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1.
Carmen García-Limones Ana Hervs Juan A. Navas-Corts Rafael M. Jimnez-Díaz Manuel Tena 《Physiological and Molecular Plant Pathology》2002,61(6):325
To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H2O2-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction. 相似文献
2.
The bean pathogen, Pseudomonas syringae B728a, may require iron-superoxide dismutase (FeSOD) and manganese–superoxide dismutase (MnSOD) activities in protection against reactive oxygen species (ROS) generated in planta. Genes encoding FeSOD or MnSOD of P. syringae B728a were cloned by hybridization with specific PCR probes amplified from P. syringae genomic DNA. The sodB gene was monocistronic whereas the sodA gene was transcribed as part of a 1.4 kb polycistronic operon, consisting of orfX-sodA. A putative Fur consensus sequence was located upstream of the orfX-sodA operon. The sodB (FeSOD) gene was expressed throughout growth, but was down regulated under iron deficient conditions. In contrast, the sodA (MnSOD) gene was expressed only under iron deficient conditions. Mutants defective in sodA and sodB genes were generated by marker exchange mutagenesis. Unlike other bacterial SOD deficient mutants, the P. syringae B728a sodAsodB mutant was not impaired in growth on rich or minimal medium, but it was more sensitive to paraquat than wild-type. The P. syringae B728a SOD deficient mutants caused bacterial brown spot disease on bean pods or leaves to the same extent as wild-type. Thus, these superoxide dismutases may not be key enzymes for aerobic metabolism and pathogenicity in P. syringae B728a. 相似文献
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Chen-Fang Wang Li-Li Huang Heinrich Buchenauer Qing-Mei Han Hong-Chang Zhang Zhen-Sheng Kang 《Physiological and Molecular Plant Pathology》2007,71(4-6):230-239
The generation and accumulation of reactive oxygen species (ROS), superoxide anion (O2−) and hydrogen peroxide (H2O2), were studied in the interaction between wheat cv. ‘Suwon 11’ and two races of Puccinia striiformis f. sp. tritici (avirulent and virulent). Generation of O2− and H2O2 was analyzed histochemically using nitroblue tetrazolium (NBT) and 3,3-diamino-benzidine (DAB), respectively. At the pre-penetration stage during appressorium formation both stripe rust races induced H2O2 accumulation in guard cells. In the incompatible interaction, a rapid increase of O2− and H2O2 generation at infection sites was detected. The percentage of infection sites showing NBT and DAB staining was 36.1% and 40.0%, respectively, 12 h after inoculation (hai). At extended incubation time until 24 hai, percentage of infection sites showing H2O2 accumulation further increased, whereas those exhibiting O2− accumulation declined. The early infection stage from 12 to 24 hai coincided with primary haustoria formation in mesophyll cells. In contrast, in the compatible interaction, O2− and H2O2 generation could not be detected in most of the infection sites. In the incompatible interaction, intensive DAB staining was also determined in mesophyll cells, especially in cell walls, surrounding the infected cells 16–24 hai; thereafter, these cells contained fluorescing compounds and underwent hypersensitive response (HR). The number of necrotic host cells surrounding the infection sites increased continuously from 20 to 96 hai. It might be concluded that H2O2 accumulation during the early infection stage is associated with the occurrence of hypersensitive cell death and that resistance response is leading to arrest the avirulent race of the obligate stripe rust pathogen. In the compatible interaction at 96 hai, H2O2 accumulation was observed in mesophyll cells surrounding the rust lesion. 相似文献
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Yeon Kyeong Lee Jeum Kyu Hong Sigrun Hippe-Sanwald Byung Kook Hwang 《Physiological and Molecular Plant Pathology》2000,57(6)
The compatible interaction of pepper stems with Phytophthora capsici showed more rapid and severe disease development than did the incompatible interaction, although pathogen penetration styles of host cells in compatible and incompatible interactions were similar to each other. Treatment with
-β-amino- n -butyric acid (BABA) protected the pepper plants against P. capsici infection. Reduced hyphal growth and sporangial formation were found after P. capsici infection in BABA-induced resistant and incompatible reactions. One of the most noticeable ultrastructural features of the BABA-induced resistant reaction was the formation of electron-dense wall appositions. The thick and dense wall appositions that encased the haustoria restricted haustorial development, thus leading to limitation of further pathogen penetration into inner plant tissues. A main host response in the incompatible interaction was the occlusion of cortical cells with an amorphous material. Plugging of the intercellular spaces in the cortical cells with electron opaque material was frequently observed in the incompatible interaction, but not in the compatible interaction. Another common feature of the BABA-induced resistant and incompatible reactions was degeneration of mitochondrial structure within penetrating hyphal cytoplasm. The mitochondrial structure in the BABA-induced resistant or incompatible reactions had no distinct double membrane layer and well-shaped cristae. 相似文献
6.
