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1.
Accumulation of pathogenesis-related proteins is thought to play a role in pathogen-induced plant defense responses. Although early accumulation of hydrolytic enzymes such as chitinase and β-1,3-glucanase has been associated previously with genetically-inherited and induced systemic resistance, their role in resistance in tomato(Lycopersicon esculentum)to the phytopathogenic fungusAlternaria solaniis not yet understood. Here we describe the accumulation patterns of specific isozymes of pathogenesis-related proteins in the resistant tomato genotypes 71B2, NC EBR-1, NC EBR-2 and the susceptible cultivar Piedmont. Western blot analysis demonstrated that four isozymes of chitinase (26, 27, 30, and 32kDa) were induced in all genotypes upon challenge withA. solani,but only resistant lines had significantly higher constitutive levels of the 30kDa isozyme as well as total chitinase activity. In addition, the 30kDa chitinase isozyme was found to accumulate to significantly higher levels in resistant lines during pathogenesis than the susceptible genotype. Two isozymes of β-1,3-glucanase (33 and 35kDa) were detected in all genotypes, but a slightly higher constitutive level was detectable in all resistant lines when compared to the susceptible. Similar accumulation patterns of these isozymes were observed in all genotypes during the course of pathogenesis. Purified preparations of acidic and basic tomato chitinase and β-1,3-glucanase isozymes were tested for their antifungal activity againstA. solani in vitro.Results presented in this study indicate that only basic isozymes of chitinase and β-1,3-glucanase were inhibitory toA. solaniwhereas, no inhibitory activity was observed with the acidic isozymes. The results of this study suggest that a higher constitutive level of chitinase and β-1,3-glucanase and the induction pattern of a 30kDa chitinase isozyme in early blight resistant breeding lines is related to genetically-inherited resistance of tomato toA. solani.  相似文献   

2.
Benzothiadiazole (BTH) and DL-β-aminobutyric acid (BABA) induced systemic resistance was investigated in susceptible and resistant pea genotypes against Uromyces pisi. Resistance was characterized by reduced infection frequency mainly due to decreases in appressorium formation, stomatal penetration, growth of infection hyphae and haustorium formation. Changes in β-1,3-glucanase, chitinase, phenylalanine ammonia-lyase and peroxidase activities and in total phenolics content, demonstrate that U. pisi resistance is induced by BTH and BABA treatments at early and late stages of the fungal infection process, but that the chemicals operate via different mechanisms. In fact, our study showed that BTH treatment primed the activity of pathogenesis related-proteins such as β-1,3-glucanase, chitinase and peroxidase in both susceptible and resistant genotypes. On the other hand, BABA treatment did not increase the enzymatic activities in the studied genotypes, but significantly increased their total phenolic contents.  相似文献   

3.
Dipping leaf strips of greenhouse or field-grown tobacco (Nicotiana tabacum L., cv. Ky-14) plants into acetone for 1 s, prior to inoculation with sporangia of Peronospora tabacina Adam, increased their susceptibility to blue mold. Disease severity and sporangial production on leaf discs from acetone-treated leaves were markedly increased compared to those on discs from untreated leaves. Treatment with acetone also decreased variation in susceptibility of leaves from plants of various ages. Disease severity on discs obtained from attached leaves which were dipped in acetone for 1 s was three times greater up to 15 days after dipping than on discs from leaves that were not dipped. TLC and GLC analyses of the acetone extracts indicated that 95% or more of the major cuticular diterpenoids, α- and β-4,8,13-duvatriene-1,3-diols (DVT), were removed from the surface by dipping for 1 s. These compounds had not reappeared on the leaf surface 15 days after leaves were dipped in acetone for 1 s. Aqueous suspensions of the acetone-soluble constituents as well as authentic DVT, inhibited sporangial germination of P. tabacina (ED100 = 25 ppm) and the antifungal activity was accounted for by DVT. When DVT was removed from a leaf surface and added back to the same leaf strip, the resistance of the leaf tissue was restored. As tobacco plants aged, their susceptibility to blue mold decreased and the quantity of DVT on the leaf surface increased. The data support a role for DVT in the resistance of tobacco against blue mold.  相似文献   

