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Phenotypic and genotypic identification and phylogenetic characterisation of Taphrina fungi on alder
Abstract and Figures
All Taphrina species are dimorphic with a mycelium stage biotrophic on vascular plants and a saprophytic yeast stage. European species
of Taphrina on Alnus species (Betulaceae) were identified using morphological, physiological and molecular characteristics, the latter including
determination of PCR fingerprints and of nucleotide sequences from selected nuclear ribosomal DNA regions. PCR fingerprinting
gives a good overview of species identification, as do nucleotide sequences, which in addition, help to clarify phylogenetic
relationships. Taphrina alni is a homogeneous species that exhibited more than 50% similarity in PCR fingerprinting with three different primers. Morphologically,
it produces tongue-like outgrowths from female catkins of Alnus incana. Taphrina robinsoniana from A. rugosa and A. serrulata in North America is phylogenetically closely related to T. alni, but the two species could be separated by their PCR fingerprints, partial sequences of 26S rDNA (D1/D2) and ITS1/ITS2 sequences.
T. epiphylla and T. sadebeckii are two phylogenetically closely related species. T. epiphylla causes witches brooms in crowns of A. incana. In addition, T. epiphylla forms slightly yellow white-grey leaf spots in midsummer on A. incana. Yellow white-grey leaf spots up to 10 mm on A. glutinosa are characteristic for T. sadebeckii. Both species can be separated well by PCR fingerprinting. Different from T. epiphylla, T. sadebeckii is genotypically more heterogeneous. Only two out of three different primers showed similarity values above 50% in different
European strains of T. sadebeckii. Although genetic variability was not detected in complete sequences of the 18S ribosomal DNA of T. sadebeckii, ITS1/ITS2 sequences appeared to be more heterogeneous, too. Taphrina tosquinetii is a genotypically homogeneous species causing leaf curl on Alnus glutinosa. It was not possible to distinguish the yeast phases from different Taphrina species on Alnus using morphological and physiological characteristics only.
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... (T. deformans) described in this study were isolated from infected fruits using spore-fall methods [14]. Wet infected fruit tissue with mature asci was attached to the lids of a petri dish, thereby allowing the ascospores to be ejected from asci onto the potato dextrose agar (PDA) plates supplemented with 0.5 g L À1 streptomycin sulfate (PDAS). ...
... The morphological characteristics including asci, ascospores, and yeast cell size of T. deformans isolates from plum were consistent with those mentioned in previous reports for Taphrina spp. [9,14,15]. ...
... Koch's postulates were fulfilled by inoculating yeast cell suspensions of the obtained fungal isolates, on an attached, unopened flower bud; therefore, this study successfully confirmed that T. deformans was the causal agent of plum pocket disease. T. deformans is well known causal agent of peach leaf curl disease [14,15]. However, there is no report on plum pocket caused by T. deformans and to the best of our knowledge, this is the first report. ...
Plum pocket caused by the dimorphic ascomycetous fungi, Taphrina spp., results in unsightly malformations and crop loss. In 2016, Japanese plums (Prunus salicina Lindl.) with plum pocket symptoms were found in Gimcheon. Three isolates were collected from symptomatic P. salicina fruits and identified as Taphrina deformans based on morphological characteristics and molecular sequence analysis of including internal transcribed space (ITS) and the mitochondrial small ribosomal subunit (SSU) regions of the three isolates. Pathogenicity test on plum fruits confirmed that, the present T. deformans isolates are causal agent of plum pocket. To the best of our knowledge, this is the first report of plum pocket caused by T. deformans in South Korea.
... In order to identify Taphrina spp. molecular methods were used by Sjamsuridzal et al. (1997), Rodríguez and Fonseca (2003), Bacigálová et al. (2003), Tavares et al. (2004), Petrýdesová et al. (2013), Isheng et al. (2014) and Selbmann et al. (2014). The most important species include: T. deformans causes deformation of leaves, fruits and flowers and the defoliation of Prunus persica and Prunus dulcis (Mill.) ...
... Las especies de Taphrina se identificaron con base a la distribución geográfica, el sitio, el tipo de infección, los síntomas, la localización del micelio, la morfología de las estructuras fúngicas y el rango del huésped (Mix, 1949;Taylor y Birdwell, 2000;Rodríguez y Fonseca, 2003). Métodos moleculares se usaron por Sjamsuridzal et al. (1997), Rodríguez y Fonseca (2003), Bacigálová et al. (2003), Tavares et al. (2004), Petrýdesová et al. (2013), Isheng et al. (2014) y Selbmann et al. (2014) para identificar especies de Taphrina. Las especies más importantes incluyen: T. deformans, que causa deformación de hojas, frutos y flores y la defoliación de Prunus persica y Prunus dulcis (Mill.) ...