Clovis S. Palmer Jennifer A. Saleeba Bruce R. Lyon 《Physiological and Molecular Plant Pathology》2005,67(6):427
A phytotoxic protein that evokes the typical symptoms of Verticillium wilt disease in seedlings of Gossypium hirsutum L. (Upland cotton) was isolated from culture filtrates of Verticillium dahliae. The protein was purified by ammonium sulfate precipitation, Sephadex-G100 fractionation, and native PAGE. The 18.5 kDa protein, designated VD18.5, appears to be a single subunit protein with an isoelectric point between 3 and 5. VD18.5 induces symptoms of leaf dehydration, chlorosis, necrosis and stem discoloration in seedlings of the disease susceptible cotton cultivar Siokra 1–4. The LD50 of VD18.5 on protoplasts of Siokra 1–4 was 18 μg mL−1. VD18.5 had no noticeable effect on Pima S-7, which is a disease resistant cultivar. Phytotoxic activity was partially destroyed at high temperature and was abolished by digestion with proteinase K. Mass spectrometry fingerprinting and protein sequence data from VD18.5 yielded no significant matches when submitted to the Mascot search engine and NCBI non-redundant protein databases, respectively. These results suggest that VD18.5 is a novel protein that may be involved in the development of some of the symptoms associated with Verticillium wilt disease in the cotton plant. 相似文献
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Tika B. Adhikari Boovaraghan Balaji Jill Breeden Stephen B. Goodwin 《Physiological and Molecular Plant Pathology》2007,71(1-3):55-68
Large-scale cDNA-AFLP profiling identified numerous genes with increased expression during the resistance response of wheat to the Septoria tritici blotch fungus, Mycosphaerella graminicola. To test whether these genes were associated with resistance responses, primers were designed for the 14 that were most strongly up-regulated, and their levels of expression were measured at 12 time points from 0 to 27 days after inoculation (DAI) in two resistant and two susceptible cultivars of wheat by real-time quantitative polymerase chain reaction. None of these genes was expressed constitutively in the resistant wheat cultivars. Instead, infection of wheat by M. graminicola induced changes in expression of each gene in both resistant and susceptible cultivars over time. The four genes chitinase, phenylalanine ammonia lyase, pathogenesis-related protein PR-1, and peroxidase were induced from about 10- to 60-fold at early stages (3 h–1 DAI) during the incompatible interactions but were not expressed at later time points. Nine other genes (ATPase, brassinosteroid-6-oxidase, peptidylprolyl isomerase, peroxidase 2, 40S ribosomal protein, ADP-glucose pyrophosphorylase, putative protease inhibitor, methionine sulfoxide reductase, and an RNase S-like protein precursor) had bimodal patterns with both early (1–3 DAI) and late (12–24 DAI) peaks of expression in at least one of the resistant cultivars, but low if any induction in the two susceptible cultivars. The remaining gene (a serine carboxypeptidase) had a trimodal pattern of expression in the resistant cultivar Tadinia. These results indicate that the resistance response of wheat to M. graminicola is not completed during the first 24 h after contact with the pathogen, as thought previously, but instead can extend into the period from 18 to 24 DAI when fungal growth increases dramatically in compatible interactions. Many of these genes have a possible function in signal transduction or possibly as regulatory elements. Expression of the PR-1 gene at 12 h after inoculation was much higher in resistant compared to susceptible recombinant-inbred lines (RILs) segregating for the Stb4 and Stb8 genes for resistance. Therefore, analysis of gene expression could provide a faster method for separating resistant from susceptible lines in research programs. Significant differential expression patterns of the defense-related genes between the resistant and susceptible wheat cultivars and RILs after inoculation with M. graminicola suggest that these genes may play a major role in the resistance mechanisms of wheat. 相似文献
10.
