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1.
When the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme) were grown in liquid stationary cultures, it was seen that they were able to live in media containing resveratrol (RES) or tannic acid (TA) as the sole carbon source and that the fungi were able to convert both compounds. Particular attention is paid here to detecting RES and TA conversion. Pch, Tmi and Fme were partially inhibited by RES or TA. Pch, Tmi and Fme produced extracellular tannase, laccase and peroxidase enzymes in liquid or agarized cultures, whether glucose was present or not. When colonies of Pch, Tmi and Fme were confronted, they showed spatially and temporally heterogeneous patterns of laccase and peroxidase activity. The results indicate the non-synergistic, competitive association of Pch and Tmi and the inhibition of Fme growth. Muconic acid, a well-known intermediate in a large number of lignin and phenol oxidative processes, can partly or completely inhibit the lignolytic agent Fme, but is tolerated by Pch and Tmi. An explanation for wood pigmentation patterns by Pch, Tmi and Fme is given.  相似文献   

2.
Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) produced scytalone, isosclerone and pullulans in liquid cultures, as well as in calli. Secondary metabolites and host defense compounds were shown to occur in calli of Vitis vinifera cv. Italia and cv. Matilde infected by the esca-associated fungi Pch, Tmi and Fomitiporia mediterranea (Fme). Calli of both cvs. were grown as dual cultures with Pch, Tmi and Fme. The fungi grew well in the presence of calli of both cvs., but callus growth was reduced. Accumulation and changes of total phenolics and recurring phenolics, and of two phytotoxic pentaketides and pullulans were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that concentrations of phenolics are strongly related to the cv. The paper discusses the possible relationship between melanin biosynthesis in Pch and Tmi, which utilize pentaketide metabolites as intermediates and their pathogenicity related to phytotoxity of scytalone and isosclerone.  相似文献   

3.
Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) produced scytalone, isosclerone and pullulans in liquid cultures, as well as in calli. Secondary metabolites and host defense compounds were shown to occur in calli of Vitis vinifera cv. Italia and cv. Matilde infected by the esca-associated fungi Pch, Tmi and Fomitiporia mediterranea (Fme). Calli of both cvs. were grown as dual cultures with Pch, Tmi and Fme. The fungi grew well in the presence of calli of both cvs., but callus growth was reduced. Accumulation and changes of total phenolics and recurring phenolics, and of two phytotoxic pentaketides and pullulans were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that concentrations of phenolics are strongly related to the cv. The paper discusses the possible relationship between melanin biosynthesis in Pch and Tmi, which utilize pentaketide metabolites as intermediates and their pathogenicity related to phytotoxity of scytalone and isosclerone.  相似文献   

4.
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   

5.
A vineyard of Vitis vinifera cv. Sangiovese was surveyed for incidence of esca and xylem sap collection. Sap samples were collected from healthy vines and from those with dual infection by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) or triple infection by Pch, Tmi and Fomitiporia mediterranea (Fme), during each early spring in a 3-year period (2001–2003). In order to analyse the possible trends in the climatic data, temperature and rainfall were assessed. At sap harvesting, aliquots of sap were assayed for phytotoxicity and extracted with ethyl acetate for phytotoxin recovery. Moreover, the exopolysaccharide (EPS) content was evaluated on several sap samples during the bleeding period. Conidia of Pch and Tmi, mycelium of Fme and their secondary metabolites were found in the sap of the esca-affected vines, indicating that the pathogens and their by-products together with some defence substances were accumulated and then translocated. Bioactivity tests showed toxicity of the sap from esca-affected vines to healthy detached leaves of cv. Sangiovese. The daily amount of sap, the pH, and the volume (Jv) and solute (Js) fluxes were analysed as a function of the infecting fungi. Pullulan, glucogalactomannan(s) and arabinogalactan(s) are the main EPS in the esca-infected vines, whereas in the sap of healthy vines no traces of pullulan were found. Scytalone and isosclerone usually produced in vitro by Pch and Tmi were also detected in the sap of vines infected by Pch and Tmi or by Pch, Tmi and Fme. The endogenous phytohormone content of healthy vines evaluated by the cutting bioassay was different from that of infected vines. Four phenolics belonging to three classes e.g., benzoic acid derivatives, stilbenes and flavonol-glycosides were separated and identified by HPLC.  相似文献   