... Method 3. The blastospore fall method. Leaf tissue of Q. eduardii bearing asci of T. caerulescens was fastened to Petri dish lids over PDA (Taylor and Birdwell, 2000;Bacigálová et al., 2003). ...
Taphrina caerulescens exclusively affects plants of the Quercus
genus. The identification and isolation of this fungus is difficult
due to its dimorphic nature and extremely slow growth habit
in artificial growth media. The objective of this research was to
isolate and identify the fungal pathogen T. caerulescens. Three
methods were used to isolate the fungus, however, only the spore
fall method was successful. In order to identify the fungus, a
visual inspection of the host plants infected leaves was carried
out. The symptoms and signs of infection were noted. In
addition to the typical blister formation on the infected leaves,
an atypical symptom comprising of chlorotic spots covered with
asci was also observed. Three biochemical tests were carried
out to prove that the pathogen was in the genus Taphrina: The
first, tested the ability to utilize urea, the second, its capacity to
produce extra cellular DNase, and the third tested its reaction to
Diazonium blue B (rapid blue salts). For species identification
the following analyses were carried out: 1) carbon assimilation
tests using the glucose, saccharose, D-xylose, raffinose, trehalose
and sorbitol, as carbon sources; 2) molecular analysis tools were
utilized to extract and sequence the genomic DNA. The resulting
sequences when analysed with the NCBI Nucleotide blast tool
confirmed, with 99% similitude, that the isolate was similar in
nature to T. caerulescens strain CBS 351.35 ascension number
AF492081.1. Pathogenicity test was also carried out and resulted
in the infection of 25 % of the inoculated plants.
... Species of the genus Taphrina have been recognized especially by symptoms caused to their host plants, anatomical and morphological features of their sexual stage in the infected tissues and by their geographic distribution (Mix, 1949;Gjaerum, 1964;Salata, 1974;Sadebeck, 1893). They are also distinguished by a unique combination of physiological and biochemical characteristics as well as by the carbohydrate composition of their cell walls (Prillinger et al., 2000;Bacig alov a et al., 2003). However, molecular biological Abbreviations: BI, Bayesian inference; ITS, internal transcribed spacer; MP, maximum-parsimony; rns, mitochondrial small subunit rRNA gene. ...
... The GenBank/EMBL/DDBJ accession numbers of the sequences of the ITS region and the rns gene of the holotype of T. gei-montani CCY 058-007-001 T determined in this study are KU134800 and KU134828, respectively. Other sequence data determined in this study are detailed in Table 1 techniques involved in yeast identification have led to a revaluation of validity of the species described hitherto (Nishida & Sugiyama, 1994;Sugiyama, 1998;Prillinger et al., 2000;Bacig alov a et al., 2003;. ...
... The infected plants were collected during the vegetation periods of 2013 and 2014 from mountain localities in the Western Carpathians (Vysok e Tatry Mts, Slovakia) ( Table 1). The yeasts in their anamorphic stage were isolated following the procedures described by Bacig alov a et al. (2003). A teleomorphic stage was characterized according to the previous morpho-anatomical investigations (Bacig alov a, 1992;Bacig alov a et al., 2014). ...
Five strains (CCY 058-007-001T, CCY 058-007-002, CCY 058-007-003, CCY 058-007-004 and CCY 058-007-005) of a novel parasitic yeast belonging to the genus Taphrina were isolated from leaf tissues of Geum montanum L. (Rosaceae), collected from the Vysoké Tatry Mts., Slovakia. Genetic analyses revealed that these isolates differ by 15 unique substitutions in the ITS region and by 6 substitutions in the rns gene from all other Taphrina species analysed hitherto. The novel strains are also distinguished from all other Taphrina species by their morphology, biochemical properties and ecology. These strains represent a new species, for which the name Taphrina gei-montani sp. nov. is proposed. The type strain is CCY 058-007-001T (= CBS 14159 = BU001). The Mycobank number is MB815677. The present study also demonstrates that two distinct Taphrina species parasitize the herbaceous Rosaceae: Taphrina gei-montani on Geum montanum and Taphrina tormentillae on Potentilla species.
