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1.
Verticillium wilt caused by Verticillium dahliae is one of the most threatening diseases of olive worldwide. For pre‐planting and post‐planting control of verticillium wilt in olive trees, availability of a rapid, reliable and non‐destructive method for detection of V. dahliae is essential. For such a method, suitable and easily performed sampling and efficient processing of samples for extraction of DNA are necessary. In this study, the suitability of young twig and leaf samples of olive trees, which are easy to collect and extract DNA from, were assessed for the detection of V. dahliae in routine procedures. The lower (about 50 cm from the tip) and top parts (about 5 cm from the tip) of twigs, as well as leaves from infected olive trees were screened for V. dahliae infection and distribution using real‐time PCR. The biomass of V. dahliae detected in individual twigs was highly variable, but there was no significant difference between mean quantities of V. dahliae DNA detected in top and lower parts of twigs. Furthermore, it was demonstrated that analysis of combined samples containing DNA extracted from five twigs of an infected tree accurately detected the presence of the pathogen. Similarly, testing combined samples of 5–10 leaves enabled reliable detection of the pathogen in an infected tree. The development of this assay enables reliable detection of V. dahliae in infected olive trees that can aid in management decisions for the implementation of integrated disease management.  相似文献   

2.
Natural recovery from wilt disease symptoms was evaluated in young olive trees root dip inoculated with Verticillium dahliae in a growth chamber over a 12 week period and, later on, when the trees were transplanted in a V. dahliae-free soil in a lathhouse during a period of 127 weeks. Recovery in an individual tree was considered when a plant showed symptom remission after having reached a maximum value of symptom severity. Recovery accounted for 53% of 464 trees that showed wilt symptoms during observations in the two environments. The remaining trees died. Recurrent wilt symptoms were not observed in recovered trees, and recovery was usually accompanied by the production of new green tissues. Recovery was clearly higher in trees inoculated with a non-defoliating (ND) isolate (86.4%) of the pathogen than in those inoculated with a defoliating (D) isolate (23.9%). The percentage of recovery and the level of resistance were significantly correlated. Recovery accounted for 92.1% of the cases in resistant and moderately susceptible cultivars, reaching 100% in plants inoculated with the ND isolate (Table 2); meanwhile it was three times lower (30.1% of the plants) in susceptible and extremely susceptible diseased trees. In the lathhouse, periodical tissue isolations for monitoring the progress of infections over a period of 127 weeks in recovered trees, showed that the pathogen could only be isolated from trees 19 weeks after inoculation. Pathogen isolation was significantly higher from susceptible and extremely susceptible cultivars (84.6%) than from resistant and moderately susceptible ones (33.3%). Results showed that if a tree overcomes infection by pathogen from a single inoculation, and it is able to begin a recovery process, it will not express wilt symptoms again in a pathogen-free environment. The pathogen remained inactive or dead over time in recovered trees. Thus, new infections from rootlets would be necessary for new symptom expression. Recovery from Verticillium wilt is an important natural mechanism that occurs in a high percentage of infected olive trees, and can complement the resistance of the cultivar, particularly in conditions of low inoculum densities of low virulence isolates of the pathogen in the soil.  相似文献   

3.
Talaromyces flavus, a fungal antagonist of Verticillium dahliae, naturally occurring in clay loam artichoke fields or sandy loam olive groves, is able to survive following application of soil solarization. Survival was almost always linked to an increase in T. flavus populations detected in the rhizosphere of artichoke plants or olive trees with a verticillium wilt history as compared with the untreated control soils. It was evident that soil solarization resulted in the control of the disease in artichoke fields and the recovery of olive trees from V. dahliae infection. It was furthermore proved that solarization had a beneficial long-term effect in controlling V. dahliae for a period of 2 or 3 consecutive years. This could at least partially be attributed to the activity of T. flavus in inhibiting the germination of microsclerotia or causing their death. Aspergillus terreus, another potential V. dahliae antagonist, was also found to survive and occasionally increase following the application of the technique.  相似文献   

