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1.
Y. Elad A. Kapat 《European journal of plant pathology / European Foundation for Plant Pathology》1999,105(2):177-189
The role of protease of Trichoderma harzianum in the biocontrol of Botrytis cinerea was examined. Two isolates of T. harzianum were compared for their ability to produce protease in liquid culture medium and on the surface of bean leaves. The biocontrol agent T. harziaum T39 produced 58mU/ml of protease and T. harzianum NCIM1185 produced 54mU/ml on the 5th day of growth in liquid culture medium. On bean leaves, combinations of B. cinerea and T. harzianum isolates were examined for the synthesis of protease. The protease activities were 0.9 and 0.6mU/ml for T. harzianum T39 and NCIM1185, respectively, and 0.5mU/ml for B. cinerea alone after 48h of incubation. In the presence of T. harzianum T39 culture liquid containing protease, a 55% reduction in B. cinerea germination and a 80% reduction in the germ tube length were observed after 17h of incubation in vitro. When T. harzianum isolates were added to B. cinerea on bean leaves, increased synthesis of protease was observed (1.0 and 1.2mU/ml for T39 and NCIM1185, respectively). In the presence of T. harzianum NCIM1185 protease, although the rate of germination was reduced, B. cinerea attained 98% germination after 17h of incubation. The hydrolytic enzymes produced by B. cinerea, endo-polygalacturonase (PG) and exoPG were partially deactivated by protease from the T. harzianum isolates. Carboxymethyl cellulase was deactivated only by protease of NCIM1185. On the surface of bean leaves, the protease (obtained from liquid culture medium of T. harzianum isolates) resulted in 56–100% reduction of disease severity. The culture liquid containing protease synthesized on the surface of bean leaves treated with B. cinerea and with T. harzianum was collected and added to fresh leaves infected by B. cinerea. There was 56–100% and 30–75% reduction of disease severity with liquid droplet collected from the leaves treated with T. harzianum T39 and NCIM1185, respectively. Increased control of disease was obtained by combining the conidia of T. harzianum isolates with protease obtained from culture media. Protease inhibitors, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), antipain hydrochloride, and a mixture of inhibitors, but not pepstatin A, fully or partially nullified the biocontrol effect of T39. T39 was found to be a poor producer of chitinase and -1,3-glucanase in vitro. These enzymes were not detected on leaves treated with T39. Involvement of protease in biocontrol of B. cinerea is suggested. 相似文献
2.
ABSTRACT Previous studies have shown that the extracellular proteins of Trichoderma harzianum ETS 323 grown in the presence of deactivated Botrytis cinerea in culture include a putative l-amino acid oxidase and have suggested the involvement of this enzyme in the antagonistic mechanism. Here, we hypothesized that the mycoparasitic process of Trichoderma spp. against B. cinerea involves two steps; that is, an initial hyphal coiling stage and a subsequent hyphal coiling stage, with different coiling rates. The two-step antagonism of T. harzianum ETS 323 against B. cinerea during the mycoparasitic process in culture was evaluated using a biexponential equation. In addition, an l-amino acid oxidase (Th-l-AAO) was identified from T. harzianum ETS 323. The secretion of Th-l-AAO was increased when T. harzianum ETS 323 was grown with deactivated hyphae of B. cinerea. Moreover, in vitro assays indicated that Th-l-AAO effectively inhibited B. cinerea hyphal growth, caused cytosolic vacuolization in the hyphae, and led to hyphal lysis. Th-l-AAO also showed disease control against the development of B. cinerea on postharvest apple fruit and tobacco leaves. Furthermore, an apoptosis-like response, including the generation of reactive oxygen species, was observed in B. cinerea after treatment with Th-l-AAO, suggesting that Th-l-AAO triggers programmed cell death in B. cinerea. This may be associated with the two-step antagonism of T. harzianum ETS 323 against B. cinerea. 相似文献
3.