4.
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5.
A two- to threefold increase in total soluble sugar content was detected in the leaves of tobacco plants(Nicotiana tabacum L.) 24 days after stem inoculation with conidia ofPeronospora tabacina (=Peronospora hyoscyami f. sp.tabacina), relative to leaves of non-inoculated control plants. Glucose and fructose accounted for most of this increase. The leaves of the inoculated plants were also protected against blue mold incited byP. tabacina. The increased sugar content may be due to the three- to fourfold increase in activity of (β-1,3 glucanase in the leaves and/or to the threefold increase in activities of β-1,3 glucanase, invertase and amylase in the inoculated sems. Tobacco leaf discs floated on glucose or other mono- and disaccharides at 55.5 mM, but not on polyethyleneglycol at a similar osmotic potential, reacted hypersensitively upon challenge withP. tabacina.  相似文献   

6.
Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.  相似文献   

7.
Two bioassay methods are described which use detached tobacco leaves to measure the sensitivity of Peronospora tabacina to systemic fungicides. Tobacco leaves (13–15 cm2), treated with fungicides before or after detachment from the plant, were inoculated with sporangia in water drops and, after incubation in beakers and Petri plates, the disease severity and/or production of sporangia was determined 4–7 days after treatment with the fungicides. Of 15 systemic fungicides applied to detached leaves, eight N-phenylamides at 0.066?1.0 μg ml?1 controlled blue mould; metalaxyl was the most effective fungicide. Isolates of P. tabacina, collected in the field from tobacco plants grown in soil treated with metalaxyl, were not resistant to the fungicide applied to detached leaves prior to inoculation. The fungicide, applied to leaves before detachment, was used to measure the efficacy of five systemic N-phenylamide fungicides sprayed on the basal and unsprayed distal portions of the leaves. Blue mould was controlled on the basal portion of the leaf by all the fungicides at 0.66?1.0 μg ml?1, but it required the application of 3–30 times more chemical on the basal portion to achieve comparable blue mould control on the distal part of the leaf.  相似文献   

8.
Two antisera raised against acidic β-1,3-glucanase and acidic chitinase from tobacco were used to investigate the subcellular localization of the two enzymes in Fusarium culmorum -infected wheat spike by means of the immunogold labelling technique. The studies demonstrated that the distribution of β-1, 3-glucanase and chitinase were very similar in the uninoculated healthy and infected wheat spikes. The enzymes were localized mainly in the cell walls of different tissues including the lemma, ovary and rachis of the wheat spike, while the cytoplasm and organelles of cells in these tissues showed almost no labelling. However, the accumulation of β-1,3-glucanase and chitinase in the infected wheat spikes differed distinctly between resistant and susceptible wheat cultivars. The labelling densities for the two enzymes in the infected lemma, ovary and rachis of the susceptible cultivar Agent increased only slightly as compared to the corresponding uninoculated healthy tissues, whereas higher labelling densities of β-1,3-glucanase and chitinase were found in the infected tissues of wheat spikes from the resistant cultivar Arina compared to the corresponding uninoculated healthy tissues. Furthermore, the labelling of β-1,3-glucanase and chitinase also occurred over the cell walls of the hyphae in the infected wheat spike, but not over the hyphal cytoplasm. In addition, labelling for the two enzymes was often detected over the cell wall appositions and the electron-dense material located between the host cell and the hyphal cell in the infected tissues of the resistant wheat cultivar. The findings reported in the present study indicate that β-1,3-glucanase and chitinase accumulation in the F. culmorum -infected wheat spike may be involved in resistance to pathogen spread in the host tissue.  相似文献   