4.
A study was conducted during 2 years to determine the effects of three different control measures on the development of Verticillium wilt on olive trees cv. ‘Nabali’ at Al-Hallabat, Jordan. The causal agent of the wilt wasVerticillium dahliae Kleb. Treating diseased trees with Cryptonol (8-hydroxyquinoline sulfate) in a soil drench, or covering trees with a solar chamber for 15 days, was effective in suppressing disease development. The fertilizer treatment (NPK, 15:15:30) decreased disease severity and percent infection. All decreases are in comparison with the untreated control, and as recorded during the active phase of the pathogen. The treatments did not differ significantly from each other, and disease incidence in treated trees remained lower than in the control throughout the examined period.  相似文献   

5.
Verticillium wilt can cause high losses in tree nurseries. To be able to predict disease and unravel disease dynamics over time and space, the relationship between verticillium wilt and soil inoculum densities of Verticillium dahliae and the nematode Pratylenchus fallax was studied in two 4-year field experiments with Acer platanoides and Catalpa bignonioides in the Netherlands. Best-fit regression equations showed that pre-planting inoculum densities of V. dahliae can be used to predict verticillium wilt over a period of at least 4 years. Pratylenchus fallax contributed significantly to disease severity in A. platanoides in some years. Disease can already occur at the detection limit of the pathogens. The 5% infection thresholds for V. dahliae were at 1 (A. platanoides) vs. 3 (C. bignonioides) colony-forming units (CFU) g−1 soil. Analysis of spatial relationships indicated that diseased plants had a higher influence on neighbouring plants at low V. dahliae inoculum densities (<5 CFU g−1 soil) than at high densities (≥5 CFU g−1 soil). Seventy-four percent of the diseased plants recovered during the following year. After that year, recovered plants had a significantly higher probability of becoming diseased again than plants that were healthy during the two previous years, at high inoculum densities of V. dahliae, indicating that inoculum density in the soil, rather than incomplete recovery, was the most important factor for disease development.  相似文献   

6.
In a study of vegetative compatibility in Verticillium dahliae in the Netherlands, a collection of 45 isolates including representatives from woody hosts, several horticultural crops and from the soil of potato fields was examined. In addition an effort was made to compare vegetative compatibility groups (VCGs) from different countries. The results of this study indicate that VCG diversity in V. dahliae in the Netherlands is limited. Only two VCGs were detected: VCG NL-I and VCG NL-II. The former is the predominant VCG for isolates from tree hosts. However, Verticillium wilt in trees can be caused by isolates from both VCGs. It is suggested that the predominance of VCG NL-I in tree hosts is the result of the origin of the tree and the cropping history of its growing site, rather than trees being preferential hosts for isolates from this VCG. Comparison of VCG testers from the Netherlands, from several other European countries and from the USA show that in Europe two major VCGs are present. The first one, including NL-I, is compatible with USA VCG 3 and VCG 4, whereas the second one, including NL-II, is compatible with USA VCG 1 and VCG 2. These groups are not completely separated; in some cases, testers formed heterokaryons with VCG testers from both main groups. Because of the presence of these bridge isolates and because mutants from the same isolate differ in ability to form heterokaryons, it is emphasised that careful selection of isolate testers is an essential step to get a clear picture of VCG diversity.  相似文献   

7.
Powdery mildew is an important disease of rubber trees worldwide. To assess the effects of temperature and leaf age on conidial germination and disease development, conidia were inoculated onto rubber tree seedlings with leaves at three phenological stages (copper bronze, colour-changing, and light green) and then incubated at six constant temperatures (10, 15, 20, 25, 30, and 35°C). Leaf age did not affect conidial germination (p = .296) whilst temperature did (p < .0001), although conidia were able to germinate at all tested temperatures. The estimated optimal temperature for conidial germination was 23.2°C. Leaf age, temperature, and their interactions had significant effects on conidial infection and hypha number (p < .0001). At 10 and 35°C, more than 2 and 4 days were needed for infection to complete, respectively, compared to <2, 1, 0.5, and 0.5 days for 15, 20, 25, and 35°C, respectively. Sporulation and mildew symptoms were only observed on those inoculated leaves of all stages at 20 and 25°C, and at the copper bronze stage only at 15°C. The latent period on the copper bronze leaves at 15°C was longer (9 days) than at 20 and 25°C (4 days). The latent period at 20 and 25°C increased from 4 to 7 days as the leaf development stage increased from copper bronze to light green. Therefore, temperature affected germination and postgermination growth of rubber tree powdery mildew, whereas leaf age primarily affected postgermination growth of the pathogen.  相似文献   