... In addition, Dothideomycetes have also been found in decaying leaves [69], and a previous study showed that Dothideomycetes from living leaves grew faster than those isolated from non-living leaves [70], resulting in a greater proportion of Dothideomycetes in the healthy leaves. Taphrinomycetes were more abundant in nonhealthy leaves, possibly because members of Taphrinomycetes are generally recognized as phytopathogens in higher plants [71]. ...
The rare and endangered plant species Abies fanjingshanensis, which has a limited habitat, a limited distribution area, and a small population, is under severe threat, particularly due to poor leaf health. The plant endophytic microbiome is an integral part of the host, and increasing evidence indicates that the interplay between plants and endophytic microbes is a key determinant for sustaining plant fitness. However, little attention has been given to the differences in the endophytic microbial community structure, network complexity, and assembly processes in leaves with different leaf health statuses. Here, we investigated the endophytic bacterial and fungal communities in healthy leaves (HLs) and non-healthy leaves (NLs) of A. fanjingshanensis using 16S rRNA gene and internal transcribed spacer sequencing and evaluated how leaf health status affects the co-occurrence patterns and assembly processes of leaf endophytic microbial communities based on the co-occurrence networks, the niche breadth index, a neutral community model, and C-score metrics. HLs had significantly greater endophytic bacterial and fungal abundance and diversity than NLs, and there were significant differences in the endophytic microbial communities between HLs and NLs. Leaf-health-sensitive endophytic microbes were taxonomically diverse and were mainly grouped into four ecological clusters according to leaf health status. Poor leaf health reduced the complexity of the endophytic bacterial and fungal community networks, as reflected by a decrease in network nodes and edges and an increase in degrees of betweenness and assortativity. The stochastic processes of endophytic bacterial and fungal community assembly were weakened, and the deterministic processes became more important with declining leaf health. These results have important implications for understanding the ecological patterns and interactions of endophytic microbial communities in response to changing leaf health status and provide opportunities for further studies on exploiting plant endophytic microbes to conserve this endangered Abies species.
... (Nishida et al., 1993). Molecular tools used in phylogenetic studies of Ascomycota (Sugiyama et al., 2006) have included large and small subunits of ribosomal RNA genes and sequence analysis of the Internal Transcribed Spacer (ITS) regions of the rRNA genes (Bacigalova et al., 2003). ...
Coriander (Coriandrum sativum L.) is an important highly nutritive crop belonging to family Apiaceae. To meet increasing global demand and consumption, coriander productivity must be enhanced. However, biotic stresses such as diseases have impeded coriander cultivation both in the tropics and subtropics. Of them, stem gall of coriander, incited by the fungus Protomyces macrosporus Unger, is one of the most common, widespread and serious diseases throughout the world and causes significant yield losses. Symptomatology, biology, and etiology of stem gall and its control through fungicides, bio-product and bio-control have not been widely studied, and substantial information is still required on the inoculum potential, growth models and epidemiological parameters of stem gall, the influence of changing the climate, impact of extensive use of fungicides and disease resistance. These critical factors may influence the development and emergence of stem gall. Because of the increasing demands of coriander in the world, control of stem gall is gaining importance. The present review deals briefly with different aspects of stem gall disease with major emphasis on its ecology, pathology, epidemiology, and management. Some new approaches such as biological control, integrated management strategies, and some other aspects are also discussed.
... Maire occurs on green alder (A. viridis (Chaix) DC.) (Bacigálová et al., 2003). ...
... No inoculation experiments have been done in any other countries, and the diameter of the ascogenous cells (Table 3) does not support the creation of a new species, so we accepted the name Protomyces macrosporus. The tools of molecular biology should clarify many phylogenetic and taxonomic problems (including species concept); Sjamsuridzal et al. (1997), for example, used them to construct a phylogenetic hypothesis on evolutionary relationships within Taphrinales, and Bacigálová et al. (2003) to characterize Taphrina species infecting alder. ...
Two interesting species of Protomyces Unger collected recently in the Tatra Mts are described, illustrated in detail and compared with similar taxa. Protomyces crepidis-paludosae Büren on Crepis paludosa (L.) Moench is a new species for Slovakia. Protomyces macrosporus Unger on Laserpitium latifolium L. is a new fungus/host combination in the Carpathians. Protomyces macrosporus Unger on Carum carvi L. is reported from the fi rst locality in the Slovak part of the Tatra Mts.
