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1.
Berto P  Jijakli MH  Lepoivre P 《Phytopathology》2001,91(11):1030-1036
ABSTRACT Ulocladium atrum (strain 385) consistently reduced Botrytis cinerea sporulation on necrotic fragments of strawberry leaves. On these tissues, two strains of U. atrum (isolates 18558 and 18559) showed lower antagonistic activities than the reference strain 385. Colonization of strawberry leaflets by the three U. atrum strains appeared similar in the absence of B. cinerea, whether quantified by chitin or immunological assays. The second method (based on anti-U. atrum antibodies) revealed that strawberry leaflet colonization by U. atrum 385 was better than by the other U. atrum strains in the presence of B. cinerea. An immunoassay using anti-B. cinerea antibodies revealed that the colonization of B. cinerea in tissues was lower in the presence of U. atrum 385 than with the two other U. atrum strains. The enzymatic activities produced by U. atrum 385 during the colonization phases of necrotic tissues were compared to B. cinerea and U. atrum strains 18558 and 18559. U. atrum 385 had the highest lipase, pectate lyase, and cellobiase activities while B. cinerea had the highest endo-beta-1,4-glucanase activity. The study of lytic activities hydrolyzing the fungal cell wall revealed higher beta-1,3-glucanase activity with U. atrum 385, which was stimulated by B. cinerea on necrotic strawberry leaflets. These results suggest that plant and fungal cell wall-degrading enzymes produced by U. atrum 385 may play a complementary role in the competitive colonization of dead strawberry leaves against B. cinerea.  相似文献   

2.
北京地区草莓灰霉病菌的转座子及其分布频率   总被引:1,自引:0,他引:1  
张佳  张晓歌  张璨  张国珍 《植物保护》2016,42(2):177-181
为了解北京地区草莓灰霉病菌的转座子类型及其分布,本研究用转座子Boty和Flipper的特异性引物对北京地区2012-2013年从12个草莓园采集和分离的60株草莓灰霉病菌进行PCR扩增。结果表明,北京地区草莓灰霉病菌群体中共存在3种转座子类型:transposa型、Boty型和Flipper型。其中,以transposa型菌株最多,占供试菌株的63.3%,Boty型菌株占供试菌株的28.3%,Flipper型菌株最少,仅占8.4%,未检测到vacuma型菌株。选取属于不同转座子类型的18株菌株测定其对草莓叶片的致病力,结果显示Boty型菌株所致病斑的平均直径显著大于Flipper型。草莓灰霉病菌转座子类型与致病力的关系有待进一步研究。转座子类型的检测为进一步研究灰葡萄孢的遗传多样性及遗传变异提供了基础。  相似文献   

3.
A serological test has been developed that allows the early detection of infection of young petals by Sclerotinia sclerotiorum , an important pathogen of rapeseed. Two steps were required to obtain an antiserum sufficiently specific for S. sclerotiorum. Soluble mycelial extracts of S. sclerotiorum were used to produce the first generation polyclonal antiserum. This was not specific for S. sclerotiorum in double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and allowed the screening of cross-reacting fungal species such as Botrytis cinerea , a pathogen commonly present on rapeseed petals. Using a polyclonal anti- B. cinerea serum enabled the absorption, by serial cycles, of S. sclerotiorum antigens common to B. cinerea. Residual antigens were then used as immunogens for the production of two second generation antisera (S1 and S2) which were then tested by DAS-ELISA. Cross-reactions with B. cinerea decreased with purification cycles of the immunogen whereas Cross-reactions with some unrelated fungi slightly increased. S. sclerotiorum and B. cinerea were distinguishable using antiserum S2.  相似文献   

4.
5.
Cheng CH  Yang CA  Peng KC 《Phytopathology》2012,102(11):1054-1063
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   