Jaqueline Wolfe Carolyn J. Hutcheon Verna J. Higgins Robin K. Cameron 《Physiological and Molecular Plant Pathology》2000,56(6):253
An early event correlated with the gene-for-gene hypersensitive response (HR) is the accumulation of active oxygen species (AOS), also known as the oxidative burst. We present data that genetically demonstrates that the oxidative burst is a downstream component of the RPS2- avrRpt2gene-for-gene signal cascade. An in planta AOS assay using the fluorescent probe 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) was modified for use with the Arabidopsis thaliana / Pseudomonas syringae pv.tomato (P. syringae pv. tomato) model system. An oxidative burst occurred between 8 and 15 hpi with avirulent P. syringae pv. tomato(avrRpt2), but not with virulent P. syringae pv. tomato. This burst preceded cell death and was not observed in the RPS2 Arabidopsis mutantsrps2-101C and rps2-201 inoculated with avirulent P. syringae pv. tomato. An HR-like response has been observed when plants undergoing a systemic acquired resistance (SAR) response are challenged with a normally virulent pathogen (manifestation stage of SAR), however an HR-like oxidative burst was not detected by the in planta AOS assay during this stage of SAR. 相似文献
11.
Roxana Y. Parada Kumiko Oka Daisuke Yamagishi Motoichiro Kodama Hiroshi Otani 《Physiological and Molecular Plant Pathology》2007,71(1-3):48-54
It has been reported that Alternaria brassicae, the causal agent of gray leaf spot in Brassica plants, produces a host-specific or host-selective toxin (HSTs) identified as destruxin B. In this study, the role of destruxin B in infection of the pathogen was investigated. Destruxin B purified from culture filtrates (CFs) of A. brassicae induced chlorosis on host leaves at 50–100 μg ml−1, and chlorosis or necrosis on non-host leaves at 250–500 μg ml−1. Destruxin B was detected in spore germination fluids (SGFs) on host and non-host leaves, but not in a sufficient amount to exert toxicity to host plants. When spores of non-pathogenic A. alternata were combined with destruxin B at 100 μg ml−1 and inoculated on the leaves, destruxin B did not affect the infection behavior of the spores. Interestingly, SGF on host leaves allowed non-pathogenic spores to colonize host leaves. Moreover, a high molecular weight fraction (>5 kDa) without destruxin B obtained by ultrafiltration of SGF had host-specific toxin activity and infection-inducing activity. From these results, we conclude that destruxin B is not a HST and does not induce the accessibility of the host plant which is essential for colonization of the pathogen. In addition, the results with SGF imply that a high molecular weight HST(s) is involved in the host–pathogen interaction. 相似文献
12.