8.
Hot water was dripped into the rhizosphere of Japanese pear trees (Pyrus serotina Rehd. grafted on P. betulifolia Bunge.) infested with the white root rot fungus Rosellinia necatrix Prillieux, to destroy the fungus. Isolates of R. necatrix from diseased roots of Japanese pear were vulnerable to water at temperatures above 35°C, and the fungus was eradicated from the colonized substrate when water at 35°C was provided for 3 days. The time required to eradicate R. necatrix decreased exponentially with increasing temperature. Japanese pear trees tolerated a temperature of 45°C without reduction in vigor. Field experiments demonstrated the practical use of hot water drip irrigation (HWD). HWD at 50°C completely destroyed white root rot mycelia on diseased roots, and many rootlets grew after the treatment. HWD at this temperature caused no injury to the trees. HWD of diseased orchard trees was assessed in Takamori and Iida in southern Nagano, Japan. The fungus recurred in two of four trees 28 months after treatment in Takamori and in two of ten trees 16 months after treatment in Iida. The new mycelia emerged on thick roots deep within the soil. Although there is a possibility of recurrence, HWD treatment is a practical control measure for white root rot.  相似文献   

9.
The application of disinfectants through drip irrigation could be a feasible practice against verticillium wilt (Verticillium dahliae) of olive. OX-VIRIN (activated peroxide) and OX-AGUA AL25 (quaternary ammonium compounds) are two disinfectants that have shown efficacy against V. dahliae in irrigation water and potential for reducing the disease in young olive plants. In this work, various post-planting application strategies incorporating OX-VIRIN (once a month, or twice a month on alternate or successive weeks) or OX-AGUA AL25 (once a month, or twice a month on alternate weeks) were assessed for their effect on V. dahliae in soil, disease in olive trees, and olive yield, in a 2-year pot-experiment under natural environmental conditions. The disinfectants were injected via metering pumps into a drip irrigation system that irrigated olive trees planted in V. dahliae-inoculated soil. All the application strategies significantly reduced the total inoculum density in soil compared to controls with no disinfectants and noninoculated soil. The microsclerotia density was also significantly reduced in disinfested soils by 73.6–86.8%, depending on the strategy. The symptoms and infection incidence were always lower in treatments subjected to disinfestation. The treatment with OX-AGUA AL25 applied twice a month on alternate weeks most reduced the symptoms (by 53.0%) and colonization index (by 70.8%) with respect to untreated water control. This soil disinfestation also significantly strengthened the symptom remission. Tree growth and production were negatively affected by soil inoculation (reduced by 45.6% and 88.7%, respectively), but not so by disinfectants, which even relieved the reduction in inoculated soils, especially when OX-AGUA AL25 was applied.  相似文献   

10.
Microcosm studies were carried out to test the survival of Ralstonia solanacearum biovar 2 (race 3) in soil at the permanent wilting point (wp) water content and at field capacity (fc) water content and on various material. Soils were placed at permanent ?5°C, 4°C, 15°C and 20°C and weekly fluctuating ?10/0/+10°C and the material at 5, 15 °C, 20°C with relative humidity (rh) uncontrolled or at constant 10% or 90%. In soil, survival was clearly dependent on temperature independent of water content. At 20°C Ralstonia solanacearum could be reisolated up to 364 days, at 15°C up to 290 days, at 4°C up to 209 days and at fluctuating temperatures (?10/0/+10°C) only up to 18 days. The lower the temperature, the more the population declined. At 15°C and 20°C appr. 107 cfu/g soil were detected after 100 days, whereas at ?5°C only 102 cfu/g soil were detected after only 18 days. The pathogen was longer detectable in sandy-clay loam than in lighter sandy soil. It could be longer reisolated at wilting point and the populations did not decline as rapidly as at field capacity. Ralstonia solanacearum could best survive on material surfaces like rubber, plastic and varnished metal with maximum survival of 40 days at 5°C and 10% rh. In general there is a low risk of Ralstonia solanacearum overwintering under European climatic conditions when the fields are cleared of plant debris and the soil is frozen. Contamined material surfaces pose the risk of pathogen transmission to healthy tubers.  相似文献   