Comparison of Antagonists of Botrytis cinerea in Greenhouse-grown Cucumber and Tomato under Different Climatic Conditions 总被引：1，自引：0，他引：1
A.J. Dik Y. Elad 《European journal of plant pathology / European Foundation for Plant Pathology》1999,105(2):123-137
The efficacy of Trichoderma harzianum T39 and the yeasts Aureobasidium pullulans and Cryptococcus albidus against Botrytis cinerea in cucumber and tomato was compared with chemical control. Four experiments were conducted in cucumber grown under different climatic conditions in The Netherlands, and two experiments were done in tomato both in the Netherlands and in Israel. T. harzianum and A. pullulans showed the most consistent control of B. cinerea, reducing stem lesions and death of plants by 40–100% in most cases. Control of stem lesions and subsequent wilting was generally better than control of symptoms on fruits. In some cases, the biocontrol agents were more effective than the broad-spectrum fungicide tolylfluanid and the selective fungicide iprodione. The climatic conditions did not strongly influence the efficacy of the biocontrol agents, but regression analysis showed that high temperature during the day and high vapour pressure deficit during the night reduced biocontrol efficacy. From the results, prospects for biocontrol of B. cinerea in greenhouse vegetables appear good under a range of conditions. 相似文献
4.
一株具有诱导抗性木霉菌株的筛选及其对黄瓜灰霉病诱导抗性的初步研究 总被引：3，自引：0，他引：3
从植物根部土壤中分离到一株能够诱导黄瓜产生系统抗性的木霉Tr-92,通过形态观察和18SrDNA序列分析对其进行了分类学鉴定,并进行了诱导黄瓜产生系统抗性能力的初步研究.结果表明,菌株Tr-92的菌落形态和显微形态与木霉一致,18SrDNA序列与哈茨木霉的相似性为99％,初步鉴定为哈茨木霉.利用该菌株孢子悬浮液对5叶期黄瓜苗进行根部诱导接种,24 h后灰霉病菌挑战接种,对灰霉病的防效达56％,叶片中抗性相关酶过氧化物酶、多酚氧化酶、β-1,3葡聚糖酶、苯丙氨酸解氨酶、几丁质酶的酶活性显著上升.表明该菌在黄瓜根部灌施可以诱导叶部系统抗性相关酶的活性升高,可为植物病害的生物防治提供适宜的菌种资源. 相似文献
5.
The salicylic acid-dependent defence pathway is effective against different pathogens in tomato and tobacco 总被引：4，自引：0，他引：4
The role of salicylic acid (SA) was investigated in basal defence and induced resistance to powdery mildew ( Oidium neolycopersici ) and grey mould ( Botrytis cinerea ) in tomato ( Lycopersicon esculentum ) and tobacco ( Nicotiana tabacum ). A comparison of NahG transgenic tomato and tobacco (unable to accumulate SA) to their respective wild types revealed that in tomato, SA was not involved in basal defence against O. neolycopersici but NahG tobacco showed an enhanced susceptibility to O. neolycopersici infection, the effect becoming more obvious as the plants grew older. In contrast, SA played no role in the basal defence of tobacco against B. cinerea , but seemed to contribute to basal defence of tomato against B. cinerea. Activation of the SA-dependent defence pathway via benzothiadiazole (BTH) resulted in induced resistance against O. neolycopersici in tobacco but not in tomato. Microscopic analysis revealed that BTH treatment could prevent penetration of the Oidium germ tube through tobacco leaves, whereas penetration was successful on tomato leaves irrespective of BTH treatment. In contrast, soil or leaf treatment with BTH induced resistance against B. cinerea in tomato but not in tobacco. It is concluded that the SA-dependent defence pathway is effective against different pathogens in tomato and tobacco. 相似文献
6.