Atsunori Isshiki Kazuya Akimitsu Hideo Ishii Hiroyuki Yamamoto 《Physiological and Molecular Plant Pathology》2000,56(6):263
An exopolygalacturonase and three endopolygalacturonases were purified from mycelia of pear scab pathogens, Venturia pirina and Venturia nashicola. The molecular weight of the isolated exoPG from V. pirina was 43 kDa, and the endoPGs from V. nashicola were 42 kDa as estimated by SDS–polyacrylamide gel electrophoresis. The pH optimum of the exoPG activity from V. pirina was 5.0. TheKm and Vmaxvalues of the exoPG were 0.08 mg ml−1and 4.44 × 10−3 mmol reducing group min−1 mg protein−1. The N-terminal amino acid sequence of the exoPG from V. pirina was similar to that of the exoPG from Fusarium oxysporum f. sp. melonis, and the N-terminal amino acid sequences of the three endoPGs fromV. nashicola races 1, 2 and 3 were similar to other fungal endoPGs with a conserved motif of ASxxxTFTxAAAxxxG. 相似文献
13.
Fabio Rossi Cavalcanti Mario Lucio Vilela Resende Joo Paulo Matos Santos Lima Joaquim Albenisio Gomes Silveira Jos Tadeu Abreu Oliveira 《Physiological and Molecular Plant Pathology》2006,68(4-6):198-208
The activities of antioxidant enzymes and photosynthetic responses were investigated in tomato (Lycopersicon esculentum L. var.) pre-treated by plant activators and inoculated by Xanthomonas vesicatoria. Plants were sprayed with acibenzolar-S-methyl, ASM [Bion® 50 WG (0.2 g l−1)] and aqueous extract from dry necrotic tissue flour (VLA) of ‘Lobeira’ (Solanum lycocarpum) bush. Four days later, the plants were challenged with a virulent strain of X. vesicatoria, under greenhouse conditions. Tomato leaves were then assessed to determine the activities of the main antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX). A reduction of bacterial leaf spot severity was observed in plants treated by ASM (49.3%) and VLA (31.4%), without any in vitro inhibitory activity over the pathogen. Controls showed decreases in CO2 assimilation, transpiration, photosynthetic rates, and stomatal conductance. Water use efficiency and carboxylation efficiency were strongly affected in ASM- and VLA-treated, in comparison to controls and healthy plants. The tested substances induced increases in SOD and CAT activities in a delayed enzymatic response typical in compatible plant–pathogen interactions. Measured at daily intervals, activities of APX and POX were significantly (ρ0.05) higher in treated plants than in controls, except for APX in ASM-treated plants where no difference was found when compared to controls. Only POX was clearly induced at the earlier stages after spraying the tomato plants with ASM or VLA. Our results suggest that late increases in antioxidant enzyme activities may play a role in mitigating oxidative damage in restoring the photosynthetic imbalance imposed by the expansion of bacterial lesions. 相似文献
14.
Iwona Morkunas Waldemar Bednarski Ma&#x;gorzata Kopyra 《Physiological and Molecular Plant Pathology》2008,72(4-6):167-178
The aim of this study was to compare the defense responses of embryo axes of Pisum sativum L. cv. Kwestor with different sucrose levels to pathogenic fungi, i.e. systemic acting Fusarium oxysporum f. sp. pisi and locally acting Ascochyta pisi. Embryo axes were cultured on Heller medium for 96 h. Four variants were compared: these included inoculated embryo axes cultured with or without 60 mM sucrose (+Si and −Si) and non-inoculated embryo axes cultured with or without 60 mM sucrose (+Sn and −Sn). After inoculation of the pea embryo axes with pathogenic fungi a generally higher concentration of free radicals was detected by electron paramagnetic resonance (EPR), in comparison to non-inoculated embryo axes. The inoculation with F. oxysporum caused stronger generation of free radicals in −Si than in +Si embryo axes. A different response was observed after inoculation with A. pisi; starting from 48 h, the concentration of free radicals in +Si axes was found to be 1.5 times higher than in −Si embryo axes. The values of spectroscopic splitting coefficients for these radicals suggest that they are semiquinone radicals. The EPR method