11.
 棉花黄萎病是一种极难防治的土传性真菌病害,研究病原菌侵染棉花的组织学过程对致病机理解析和抗病资源利用具有重要意义。本研究利用绿色荧光蛋白标记的大丽轮枝菌系统研究了其对抗病棉种海岛棉7124和三裂棉、感病棉种军棉1号和戴维逊棉的侵染过程。结果表明,大丽轮枝菌对抗/感棉种的初始侵染没有明显差异,接菌5 h后,分生孢子均能吸附在感病和抗病棉种的根表面。但侵染过程存在显著差异,侵染感病棉种中病原菌3~5 d到达皮层,5~7 d达到维管束,随后迅速扩展繁殖,侵染14 d后即完成系统侵染,并开始产生黄萎病症状;而病原菌侵染抗病棉种,5~7 d才侵入皮层,7~10 d到达维管束,14 d后仍无法扩展,病原菌的定殖与发展受到限制,无法形成系统侵染,较少形成黄萎病症状。本研究通过绿色荧光蛋白标记大丽轮枝菌对抗/感棉种的侵染过程研究,为大丽轮枝菌致病机理研究和抗性资源利用提供了强有力的理论依据。  相似文献   

12.
Agricultural soil samples containing survival structures of the fungal crop pathogens Verticillium dahliae, Sclerotinia sclerotiorum, Sclerotium cepivorum, Pythium ultimum, potato cyst nematodes Globodera rostochiensis and G. pallida and weeds Chenopodium album and Agropyron repens [Elymus repens] were treated in the laboratory with aerated steam at temperatures ranging from 40 to 80°C in a specially constructed apparatus. Steaming at 50 or 60°C for 3min, followed by an 8-min resting period in the steamed soil and immediate removal from the soil thereafter, resulted in 100% kill of all weeds, diseases and nematodes. When steamed at 45°C, there was a small but significant reduction in the survival of V. dahliae microsclerotia but no reduction in survival of S. cepivorum.  相似文献   

13.
Heat treatment of dates at the time of harvest, was examined as an alternative to fumigation with methyl bromide, which has been phased out in Israel under the Montreal Protocol. In laboratory studies, the influence of 40°, 45°, 50° and 55°C on the levels of disinfestation and mortality ofCarpophilus hemipterus larvae was examined over a 2-h exposure period. The ratio of the number of insects found outside the feeding sites to the total number of insects, was greatest at 50°C (92.3%), significantly greater than at 40° and 55°C. At 50° and 55°C, 100% mortality was obtained. Since drying temperatures for most date varieties is between 45° and 50°C, and because percent disinfestation and control was most effective at 50°C, these findings were examined under field conditions. Field trials were carried out at a commercial drying station where the crated dates are placed on the ground inside a plastic-clad hothouse. The stacks were covered with plastic liners to form ‘drying ducts’ through which heated air was sucked using fans positioned at the opposite end of the ducts. It was shown that from 1 to 2 h were required for the dates to reach the set temperature of 50°C. During the following 2-h aeration, the dates were exposed to 50°C heated air, after which an examination of infested dates inserted into the drying ducts, and natural infestations showed that successful control and emigration were obtained. This method produced results comparable to those obtained with methyl bromide fumigation, and was suitable as a replacement technology for infestation control. http://www.phytoparasitica.org posting Nov. 7, 2005.  相似文献   

