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1.
Leaf tissue harvested from cucumber plants (Cucumis sativus L.) expressing induced resistance against the powdery mildew fungus Podosphaera xanthii (syn. Sphaerotheca fuliginea, Castagne; Braun and Shishkoff) was extracted and analyzed for phytoalexin compounds. Fluorescence microscopy was then used to observe the production of these compounds in planta, and laser scanning confocal microscopy observations were made to locate the subcellular sites of phytoalexin accumulation. Phytochemical analyses and fluorescence microscopy observations revealed the production of autofluorescent C-glycosyl flavonoid phytoalexins within the epidermal tissues of disease-resistant plants undergoing fungal ingress. Phytoalexin production was triggered by the combination of an eliciting/inoculation treatment, and tissue autofluorescence of color characteristic of the phytoalexins reached a maximum 48 h after elicitation prior to subsiding following the collapse of the pathogen. After a second eliciting treatment, disease-resistant plants produced phytoalexins more rapidly in response to fungal challenge. At the cellular level, autofluorescent C-glycosyl flavonoid phytoalexins were observed associated with the plasma membrane of infected epidermal cells immediately following elicitation. In the hours that preceded the collapse of conidial chains, phytoalexins accumulated inside the haustorial complexes of the pathogen within the epidermal cells of disease-resistant plants. Taken together, the results of this study show the timely synthesis of C-glycosyl flavonoid phytoalexins at precise subcellular locations as a key defense reaction used by cucumber to create incompatible interactions with powdery mildew.  相似文献   

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为高效精准防治柑橘溃疡病,本研究对不同成熟度柑橘叶片接种溃疡病菌Xanthomonas citri subsp.cirri后的发病情况进行观察,并在显微镜下观察溃疡病菌侵染叶片的过程,并对不同浓度、不同施用方法下噻森铜的室内防治效果和田间防治效果进行测定,对施用噻森铜后土壤、叶片和果实中的残留进行测定.结果 表明,溃疡...  相似文献   

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The 3-deoxyanthocyanidin phytoalexins produced in sorghum leaves in response to Colletotrichum sublineolum have chemical structure similarities to the 3-deoxy flavonoids that are precursors of phlobaphene pigments. Phlobaphenes are commonly observed in the pericarp of mature sorghum grains, while synthesis of 3-deoxyanthocyanidin phytoalexins is a site-specific response to infection with C. sublineolum. We have taken a genetic approach to investigate the possible overlap between the two sub-branches of flavonoid biosynthesis in sorghum that lead to phlobaphenes and 3-deoxyanthocyanidin phytoalexins. A sorghum line with a functional y1 gene synthesizes 3-deoxyanthocyanidins as well as phlobaphenes. However, a progenitor line with the mutable Y1-candystripe (Y1-cs) allele shows variable levels of biosynthesis of these compounds. The Y1-cs allele carries a copy of the Candystripe1 (Cs1) transposable element in the y1 gene. We demonstrate here that the variability in the expression of 3-deoxyanthocyanidins produced in individual mesocotyls of hyper-mutable Y1-cs plants is a function of the activity of the y1 gene. TheCs1 insertion in the Y1-cs allele blocks y1 function, while excision of Cs1 out of they1 locus restores the gene to a functional state. The combined molecular and biochemical characterization of sibling plants confirms that the allelic state of the y1 gene is completely correlated with the production of phytoalexins in response to fungal infection. These results provide support for the idea that the y1 gene regulates the biosynthesis of both 3-deoxyanthocyanidin phytoalexins and phlobaphene pigments in sorghum.  相似文献   

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Four carnation cultivars, Novada (resistant to races 1 and 2 ofFusarium oxysporum f.sp.dianthi), Elsy (susceptible to race 1), Lena (susceptible to race 2) and Sam's Pride (susceptible to both races), were selfed and crossed. When three months old, the seedlings were inoculated via the roots or via the stems, after which wilting was recorded weekly according to a 5-point ordinal scale.Analyses were carried out on the proportions of diseased plants. For race 1 variation between the progenies could be described by means of general combining abilities only; GCA values were not affected by the inoculation method used. Also for race 2 GCAs were most important but the GCA values appeared different for the two inoculation methods. It is concluded that resistance to both races is inherited in an additive way.Indications for independently inherited root-specific resistance components (extravascular resistance) were only found with race 2. With both races, the ability to confine the pathogen at the infection site appeared the most important resistance component. Resistant progenies were also characterized by longer latent periods and lower wilting rates.Both race 1 and race 2 induced the accumulation of the phytoalexins dianthalexin and methoxydianthramide S, but race 2 induced higher amounts than race 1. The accumulation of phytoalexins was positively correlated to the resistance level of the progenies against the respective races. The progenies of the double-resistant cultivar Novada appeared to produce particularly high levels of phytoalexins.  相似文献   