6.
A vineyard of Vitis vinifera cv. Sangiovese was surveyed for incidence of esca and xylem sap collection. Sap samples were collected from healthy vines and from those with dual infection by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) or triple infection by Pch, Tmi and Fomitiporia mediterranea (Fme), during each early spring in a 3-year period (2001–2003). In order to analyse the possible trends in the climatic data, temperature and rainfall were assessed. At sap harvesting, aliquots of sap were assayed for phytotoxicity and extracted with ethyl acetate for phytotoxin recovery. Moreover, the exopolysaccharide (EPS) content was evaluated on several sap samples during the bleeding period. Conidia of Pch and Tmi, mycelium of Fme and their secondary metabolites were found in the sap of the esca-affected vines, indicating that the pathogens and their by-products together with some defence substances were accumulated and then translocated. Bioactivity tests showed toxicity of the sap from esca-affected vines to healthy detached leaves of cv. Sangiovese. The daily amount of sap, the pH, and the volume (Jv) and solute (Js) fluxes were analysed as a function of the infecting fungi. Pullulan, glucogalactomannan(s) and arabinogalactan(s) are the main EPS in the esca-infected vines, whereas in the sap of healthy vines no traces of pullulan were found. Scytalone and isosclerone usually produced in vitro by Pch and Tmi were also detected in the sap of vines infected by Pch and Tmi or by Pch, Tmi and Fme. The endogenous phytohormone content of healthy vines evaluated by the cutting bioassay was different from that of infected vines. Four phenolics belonging to three classes e.g., benzoic acid derivatives, stilbenes and flavonol-glycosides were separated and identified by HPLC.  相似文献   

7.
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   

8.
Two naphthalenone pentaketides (scytalone and isosclerone) and α-glucans (pullulan) are produced in vitro and in planta by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi), two tracheiphilous fungi associated with the ‘esca’ disease of grapevines. The possible role of such fungal metabolites in inducing symptoms on leaves and berries was studied in a vineyard of Vitis vinifera cv. Italia located in southern Italy. During early spring, pruning off two to four branches per vine allowed samples of xylem sap to be collected. Vine bleeding, assessed as ml day−1 vine−1, reached its maximum at bud burst and stopped within 28 days. The total amount of sap collected from healthy vines was about a quarter of that from esca-affected vines. During the same period, the leaf water potential of diseased vines increased progressively (i.e. showed less negative values), indicating a dysfunction in water and nutrient supply to the new growth. Both fungi were isolated from the xylem sap and from the woody tissue of branches and the trunk of diseased vines. Conidia isolated from the sap showed a high germination rate (>90%). Bioactive concentrations of the two pentaketides were detected in xylem sap, leaves and berries at various stages of seasonal growth. Exopolysaccharides, including pullulan, were found in the xylem sap. Absorption of culture filtrates of Pch and Tmi, as well as weak solutions of purified preparations of scytalone, isosclerone or pullulan, on detached leaves and berries caused symptoms similar to those shown by the esca-affected vines in the field.  相似文献   

9.
In recent years an increasing number of species of Botryosphaeriaceae have been associated with grapevine decline worldwide. Five species isolated from declining grapevines in Spain (Botryosphaeria dothidea, Diplodia seriata, Dothiorella viticola, Neofusicoccum luteum and N. parvum) were checked for toxin production in liquid cultures. Cultural conditions for all fungi were adjusted to obtain optimal production of phytotoxic culture filtrates, by growing the fungi in steady liquid cultures of Czapek–Dox broth for different time intervals. Phytotoxicity of D. seriata and N. parvum reached a maximum after 14 days while the remaining species showed the highest phytotoxicity levels after 21 days in culture. All fungi produced hydrophilic high-molecular weight compounds with phytotoxic properties. In addition, N. luteum and N. parvum produced lipophilic low-molecular weight phytotoxins, not detected consistently among the remaining species. This led to a more exhaustive study on the phytotoxicity of N. luteum and N. parvum. Culture filtrates and corresponding extracts of both species were consistently highly phytotoxic in different assays. The gas-chromatography analysis of the acetylated O-methyl glycosides of the phytotoxic exopolysaccharides produced by N. parvum showed these substances to be composed mainly of glucose, mannose and galactose. Results suggest that phytotoxic metabolites could be involved in the virulence of both species in planta.  相似文献   

10.
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11.
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   