9.
 为了明确β-1,3-葡聚糖酶和几丁质酶与大豆抗疫霉根腐病的关系,测定了不同抗性大豆品种接种大豆疫霉菌后β-1,3-葡聚糖酶和几丁质酶活性的变化情况和2种酶对大豆疫霉菌的抑制作用。结果表明,大豆疫霉菌能诱导大豆β-1,3-葡聚糖酶和几丁质酶的活性增强,但2种酶在积累速度和幅度上,抗病品种和感病品种有显著的差异。与感病品种"857-1"相比,抗病品种"垦农4号"2种酶活性不仅升高的速度快、幅度大,且高活性维持的时间长。β-1,3-葡聚糖酶和几丁质酶混合液对大豆疫霉菌的菌丝生长、孢子囊形成和孢子萌发的抑制作用明显,其次是β-1,3-葡聚糖酶,而几丁质酶对大豆疫霉菌的抑制作用不明显。表明大豆对大豆疫霉根腐病的抗性与β-1,3-葡聚糖酶和几丁质酶的活性呈正相关关系。β-1,3-葡聚糖酶和几丁质酶混合对大豆疫霉菌的抑制具有协同增效作用。  相似文献   

10.
Disease symptoms typical of anthracnose were observed in cucumber, pumpkin and squash after infiltrating leaves with a conidial suspension ofColletotrichum lagenarium, but symptoms developed only in cucumber when droplets of the conidial suspension were applied to the leaf surface. There was no difference in the germination of conidia or in appressorium formation on leaf surfaces of cucumber, pumpkin or squash plants; however, penetration was markedly reduced into pumpkin and squash with or without systemic acquired resistance (SAR) and into cucumber with SAR. Little β-1,3-glucanase and chitinase activities were detected in challenged pumpkin and squash leaves without symptoms even 5 days after inoculation in leaf surfaces. However, the enzymes were detected in pumpkin and squash leaves with symptoms, and activities of the enzymes were greater than those in cucumber. These results suggest that β-1,3-glucanase and chitinase activities are not primary initial defence compounds associated with non-host resistance of pumpkin and squash toC. lagenarium.  相似文献   

11.
Infection of groundnut leaves with the early leaf spot pathogen Cercospora arachidicola leads to a marked increase in extracellular 1,3-β-glucanase activity, limited to the infected tissue. Three isoforms of low molecular weight and extreme pI values, typical of pathogenesis-related proteins, were induced. These β-glucanases, when acting together, were capable of degrading the pathogen cell wall in vitro. Glucanases from homogenates of infected leaf tissue were partially purified by ion-exchange chromatography to give enzymes with molecular weights of 35, 32 and 20 kDa and pI values of 3·8, 3·6 and > 9, respectively. They were electrophoretically identical to the β-glucanases found in the intercellular washing fluid. Treatment of groundnut plants with 200 μM mercuric chloride induced the accumulation of identical extracellular β-glucanases. During the course of the infection an increase in peroxidase activity was also observed, but chitinase activity remained more or less constant.  相似文献   

12.
In this study, the biocontrol ability of seven grapevine-associated bacteria, previously reported as efficient against Botrytis cinerea under in vitro conditions, was evaluated in two vineyard orchards with the susceptible cv. Chardonnay during four consecutive years (2002–2005). It was shown that the severity of disease on grapevine leaves and berries was reduced to different levels, depending on the bacterial strain and inoculation method. Drenching the plant soil with these bacteria revealed a systemic resistance to B. cinerea, even without renewal of treatment. Accordingly, this resistance was associated with a stimulation of some plant defense responses such as chitinase and β-1,3-glucanase activities in both leaves and berries. In leaves, chitinase activity increased before veraison (end-July) while β-1,3-glucanase reached its maximum activity at ripening (September). Reverse patterns were observed in berries, with β-1,3-glucanase peaking at full veraison (end-August) and chitinase at a later development stage. Highest activities were observed with Acinetobacter lwoffii PTA-113 and Pseudomonas fluorescens PTA-CT2 in leaves, and with A. lwoffii PTA-113 and Pantoea agglomerans PTA-AF1 in berries. These results have demonstrated an induced protection of grapevine against B. cinerea by selected bacteria under field conditions, and suggest that induced resistance could be related to a stimulation of plant defense reactions in a successive manner.  相似文献   