also revealed Mn2+ ion accumulation after 24 h of culture. Over time, high levels of these ions were recorded in +Si embryo axes inoculated with F. oxysporum, while in +Si embryo axes inoculated with A. pisi they decreased. Up to 48 h after inoculation with the pathogenic fungi, Mn2+ ion levels were higher in +Si embryo axes than in +Sn axes. The activity of superoxide dismutase (SOD, EC 1.15.1.1) increased in +Si embryo axes up to 72 h after inoculation with pathogenic fungi; however, it was generally lower than in +Sn axes. Catalase activity (CAT, EC 1.11.1.6) increased up to 72 h after inoculation with F. oxysporum and the values were higher than in the non-inoculated tissue. Especially high activity of this enzyme was noted in −Si embryo axes after inoculation with either F. oxysporum or A. pisi. Peroxidase activity (POX, EC 1.11.1.7) towards pyrogallol in embryo axes increased during culture; however, it was lower or similar to that in non-inoculated embryo axes. SOD, CAT and POX zymograms showed that the synthesis of new isoforms was induced after inoculation with pathogenic fungi. Peroxidase isozymes detected by the reaction with diaminobenzidine in native PAGE were intensely stained in +Si embryo axes after inoculation with pathogenic fungi. Respiratory activity of the inoculated tissues was considerably higher than in non-inoculated tissues. The respiration rate was generally much higher in +Si than in −Si embryo axes. Growth of −Si embryo axes was more significantly retarded as a consequence of inoculation than that of +Si embryo axes.These results indicate that, depending on the manner of influence of a pathogenic fungus, both similar and differing defensive strategies may be initiated and a raised sugar levels in pea tissues limit the development of F. oxysporum and A. pisi. 相似文献
15.
Grisel Ponciano Masayasu Yoshikawa Jamie L. Lee Pamela C. Ronald Maureen C. Whalen 《Physiological and Molecular Plant Pathology》2006,69(4-6):131-139
Disease resistance mediated by the resistance gene Xa21 is developmentally controlled in rice. We examined the relationship between Pathogenesis Related (PR) defense gene expression and Xa21-mediated developmental disease resistance induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1a, OsPR1b, and OsPR1c genes were cloned and their induction was analyzed, in addition to the OsPR10a gene, at the juvenile and adult stages in response to a wildtype Xoo strain that induces a resistance response (incompatible interaction) and an isogenic mutant Xoo strain that does not (compatible interaction). We found that the adult stage leaves are more competent to express these OsPR1 genes and that the Xa21 locus is required for the highest levels of induction. 相似文献
16.
Lin He Xiwu Gao Jinjun Wang Zhimo Zhao Nannan Liu 《Pesticide biochemistry and physiology》2009,95(3):147-151
The carmine spider mite is the most serious crop mite pests in China. Abamectin has been used to control insects and mites worldwide but carmine spider mites, Tetranychus cinnabarinus, had developed resistance to it. Genetic research on insecticide resistance has been fundamental for understanding the resistance development, studying resistance mechanisms, and designing appropriate resistance management strategies to control insect pests. A resistant colony of T. cinnabarinus, RRG42, was established to examine the inheritance of abamectin resistance in T. cinnabarinus. The females of T. cinnabarinus were selected for bioassay using a slide dip method. After 42 generations of selection, the RRG42 strain was 8.7-fold resistant to abamectin compared with the susceptible strain (SS). The logarithm (log) concentration–probit response curve for F1s from reciprocal crosses, of F1RS and F1SR, were inclined to that for SS and the degree of dominance (D) values for F1s were −0.81 and −0.17. There was a significant difference in values of LC50 and slope of log concentration–probit lines between F1RS and F1SR. The observed mortalities of BC1 (F1RS♀ × RRG42♂) and BC1′ (F1SR♀ × SS♂) were significantly different from the expected mortalities based on a monogenic resistance in the chi-square tests. The inheritance of abamectin resistance in T. cinnabarinus is incompletely recessive and may be controlled by more than one gene. The maternal or cytoplasmic effect may exist in the inheritance of resistance to abamectin in T. cinnabarinus. 相似文献
