358 research outputs found
Evaluating the expression profile in plane leaves treated with cerato-platanin and C. fimbriata f. sp. platani conidia
No full textCerato-platanin (CP) is produced from the Ascomycete Ceratocystis fimbriata (Ell. and Halst.) Davidson f. sp. platani Walter (Cfp) that causes the canker stain of the plane trees (Boddi et al. 2004, Pazzagli et al. 1999 and 2006a). In in vitro experimental conditions CP self-assembles and interacts with the host plane leaves by eliciting phytoalexin synthesis, cell plasmolysis and necrosis and restriction of Cfp growth (Bennici et al. 2005, Pazzagli et al 2006b, Santini et al 2006, Scala et al. 2004). We have applied the suppression-subtractive hybridisation (SSH) methodology for isolation and characterisation of genes induced after CP treatment for 48 hours. The data show an intense metabolic activity and many genes differentially expressed, the most part of which code for: 1) defence and/or stress related proteins; 2) proteins involved in the protein synthesis/turnover; 3) proteins belonging to cell primary metabolism and 4) proteins involved in the signalling pathway s. We have analysed some of them by relative PCR using total RNA from treated leaves with CP and Cfp conidia at different times. From the results the CP has been shown to stimulate defence responses
Early gene expression during the experimental induction of plane canker disease
No full textCerato-platanin (CP), a 120 amino acids protein purified from the culture filtrate of Ceratocystis fimbriata (Ell. and Halst.) Davidson f. sp. platani Walter (Cfp), is the causal agent of the canker stain of the plane trees (Pazzagli et al., 1999). Cfp belongs to Ascomycetes family and attacks Platanus occidentalis, P. orientalis and their hybrid Platanus acerifolia that is the most susceptible. The CP is produced in the first steps of the Cfp growth and it is located in the Cfp cell wall of hyphae, conidia and ascospores (Sereni et al., 2002). Pazzagli et al. (1999) suggested the potential role of CP as a signal molecule in the induction of plant defence mechanisms. In in vitro experimental conditions CP self-assembles and interacts with the host plane leaves by eliciting phytoalexin synthesis, and inducing extended cell plasmolysis and abundant starch accumulation in the chloroplasts (Bennici et al., 2005; Boddi et al., 2004; Pazzagli et al., 2005; Scala et al., 2004). Many defence techniques been used (chemical methods, genetic improvement, biological struggle) against this pathology, but they were ineffective. In consequence of the absence of defence methods it is very important to improve the knowledge of the plant-host interaction and of the fungus factors involved in the development of new defence strategies. For this purpose we have applied the suppression-subtractive hybridisation (SSH) methodology for isolation and further characterisation of pathogen induced genes. We have isolated many clones and we have analysed some of them by relative PCR using total RNA extracted from treated leaves with fungus conidia and cerato-platanin for 6, 24 and 48 hours after treatments in order to investigate possible differences in gene expression during CP/plant and fungus/plant interactions. The data show an intense activity in consequence of CP and fungus treatments: in fact, the treatments seem to increase the cell primary metabolism (particularly, the photosynthesis, the pentose phosphate cycle and the ammonia assimilation pathway), the signalling, the protein synthesis/turnover and the defence and/or stress related protein. These results show the CP ability to stimulate defence responses and to act like the fungus
Gene induction in Platanus acerifolia after treatments with cerato-platanin and conidia of Ceratocystis fimbriata sp. Platani