ABSTRACT A strategy for integrated biological and chemical control of Botrytis cinerea in nonheated greenhouse vegetables was developed. The biocontrol agent used was a commercial preparation developed from an isolate of Trichoderma harzianum, T39 (Trichodex). Decisions concerning whether to spray the biocontrol agent or a fungicide were made based on a weather-based disease warning system. The integrated strategy (BOTMAN [short for Botrytis manager]) was implemented as follows: when slow or no disease progress was expected, no spraying was needed; when an outbreak of epidemics was expected, use of a chemical fungicide was recommended; in all other cases, application of T. harzianum T39 was recommended. Future weather information (a 4-day weather forecast provided by the Israel Weather Forecast Service) was more useful for disease warnings than immediate past weather. The integrated strategy was compared with weekly applications of fungicide in 11 experiments conducted over 3 years in greenhouse-grown tomato and cucumber. Disease reduction in the integrated strategy (63.9 +/- 3.0%) did not differ significantly (P < 0.05) from the fungicide-only treatment (70.1 +/- 3.6%). The number of fungicide sprays in the integrated strategy ranged from 2 to 7 (mean 4.2) compared to 7 to 13 (mean 10.5) in the fungicide treatment. The integrated strategy averaged 5.9 sprays of T. harzianum T39. For the integrated strategy, one treatment omitted use of T. harzianum T39 to estimate the contribution of this agent to disease control. Disease reduction in that treatment (49.1 +/- 4.8%) was significantly (P < 0.05) inferior to the combined chemical and biological strategy, indicating that the T. harzianum T39 sprays had a measurable effect on disease control. 相似文献
7.
哈茨木霉SH2303防治玉米小斑病的初步研究 总被引：2，自引：0，他引：2
哈茨木霉SH2303是本实验室分离获得的1株具有生防效果的菌株,该菌株能够较好的防治玉米小斑病。通过离体叶片试验确定哈茨木霉SH2303诱导玉米抗小斑病的持效期达15 d。盆栽及大田试验表明,防治玉米小斑病防效分别达到78.1%和56.3%。盆栽试验表明,哈茨木霉 SH2303处理的叶片在挑战接种小斑病菌后第36 h,玉米体内防御反应酶系PAL和SOD活性达到峰值。同时,防御基因Pal和Sod的表达水平也明显上升。综合分析表明,哈茨木霉SH2303的诱导抗性作用是防治玉米小斑病的重要机制之一。 相似文献
8.
9.
Membrane damage caused by the non-specific fungal toxin fusaric acid was less on pretreated than on control leaves when tobacco leaves were pretreated with anti-senescence plant hormones, such as kinetin, benzyladenine or the anti-ozonant N-[2-(2-oxo-1-imidazolidinyl)]ethyl-N'-phenylurea. Similarly, the necrosis caused by mercuric chloride was reduced by the above anti-senescence agents. In addition, in in vitro tests, leaves from selected paraquat-tolerant tobacco plants were less sensitive to Alternaria alternata (Fr) Keissler infection than those of the control paraquat-sensitive tobacco leaves. Paraquat-tolerant Conyza canadensis (L) Cronq weeds naturally selected in vineyards in Hungary showed similar inhibition of senescence to paraquat-tolerant tobacco, expressed as more green leaves and slower development. In accordance with this, the paraquat-tolerant Conyza leaves remained almost symptomless, while paraquat-sensitive plants showed severe symptoms after infection with Botrytis cinerea Pers. Oxidative burst (accumulation of hydrogen peroxide) was attenuated in TMV-infected leaves of Xanthi-nc tobacco as a result of treatment with salicylic acid or in leaves where systemic acquired resistance (SAR) had been induced by a previous TMV infection. Accordingly, superoxide dismutase (SOD) activity was higher in Xanthi tobacco leaves with SAR than without SAR. However, in NahG tobacco, in which SAR cannot develop, there was no augmentation of SOD activity. All the above data support the significance of delayed senescence and antioxidants in the resistance of plants to biotic and abiotic necrosis-inducing agents. 相似文献
10.
为验证哈茨木霉菌与啶酰菌胺联用对番茄灰霉病菌的增效作用,采用显微镜观察法观察了菌药联用后哈茨木霉菌和番茄灰霉病菌的菌丝形态,用菌丝生长速率法和圆盘滤膜法测定了菌药联用后哈茨木霉菌挥发性物质、非挥发性物质和粗酶液对番茄灰霉病菌的抑菌活性.结果显示:啶酰菌胺对哈茨木霉菌的菌丝基本无影响,但能使灰霉病菌的菌丝发生间断性溶解;菌药联用后哈茨木霉菌挥发性物质对灰霉病菌的最高抑菌活性为61.17％,而哈茨木霉菌和啶酰菌胺单独对灰霉病菌的抑菌活性分别为48.79％、0.97％,表现增效作用;菌药联用后哈茨木霉菌非挥发性物质和粗酶液对番茄灰霉病菌的抑菌活性未明显增强.研究表明,啶酰菌胺与哈茨木霉菌联用的增效机制是啶酰菌胺导致灰霉病菌的菌丝部分溶解,同时增强了哈茨木霉菌挥发性物质对灰霉病菌的抑菌活性. 相似文献
11.