6.
ABSTRACT The effect of 2-deoxy-D-glucose on major postharvest pathogens was investigated at the ultrastructural and cytochemical level. Hyphae of Botrytis cinerea, Penicillium expansum,, and Rhizopus stolonifer grown in the absence of 2-deoxy-D-glucose were normal and showed no apparent cytological alterations. In the presence of 2-deoxy-D-glucose, however, these fungi exhibited severe cellular injuries ranging from cell wall disruption to cytoplasm disintegration. Although 2-deoxy-D-glucose caused cytoplasmic degeneration in the three fungi tested, cell wall alterations were exhibited only by B. cinerea and R. stolonifer. In the latter, the retraction of degenerated cytoplasm was often accompanied by the deposition of amorphous material in paramural spaces. Cytochemical study of fungal cell wall components showed that 2-deoxy-D-glucose caused a marked increase of chitin- and beta-1,3-glucan-labeling in R. stolonifer and B. cinerea, indicating an interference of 2-deoxy-D-glucose with fungal wall biosynthesis. The observed cellular alterations indicate that 2-deoxy-D-glucose may also have affected other metabolic processes.  相似文献   

7.
Monoclonal antibodies have been raised against an haustorium-enriched sample prepared from flax leaves infected with the biotrophic flax rust pathogen Melampsora lini. The monoclonal antibodies were produced following conventional and co-immunisation procedures and the range of antibody specificities was compared. The preparation used as immunogen for the conventional protocol was a crude isolate of haustoria consisting of approx. 65% fungal haustoria, the other components being mainly mesophyll cells or cell wall and chloroplast fragments. Following hybridoma production, 40% of positive cell lines produced antibodies that reacted with haustoria and other fungal cells, but 60% bound to plant cells in the infected leaves. For the co-immunisation protocol, the preparation used for immunisation consisted of the crude isolate of haustoria mixed with serum raised against an haustorium-depleted leaf homogenate. In two fusions, 92-94% of the antibodies reacted with fungal cells, including 3 cell lines that localised specifically to the cell wall of haustoria. Only 6-8% of the antibodies produced via co-immunisation reacted with plant cells. The antigens targeted by the three haustorium-specific monoclonal antibodies are incorporated into the wall at early stages of haustorium development, remain in the wall throughout haustorium maturation, and are present in both compatible and incompatible interactions. The epitopes recognised by the monoclonal antibodies are oligosaccharide in nature and the antigens are highly resistant to extraction from the wall. These results highlight the value of the co-immunisation protocol for the production of monoclonal antibodies to specific components in an impure preparation and provide direct evidence for molecular differentiation within the wall of the haustorium of M. lini.  相似文献   

8.
Cook DW 《Phytopathology》2002,92(12):1293-1299
ABSTRACT A mechanism that could contribute to the suppression of Botrytis cinerea during pathogen sporulation was examined in this study. Yeasts capable of binding to B. cinerea were formulated with a cellulose carrier and applied to sporulating colonies of the pathogen. The particles from this yeast/cellulose product attached to B. cinerea conidia in the sporulating colony. Inoculum from treated colonies was harvested and applied to tomato stem tissue to test for subsequent pathogenicity. Disease development from inoculum obtained from cultures that had been treated with Trichosporon pullulans was significantly retarded (P = 0.0001) compared with cellulose-only controls. However, between 5 and 11% of conidia applied were attached to yeast cells. The removal of conidia not attached to yeast resulted in inoculum composed of >90% of conidia attached to yeast, and from this inoculum, disease development was significantly retarded (P < 0.05). When inoculum from treated B. cinerea colonies was applied to nutrient limiting agar and then incubated, the B. cinerea conidia germinated, and yeast cells infested the new hyphal growth. Constraints of the formulation of the yeast used in this study, and the implications of this vectoring approach for the suppression of B. cinerea during pathogen sporulation are discussed.  相似文献   

