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Dieback of apricot plantations caused by 'Ca. Phytoplasma prunorum' in Borsod-Abaúj-Zemplén county (Northern-Hungary)
34-41Views:119Plant diseases caused by phytoplasmas have increasing importance in all over the world for fruit growers. Lately, phytoplasma diseases occur on many fruit varieties and responsible for serious losses both in quality and quantity of fruit production. In the long-run these diseases cause destruction of fruit trees. The apricot phytoplasma disease (Ca. Phytoplasma prunorum) was first reported in Europe in 1924 from France. In 1992 the disease has also been identified in Hungary. On the base of growers' signals serious damages of "Candidatus Phytoplasma prunorum" Seemüller and Schneider, 2004 (formerly: European stone fruit yellows phytoplasma) could be observed in different stone fruit plantations in the famous apricot-growing area nearby Gönc town, Northern-Hungary. Field examinations have been begun in 2009 in several stone fruit plantations in Borsod-Abaúj-Zemplén County mainly in Gönc region which is one of the most important apricot growing regions in Hungary, named “Gönc Apricot Growing Area”. Our goals were to diagnose the occurrence of Ca. Phytoplasma prunorum on stone fruits (especially on apricot) in the North-Hungarian growing areas by visual diagnostics and confirm data by laboratory PCR-based examinations. All the 28 collected samples were tested in laboratory trials and at 13 samples from apricot, peach, sour cherry and wild plum were confirmed the presence of phytoplasma (ESFY). On the base of observations it seems evident that the notable losses caused by "Ca. Phytoplasma prunorum" is a new plant health problem to manage for fruit growers, especially apricot producers in Hungary.
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Molecular studies on Cryphonectria parasitica isolates from Carpathian-basin
76-80Views:96Chryphonectria parasitica, the casual agent of chestnut blight, is one of the most important fungal pathogens of chestnut (Castanea spp.) in Europe and Hungary. In this study we analyzed the diversity of 14 Cryphonectria parasitica strains isolated from different location of Carpathian-Basin. For the analyses we used the partial sequences of the translation elongation coding gene, tef1. Our results showed that the tef1 gene, contrary to other fungal species, is not suitable for the molecular analyses of C. parasitica. In the future, for the molecular studies of C. parasitica, we need to use other molecular markers like microsatellites.
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Studies of Cryphonectria parasitica (MURR.) BARR subpopulations on Quercus petraea in Ukraine
70-75Views:122Chryphonectria parasitica (Murr.) Barr, the casual agent of chestnut blight disease, which is one of the most important fungal pathogens of chestnut (Castanea sativa). The disease seriously affected the chestnut in Northern-America and in Europe as well. It is important to mention that the pathogen does not only infect the chestnut but oak species (Quercus spp.) also. In the Carpathian-Basin, the chestnut is endemic in the Mecsek mountains, in Zala, in Somogy counties but it also can be found in the Danube-Bend. In the Carpathian-Basin (outside Hungary) the chestnut is found in Slovakia, Ukraine, Romania. In our study bark samples infected by Cryphonectria parasitica were collected from Bobovyshche, Serednje and Rostovjatica (Ukraine). The rate of infected chestnut tree were higher than 90% around Bobovyshche and beside chestnut, the symptoms were detected on oak trees as well. We collected bark samples from chestnut and oak as well and then we isolated the pathogen Cryphonectria parasitica in the lab of University of Debrecen. Symptomatological observations, laboratory examinations on fungus morphology, as well as comparisons of ITS sequency homology were made and approved that the causal agent of new disease was Cryphonectria parasitica. Our results proved that the Cryphonectria. parasitica infects oak trees beside chestnut in the Carpathian-Basin. Further studies are needed to determine the VCG (Vegatative Compatibility Group) group of the Cryphonectria parasitica found on oak trees.
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Laboratory diagnoses of the isolates of chestnut blight disease fungus Cryphonectria parasitica (MURR. BARR)
45-52Views:100Chryphonectria parasitica, the casual agent of chestnut blight, is one of the most important fungal pathogens of chestnut (Castanea spp.) in Europe and Hungary. In this study, we analyzed the ITS region of five Cryphonectria parasitica strains isolated from different location of Hungary. The differences among the Cryphonectria parasitica isolates were not insignificant because only two sites were considered as informative for the parsimony analysis. As the differences among geographically different isolates were insignificant, we mean that the evolutionary distance by ITS sequences within Hungarian Cryphonectria parasitica isolates is too small to get well based consequences for the phylogenetic relationships.
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Phylogenetic studies of soybean pathogen Phoma species by Bayesian analysis
53-61Views:137We carried out phylogenetic study analyzing sequences of genetic markers in the taxonomy of Phoma and Phoma-like fungi. Different species of Phoma and Phoma-like fungi occurring on soybean (Phoma pinodella, Phoma sojicola, Phyllosticta sojicola, Phoma exigua var. exigua) are difficult to identy because of their high morphological and symptomatic similarities.
Twenty-two isolates of nine different Phoma species were obtained from reference culture collections. Seven of them were isolated from soybean, the others were collected from different hosts.
The Phoma isolates were firstly characterised by morphologically, and then we employed a part of the gene responsible for the synthesis of translation elongation factor 1 subunit alpha protein (tef1), ITS region, as well as β-tubulin partial sequences as potential genetic markers to infer
phylogenetic relationships among different Phoma species..Finally, their ITS and tef1 sequences were sequenced and analysed by Bayesian approaches.
According to phylogenetic trees inferred by Bayesian analysis of tef1, ITS and β-tubulin sequences, different Phoma species can be separated proving that these phylogenetic markers are well suited for phylogenetic studies of Phoma species. However, the phylogenetic tree does not support the traditional Phoma sections based on morphological characterization.
