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Phylogenetic studies of soybean pathogen Phoma species by Bayesian analysis
53-61Views:119We 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. -
Phylogenetic analysis of Phoma species
100-107Views:92The 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:104The 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. -
Examining reaction of sunflower genotypes on planting time on chernozem soil
93-99Views:103We studied the effect of planting time on plant pathological factors, leaf area index and yield production by applying various fungicid treatments on two different sunflower genotypes in 2013.
By delaying planting time, both the extent of Diaporthe, Alternaria and Phoma infections decreased. The differences between the volume of infections were significant in the case of the early and late sowing time results. The application of fungicide treatments induced a notable decrease in the extent of infections for all three pathogens examined. The LAI-values varied between 0.3 and 5.6 m2/m2 in 2013 depending on the hybrid, sowing time and treatment. Stocks planted at distinct times reached maximum leaf coverage at different times. The planting time and the fungicide treatment had a significant effect on the formation of the leaf area. In the case of average and late planting times, fungicide treatments elongated the preservation of the green leaf area.
With respect to the yield amount, average planting time (27 April, 2013) turned out to be optimal in 2013 (control – NK Ferti: 4.621 kg ha-1, PR64H42: 4.196 kg ha-1; double-treated: NK Ferti: 5.282 kg ha-1, PR64H42: 5.090 kg ha-1). Fungicide treatments resulted in significant yield growth in all cases during our research.
We applied Person correlation analysis to evaluate the hybrids’ sensibility to infections and our results varied in the case of Diaporthe and Phoma (r=-0.343*, -0.379**). Infections of the three pathogens were significantly reduced by delaying the planting time and applying fungicides. Late sown stocks preserved the green leaf area for a longer period. Besides, the application of the fungicide treatment and the hybrid itself also led to the preservation of the green leaf area. However, pathogens examined notably decreased the leaf area by the end of the growing year. The fungicide treatment had a remarkable effect on yield growth (r=0.603**). Furthermore, the presence of higher LAI-values in the period prior to August also induced higher yields.
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Management of phytopathogens by application of green nanobiotechnology: Emerging trends and challenges
15-22Views:250Nanotechnology is highly interdisciplinary and important research area in modern science. The use of nanomaterials offer major advantages due to their unique size, shape and significantly improved physical, chemical, biological and antimicrobial properties. Physicochemical and antimicrobial properties of metal nanoparticles have received much attention of researchers. There are different methods i.e. chemical, physical and biological for synthesis of nanoparticles. Chemical and physical methods have some limitations, and therefore, biological methods are needed to develop environment-friendly synthesis of nanoparticles. Moreover, biological method for the production of nanoparticles is simpler than chemical method as biological agents secrete large amount of enzymes, which reduce metals and can be responsible for the synthesis and capping on nanoparticles.
Biological systems for nanoparticle synthesis include plants, fungi, bacteria, yeasts, and actinomycetes. Many plant species including Opuntia ficus-indica, Azardirachta indica, Lawsonia inermis, Triticum aestivum, Hydrilla verticillata, Citrus medica, Catharanthus roseus, Avena sativa, etc., bacteria, such as Bacillus subtilis, Sulfate-Reducing Bacteria, Pseudomonas stutzeri, Lactobacillus sp., Klebsiella aerogenes, Torulopsis sp., and fungi, like Fusarium spp. Aspergillus spp., Verticillium spp., Saccharomyces cerevisae MKY3, Phoma spp. etc. have been exploited for the synthesis of different nanoparticles. Among all biological systems, fungi have been found to be more efficient system for synthesis of metal nanoparticles as they are easy to grow, produce more biomass and secret many enzymes. We proposed the term myconanotechnology (myco = fungi, nanotechnology = the creation and exploitation of materials in the size range of 1–100 nm). Myconanotechnology is the interface between mycology and nanotechnology, and is an exciting new applied interdisciplinary science that may have considerable potential, partly due to the wide range and diversity of fungi.
Nanotechnology is the promising tool to improve agricultural productivity though delivery of genes and drug molecules to target sites at cellular levels, genetic improvement, and nano-array based gene-technologies for gene expressions in plants and also use of nanoparticles-based gene transfer for breeding of varieties resistant to different pathogens and pests. The nanoparticles like copper (Cu), silver (Ag), titanium (Ti) and chitosan have shown their potential as novel antimicrobials for the management of pathogenic microorganisms affecting agricultural crops. Different experiments confirmed that fungal hyphae and conidial germination of pathogenic fungi are significantly inhibited by copper nanoparticles. The nanotechnologies can be used for the disease detection and also for its management. The progress in development of nano-herbicides, nano-fungicides and nano-pesticides will open up new avenues in the field of management of plant pathogens. The use of different nanoparticles in agriculture will increase productivity of crop. It is the necessity of time to use nanotechnology in agriculture with extensive experimental trials. However, there are challenges particularly the toxicity, which is not a big issue as compared to fungicides and pesticides.
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Mass occurrence of a Phoma-like fungus on common ragweed (Ambrosia artemisiifolia) in Hajdúság region, East Hungary
55-60Views:206Common ragweed (Ambrosia artemisiifolia L.) is one of the most important, allergenic weed species in Hungary. A. artemisiifolia invades both a broad range of often disturbed areas (brownfields) and either undisturbed ones like waste lands, roadsides, riverbanks and railway tracks. In field crops it can cause considerable yield losses mainly in sunflower, maize, cereals and soybean. In Hungary many inhabitants suffer from allergy caused by Ambrosia pollen which results a serious human-health risk. The aim of the control is to prevent flowering and seed propagation of A. artemisiifolia. Until now the occurrence of numerous pathogenic fungi which attack common ragweed plants have been identified in Hungary, however there is not yet available biological weed control program because of shortage in acceptable effectiveness, and endangering cultural plant species. During our weed surveys in the region of Hajdúság (East-Hungary) we found numerous common ragweed plants showing heavy necrotic lesions on leaves and stems. The objective of this study was to identify the fungus which was isolated from diseased tissues of common ragweed (A. artemisiifolia). The identification of fungus based on morphological characters of colonies and features of conidia and chlamydospores developed on malt extract agar (MEA) plates. After examination of axenic cultures we revealed that the fungus isolated from the leaves ands stems of common ragweed was a Phoma-like species.