9.
为了探明生防酵母菌菌株C410产生的挥发性物质对灰霉菌的抑制作用机理,本文以挥发性物质的主要成分之一环辛四烯为研究对象,测定其对灰霉菌的糖、蛋白、DNA、RNA、细胞的形态及细胞膜通透性的影响。结果表明,在16.5μL/L环辛四烯的作用下,灰霉菌生长速率显著降低,总糖含量基本不变;还原糖含量显著降低;蛋白含量变化不显著;RNA的完整性显著降低。在49.5μL/L环辛四烯的作用下,灰霉菌的生长几乎完全停止,但细胞通透性不受影响,不导致细胞死亡。环辛四烯孵育下灰霉菌DNA分子没有被打断。由此推测还原糖和RNA代谢过程是环辛四烯作用于灰霉菌的重要靶标,抑制其生长及侵染。  相似文献   

10.
ABSTRACT The induction of resistance to Botrytis cinerea in carrot roots by UV radiation, a possible means for controlling storage diseases, was compared with systemic resistance induced by inoculation with the pathogen. UV radiation did not have any systemic effect, and disease resistance was induced only in tissues directly exposed to the radiation. Although UV radiation induced a local accumulation of 6-methoxymellein (6-MM), inoculation with B. cinerea caused 6-MM to accumulate systemically, away from the inoculation site. Because of amounts near or higher than the ED(50) (50% effective dose) for inhibiting B. cinerea found in both UV-treated and preinoculated roots at the time of challenge, 6-MM could be involved in both types of resistance. Freshly harvested carrots had a number of constitutive chitinases and beta-1,3-glucanases, which were not affected by UV radiation or inoculation. When challenged with B. cinerea, the induction of a 24-kDa chitinase was enhanced in UV-treated and preinoculated roots. Again, UV radiation had only a local effect in priming this chitinase response. Although UV- and pathogen-induced resistance in carrot may involve the same defenses, the responses are probably mediated differently, because UV radiation has an essentially local effect.  相似文献   

11.
Y. ELAD 《Plant pathology》1992,41(1):47-54
Isolates of Botrytis cinerea having reduced sensitivity to the sterol biosynthesis-inhibiting (SBI) fungicides fenetrazole and fenethanil were obtained from one out of four sites from which isolates were tested. Reduced sensitivity was associated with poor disease control by fenetrazole, which had been applied with dichlofluanid. Conidial germination and hyphal growth of B. cinerea from the four sites were tested in vitro on media amended with the fungicides. Following fenetrazole or fenethanil treatment, at the site where control had failed, populations of B. cinerea were detected with higher EC50 and EC90 values than at the three other sites. Germination of conidia of B. cinerea was markedly inhibited by 1.0 μg/ml fenetrazole and 0.5–1.0 μg/ml fenethanil. The frequency of isolates insensitive to 1.0 μg/ml fenetrazole or to 0.5 μg/ml fenethanil was 3.4 and 1.8 times higher, respectively, at the site where control had failed, compared with another site where SBI fungicides had never been applied to control grey mould. Grey mould caused by selected isolates of B. cinerea which expressed the phenotype of low sensitivity to SBI fungicides in leaves of tomato, pepper and Senecio cineraria was not controlled by either fenetrazole or fenethanil (1 .5–3.0 μg/ml). However, up to 100% disease reduction was obtained when leaves infected by sensitive isolates were treated with the fungicides.  相似文献   

12.
ABSTRACT The exo-beta-1,3-glucanase (EC 3.2.1.58) activity of Pichia anomala strain K, an antagonistic yeast of Botrytis cinerea on postharvest apples, was studied in a synthetic medium supplemented with laminarin, a cell wall preparation (CWP) of B. cinerea, or glucose. The highest enzyme activity was detected in culture media containing a CWP of B. cinerea as the sole carbon source, whereas the lowest activity was observed in culture media supplemented with glucose. Exoglc1, an exo-beta-1,3-glucanase, was purified to homogeneity from culture filtrates of strain K containing a CWP. The molecular mass of exoglc1 was estimated to be under 15 kDa. Optimum activity of exoglc1 was recorded at 50 degrees C and pH 5.5. The exoglc1 K(m) value was estimated at 22.4 mg/ml. Exoglc1 showed in vitro a stronger inhibitory effect on germ tube growth of B. cinerea than on conidia germination and caused morphological changes such as leakage of cytoplasm and cell swelling. Exo-beta-1,3-glucanase activity was detected on apples treated with strain K and was similar to exoglc1 on the basis of activity on native gel. Moreover, the addition of a CWP to a suspension of P. anomala stimulated both in situ exo-beta-1,3-glucanase activity and protective activity against the pathogen, strengthening the hypothesis that exo-beta-1,3-glucanase activity is one of the mechanisms of action involved in the suppression of B. cinerea by P. anomala strain K.  相似文献   