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Role of stilbenes in the resistance of grapevine to powdery mildew   总被引:1,自引:0,他引:1  
Stilbene phytoalexins are identified as defence response in pathogen–grapevine interactions, but little information is available on the role of stilbenes on Erysiphe necator, causal agent of grapevine powdery mildew. Analysis of stilbenes in artificially infected leaf discs from susceptible to highly resistant cultivars was performed and compared to the development of the pathogen. Results indicate that stilbene synthesis is confined in infected cells, penetrated by an appressorium–peg. Stilbene amounts expressed by infection site allow discriminating susceptible and resistant cultivars. Highest viniferins concentrations on resistant cultivars are in correspondence with the observed inhibition of the pathogen growth. The analysis of stilbenes at the infection site and viniferins accumulation in grapevine defence reaction is discussed.  相似文献   

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Susceptibility to Xanthomonas axonopodis pv. citri of a citrus cybrid, in which the nuclear and cytoplasmic genomes were derived from Citrus sinensis and C. unshiu, respectively, was evaluated. Bacterial growth in the leaves of the cybrid was similar to that in C. sinensis after pin-prick inoculation throughout the experiment, whereas growth was greater than that in C. unshiu from 8 days after inoculation. Lesion expansion and pustules that later developed from the lesions on the cybrid and on C. sinensis also appeared to be greater than those on C. unshiu. The incidence of citrus canker disease caused by the bacteria on the cybrid trees was in the field was equivalent to that on C. sinensis but was severer than on C. unshiu. These results indicate that the nuclear genome of the cybrid plays a critical role in susceptibility to citrus canker disease. However, the pathogenicity gene pthA of the bacteria is not likely to be involved in the difference in susceptibility to the bacteria between C. unshiu and C. sinensis because their susceptibility to a pthA-deficient mutant of the bacteria also differs.  相似文献   

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When lower leaves of rice plants were inoculated with powder formulation of a saprophytic strain ofPseudomonas fluorescens, Pfl, upper leaves, in addition to the inoculated lower leaves, showed resistance to the rice bacterial blight pathogenXanthomonas oryzae pv.oryzae. When the leaves were challenge-inoculated withX. oryzae pv.oryzae 4 days afterP. fluorescens application on lower leaves, the disease intensity in upper leaves decreased from 6.7 to 1.1. When rice seeds were treated with the formulation ofP. fluorescens Pfl and sown, 30-day-old seedlings showed resistance toX. oryzae pv.oryzae and the disease intensity decreased from 6.8 to 1.2. The induced resistance was transient; leaves sprayed withP. fluorescens Pfl at 30 days after treatment and leaves of 60-day-old seedlings fromP. fluorescens-treated seeds did not show resistance to the pathogen. In field trials, seed treatment followed by foliar application of the powder formulation ofP. fluorescens Pfl effectively controlled rice bacterial blight and increased the yield. In the induced resistant leaves a sharp increase in lignification and activities of peroxidase, phenylalanine ammonia-lyase and 4-coumarate: CoA ligase was observed when the leaves were challenge-inoculated withX. oryzae pv.oryzae. An approximately threefold increase in lignin content, peroxidase activity and phenylalanine ammonia-lyase activity and a fivefold increase in 4-coumarate: CoA ligase activity were observed 5 days after challenge inoculation withX. oryzae pv.oryzae in rice leaves pretreated withP. fluorescens for 5 days. A similar increase in defense-related activities was not observed in susceptible interactions or inP. fluorescens-treated plants at later stages of interactions when no resistance to the pathogen was observed.  相似文献   

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Effects of crop rotation on the incidence of soil-borne pathogens and on the performance of potato were investigated in five field experiments. Rotations differed in cropping frequency of potato and in crop sequence.Incidence of stem canker caused byRhizoctonia solani was strongly influenced by the cropping frequency of potato and not by crops with which the potato was alternated in the rotation. Cropping frequency of potato also affected the occurrence of black scurf, but less pronounced than for stem canker. The antagonistVerticillium bigutatum slightly reducedR. solani (black scurf) in plots on sandy soil continuously cropped with potato. Incidence of stem canker was also strongly affected by granular nematicides applied to the soil, nitrogen level and the cultivar grown.  相似文献   

