahliae together in field ‘micro-plots’ the nematode increased the rate of wilt development except when the concentration of V. dahliae microsclerotia in the soil was very low. In a pot experiment, under conditions not conducive to symptom expression, the incidence of infection was increased by nematodes in Cambridge Favourite, Cambridge Vigour and Redgauntlet; the incidence was least in Red-gauntlet, the most resistant cultivar. The nematode appeared to cause local changes in the root cortex which aided hyphal penetration of the adjacent tissues. Growth of strawberry plants in the absence of V. dahliae was not significantly affected even by large populations of the nematode, but growth was reduced by V. dahliae at all rates of inoculation in the field. Infection by Verticillium did not affect the total numbers of nematode per plant at the end of the experiments, although the number per g fresh weight was greater on the small amount of white root on severely diseased plants than on the larger amount on healthy or mildly diseased plants.     

Growth and wilt (Verticillium dahliae) development in strawberry cultivars over a transition between two soil series

On a site infested with V. dahliae the incidence of wilt was low consistently in strawberry cvs Talisman and Redgauntlet, high in Merton Herald and Gorella, and very high in Merton Ruby, Cambridge Vigour, Prizewinner, Templar and Crusader. Changes in mean volume per plant for the whole population of each cultivar were closely related to disease incidence and therefore differed between high-resistance and low-resistance cultivars. Height and spread measurements of diseased and symptomless plants in certain cultivars showed differences in the severity of the effect of the pathogen on growth that were not closely related to wilt incidence. Gradients of disease incidence and plant growth in 1966 and 1967 were associated with a transition from one soil series to another. Review of two earlier experiments on the same site revealed a similar gradient of disease incidence in 1963-64, but a reversed gradient in 1960-62. Variations related to soil conditions were smaller in the earlier experiments than in 1966-67.     

Variation in the virulence of Verticillium dahliae from strawberry

Isolates of V. dahliae from strawberry, tested in 1968 against cv. Cambridge Vigour, showed differences in virulence; in cvs Talisman and Cambridge Favourite there were no significant differences between plants that were inoculated and those that were not. Evidence from this experiment, and from an earlier non-replicated experiment with Cambridge Favourite, in which differences of virulence had been apparent, showed that environmental factors can cause variations in disease development that are of similar magnitude to those attributable to genetic variations in host and parasite.     

Soil inoculum density of Verticillium dahliae and Verticillium wilt of olive in Lebanon

In the Mediterranean basin, Verticillium Wilt of Olive (VWO) is diffused throughout its range of cultivation, causing severe yield losses and tree mortality. The disease was reported in almost all the Mediterranean and Middle East countries, and in Lebanon it is of increasing significance also on many valuable crops. The disease has already been reported on potato, peach and almond in the Bekaa valley; however, to date no information is available about the incidence of VWO and the inoculum density of Verticillium dahliae microsclerotia in soil of the main agricultural areas of Lebanon. Results from the present investigations demonstrate a high V. dahliae frequency in soils (75.3%), coupled with a mean soil inoculum density of 17.0 MS g^?1, clearly indicating a great impact on the production of susceptible hosts in Lebanon, mainly in Bekaa region. Molecular method to assess the microsclerotia inoculum density in soil allowed the detection of a higher frequency of infested soils, as compared with the traditional plating, thus confirming its higher sensitivity. The overall Verticillium wilt prevalence in the inspected olive orchards was 46.2%, and the frequency of V. dahliae‐infected trees was 25.7%. The widespread presence of V. dahliae in all olive growing areas of Lebanon enforces the adoption of measures aimed at reducing the soil inoculum density before any new olive plantation, and the use of strong phytosanitary regulations to improve the certification schemes of propagating material.     