Bayesian analyses of the three sequences confirmed that the Phyllosticta sojicola species is clustered with the Phoma exigua var. exigua group and the Phoma sojicola is grouped with Phoma pinodella group. The molecular data provide evidence for reclassification of formerly mentioned soybean pathogens. -
Cytochrome b diversity of Hungarian Botrytis cinerea strains
18-21Views:103In the mitochondrion of eukaryotes, cytochrome b is a component of respiratory chain complex III. Cytochrome b is encoded by the
cytochrome b (CYTB) gene located in the mitochondrial genome. The fungicidal activity of QoIs relies on their ability to inhibit mitochondrial respiration by binding at the so-called Qo site (the outer quinol-oxidation site) of the complex III. Since their introduction, QoIs (like azoxystrobin) have become essential components of plant disease control programs because of their wide-ranging efficacy against many agriculturally important fungal diseases like grey mould on various crops. QoI resistance primarily arises from a target-site-based mechanism involving mutations in the mitochondrial CYTB. As the management of grey mould is often dependent on chemicals, the rational design of control programs requires the information about the diversity of genes connected with resistance in field populations of the pathogen.
Monospore B. cinerea field isolates has been collected during 2008-2009 from different hosts in Hungary. PCR fragment length analysis
indicated the high frequency presence of type large intron in the isolates while in a few strains G143A substitution could also be detected.
These results indicated the heterogeneity of CYTB in the Hungarian B. cinerea populations, which possibly involve the heteroplasmy of this
mitochondrial gene, moreover indicates the existence op azoxystrobin resistant populations in Hungary.
This work was supported by NKFP-A2-2006/0017 grant. Erzsébet Fekete is a grantee of the János Bolyai Scholarship (BO/00519/09/8). -
Studies of Expression of Peptaibol Synthetase of Trichoderma reesei
188-190Views:130Because of the potential importance of peptaibols in the biological control of plant diseases, a transgenic, a T. reesei strain carrying a tex1-promoter: goxA fusion plasmid was constructed for furthur studies. The peptaibol synthetase gene (which is highly similar to T. virens tex1) was identified in the genome sequence of T. reesei. A 900 bp 5’ upstream noncoding fragment, presumed to include the promoter region of tex1, was cloned into the pSJ3 plasmid (which contains the Aspergillus niger goxA gene encoding glucose oxidase). Finally, we transformed T. reesei with the tex1-promoter: goxA fusion containing pSJ3 plasmid.
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Phylogenetic analysis of Phoma species
100-107Views:111The cosmopolitan Phoma genus contains mainly phytopathogenic, opportunistic parasites, and saprophyte fungal species. Up to now, the characterization of Phoma species and other taxa of Phoma has been determined on the basis of morphology on standardized media, and gene sequence analysis was only used as a confirmative or distinctive complement.
In this study, we tried to find molecular markers which can be used as phylogenetics markers in the molecular based classification in the Phoma genus.
We employed a part of the translation elongation factor 1 subunit alpha (EF-1α=tef1) containing both introns and exons and ITS region containing the internal transcribed spacer regions 1 and 2 and the 5.8S rDNA, as potential genetic markers to infer phylogenetic relationships among different Phoma taxa. Twelve different Phoma species sequences were analysed together with the closely related Ascochyta ones. The constructed phylogenetic trees, based on tef1 and ITS sequences, do not support the traditional Phoma sections based on morphological characterization. However, we managed to distinguish between the Phoma strains and Ascochyta species by comparing their tef1 sequences through parsimony analysis. We proved that a tef1 can be a useful phylogenetic marker to resolve phylogenetic relationships at species level in Phoma genus.
Both parsimony sequence analyses confirmed that the Phyllosticta sojicola species is identical to the Phoma exigua var. exigua species as Kövics et al. (1999) claimed. However, the evolutionary distance by ITS sequences within Phoma species is too small to get well based consequences for the phylogenetic relationships of Phoma genus.
Further investigations would be necessary to clarify whether the tef1 and ITS sequences as phylogenetic molecular markers are well suited for the classification of Phoma species. -
Phylogenetic studies of Phoma species by maximum likelihood analysis
37-46Views:116The cosmopolitan Phoma genus contains mainly phytopathogenic, opportunistic parasite, and saprophyte fungal species. Up to now the characterization of Phoma species and other taxa of Phoma has so far been determined on the basis of morphology on standardized media, and gene sequence analysis was only used as a confirmative or distinctive complement.
In this study we have tried to study phylogenetic relationships by maximum likelihood method in the Phoma genus. We employed a part of the gene responsible for the synthesis of translation elongation factor 1 subunit alpha protein (tef1) containing both introns and exons, a part of the gene responsible for synthesis of tubulin protein and ITS region containing the internal transcribed spacer regions 1 and 2 and the 5.8S rDNA as potential genetic markers to infer phylogenetic relationships among different Phoma taxa. Twenty-four isolates of eleven different Phoma species were firstly characterised by morphologically, and then their tef1, tubulin and ITS sequences were sequenced and analysed by maximum likelihood method carried out by PAUP*4.0b program. According to constructed phylogenetic trees, the different Phoma taxons are well separated. However these trees do not support the traditional Phoma sections based on morphological characterization.
The maximum likelihood analyses of all three sequences confirmed that the Phyllosticta sojicola species is clustered with the Phoma exigua var. exigua group and the Phoma sojicola is grouped with Phoma pinodella group. The experienced molecular evidences initiate the demand of reclassification of formerly mentioned soybean pathogens.