Prior infection of faba bean with the viruses bean yellow mosaic and bean leaf roll increased host susceptibility to subsequent infection by Botrytis fabae and B. cinerea. Cell necrosis beneath inoculum droplets, rate and extent of lesion spread and sporulation of B. fabae were all increased on detached leaves from virus-infected compared with healthy plants. Changes were most marked in young leaves showing conspicuous symptoms of systemic virus infection and in plants virus-infected for at least 2-4 weeks. Localized lesions produced by B. cinerea or a low concentration of B. fabae conidia (103 spores/ ml) showed increased cell necrosis but were not transformed into aggressive, spreading lesions on virus-infected leaves. 相似文献
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13.
Biosuppression of Botrytis cinerea in grapes 总被引：2，自引：0，他引：2
There is increasing interest in the use of biological control agents (BCAs) and plant resistance stimulants to suppress botrytis bunch rot in grapes, caused by Botrytis cinerea . Numerous different filamentous fungi, bacteria and yeasts have been selected as potential BCAs for control of grey mould based upon demonstrated antagonism towards B. cinerea. Biological suppression of the pathogen arises via competition for nutrients and space, the production of inhibitory metabolites and/or parasitism. Preformed and inducible grapevine defence mechanisms also contribute to disease suppression by preventing or delaying pathogenic infection. Furthermore, various biotic and abiotic agents can stimulate grapevine defence mechanisms and so elevate resistance to B. cinerea infection. Biosuppression of B. cinerea in vineyards, using BCAs and resistance stimulants, has been inconsistent when compared with that observed in controlled glasshouse or laboratory conditions. This may be attributable, in part, to the innate variability of the field environment. Research to improve field efficacy has focused on formulation improvement, the use of BCA mixtures and combinational approaches involving BCAs and plant resistance stimulants with complementary modes of action. 相似文献
14.
In a previous study, we isolated a new harpin protein, PopW, from the bacterium Ralstonia solanacearum ZJ3721 that can induce a hypersensitive response in tobacco, Nicotiana tabacum, leaves. In the current study, we demonstrate that, in a greenhouse experiment, PopW induced tobacco-acquired resistance against the Tobacco mosaic virus (TMV) with a biocontrol efficacy of 80.9 to 97.4% at a concentration as low as 25 μg/ml in both PopW-treated and neighboring leaves. The resistance induced by PopW is systemic acquired resistance mediated by salicylic acid, which was certified by the development of resistance being accompanied by the expression of the pathogenesis-related-1 gene (PR1) 8 h after PopW was sprayed onto the tobacco leaves. In addition, hydrogen peroxide began to accumulate 10 h after PopW spraying, peaking at 24 h with a maximum concentration of 1.97 μM/g fresh weight. The activities of phenylalanine ammonia lyase (EC4.3.1.5), polyphenoloxidase (EC1.14.18.1), and peroxidase (EC1.11.1.7) also increased, peaking at different times in the PopW-treated tobacco leaves. PopW also reduced the level of TMV disease in field trials with a biocontrol efficacy of 45.2%. Furthermore, PopW both increased tobacco yield (by 30.4 more than in control plants) and improved tobacco foliar quality, with an increase of 50.2% in the number of first-class tobacco leaves from treated compared with untreated plants. All of these results indicate that the new harpin protein PopW has the potential to be an effective biocontrol agent against TMV in tobacco. 相似文献
15.