12.
Polygalacturonase inhibiting proteins (PGIPs) are leucine-rich repeat glycoproteins, localized in the cell wall of most plant species, capable of countering the activity of endo-polygalacturonases (endo-PGs) produced by phytopathogenic fungi. The PGIP from Allium porrum leaves was analysed to ascertain the presence of different molecular forms of PGIP. Leek PGIP was separated into two fractions: a soluble and an ionically wall-bound PGIP, each of which was then purified by cation-exchange chromatography. Two and three peaks of PGIP activity were obtained, respectively. PGIP isoforms contained in each peak were separated by isoelectrofocusing (IEF) on a polyacrylamide gel. Following the separation, the gel was first overlaid with sodium polygalacturonate and then treated with the endo-PG from either Sclerotinia sclerotiorum, Fusarium moniliforme or Botrytis aclada. The endo-PG(s) hydrolyse the overlaid substrate except where active inhibitors are present. The presence of PGIPs is revealed by ruthenium red staining of the nonhydrolysed substrate. Each PGIP peak following IEF separation revealed several PGIP isoforms with pIs between 5.0 and 7.0. More than 20 isoforms were detected in total, with considerable differences in their inhibitory activity. While similar PGIP isoform patterns were obtained by developing the IEF gels with the endo-PGs of S. sclerotiorum and B. aclada, less intense PGIP bands were observed with the endo-PG from B. aclada, consistent with inhibition assays performed in solution. The endo-PG from F. moniliforme, which is not inhibited at all by leek PGIP in solution, consistently showed no PGIP band on the gel assay.  相似文献   

13.
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   

14.
Elicitin and a new protein 75 kDa elicitor were purified from the culture filtrate of Phytophthora palmivora, a pathogen of Hevea brasiliensis (rubber plant). Elicitin was obtained by using a one step of DEAE cellulose chromatography and the new elicitor was obtained by two steps of chromatography: a DEAE cellulose column followed by a hydrophobic column. Both elicitors were stable to heat and a wide range of pH values, but were sensitive to ProteaseK. Both elicitors induced scopoletin, peroxidase isozymes (with substrate o-dianisidine and scopoletin) and total phenolic compounds in cell suspension of H. brasiliensis with similar kinetics. In addition, both elicitors induced peroxidase enzyme (o-dianisidine), total phenolic compounds and enhanced local resistance against P. palmivora on young rubber tree seedlings. However, the increase of peroxidase enzyme and total phenolic compounds in rubber tree seedlings was different from those in cell suspension. Furthermore, during the expression of local resistance the zoospore of P. palmivora induced the peroxidase enzyme (o-dianisidine) more rapidly and with higher level than the control plants. H. brasiliensis is more responsive to the new elicitor than elicitin in triggering defense responses. That is the new elicitor was active at a concentration lower than those required for elicitin, about a 30-fold decrease for activation defense responses in cell suspension. For induction of peroxidase enzyme (o-dianisidine), phenolic compounds and local resistance of rubber plants against P. palmivora, the 75 kDa protein was active at about a 2-fold lower concentration when compared to elicitin.  相似文献   

15.
宁夏地区苦豆子内生真菌的分离与初步鉴定   总被引：1，自引：0，他引：1  
为了探讨宁夏地区苦豆子内生真菌类群的多样性,以宁夏地区苦豆子草为试验材料,采用组织分离法从其根、茎、叶中分离内生真菌,依据形态进行初步鉴定。结果表明,从宁夏地区苦豆子875个表面消毒的根、茎、叶组织块中,共分离出内生真菌115株。其中根中分离出18株,占15.7%,茎中分离出41株,占35.7%,叶中分离出56株,占48.6%,分布规律为叶、茎、根依次递减,叶组织中的内生真菌多样性较高。对产孢的113株进行初步鉴定,分类为2纲3目5科7属,其中青霉属(Penicillium)3株,占2.6%,黑葱花霉属(Periconia)4株,占3.6%;链格孢属(Alternaria)45株,占39.1%;镰孢霉属(Fusarium)9株,占7.8%,曲霉属(Aspergillus)10株,占8.7%,枝梗茎点霉属(Dendrophoma)30株,占26.1%,根霉属(Rhizopus)9株,占7.8%;其中链格孢属(Alternaria),枝梗茎点霉属(Dendrophoma)为优势种群。宁夏地区苦豆子内生真菌多样性丰富,同时不同部位内生菌的数量、种类及分布存在差异。  相似文献   

16.
Effects of paraquat dichloride (PQ) on the laccase (LAC) activity and some biochemical parameters of Trametes versicolor and Abortiporus biennis strains belonging to white rot Basidiomycetes fungi were examined. PQ water solution was added to 10-day-old stationary cultures cultivated on a liquid medium. Having measured the activity of extracellular laccase during the first 120 h, we found that the addition of 25 μM paraquat to T. versicolor and 20 μM paraquat to A. biennis cultures significantly stimulated the LAC activity in comparison to the control value (without PQ). Native PAGE gel analysis demonstrated that no new isoforms of laccase appeared in the presence of PQ stress. The increase of LAC activity was connected with dry weight loss. Enhanced activity of extracellular superoxide dismutase was observed during the first 48 h after PQ application in both investigated strains. The PQ-treatment also caused an evident increase of catalase activity, formaldehyde level and depletion of glutathione in T. versicolor as well as in A. biennis mycelia.  相似文献   