13.
Abiotic stress may affect plant response to pathogen attack through induced alterations in growth regulator and gene expression. Abscisic acid (ABA) mediates several plant responses to abiotic stress. The effects of drought, salt stress and ABA on the interaction of tomato ( Lycopersicon esculentum ) with the biotrophic fungus Oidium neolycopersici and the necrotrophic fungus Botrytis cinerea were investigated. Drought stress resulted in a twofold increase in endogenous ABA as well as a 50% reduction in B. cinerea infection and a significant suppression of O. neolycopersici on tomato cv. Moneymaker. Salt stress did not affect B. cinerea infection, but significantly reduced infection by O. neolycopersici , with no obvious increase in endogenous ABA. Compared with the wild type, the ABA-deficient sitiens mutant was more resistant to O. neolycopersici and B. cinerea . Exogenous ABA resulted in increased susceptibility of sitiens to both pathogens, but did not increase the basal susceptibility of wild-type plants. It is concluded that, in tomato, drought and salt stress stimulate different, but possibly overlapping, pathogen-defence pathways which may not necessarily involve ABA. Meanwhile, basal endogenous ABA levels suppress the resistance of tomato to O. neolycopersici and B. cinerea , but an ABA increase above the basal level, resulting from exogenous application, does not increase susceptibility to these pathogens.  相似文献   

14.
丁香酚对灰葡萄孢的抑制作用研究   总被引:8,自引:0,他引:8  
丁香酚是从植物中提取的天然化合物。采用菌丝生长速率法测定了其对多种植物病原真菌的抑制作用以及田间分离得到的7株灰葡萄孢Botrytis cinerea对丁香酚的敏感性,同时测定了丁香酚对B.cinerea 03 孢子和菌核的抑制活性。结果显示,丁香酚对多种植物病原真菌均有不同程度的抑制作用;灰葡萄孢不同菌株对丁香酚的敏感性不同,EC50值在29.97 ~83.62 μg/mL之间;丁香酚可抑制灰葡萄孢产孢,但对孢子萌发无影响,可抑制其菌核的产生和萌发。荧光染色结果表明丁香酚能破坏灰葡萄孢菌丝细胞膜。利用原子吸收光谱法和紫外分光光度法分别测定丁香酚处理前后菌丝培养液中K+浓度和OD260值变化,结果显示处理后K+浓度和OD260值均升高。综上所述,细胞膜可能是丁香酚对灰葡萄孢菌丝的作用位点。  相似文献   

15.
ABSTRACT Botrytis cinerea is an economically important pathogen. Epidemiological studies are difficult because of the genetic variability within this species. The objectives of this work were to study the variability and to compare the inhibitory effects of Ca on three isolates of B. cinerea from decayed apple (B) and grape (C and C77:4). Among these isolates, B had the least radial growth but had a sporulation rate 40% higher than that of both C77:4 and C. In situ, isolate C incited the largest decay area in the fruit of two of four apple cultivars examined and had the highest polygalacturonase activity in vitro. Maximum mycelial growth was reached with CaCl(2) at 1 g liter(-1) for isolates B and C77:4 and at 4 g liter(-1) for isolate C. Calcium (CaCl(2)) inhibited polygalacturonase activity at 1 g liter(-1) for C and C77:4 and at 16 g liter(-1) for B. Calcium infiltration reduced decay caused by all three isolates by three to five times. Mycelial DNA analysis showed that 42% of the character loci scored were polymorphic and the greatest similarities were found between B and C77:4. These results support the evidence that the biological and statistical variability in research can be affected by the B. cinerea isolate selected. Despite this variation, Ca treatment of apples reduced decay caused by all three Botrytis cinerea isolates.  相似文献   