甘肃省甜瓜黄萎病的病原鉴定

【背景】2013年11月在甘肃省兰州市皋兰县的日光温室秋冬茬甜瓜种植棚发现黄萎症状的甜瓜植株,病株率约为1%。【目的】明确甜瓜黄萎病的病原。【方法】采用组织分离法进行病原菌分离;通过科赫氏法则(Koch’s法则)明确分出病菌的致病性;采用形态学和分子生物学方法对病原菌进行种类鉴定。【结果】分离得到轮枝菌属真菌8株,轮枝菌属真菌的病株分出率达100%;2个代表性菌株GLTG-2和GLTG-5(显微特征相似但菌落形态和生长速率不同),在温度18-24℃及昼/夜光周期为11.5 h/12.5 h的试验条件下,人工接种可引起甜瓜苗矮化、枯萎;接种后40 d,枯死株率分别为70%和40%;BLASTn分析结果显示,菌株GLTG-2的rDNA-ITS序列与Verticillium dahliae菌株MRHf7的序列相似性达99.78%,菌株GLTG-5的rDNA-ITS序列与V.dahliae菌株MRHf7和Vd414的序列相似性达100.00%。【结论】引起甜瓜黄萎病的病原菌被鉴定为大丽轮枝菌(V. dahliae),这是大丽轮枝菌引起甜瓜黄萎病在我国和亚洲地区的首次报道。     

Verticillium wilt of cacao in Uganda: the relationship between Verticillium dahliae and cacao roots

Isolates of Verticillium dahliae Kleb. from wilted cacao (Theobroma cacao L.), cotton (Gossypium hirsutum L.), and okra (Abelmoschus esculentus Medik.) penetrated all regions of living cacao tap and lateral roots and progressed intracellularly from the epidermis to the xylem in 4–6 days. The hypocotyl and tissues of the unerupted lateral roots beneath the epidermis resisted invasion. Host reactions included browning of extensively colonized cells, alteration (with apparent granulation) of the cytoplasm, and accumulation of materials in the lumina of endodermal cells. Resistance in the hypocotyl was associated with occasional thickening of inner tangential walls of colonized epidermal cells. The fungus formed conidia, microsclerotia, and narrow and wide hyphae within root tissues. The narrow hyphae predominated at the front of mycelial invasion of tissues while the broad hyphae developed behind this front. Limited studies under non-sterile conditions indicated that the apparent host-parasite interactions were similar to those observed with sterile roots and cultures of V. dahliae.     

Duration of effects of benomyl on growth and wilt (Verticillium dahliae) development in the strawberry, cv. Cambridge Vigour, in relation to time of inoculation

Single doses of benomyl, 0–4 g/plant in 1972 or 0–04 g in 1973, were applied in 100 ml water to the surface of the soil in 12-5 cm pots containing Cambridge Vigour strawberry plants, either before inoculation with Verticillium dahliae or at various times up to 56 days afterwards. Pre-inoculation treatments were terminated by washing the potting medium from the roots at the time of inoculation and their effects on pathogenesis and growth were relatively small. Post-inoculation treatments with benomyl prevented or suspended pathogenesis for at least as long as fungistatic activity could be detected in petiole segments by bioassay; this was for more than 150 days after the larger dose, 50–60 days after the smaller dose applied 7–28 days after inoculation and 30 days when applied 56 days after inoculation. There were no consistent differences in the uptake and persistence of benomyl in inoculated and non-inoculated plants. Early post-inoculation treatment caused some retardation of plant growth, but benomyl-treated inoculated plants were generally comparable in size with similarly treated non-inoculated ones, and much larger than untreated inoculated plants. Increasing the dose of benomyl applied to the soil apparently had little influence on the intensity of its effects but greatly increased their duration, probably because of the low solubility of the systemic chemical.     