17.
The phenolic composition of olive roots and stems was studied by high performance liquid chromatography–mass spectrometry. The in vivo levels of the principal phenolic compounds found in olive plants infected by Phytophthora megasperma Drechsler and Cylindrocarpon destructans (Zinssm.) Scholten differed from the levels observed in non-infected plants. When the antifungal activity of these compounds against both fungi was studied in vitro, the most active were quercetin and luteolin aglycons, followed by rutin, oleuropein, p-coumaric acid, luteolin-7-glucoside, tyrosol and catechin. Microscopic study showed that these phenolic compounds affected the growth, morphology and ultrastructure of the fungi. Taken together, these findings suggest that the phenolic compounds present in olive plants play an active role in the protection against pathogen attack.  相似文献   

18.
We have previously shown that flagellin of Pseudomonas syringae pv. tabaci is an elicitor that induces a hypersensitive reaction (HR) in nonhost tomato cells. Flagellin is the major HR elicitor produced by this pathogen, as shown by the inability of a flagellin-defective mutant, ΔfliC, to induce HR. Also, a ΔfliD mutant that secretes large amounts of monomer flagellins induces a strong HR in tomato. In this study, the possible involvement of an Hrp type III secretion system (TTSS) in flagellin-induced HR was investigated using flagella-defective mutants or Hrp TTSS-defective mutants. The hrcC gene encodes HrcC protein, which is required for Hrp pilus formation in the outer membrane. An hrcC mutation, introduced into the wild-type, ΔfliC, and ΔfliD mutants of P. syringae pv. tabaci did not affect swimming motility or flagellin secretion, whereas all ΔhrcC, ΔfliC, and ΔfliD mutants lost the ability to cause disease on host tobacco leaves. However, the ΔhrcC mutant and the ΔfliD/ΔhrcC double mutant were still able to induce HR cell death, expression of one of the defense-related genes hsr203J, and the generation of hydrogen peroxide in nonhost tomato cells. Thus, flagellin is required for both pathogenicity in host tobacco and HR in nonhost tomato. On the other hand, hrp TTSS is necessary for pathogenicity on host tobacco but is not indispensable to induce HR in nonhost tomato. These results clearly show that flagellin-induced HR is hrp-independent in tomato.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB049570  相似文献   

19.
Fusarium graminearum, Fusarium culmorum and Fusarium cerealis are major causal agents of Fusarium Head Blight (scab) which is a disease of global significance in all cereal growing areas. These fungi produce trichothecene mycotoxins, principally nivalenol (NIV) and deoxynivalenol (DON). Genes Tri13 and Tri7 from the trichothecene biosynthetic gene cluster convert DON to NIV (Tri13) and NIV to 4-acetyl-NIV (Tri7). We have developed positive–negative PCR assays based on these two genes, which accurately indicate a DON or NIV chemotype in F. graminearum, F. culmorum and F. cerealis. These assays are useful in assessing the risk of trichothecene contamination, and can be informative in epidemiological studies. All NIV chemotype isolates studied have functional copies of both Tri13 and Tri7, and all DON-producing isolates have both genes disrupted or deleted. We have identified several mutations in these genes, which are conserved across F. graminearum lineage, RAPD and SCAR groupings and between the three species. There appears to be evidence of inter-species hybridisation within the trichothecene biosynthetic gene cluster.  相似文献   

20.
为了解新疆天山野果林中塞威士苹果Malus sieversii与其林下伴生植物短距凤仙花Impatiens brachycentra两种植物叶斑病病原菌的多样性及同源性,采用组织分离法获得病原菌,基于rDNA-ITS序列构建系统发育树,并进行ITS序列BLAST同源性比对,对病原菌进行鉴定分类,并依据科赫氏法则测定致病性。结果显示,从新疆新源县天山野果林中的塞威士苹果及短距凤仙花病叶上共分离得到18株菌落形态各异的病原菌,分属于2属4种,绝大多数属于半知菌亚门。其中,链格孢属Alternaria sp.为塞威士苹果和短距凤仙花叶斑病病原真菌中的优势菌群。致病性测定结果显示,其中11株病原真菌对塞威士苹果具有致病性,7株病原真菌对短距凤仙花具有致病性;塞威士苹果所有的病原真菌对短距凤仙花均有致病性,而且短距凤仙花所有的病原真菌对塞威士苹果也有致病性。推测2种植物叶斑病可能由相同来源的病原真菌引发,短距凤仙花染病加剧了塞威士苹果叶斑病暴发,可能是野果林退化的重要原因。  相似文献   

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