Peach leaf curl disease (PLCuD) is a devastating fungal disease that poses a significant threat to peach production globally, and its impact is also evident in Pakistan's fruit industry. The disease is caused by phyto-parasitic fungi from the Taphrinaceae family, primarily the genus Taphrina. Chemical fungicides are the most common method for controlling the disease; however, their use presents health and environmental concerns and can lead to resistance over time. As a result, alternative control strategies that are both cost-effective and environmentally friendly are essential. Unfortunately, there is a significant gap in knowledge about PLCuD in Pakistan, and the actual cause of the disease remains unidentified, making management impossible. Therefore, discovering novel biological control methods is crucial to combat this disease effectively. Recent advances in biotechnology have shown promising results in developing biological control agents, such as microbial inoculants, bio-fungicides, and bio-pesticides. These agents can specifically target the fungi responsible for the disease, leading to more efficient and sustainable control measures. The review highlights the urgent need to develop sustainable and effective biological control methods to combat PLCuD in Pakistan and to provide innovative and eco-friendly solutions for managing this devastating disease and ensuring the long-term sustainability of the peach industry in Pakistan.
The early diverging Ascomycota lineage, detected primarily from nSSU rDNA sequence-based phylogenetic analyses, includes enigmatic key taxa important to an understanding of the phylogeny and evolution of higher fungi. At the moment six representative genera of early diverging ascomycetes (i.e. Taphrina, Protomyces, Saitoella, Schizosaccharomyces, Pneumocystis and Neolecta) have been assigned to “Archiascomycetes” sensu Nishida and Sugiyama (1994) ———, ———. 1994a. Phylogeny and molecular evolution among higher fungi. Nippon Nogeikagaku Kaishi 68:54–57. (In Japanese.) or the subphylum “Taphrinomycotina” sensu Eriksson and Winka (1997) ———, Winka K. 1997. Supraordinal taxa of the Ascomycota. Myconet 1:1–16. . The group includes fungi that are ecologically and morphologically diverse, and it is difficult therefore to define the group based on common phenotypic characters. Bayesian analyses of nSSU rDNA or combined nSSU and nLSU rDNA sequences supported previously published Ascomycota frameworks that consist of three major lineages (i.e. a group of early diverging Ascomycota [Taphrinomycotina], Saccharomycotina and Pezizomycotina); Taphrinomycotina is the sister group of Saccharomycotina and Pezizomycotina. The 50% majority rule consensus of 18 000 Bayesian MCMCMC-generated trees from multilocus gene sequences of nSSU rDNA, nLSU rDNA (D1/D2), RPB2 and β-tubulin also showed the monophyly of the three subphyla and the basal position of Taphrinomycotina in Ascomycota with significantly higher statistical support. However to answer controversial questions on the origin, monophyly and evolution of the Taphrinomycotina, additional integrated phylogenetic analyses might be necessary using sequences of more genes with broader taxon sampling from the early diverging Ascomycota.
The ascomycetous genus Taphrina Fries originally contained more than 90 phytopathogenic microscopic dimorphic ascomycetes causing specific infections in different vascular plants. Although species have mainly been identified historically according to their host and morphological and physiological traits, they can be studied and preserved in the form of yeasts arising from germinating ascospores. Due to low DNA sequence divergence and the lack of available strains, the number of accepted Taphrina species has currently been reduced to twenty-eight. The aim of this study is the description of three previously accepted species. Taphrina bullata, Taphrina insititiae and Taphrina rhizophora, which have been omitted from a recent key (Fonseca & Rodrigues, 2011). The host range, the divergence of the 26S rDNA sequence, internal transcribed spacers (ITS) and the mitochondrial small ribosomal subunit (rns) sequence strongly support their reassignement as a new species.
To study the phylogeny and evolution of archiascomycetes, we determined the full sequence of the nuclear 18S rRNA gene from
14Taphrina species and 2Protomyces species, and the partial sequence ofSchizosaccharomyces japonicus var.japonicus. The sequences were phylogenetically analyzed by the neighbor-joining, maximum parsimony, and maximum-likelihood methods.