The plant hormone ethylene regulates fruit ripening, other developmental processes, and a subset of defense responses. Here, we show that 1-aminocyclopropane-1-carboxylic acid synthase (ACS)-silenced apple (Malus domestica) fruit that express a sense construct of ACS were more susceptible to Botrytis cinerea than untransformed apple, demonstrating that ethylene strengthens fruit resistance to B. cinerea infection. Because ethylene response factors (ERFs) are known to contribute to resistance against B. cinerea via the ethylene-signaling pathway, we cloned four ERF cDNAs from fruit of M. domestica: MdERF3, -4, -5, and -6. Expression of all four MdERF mRNAs was ethylene dependent and induced by wounding or by B. cinerea infection. B. cinerea infection suppressed rapid induction of wound-related MdERF expression. MdERF3 was the only mRNA induced by wounding and B. cinerea infection in ACS-suppressed apple fruit, although its induction was reduced compared with wild-type apple. Promoter regions of all four MdERF genes were cloned and putative cis-elements were identified in each promoter. Transient expression of MdERF3 in tobacco increased expression of the GCC-box containing gene chitinase 48. 相似文献
16.
ABSTRACT The effect of preliminary nutrient activation on the ability of conidia of the antagonist Trichoderma harzianum (atroviride) P1 to suppress Botrytis cinerea was investigated in laboratory, greenhouse, and field trials. Preliminary nutrient activation at 21 degrees C accelerated subsequent germination of the antagonist at temperatures from 9 to 21 degrees C; at >/=18 degrees C, the germination time of preactivated T. harzianum P1 conidia did not differ significantly from that of B. cinerea. When coinoculated with B. cinerea, concentrated inocula of preactivated but ungerminated T. harzianum P1 conidia reduced in vitro germination of the pathogen by >/=87% at 12 to 25 degrees C; initially quiescent conidia achieved this level of suppression only at 25 degrees C. Application of quiescent T. harzianum P1 conidia to detached strawberry flowers in moist chambers reduced infection by B. cinerea by >/=85% at 24 degrees C, but only by 35% at 12 degrees C. Preactivated conidia reduced infection by >/=60% at 12 degrees C. Both quiescent and preactivated conidia significantly reduced latent infection in greenhouse-grown strawberries at a mean temperature of 19 degrees C, whereas only preactivated conidia were effective in the field at a mean temperature of 14 degrees C on the day of treatment application. An antagonistic mechanism based on initiation of germination in sufficiently concentrated inocula suggests that at suboptimal temperatures the efficacy of Trichoderma antagonists might be improved by conidia activation prior to application. 相似文献
17.
木霉菌REMI转化体对番茄灰霉病的防治及其机理的研究 总被引：3，自引：1，他引：2
研究不同木霉菌转化体对番茄灰霉病防治效果及机理,为木霉菌生物防治的合理利用奠定基础。利用限制性内切酶介导基因整合技术(restriction enzyme-mediated integration,REMI),通过插入线性化质粒DNA获得了生物防治番茄灰霉病(Botrytis cinerea)效果优于出发菌T21菌株(出发菌)的3个木霉菌转化体Ttrm31、Ttrm34和Ttrm55,对侵染花器和叶片的灰霉病防效分别比原生物防治木霉菌株提高了16.9%和8%。木霉菌转化体的产孢能力、分生孢子的萌发率、对碳氮源的利用能力及对高温的抵抗能力都有所提高;木霉菌转化体本身产生的几丁质酶和β-1,3-葡聚糖酶的活性均比出发菌高,因此通过REMI技术可以获得新的有益木霉菌转化体,在一定程度上提高了生物菌株防治番茄灰霉病的水平。说明REMI技术可以用于改良生防木霉菌株的功能,提高生物防治效果。 相似文献
18.
D. Shtienberg Y. Elad Ariela Niv Y. Nitzani B. Kirshner 《European journal of plant pathology / European Foundation for Plant Pathology》1998,104(8):753-763
The most serious symptom of Botrytis cinerea in tomatoes grown in greenhouses is stem rotting. Lesions on the stem may result from direct infection or from progression of the rot along infected leaves, until infection approaches the stem. In a set of experiments conducted in commercial greenhouses, an experimental greenhouse and growth chambers, the significance of the two types of stem infections was studied. In non-heated greenhouses most of the stem lesions originated from progression of the pathogen along infected petioles. The rate at which B. cinerea had progressed on infected petioles was 0.3–0.5 cm/day, an average of ca. 6 weeks was needed for a leaf infection to approach the stem. Application of Trichoderma harzianum T39 extended this time by 1–2 weeks and application of chemical fungicides by 3 weeks. Influence of the environment on the progression of B. cinerea along infected petioles was then determined. Within range of 5–30 °C, the higher the temperature, the more rapid was the rate of disease progression. The fungus progressed more rapidly on tomato petioles incubated at high vapour pressure deficit (VPD) rather than at low VPD. The source-sink relationship of the plant governed the rate of B. cinerea progression along the petioles as well: it was more rapid when the source was restricted (by shading) and slower when the sink was restricted (by removal of flowers and small fruits). The possibility that sanitation of infected leaflets would reduce the incidence of stem rotting was examined in two experiments. In plots not treated with a fungicide, the sanitation treatment substantially decreased the incidence of stem lesions and this treatment was as effective as weekly application of chemical fungicides. 相似文献
19.