No full textCerato-platanin (CP) is a 120 amino acids-long protein purified from the culture filtrate of Ceratocystis fimbriata (Ell. and Halst.) Davidson f. sp. platani Walter (Cfp), the causative agent of the canker stain of the plane trees (Pazzagli et al., 1999). CP contains 4 cysteines forming two S-S bridges, Cys20-57 and Cys60-115, and has a high percentage (40%) of hydrophobic residues. It is the founder member of the cerato- platanin family, and its N-terminal region is very similar to cerato-ulmin, a class II hydrophobin involved in the pathogenesis of Dutch elm disease (Del Sorbo et al., 2002). CP is located in the Cfp cell walls, and is early-detected in Cfp culture filtrates. In in vitro experimental conditions CP self-assembles, and interacts with the host plane leaves by eliciting phytoalexin synthesis, extended cell plasmolysis and crushing, and abundant starch accumulation in the chloroplasts (Bennici et al., 2005; Boddi et al., 2004; Pazzagli et al., 2005; Scala et al., 2004). cDNA libraries were constructed from RNA extracted from leaves treated with CP (treated) and with water (control) for 48 hours using the Suppressive Subtractive Hybridisation (SSH) method, a powerful technique that enables to compare two population of mRNA and to obtain clones of genes that are expressed in one population of mRNA but not in the other. The positive clones were sequenced and the sequences were analysed using the FASTA, BLAST, ProDom, BLOCK software for their identification in GenBank, EMBL. Many clones from the forward library contain sequences involved: i) in synthesis, processing and translation of proteins, i.e., the DEAD box ATPase/RNA helicase protein, the oligouridylate binding protein, an unknown protein with the RIBOSOMAL S8E domain, the elongation factor 1 alpha, the translation factor erF3, the ribosomal protein PRPL5; ii) in signal transfer in the cell, i.e. the Pto-like serine-threonine kinase; iii) in transcription control, i.e. the histone deacetilase, the DEAD-box helicase; iv) in photosynthesis, i.e. the PSI-D subunit, the ferredoxin, the chlorophyll a/b binding protein. Someone else interesting are: the TUBBY-like protein (TULP); the Fatty Acid Elongase (FAE), in particular, a keto-acyl CoA synthase; the Inosine 5’-phosphate-dehydrogenase; the PEP-carboxyl-kinase; the alanine-amino-transferase; the selenium binding protein; the RAR1; a protein with a DUF597 domain; the thaumatin protein. Moreover, many clones were analysed by Real Time RT-PCR and/or relative PCR in order to study their regulation after treatments with CP or with conidia of Cfp wild strain. Analysis of forward clones showed that many genes are positively regulated. The knowledge of the gene expression after treatments could open the possibility of inducing resistance to Cfp in plane trees
Plasminds containing multiple competitors and their application for angiogenic factor quantification (Angiobalance)
No full textModulation of Gene Expression Induced in Platanus Acerifolia by Cerato-Platanin and Conidial Suspension of Ceratocystis Platani
No full textCerato-platanin (CP) is a small protein produced by Ascomycete Ceratocystis platani, the causal agent of plane canker stain. Cep is pathogenic to Platanus orientalis, P. occidentalis and their hybrid P. acerifolia. CP is located in the fungal cell walls, is early released in culture, and elicits defence-related structural and physiological responses in host and non-host plants, such as cell plasmolysis and death, phenolic compounds and phytoalexin accumulation (Pazzagli et al., J. Biol. Chem. 274: 24959-24964, 1999; Boddi et al., FEMS Microbiol. Letters 233: 341-346, 2004; Scala et al., J. Plant Pathol. 86: 23-29,2004; Bennici et al., Caryologia 59: 291-298, 2006). Recently, it has been demonstrated the close correlation between the CP treatment of plane leaves and the subsequent growth inhibition of C. platani on the surface of the treated leaves together with the production of phytoalexins and the over-transcription of regulatory and defense-associated genes (Fontana et al., J. Plant Pathol. 90: 293-304, 2008).
The aim of the present study was to provide further information on the modulation of gene expression that occurs in the plane tree by CP treatment in order to improve our understanding of the potential of CP to function as a PAMP (Pathogen-Associated Molecular Pattern) protein in the pathogenic process of the C. platani-P. acerifolia interaction.