Rhamnose synthase activity is required for pathogenicity of the vascular wilt fungus Verticillium dahliae

The initial interaction of a pathogenic fungus with its host is complex and involves numerous metabolic pathways and regulatory proteins. Considerable attention has been devoted to proteins that play a crucial role in these interactions, with an emphasis on so‐called effector molecules that are secreted by the invading microbe to establish the symbiosis. However, the contribution of other types of molecules, such as glycans, is less well appreciated. Here, we present a random genetic screen that enabled us to identify 58 novel candidate genes that are involved in the pathogenic potential of the fungal pathogen Verticillium dahliae, which causes vascular wilt diseases in over 200 dicotyledonous plant species, including economically important crops. One of the candidate genes that was identified concerns a putative biosynthetic gene involved in nucleotide sugar precursor formation, as it encodes a putative nucleotide‐rhamnose synthase/epimerase‐reductase (NRS/ER). This enzyme has homology to bacterial enzymes involved in the biosynthesis of the nucleotide sugar deoxy‐thymidine diphosphate (dTDP)‐rhamnose, a precursor of L‐rhamnose, which has been shown to be required for virulence in several human pathogenic bacteria. Rhamnose is known to be a minor cell wall glycan in fungi and has therefore not been suspected as a crucial molecule in fungal–host interactions. Nevertheless, our study shows that deletion of the VdNRS/ER gene from the V. dahliae genome results in complete loss of pathogenicity on tomato and Nicotiana benthamiana plants, whereas vegetative growth and sporulation are not affected. We demonstrate that VdNRS/ER is a functional enzyme in the biosynthesis of uridine diphosphate (UDP)‐rhamnose, and further analysis has revealed that VdNRS/ER deletion strains are impaired in the colonization of tomato roots. Collectively, our results demonstrate that rhamnose, although only a minor cell wall component, is essential for the pathogenicity of V. dahliae.     

Electrophoretic karyotype and gene mapping of the vascular wilt fungus Verticillium dahliae

The karyotype profile of Verticillium dahliae was resolved by pulse-field gel electrophoresis. It revealed 6 chromosomal bands that corresponded to 7 chromosomes as shown by RFLP analysis using as probe the telomeric consensus sequence (AACCCT)(5). The number of chromosomes was further verified by the sensitivity of the hybridization signals to Bal31 digestion and the exclusion of interfering mitochondrial DNA signals. The corresponding sizes of the seven separated chromosomes were 6.7, 5.6, 4.1, 3.4, 3.1, 3.1 and 2.4Mb, raising the total genomic size of the fungus to approximately 28.4Mb. Twenty five homologous V. dahliae genes obtained either from randomly sequenced clones or PCR amplification were used as hybridization probes and were located onto the seven chromosomes.     

The modes of action of two benzimidazoles and thiophanate-methyl used for the control of Verticillium wilt of strawberry

Benomyl, thiabendazole and thiophanate-methyl were found to be fungistatic against Verticillium dahliae in vitro. All materials suppressed wilt symptoms on artificially inoculated strawberry plants when applied as a root dip, soil drench or a foliar application. Quantitative bioassays of the amount of toxic residues and fungal colonization in tissues of treated plants after root treatment revealed that these materials or their toxic breakdown products were readily translocated to crown and leaves: they suppressed the internal spread of the pathogen throughout the plant. After foliar application there was no marked basipetal movement of material within the plant: fungal colonization was greatly reduced in treated leaves, but the roots and crowns were colonized to a similar extent to those of untreated plants.     

Pathogenicity of benomyl-tolerant variants of Verticillium dahliae to strawberry, cv. Cambridge Vigour

Plants of cv. Cambridge Vigour were inoculated either with a benomyl-sensitive isolate of Verticillium dahliae or with one of two variants selected from it in vitro; these variants, Ri/8 and Ri/ii, were tolerant of benomyl at concentrations of 5 and 10 ppm respectively in the culture medium. The parent isolate and Ri/8 caused only mild wilt symptoms, but Ri/n was strongly pathogenic. A single treatment with benomyl (0–04 g/plant, in 100 ml water), applied as a soil drench o, 14, 29 or 57 days after inoculation, tended to reduce the mild symptoms caused by the parent isolate; the treatments had no apparent effect on Ri/8, but they slightly increased the severity of the wilt caused by Ri/n, especially after the earlier applications.     