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Bacterial black spot disease of mango is caused by Xanthomonas campestris pv. mangiferaeindicae (Xcm), which consists of two genotypically and phenotypically distinct groups of strains. Monoclonal antibodies (MABs) were produced – 15 against CFBP 1717, a group I strain, and 9 against CFBP 2919 (yellow-pigmented), a group II strain – and were analyzed for their characteristics. On the avidin-biotin peroxidase complex enzyme-linked immunosorbent assay, the dilution limit of the MABs was between 100 and 200000 and was 10 times higher when measured on the corresponding ascitic fluid. All kinds of isotypes were represented among the MABs. All the Japanese Xcm strains, designated group I by hrp-restriction fragment length polymorphism (RFLP) analysis, reacted equally with MAB 1A7H12G3, which is the most specific for all but one worldwide group I strains, and to only one strain among group II. Also, to various extents, serological heterogeneity inside the two groups was consistently differentiated based on isozyme and RFLP analyses. MAB 1E2E1 against CFBP2919, because of its narrow specificity, and MAB 1A7H12G3 against CFBP1717, because of its broad specificity, will be useful for epidemiological studies or general control of the pathogen. 相似文献
19.
A. Ortuo I. Nemsa N. Alvarez A. Lacasa I. Porras A. Garcia Lidn J.A. Del Río 《Physiological and Molecular Plant Pathology》2008,72(4-6):162-166
The susceptibility of Fortune (Citrus clementina × Citrus reticulata), Citrus paradisi and Citrus limon fruits to Alternaria alternata pv. citri was investigated using different artificial inoculation methods. The results obtained reveal that the C. paradisi and C. limon fruits are less susceptible to A. alternata pv. citri than Fortune fruits, although all showed symptoms of Alternaria brown spot when the cuticle was broken and the flavedo or flavedo + albedo was removed. Furthermore, it was seen that susceptibility to the fungus decreased as the age of the fruit increased. There was a positive correlation between the susceptibility of the different Citrus fruits to A. alternata pv. citri and their “in vivo” ethylene levels, the most susceptible fruit (Fortune) producing more ethylene during growth than the less susceptible C. limon and C. paradisi. This suggests that ethylene may well be considered as a possible marker of Citrus fruit susceptibility to A. alternata pv. citri. Disease development increased when the Fortune fruits were treated with 1 mM ACC (a precursor of ethylene biosynthesis) or 1 mM Ethephon (an ethylene-releasing compound) prior to inoculation with A. alternata pv. citri. The role of ethylene as a factor involved in disease development is discussed. 相似文献
20.
Qiuju Zhou Junfeng Xiang Yalin Tang Jiangpeng Liao Chuyi Yu Hongyan Du Qianfan Yang Guangzhi Xu 《Pesticide biochemistry and physiology》2008,92(1):43-47
The interaction of a novel pesticide, NMD (spiro[(2R,3R,4S)-4-benzyloxy-2,3-iso-propylidenedioxy-1-oxacyclopentane-5,5′-(2-nitromethylene-1,3-diazacyclohexane)]), with bovine serum albumin (BSA) has been investigated by using absorption, fluorescence, and circular dichroism (CD) spectroscopy methods. Quenching of the fluorescence of BSA has been observed in the presence of NMD. The binding parameters were determined using Stern–Volmer equation. From the thermodynamic parameters calculated according to the van’t Hoff equation, the enthalpy change ΔH, and entropy change ΔS were found to be −2.71 kJ mol−1 and 82.56 J mol−1 K−1, respectively. These values suggested that, apart from an initial hydrophobic association, the complex is held together by van der Waals interactions and hydrogen bonding. These results provided a quantitative understanding of the binding of NMD to BSA, which is important in understanding its toxicity in vertebrates. 相似文献