病原菌与植物互作过程中分泌的细胞壁降解酶具有致病和激发植物免疫的双重功能。BcGs1是从灰葡萄孢菌代谢物中分离的葡聚糖-1,4-α糖苷酶,能激发番茄和烟草的免疫防御反应,提高抗病性。BcGs1包含糖苷水解酶家族15(GH15)和糖结合模块家族20(CBM20)两个结构域,其激发植物防御反应的作用方式及功能结构域尚不清楚。本文测定了BcGs1不同诱导时间对提高番茄抗灰葡萄孢菌的效果、体外糖苷酶活性,并比较了BcGs1不同结构域截短蛋白与全长蛋白引起细胞坏死活性的差异。结果表明,BcGs1蛋白诱导番茄72 h可将灰葡萄孢菌引起的病斑面积减少23.25%;BcGs1能水解淀粉和海带多糖,但不能水解羟甲基纤维素和微晶纤维素;BcGs1全长、GH15和CBM20截短蛋白均能在烟草细胞中正常瞬时表达,但表达GH15的烟草叶片枯斑面积明显小于表达全长蛋白BcGs1的枯斑面积,表达CBM20截短蛋白的叶片没有产生枯斑反应;为了进一步明确截短蛋白GH15和全长蛋白BcGs1激发烟草细胞坏死活性的差异,用大肠杆菌表达并获得了纯化的重组蛋白GH15,叶片浸润法测定结果表明,截断蛋白GH15的细胞坏死活性仍然低于BcGs1全长蛋白的坏死活性,说明BcGs1诱导烟草细胞坏死活性需要GH15和CBM20结构域的共同参与。本文研究结果为进一步探讨BcGs1诱导植物抗病机制奠定了基础。 相似文献
20.
Bacteria such as Pantoea agglomerans (Pa-AF2), Bacillus subtilis (Bs-271), Acinetobacter lwoffii (Al-113), and Pseudomonas fluorescens (Pf-CT2), originating from the vineyard, can induce defense responses and enhance resistance of grapevine against the fungal pathogen Botrytis cinerea. The perception of these bacteria by plant cells or tissues in relation to their activities remains unknown. In this study, we examined the relationships between the activity of each bacterium to induce or prime some defense responses, and its effectiveness to induce resistance in grapevine against B. cinerea. We showed that all selected bacteria are capable of inducing early oxidative burst and phytoalexin (trans-resveratrol and trans-ε-viniferin) production in grapevine cells and leaves. Pf-CT2 and Al-113 induced higher H(2)O(2) and trans-resveratrol accumulations, and were able to further prime plants for accelerated phytoalexin production after B. cinerea challenge. These two bacteria were also the most effective in inducing local and systemic resistance. A similar level of induced resistance was observed with live Pa-AF2 which also induced but not primed a greater accumulation of trans-resveratrol. However, Bs-271, which was less effective in inducing resistance, induced a lower trans-resveratrol synthesis, without priming activity. Treatment of grapevine cells with growing medium or crude extract of the bacteria quickly and strongly enhanced oxidative burst compared with the live bacteria. However, both treatments resulted in comparable amounts of phytoalexins and induced local and systemic resistance to B. cinerea as compared with those induced by living bacteria, with extracts from Pf-CT2 and Al-113 being the most effective. Together, these results indicate that induced resistance can be improved by treatment with bacteria or derived compounds which induced or primed plants for enhanced phytoalexin accumulation. 相似文献