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Bean hypocotyls, pea pods and tomato fruits were tested for phaseollin, pisatin and rishitin production when challenged with the phytopathogenic bacteriaErwinia carotovora, Pseudomonas phaseolicola, P. pisi andP. solanacearum, and their isolated extracellular polysaccharides. All bacteria induced phytoalexin accumulation, whereas only phaseollin and pisatin, but not rishitin, were elicited by EPS. The inhibitory effect of these three phytoalexins on bacterial growth was studied in liquid medium; whereas phaseollin and pisatin strongly inhibited growth, only a slight inhibitory effect resulted from the presence of rishitin in the medium.Samenvatting Bonehypocotylen, erwtepeulen en tomatevruchten werden onderzocht op hun vermogen tot vorming van respectievelijk faseolline, pisatine en rishitine, na inoculatie met de fytopathogene bacteriënErwinia carotovora, Pseudomonas phaseolicola, P. pisi enP. solanacearum en na behandeling met oplossingen van hun extracellulaire polysacchariden (EPS). Alle bacteriesoorten induceerden fytoalexinevorming, terwijl hun EPS wel faseolline- en pisatine-, maar geen rishitinevorming induceerden. Faseolline en pisatine remden de groei van de bacteriën in vloeibaar medium sterk; rishitine daarentegen had slechts een geringe groeiremming tengevolge.  相似文献   

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In winter oilseed rape experiments at Rothamsted in 2000/01 to 2002/03 growing seasons, the severity of phoma stem canker epidemics in summer depended on the timing of phoma leaf spot epidemics in the previous autumn, and hence on the timing of Leptosphaeria maculans ascospore release. The first major release of L. maculans ascospores was earlier in 2000 (26 September) and 2001 (18 September) than in 2002 (21 October). Consequently, the autumn phoma leaf spot epidemic was also earlier in 2000 and 2001 than in 2002. The resulting stem canker epidemics were severe by harvest (July) in 2001 and 2002 but not in 2003. No correlation was found between the severity or duration of phoma leaf spotting (lesion days or lesion °C-days) and the subsequent severity of phoma stem canker epidemics. Rates of leaf production and loss were similar in the three growing seasons. Out of ca. 25 leaves produced on plants during each season, leaf numbers 10–14 generally remained on plants for the longest. Treatment with flusilazole + carbendazim in autumn decreased the severity of phoma leaf spotting for several weeks after treatment, decreased the severity of stem canker the following summer and increased yield significantly in 2001 and 2002 but not in 2003. The most effective timings for flusilazole + carbendazim application were when leaves 7–11 were present on most plants and at least 10% of plants were affected by phoma leaf spot. Two half-dose applications of fungicide reduced phoma stem canker and increased yield more than a single full dose application when phoma leaf spot epidemics were early (<800 °C-days after sowing).  相似文献   

15.
Copper-containing bactericides have been used to control bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae. However, the efficacy of copper has been reduced by the occurrence of copper-resistant strains. Analysis of the DNA sequence of a cluster region containing the copper-resistance genes from P. syringae pv. actinidiae suggested the presence of three possible different systems for copper resistance: copper-trapping, copper-efflux and copper-transport systems. Transposon insertional inactivation analysis indicated that the copper-trapping system was essential for copper resistance.  相似文献   

16.
An inoculation method for Exobasidium splendidum and Exobasidium vaccinii was developed on the dwarf shrub Vaccinium vitis-idaea. Using inoculated ramets, we investigated whether there are differences between V. vitis-idaea populations in the susceptibility to Exobasidium infections and whether the defence reaction of V. vitis-idaea is visible at a molecular level. Sixteen V. vitis-idaea clones from four populations were propagated in tissue cultures and the ramets were inoculated with E. splendidum or E. vaccinii fungi. The expression of three flavonoid biosynthetic genes (chalcone synthase, dihydroflavonol 4-reductase and anthocyanidin synthase) and the accumulation of flavonoids and hydroxycinnamic acids were determined in response to E. splendidum infection. A pathogenesis-related (PR 4) gene was isolated and its expression was studied in host ramet leaves. To our knowledge, this was the first successful artificial infection reported with E. splendidum. Disease frequencies of the inoculated ramets were between 32% and 47% for E. splendidum and 33% for E. vaccinii, but below 10% in uninoculated control ramets. There were no differences in disease frequencies between V. vitis-idaea populations. Both symptomatic leaves and healthy leaves of diseased ramets showed activation of flavonoid biosynthesis at the gene level, whereas expression of PR 4 was observed only in symptomatic leaves. The increase of flavonoid biosynthesis in healthy leaves of diseased ramets may represent a general response to stress or a role in defence against the pathogen E. splendidium. Ability of V. vitis-idaea to defend chemically against Exobasidium fungi and the heterogeneity of genotypes, age, size, and growth rates in host plant populations might be reasons for the low infection incidence of Exobasidia in nature.  相似文献   