14.
The behaviour of the herbicide metamitron and of the main transformation product, hydroxy-chlorothalonil (HTI), of the fungicide chlorothalonil was studied to assess the risk of leaching from low-humic sandy soil. The adsorption of metamitron corresponded to a Kom value of about 60 dm3 kg−1 (moderate adsorption). The half-life of metamitron in soil at 15 °C was only three days, presumably due to adaptation of the micro-organisms. In the autumn, the residue of metamitron in the soil profiles corresponded to less than 1% of the cumulative dosage. The half-life of chlorothalonil at 15 °C was about 12 days and about 45% of it was transformed to HTI. The adsorption of HTI to the soils corresponded to a Kom value of 260 dm3 kg−1. The incubation study (15 °C) showed the transformation of HTI in the soils to be very slow. The amounts of HTI remaining in the soil profiles in the autumn corresponded to 4 and 16% of the cumulative dosage of chlorothalonil. In winter, the HTI residue decreased by 40% relative to the autumn level. Occasionally, HTI could be detected in the upper ground-water level (at a depth of about 1 m), at an average concentration of 0.1 to 0.2 µg dm−3. © 1999 Society of Chemical Industry  相似文献   

15.
Real-time PCR was used to detect and quantify Verticillium dahliae and to assess the susceptibility of four Capsicum annuum cultivars (Luesia, Padrón, SCM331 and PI201234) and the Capsicum chinense cv. C118 to this pathogen. The symptoms which developed after infection included stunting and yellowing, and were more acute in the cv. SCM331, which also suffered defoliation in later stages of the disease and in C118, which suffered severe stunting. Quantification of the pathogen DNA in roots 23 and 34 days post-inoculation (dpi) revealed that there were significantly higher amounts of Verticillium dahliae DNA in C118 than in the other cultivars, followed by SCM331, Padrón and PI201234. The lowest amounts of fungal DNA in roots were found in Luesia. In hypocotyls, the highest amounts of fungal DNA were found in SCM331, while Luesia, Padrón and PI201234 had much lower amounts, and C118 had intermediate levels. When a compatible versus an incompatible system was studied, using the near-isogenic tomato lines LA3030 (susceptible) and LA3038 (resistant to V. dahliae), we were able to detect fungal DNA in both lines. As expected, the fungus/plant DNA ratio was lower in LA3038 than in LA3030 and it decreased with time in LA3038. The amount of Verticillium dahliae DNA in the roots of LA3030 remained constant between days 23 and 34 post-inoculation, but increased 10-fold in collars. Finally, when real-time PCR was applied as a diagnostic method to samples from pepper plants, soil and water collected from farms in northwest Spain, we were able to detect V. dahliae DNA in these samples even when symptoms of the disease were not evident.  相似文献   

16.
Verticillium dahliae causes wilt disease of many crops worldwide. Microsclerotia are the main resting structure of V. dahliae in soil and can survive for more than 10 years, serving as an important source of primary inoculum. Mass production of microsclerotia in laboratory is valuable for studying various aspects of V. dahilae, such as its biology, epidemiology and control. We failed to produce a sufficient amount of microsclerotia in vitro for one strain of our interest using several previously published methods. Therefore, we developed a new protocol for mass in vitro production of viable microsclerotia. Verticillium strains were cultured in a modified basal agar medium at pH of 11.5 under 20 °C and incubated for 25 days in dark. When 16 strains were subjected to this condition, large numbers of microsclerotia were produced although varied greatly among strains, including the strain that failed to produce microsclerotia with previously published methods. Microsclerotia from 14 of the 16 strains resulted in wilt development on inoculated cotton seedlings.  相似文献   

17.
Verticillium dahliae colonizes the xylem vessels of susceptible host plants. Hence it can be expected that the distribution of the fungus as well as disease progress will be influenced by the anatomy of the xylem of that host. Here, we studied the spatial and temporal distribution of V. dahliae in relation to recovery from disease symptoms in young European ash (Fraxinus excelsior) and Norway maple (Acer platanoides) trees that differ in vascular anatomy. Quantifying the amount of V. dahliae DNA at different heights in the stem of inoculated trees at different time points after inoculation showed that, in the year of inoculation, the speed of colonization of these two species by V. dahliae was similar. Nevertheless, in the year after inoculation disease incidence and also quantities of V. dahliae detected in maple trees were significantly higher than in ash trees, suggesting that the xylem of ash trees is much less supportive for growth and survival of V. dahliae than that of maple trees. Moreover, in this second year V. dahliae could not be re-isolated from the wood of ash trees that had recovered from disease and was only rarely detected by PCR, only in xylem of the previous year and never in the current xylem. In contrast, V. dahliae easily was detected in the wood of diseased ash and maple trees. Furthermore, despite the presence of a layer of parenchyma cells between growth rings in ash trees, in symptomatic ash trees V. dahliae was present in the xylem of the new growth ring. We also observed that V. dahliae can move downward from the point of inoculation into the root collar, which possibly provides a way for infection of new growth rings by circumventing the physical barriers within the stem xylem.  相似文献   