We also looked at their principal phenotypic characters and genotypic character. Relationships within the Ascomycota are concordant
with the previously published phylogenies inferred from 18S rDNA sequence divergence and divide the archi-, hemi-and euascomycetes
into distinct major lineages. All the trees show that, within the archiascomycete lineage, 11 of the 14Taphrina species and the 2Protomyces species are monophyletic. A core groups ofTaphrina andProtomyces is always monophyletic. The evidence from molecular and phenotypic characters such as cell wall sugar composition, ubiquinone,
cell wall ultrastructure, and mode of conidium ontogeny, strongly suggests that ‘T’. californica CBS 374.39, ‘T’. maculans CBS 427.69 and ‘T’. farlowii CBS 376.39 should be excluded from the archiascomycete lineage. ‘Taphrina’ farlowii CBS 376.39 groups withCandida albicans in the Saccharomycetales, whereas ‘T’. californica CBS 374.39 and ‘T’. maculans CBS 427.69 have a basidiomycete affinity and group with Tremellalean members in the hymenomycete lineage.Schizosaccharomyces is monophyletic. The strictly anamorphic yeastSaitoella complicata groups with the apothecial ascomyceteNeolecta vitellina rather than theTaphrina/Protomyces branch.
Seventy-four isolates, representing twenty-six species and fifty-five host-forms of Taphrina, were grown in mineral solution with thirty-two different carbon compounds. All of these fungi utilized dextrose, suerose, maltose, melezitose, trehalose, dextrin, inulin, and pectin. All but one (T. polystichi) used xylose and all but two (T. polystichi and T. nana) used succinic acid. None of them used lactose, rhamnose, inositol, nor i-erythritol. With regard to eighteen compounds: galactose, levulose, mannose, raffinose, cellobiose, 1-arabinose, melibiose, 1-sorbose, d-ribose, adonitol, dulcitol, mannitol, d-sorbitol, ethanol, glycerol, salicin, soluble starch, and citric acid, each species exhibited an individual pattern of carbon utilization.
Different host-forms of Taphrina deformans, T. flavorubra, T. robinsoniana, and T. ulmi agreed as to carbon compounds utilized. Other host-forms differed from each other in this respect, sometimes as much as did individual species. There was an approach to agreement between different host-forms of T. communis and also between the fungi from Populus, T. johansonii, T. populina, and T. populi-salicis.
None of these fungi required the addition of thiamine chloride to the medium.
Fifty-seven host-forms, representing twenty-seven species of Taphrina, and thirteen asporogenous yeasts were grown in liquid media with different sources of nitrogen as follows: eight ammonium compounds, four nitrates, urea, peptone, asparagine, and seventeen amino acids.
Ammonium chloride was refused by Taphrina americana and by T. caerulescens from Quercus macrocarpa and Q. prinoides.
Ammonium nitrates was refused by Taphrina nana, T. ulmi from Ulmus rubra, Candida pseudotropicalis var. lactose and by Torulopsis sphaerica.
Urea and asparagine were used by all the fungi studied. Peptone was refused only by Taphrina americana.
All non-sporulating yeasts but Torulopsis dattila used ammonium acetate; none of the species of Taphrina studied used this compound.
All species of Taphrina showed a characteristic pattern of nitrogen utilization. Some host-forms within species agreed closely in behavior while others showed various degrees of difference.
None of the fungi studied required the addition of thiamine to the media used, though in general they made better growth with added thiamine.
Most of the asporogenous yeasts were omnivorous or nearly so with regard to the compounds offered them. Torulopsis dattila and Candida pseudotropicalis var. lactosa were highly selective.
It seems possible to differentiate species of Taphrina in culture on the basis of their use or refusal of different compounds of nitrogen.
Brief notes on the distribution of Taphrinales fungi in Slovakia during the last 90 years are presented. Possible response of the fungi to rapid climate changes in the territory of Slovakia during this period is discussed.
— We studied sequence variation in 16S rDNA in 204 individuals from 37 populations of the land snail Candidula unifasciata (Poiret 1801) across the core species range in France, Switzerland, and Germany. Phylogeographic, nested clade, and coalescence analyses were used to elucidate the species evolutionary history. The study revealed the presence of two major evolutionary lineages that evolved in separate refuges in southeast France as result of previous fragmentation during the Pleistocene. Applying a recent extension of the nested clade analysis (Templeton 2001), we inferred that range expansions along river valleys in independent corridors to the north led eventually to a secondary contact zone of the major clades around the Geneva Basin. There is evidence supporting the idea that the formation of the secondary contact zone and the colonization of Germany might be postglacial events. The phylogeographic history inferred for C. unifasciata differs from general biogeographic patterns of postglacial colonization previously identified for other taxa, and it might represent a common model for species with restricted dispersal.