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Excised twig assay and excised stem inoculation were used to evaluate nine fungicides (metalaxyl, fosetyl-Al, copper hydroxide, copper sulfate, copper oxychloride, captan, quintozene, propamocarb and chlorothalonil) againstPhytophthora cactorum andP. citrophthora associated with crown rot in peach trees. Segments were soaked in fungicide solutions at different concentrations and then inserted vertically intoP. cactorum orP. citrophthora cultures growing on cornmeal agar plus antibiotics, or inoculated by inserting a mycelium-bearing agar plug directly into the cambium. Following incubation, the bark was scraped off and length of necrosis was measured. Metalaxyl was the only fungicide that inhibited canker development on segments at the manufacturer-recommended concentration. Fosetyl-Al, captan, copper hydroxide and copper sulfate inhibited canker development at 3, 4, 4 and 8 gl-1, respectively. The other fungicides did not affect canker length significantly compared with non-treated twigs, with the exception of propamocarb, which reduced the development ofP. cactorum on excised stems. The tested methods enabled rapid and effective evaluation of a large number of chemicals to prevent crown rot diseases caused byPhytophthora in the laboratory. http://www.phytoparasitica.org posting Dec. 5, 2001.  相似文献   

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Secondary metabolites and host defense compounds were shown to occur in xylem sap, and leaves of Vitis vinifera cv. Italia and cv. Matilde naturally infected by the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme). Samples of xylem sap and leaves were collected from healthy vines and from vines showing severe symptoms of brown wood-streaking caused by Pch and Tmi, or from vines with symptoms of both brown wood-streaking and white rot caused by Fme. Xylem sap collection was carried out during the early spring of 2003 and 2004, corresponding to the phenological phases: (A) cotton bud; (B) green tip; (C) leaves out; (D) stretched out leaves; and (E) visible clusters. In the present work we have studied the accumulation of biomolecules (pentaketides and α-glucans), host defense compounds (benzaldehydes, benzoic acid and cinnamic acid derivatives, flavonols, flavanols, flavan-3-ol derivatives and stilbenes) at different stages of grapevine development. Accumulation and changes in total phenolics and recurring phenolics, and of three phytotoxic secondary metabolites (scytalone, isosclerone and pullulan) were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that phenolic concentrations are strongly related to the cv.  相似文献   

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The numbers of diseased plants could significantly be reduced when microconidia ofFusarium oxysporum f. sp.dianthi were inoculated into the stem and viable-, heat-killed cells or purified LPS of the bacteriumPseudomonas sp. strain WCS417r were applied to the roots. Because the competition betweenF. o. dianthi and the bacterium could be excluded, the disease suppression seems to be due to an induced resistance. Accumulation of phytoalexins was found in the stem segments. No accumulation of these compounds was found when the plants were bacterized but noninfected. It is concluded that cell surface components present in the lipopolysaccharides of the bacterium are the inducing factors.Increased peroxidase activity could be measured in root washes and root extracts after only bacterial preparations were added. No significant differences in peroxidase activity were found in stem extracts. The possible role of increased peroxidase activity in suppression of Fusarium wilt in carnation is discussed.  相似文献   

20.
Arabidopsis thaliana ecotype Columbia-0 was transformed with a green fluorescent protein (GFP) gene under control of a phenylalanine ammonia-lyase (PAL) promoter. PAL is a key enzyme of the phenylpropanoid pathway and is induced to high levels during plant stress. Constitutive expression of PAL1 promoter-controlled GFP occurred in vascular tissues within stems, leaves and roots and in developing flowers. PAL1 promoter–GFP expression was examined in leaves of transgenic plants subjected to an abiotic elicitor, mechanical wounding or to inoculation with the pathogens Pseudomonas syringae pv. tomato or Peronospora parasitica. Wounding of leaves and treatment with an abiotic elicitor and compatible interactions produced low to moderate levels of GFP. However, in incompatible interactions there were high levels of GFP produced. In incompatible interactions, the intensity of GFP fluorescence was similar to that produced in transgenic plants expressing GFP driven by the CaMV promoter. The bright green fluorescence produced in live cells and tissues was readily visualised using conventional fluorescence microscopy and was quantified using spectroflourometry. This is the first report of the use of GFP as a reporter of defence gene activation against pathogens. It has several advantages over other reporter genes including real time analysis of gene expression and visualisation of defence gene activation in a non-invasive manner.  相似文献   

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