13.
Systemic resistance was induced to Pyricularia aryzae in rice by inoculation of the first leaf with the hypersensitive response causing bacterium, Pseudomonas syringae pv. syringae. Lesions caused by Pyricularia oryzae were decreased in number and size by 85% and 50%, respectively, in systemically protected leaves. Increased resistance was associated with the deposition of a dark brown material around sites of Pyricularia oryzae infection. The systemically acquired resistance was not associated with an increase in the activities of phenylalanine ammonia lyase, coniferyl alcohol dehydrogenase, peroxidase, β-1, 3-glucanase or chitinase after the challenge inocculation with Pyricularia oryzae. The levels of these enzymes were elevated by local exposure to Pseudomonas syringae pv. syringae or Pyricularia oryzae, but were not systemically induced. These data suggest that the physiological changes which occur during induced resistance in rice are different from those correlated with induced resistance in tobacco or cucumber.  相似文献   

14.
ABSTRACT Treatment of peach fruit with UV-C light caused a rapid induction of chitinase, beta-1,3-glucanase, and phenylalanine ammonia lyase (PAL) activities starting 6 h after treatment and reaching maximum levels at 96 h after treatment. By 96 h after UV-C treatment, chitinase, beta-1,3-glucanase, and PAL activities in UV-C-treated fruit were over twofold above the levels observed for the control. In nontreated control fruit, no apparent increase in chitinase and beta-1,3-glucanase activities was detected but a minor increase in PAL activity was seen. The transient increase in chitinase, beta-1,3-glucanase, and PAL activities in UV-C-treated fruit was preceded by a gradual activation of the corresponding genes. UV-C-treated fruit showed an increase in accumulation of beta-1,3-glucanase and chitinase mRNAs at 3 h after treatment, which peaked approximately 96 h posttreatment. A similar induction kinetic pattern was observed for PAL mRNA in response to UV-C treatment, except the induction started 6 h after UV-C treatment. These results show that the response of peach fruit to elicitor treatment is similar to that seen in other plant-elicitors interactions and suggests the involvement of peach biochemical defense responses in UV-C-mediated disease resistance.  相似文献   

15.
Genes encoding an acidic wheat class IV chitinase (383), an acidic wheat β 1,3-glucanase (638) and a rice cationic peroxidase (POC1) were introduced into ‘Nantes Coreless’ carrot (Daucus carota) by Agrobacterium-mediated transformation. The genes were introduced singly or in various combinations followed by selection imposed by the herbicide phosphinothricin. Regenerated plantlets were screened for presence and expression of the three transgenes using PCR, Southern and Northern hybridisations. Eighteen transgenic lines expressing a single transgene and 2 lines each co-expressing 638/383 and 383/POC1 were assessed for resistance to the necrotrophic fungal pathogens Botrytis cinerea and Sclerotinia sclerotiorum. Percentage leaf area diseased was measured 4 and 7 days after inoculation (dai) and compared to non-transformed control plants. Six lines expressing β-1,3-glucanase 638 alone had no enhanced resistance to B. cinerea at 4 dai and only slight resistance to S. sclerotiorum; there was no effect at 7 dai. Two out of the six lines expressing 383 alone had enhanced tolerance to both pathogens with a 20–50% reduction in disease development at 7 dai. Two lines co-expressing 638/383 had slight reductions in disease by (10–20%) similar to that of the lines expressing chitinase 383 alone. Highest levels of disease resistance were seen in transgenic lines expressing POC1, alone or in combination with chitinase 383. Disease symptoms were slower to develop and symptoms were reduced by up to 90% for B. cinerea and 70% for S. sclerotiorum. The 383/POC1 co-expressing plants developed disease at levels similar to that of POC1 alone. Petioles of plants over-expressing POC1 had higher levels of lignin accumulation constitutively compared to control plants, which was greatly enhanced following inoculation with S. sclerotiorum. These results indicate that peroxidase over-expression can lead to significant disease reduction against necrotrophic pathogens in transgenic carrot plants.  相似文献   