16.
研究了新型环烷基磺酰胺类化合物N-(2,4,5-三氯苯基)-2-氧代环己烷基磺酰胺(简称化合物108)对灰葡萄孢菌丝生长、孢子形成和萌发以及菌核产生等不同生育阶段的抑制作用及其对菌丝致病力和形态结构的影响。结果表明:化合物108对灰葡萄孢菌丝生长和孢子形成及孢子萌发具有明显的抑制作用,其EC50值分别为6.90、4.70和4.11μg/mL;菌核形成受到明显抑制,当药剂质量浓度达20μg/mL时,无菌核产生。经化合物108处理后的灰葡萄孢菌丝致病力下降,40μg/mL处理的菌丝致病力显著低于对照。超微结构观察结果表明,化合物108能导致灰葡萄孢菌丝萎缩、塌陷和变形,菌体细胞壁增厚、皱缩及分层。  相似文献   

17.
The primary evaluation of ten fungicides for the control of cucumber powdery mildew (Sphaerotheca fuliginea), tomato leaf mould (Cladosporium fulvum) and tomato grey mould (Botrytis cinerea) indicated that nine of them were systemically active against one or more of the pathogens when applied as soil drenches. Benomyl, thiophanate, thiophanate-methyl, thiabendazole and Cela W52A showed activity against all three diseases, but dimethirimol, triarimol and dodemorph were more specific and mainly active against cucumber powdery mildew. An in vitro bioassay generally indicated that fungitoxicity was not translocated from sprayed to unsprayed leaves, and though toxicity to B. cinerea, following soil drenches of benomyl, thiophanate and thiophanate-methyl, was detected by bioassay in leaf sap and calyces it was not detected in fruit. Furthermore, extended evaluation of benomyl for the control of grey mould on cropping tomotoes showed that in the presence of abundant B. cinerea inoculum, benomyl soil drench treatment effectively controlled flower and leaf infection but not fruit ghost-spotting.  相似文献   

18.
Five stable hybridoma cell lines secreting monoclonal antibodies (Mabs) specific for GLRaV-1, one of the agents involved in the aetiology of grapevine leafroll disease, were produced by fusing a nonsecreting myeloma cell line with spleen cells from BALB/c mice immunized with purified GLRaV-1. The Mabs were characterized for their recognition of virus coat protein by DAS-ELISA and Western blotting. Mab (1G10) reacted specifically in ELISA, immuno-electron microscopy and immunoblotting with both GLRaV-1 and GLRaV-3 coat proteins. Mab 1C4 detected 25 of the 33 GLRaV-1 isolates, while Mab 1B7 reacted with 32 isolates including the eight isolates not recognized by Mab 1C4. Two of these hybridoma lines (2F11 and 2F3) are now used routinely for the immunodiagnosis of GLRaV-1.  相似文献   

19.
Most potential fungal antagonists have been identified only after exhaustive non-selective screening. Bacteria antagonistic to Botrytis cinerea were selected using a simple method based on techniques developed for trace enrichment studies, where a crude cell wall preparation from B. cinerea was used as a selective substrate and bacteria were isolated based on growth on cell wall agar media. Most of the 52 bacterial isolates that were obtained from the surface of organically grown strawberry fruit and could grow on B. cinerea wall media in culture showed some ability to inhibit the growth of the fungus. Potato dextrose agar seeded with B. cinerea spores (104ml–1) were challenged with each of these bacterial isolates at concentrations of 109 and 106 colony forming units ml–1. Eleven of the 52 isolates initially recovered, subsequently demonstrated strong antagonism in vitro and were selected for additional screening tests on strawberry fruit. All 11 isolates reduced grey mold rot incidence on fruit in storage. Three of the best isolates were tested in limited field trials, and also reduced grey mold rot on fruit under field conditions.  相似文献   

20.
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