Characterization of the glyoxalase I gene from the vascular wilt fungus Verticillium dahliae

A glyoxalase I gene homologue (VdGLO1) was identified in the vascular wilt fungus Verticillium dahliae by sequence tag analysis of genes expressed during resting structure development. The results of the current study show that the gene encodes a putative 345 amino acid protein with high similarity to glyoxalase I, which produces S-D-lactoylglutathione from the toxic metabolic by-product methylglyoxal (MG). Disruption of the V. dahliae gene by Agrobacterium tumefaciens-mediated transformation resulted in enhanced sensitivity to MG. Mycelial growth of disruption mutants was severely reduced in the presence of 5 mmol/L MG. In contrast, spore production in liquid medium was abolished at 1 mmol/L MG, although not at physiologically relevant concentrations of 相似文献   

Proteomic analysis of the sea-island cotton roots infected by wilt pathogen Verticillium dahliae

Verticillium wilt of cotton is a vascular disease mainly caused by the soil-born filamentous fungus Verticillium dahliae. To study the mechanisms associated with defense responses in wilt-resistant sea-island cotton (Gossypium barbadense) upon V. dahliae infection, a comparative proteomic analysis between infected and mock-inoculated roots of G. barbadense var. Hai 7124 (a cultivar showing resistance against V. dahliae) was performed by 2-DE combined with local EST database-assisted PMF and MS/MS analysis. A total of 51 upregulated and 17 downregulated proteins were identified, and these proteins are mainly involved in defense and stress responses, primary and secondary metabolisms, lipid transport, and cytoskeleton organization. Three novel clues regarding wilt resistance of G. barbadense are gained from this study. First, ethylene signaling was significantly activated in the cotton roots attacked by V. dahliae as shown by the elevated expression of ethylene biosynthesis and signaling components. Second, the Bet v 1 family proteins may play an important role in the defense reaction against Verticillium wilt. Third, wilt resistance may implicate the redirection of carbohydrate flux from glycolysis to pentose phosphate pathway (PPP). To our knowledge, this study is the first root proteomic analysis on cotton wilt resistance and provides important insights for establishing strategies to control this disease.     

Molecular analysis of Japanese isolates of Verticillium dahliae and V. albo-airum

Sixteen isolates of different pathogenicity groups of the plant pathogen Verticillium dahliae and four isolates of V. albo-atrum from Japan were analysed by means of an RAPD (random amplified polymorphic DNA) method using a PCR (polymerase chain reaction). Verticillium dahliae and V. albo-atrum could be distinguished by RAPD analysis. Four pathogenicity groups of V. dahliae could also be classified to a certain extent by this method. Similarities and differences in banding patterns obtained by RAPD may be a useful molecular tool in phylogenetic studies of the pathogenicity groups.     

Application of RAPD-PCR for Virulence Type Analysis within Verticillium dahliae and V. longisporum

A collection of 24 isolates of Verticillium dahliae, 11 isolates of V. longisporum and one isolate of V. albo‐atrum originating from different host plants and geographical regions was tested for genetic variability by random amplified polymorphic DNA‐polymerase chain reaction (RAPD–PCR). Based on nine primers, the three Verticillium species could be clearly differentiated. Likewise, this analysis provided a distinct separation of vegetative compatibility groups (VCG) 2B, 4A and 4B of V. dahliae by specific DNA banding patterns. Additionally, V. longisporum was found to segregate into two subgroups with only 88% similarity. This molecular‐genetic approach was used for the analysis of randomly selected Verticillium isolates from a field with high intensity of oilseed rape cultivation (33% in crop rotation). RAPD‐PCR analysis revealed that 95 of 100 isolates tested belonged to V. longisporum and 5 to VCG 2B of V. dahliae. This study demonstrates an adaptation of Verticillium soil populations to a specific cropping history.