16.
Chitinase and-1,3-glucanase activities were assayed in roots, hypocotyls and cotyledons of downy mildewsusceptible and -resistant sunflower (Helianthus annuus L.) cultivars. While the highest-1,3-glucanase activity was in roots, that of chitinase activity was in hypocotyls. Inoculation of both sunflower cultivars withPlasmopara halstedii resulted in a marked increase of chitinase and-1,3-glucanase activities. The increase was observed earlier in incompatible than in compatible reactions. Both enzymes occurred in root tissue as a complex mixture of isoenzymes. At least three different peaks with chitinase activity and three with glucanase activity could be resolved by gel filtration chromatography on Sephacryl S-100 and chromatofocusing on PBE 94 (pH 7-4). Following ammonium sulfate precipitation and ion-exchange on CM- and DEAE-Trisacryl, three glucanase and chitinase fractions, referred to as basic, neutral and acidic, were separated on the basis of their Chromatographic behaviour. A different pattern of distribution of chitinase and-1,3-glucanase fractions was observed between inoculated and non-inoculated plants in both resistant (cv. RS-105) and susceptible (cv. Peredovik) cultivars. In healthy plants-1,3-glucanase was mainly found in the basic (cv. Peredovik) and neutral (cv. RS-105) fractions, whereas chitinase was in the basic fraction for both cultivars. The neutral and acidic fractions of chitinases were induced in the compatible and incompatible reactions. Inoculation of the plants induced the neutral-1,3-glucanase fraction in resistant and susceptible cultivars and the acidic only in the susceptible one. Induction of the basic fraction of both activities was not observed in any case.  相似文献   

17.
Bioassays testing the fungicide sensitivity against metalaxyl of Peronospora tabacina isolates collected in German tobacco fields in 2005 revealed the presence of two phenotypes, resistant and sensitive. DNA fingerprints using SSR and minisatellite primers allowed separation of the samples into two groups. The differences in amplification patterns coincided with the sensitive and resistant reaction of the isolates in metalaxyl bioassays. New primers were developed which allowed PCR-based detection of P. tabacina and differentiation of the metalaxyl-sensitive and the metalaxyl-resistant phenotype, respectively. Screening of recent blue mold isolates from Germany and other European countries for metalaxyl sensitivity with leaf disk bioassays coincided completely with the PCR-based identification of the two phenotypes. In Germany, exclusively resistant isolates were found between 2002 and 2004. These still dominate. Since 2005 the co-occurrence of sensitive isolates has been shown. No similar monitoring has been done in any other European country. However, we found the resistant phenotype reaction in a French isolate of 2004 and in two out of three Italian isolates in 2007. Two isolates from Poland and Bulgaria in 2007 were sensitive to metalaxyl. For all 58 isolates tested since 2002 the metalaxyl bioassay-based resistance type and the PCR-based tests for sensitive and resistant genotype coincided. Using genotype-specific primers for future population studies may help to trace the sources of the pathogen from which yearly propagation starts.  相似文献   

18.
Inoculation of 3-4 lower leaves of tobacco with tobacco mosaic virus (TMV) increased ribonuclease (RNase) and protease activities in inoculated leaves. Little or no increase in the activities were apparent in upper noninoculated leaves prior to challenge. After challenge with TMV or Peronospora tabacina, RNase activities increased more rapidly in the upper leaves of induced plants as compared to those of noninduced plants. Protease activities in the leaves challenged with P. tabacina or TMV also increased after challenge, but little or no differences in the activities were apparent between induced and noninduced plants. The incubation of purified TMV with leaf extracts obtained from induced challenged, noninduced challenged and noninduced unchallenged plants prior to inoculation did not affect the number of local lesions formed on tobacco plants.  相似文献   

19.
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20.
知母提取物诱导马铃薯植株抗晚疫病作用机制初探   总被引：5，自引：0，他引：5  
为明确知母提取物诱导马铃薯抗晚疫病的作用,本研究测试了经知母提取物处理并接种晚疫病菌后马铃薯植株的超氧化物歧化酶（SOD）、苯丙氨酸解氨酶(PAL)、β 1,3 葡聚糖酶和几丁质酶活性的变化。结果表明,经过知母提取物处理后的马铃薯植株,在受到晚疫病菌侵染后24～96 h,SOD和PAL活性显著升高并持续处于高活性水平;在接种48 h后β 1,3 葡聚糖酶活性开始显著升高,直到接种后96 h一直维持在较高活性水平;而无论是接种晚疫病菌还是经过知母提取物处理,几丁质酶活性都没有发生显著的变化。  相似文献   

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