In the present study we have isolated others clones from a cDNA library constructed using suppression subtractive hybridization (SSH) technique (Fontana et al., J. Plant Pathol. 90: 293-304, 2008) and the putative differentially expressed genes were identified using the FASTA, BLASTN and BLASTX programs. The up-regulated clones were classified in the macro putative groups taking in account the functional categories established for Arabidopsis (The Arabidopsis Genome Initiative, Nature 408: 796-815, 2000). We have analysed, moreover, some of them by relative PCR using total RNA extracted from leaves treated with CP or fungal conidia at 6, 24 and 48 hours after treatments in order to investigate possible differences in gene expression during CP/plant and fungus/plant interactions
Cerato-platanin from C. fimbriata f. sp. platani is an host resistance inducing protein and is produced by other strains of C. fimbriata and by some other species of the genus Ceratocystis
No full textCerato-platanin (CP) is a 120 amino acids protein [1, 5], produced by the Ascomycete Ceratocystis fimbriata f. sp. platani (Cfp), the causal agent of the plane canker stain. The species C. fimbriata attacks various other plants of considerable importance in agriculture, forestry and for their ornamental value; as a rule, one fungal strain isolated from one host is not virulent on the other plant species, and conversely, susceptible hosts are resistant to C. fimbriata strains if they come from hosts other than themselves. This means that the forma specialis platani of the species C. fimbriata attacks only the trees belonging to the genus Platanus, but not the hosts of the all other formae speciales of the fungus. CP is located in the cell walls of Cfp ascospores, hyphae and conidia, and is early secreted when Cfp is grown in liquid culture [2, 3]. CP elicits phytoalexin synthesis and/or cell necrosis in host and in non-host tissues; in plane leaves the main effects of CP are to cause a great increase in primary starch and a certain degree of intercellular and intracellular disorganization of the spongy parenchyma cells and plasmolysis processes; in addition, an increase of intracellular phenolic compounds has been observed in the palisade cells [3, 4]. In the present work we report that the minimum CP concentration able to induce the decrease of the 50% Cfp growth on plane leaves is of about 5 x 10-5 M; the maximum inducing effect has been obtained 24-48 hours post treatment. At this time, numerous defense-related genes are over-expressed, as it has been shown by Suppressive Subtractive Hybridisation. Moreover, results so far obtained by immunotechnical experiments on a total of 17 strains (9 of C. fimbriata, as well as 1 isolate each of C. moniliforme, C. allantospora, C. fagacearum, C. laricicola, C. ambrosia, Microascus cirrosus, Ophiostoma ulmi and O. novo-ulmi) indicate that a CP-homologous protein occurs in all strains of C. fimbriata and in some other species of Ceratocystis. For some strains of C. fimbriata the coding sequences of the cp-hortologous genes have been obtained, and then the sequences of the deduced proteins. BIBLIOGRAFIA 1. Pazzagli L, Cappugi G, Manao G, Camici G, Santini A and Scala A, 1999. Purification of cerato-platanin, a new phytotoxic protein from Ceratocystis fimbriata f.sp. platani. Journal of Biological Chemistry 274: 24959-24964. 2. Boddi S, Comparini C, Calamassi R, Pazzagli L, Cappugi G and Scala A, 2004. Cerato-platanin protein is located in the cell walls of ascospores, conidia and hyphae of Ceratocystis fimbriata f. sp. platani. FEMS Microbiology Letters 233: 341-346. 3. Scala A, Pazzagli L, Comparini C, Santini A, Tegli S and Cappugi G, 2004. Cerato-platanin, an early-produced protein by Ceratocystis fimbriata f. sp. platani, elicits phytoalexin synthesis in host and non-host plants. Journal of Plant Pathology 86: 23-29. 4. Bennici A, Calamassi R, Pazzagli L, Comparini C, Schiff S, Bovelli R, Mori B, Tani C and Scala A, 2005. Cytological and ultrastructural responses of Platanus acerifolia (Ait.) Willd. leaves to cerato-platanin, a protein from Ceratocystis fimbriata f. sp. platani. Phytopathologia Mediterranea 44: 153-161. 5. Pazzagli L, Pantera B, Carresi L, Zoppi C, Pertinhez TA, Spisni A, Tegli S, Scala A, Cappugi G, 2006. Cerato-platanin, the first member of a new fungal protein family: cloning, expression and characterization. Cell Biochemistry and Biophysics 44: 512-521
Contesti umanitari e periferie emergenti: la cooperazione internazionale e i nomadismi della contemporaneità
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No full textEditoriale. Verso un ‘grappolo’ di storia multidisciplinare del territorio
No full textSulla storia multidisciplinare del territorio
Computerized impedance plethysmography in the diagnosis of delayed deep vein thrombosis after total hip replacement
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