18.
The growth and survival of three strawberry pathogens, Fusarium oxysporum f. sp. fragariae (FOF), Phytophthora cactorum, and Verticillium dahliae, were examined in anaerobic (anoxic) conditions at several temperatures (10–40 °C). The growth and survival of these fungi were suppressed by anaerobic conditions in comparison with those in cultured aerobically. Under anaerobic conditions at 22.5 °C, tested isolates of FOF and P. cactorum grew slightly, but V. dahliae did not grow. The three fungi survive for markedly shorter time in the anaerobic conditions compared with the aerobic conditions at all tested temperatures except 40 °C for FOF and P. cactorum. Moreover, survival periods shortened as the cultivation temperature increased. These results demonstrate that anaerobic conditions contribute to eradicating these pathogens during flooding or reductive soil disinfestation.  相似文献   

19.

The suppressive effects of soil bio-solarization, which is a new method of soil disinfestation that combines soil bio-fumigation with soil solarization, against the sclerotial viability of Stromatinia cepivora and the subsequent control of white rot disease of onion and garlic were evaluated. Soil was bio-fumigated with fresh amendments of cow manure, chicken manure, horse manure, cruciferous plant residues, or Allium waste, at 30,000 kg/ha. After bio-fumigation, the soil was irrigated and covered with a 200 μm transparent plastic sheet for 60 days. Plots that received fresh amendment and remained uncovered and untreated served as controls. Solarization alone increased the maximum soil temperature to 55.3 °C, 50.3 °C and 46.3 °C at 10, 20, and 30 cm depths, respectively, which led to significant reductions (98.0%, 89.3%, and 62.7%, respectively) in the sclerotial viability of S. cepivora. Soil bio-solarization with cruciferous plant residues or Allium waste resulted in the strongest negative effects on the sclerotial viability of S. cepivora, with reductions of 100.0%, 98.7%, and 87.3% or 100.0%, 99.3%, and 87.7%, at 10, 20, and 30 cm depths, respectively. Compared to the non-treated control, these treatments significantly reduced the incidence of white rot disease in onion and garlic, which led to increases in onion and garlic yield in fields that were heavily infested by S. cepivora.

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20.
E. C. TJAMOS 《EPPO Bulletin》1993,23(3):505-512
Control of verticillium wilt of olive currently depends on preventive measures. Since systemic fungicides are unable to prevent or control the disease, its control should primarily be based on cultural methods, including irrigation systems which restrict dissemination of Verticillium dahliae propagules by irrigation water and avoidance of intercropping with hosts susceptible to V. dahliae. Since leaves from affected olive trees contribute, through formation of microsclerotia, to the inoculum in the soil, pruning should be practised prior to branch defoliation. As for resistant olive rootstocks or cultivars, promising verticillium-wilt resistance has been found in two rootstocks selected in California (US). However, these have to be tested under local conditions before they can be released to Mediterranean growers, while further search for other resistant rootstocks is needed. Soil solarization applied to individual diseased trees in established olive groves could substantially contribute to recovery or long-lasting symptom remission in the treated trees. This effect is mainly attributed to the decrease or eradication of V. dahliae microsclerotia in the treated soil but also to heat-tolerant fungal antagonists of the pathogen. Using herbicides to control weeds, and limiting soil rotovation, can restrict symptom development. Biological control can also be considered as a promising trend in controlling the disease by searching, testing and exploiting potential fungal or bacterial antagonists.  相似文献   

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