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Morphology of the isolate CNU 060219 (Peyronellaea calorpreferens). Colonies on oatmeal agar (A) and malt extract agar (B) media for 14 days at 22℃, pycnidia (C~E), chlamydospores (F~L), conidia (M) (scale bars: C~E = 40 µm, F~L = 20 µm, M = 10 µm).
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Species of Phoma and its allies were isolated during a survey on the diversity of endophytic fungi associated with pine trees in Korea. Based on the phylogenetic analyses of internal transcribed spacer and β-tubulin gene sequences, two Phoma-like species from the needles of Pinus koraiensis were identified as Peyronellaea calorpreferens and P. glom...
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Endophytic fungi have been creating a considerable interest and curiosity among researchers since past three decades globally, owing to their recognition as an inexhaustible source of structurally and biologically novel compounds, alternative source of metabolites functionally identical to plant produced metabolites and their ability to impart resi...
Preparation of fungal material for microscopical examination of their conidiogenous structures is described below. A modification to the existing methods used coupled with a sporulation medium has proved to be simple to use and successful for the induction of sporulation and subsequent examination of conidiogenous structures in a range of xylariace...
Diversity of endophytic fungi
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... It has also been reported to be a mycoparasite of powdery mildew [85]. Didymella glomerata as P. glomerata has been recorded as an endophytic fungus from Korean pine (Pinus koraiensis) leaves [86]. It has also been associated with the pea "Ascochyta blight complex" in Australia [87]. ...
Peach (Prunus persica L.) is one of the most important and oldest stone fruits grown in China. Even though P. persica is one of the most commonly grown stone fruits in China, little is known about the biodiversity of microfungi associated with peach branch diseases. In the present study, samples were collected from a wide range of peach growing areas in China, and fungal pathogens associated with peach branch diseases were isolated. In total, 85 isolates were obtained and further classified into nine genera and 10 species. Most of the isolates belonged to Botryosphaeriaceae (46), including Botryosphaeria, Diplodia, Neofusicoccum, Phaeobotryon, and Lasiodiplodia species; Ascochyta, Didymella, and Nothophoma species representing Didymellaceae were also identified. Herein, we introduce Ascochyta prunus and Lasiodiplodia pruni as novel species. In addition, we report the first records of Nothophoma pruni, Neofusicoccum occulatum, and Phaeobotryon rhois on peach worldwide, and Didymella glomerata, Nothophoma quercina, and Phaeoacremonium scolyti are the first records from China. This research is the first comprehensive investigation to explore the microfungi associated with peach branch disease in China. Future studies are necessary to understand the pathogenicity and disease epidemiology of these identified species.
... Sample fascicles were immersed in sterile deionized water and stirred for 1 min prior to immersion for 1 min in EtOH, 3 min in NaOCl (4% available Cl), 30 s in EtOH, and 30 s in sterile deionized water following [54]. Sterilized samples were air-dried in a sterile flow-hood, and dried needles were then stored at −20 • C under sterile conditions. ...
Plant endosymbionts (endophytes) influence host plant health and express genotype-dependent ecological relationships with plant hosts. A fungal species intended to confer host plant resistance to a forest pathogen was used as inoculum to test for effects of inoculation on disease resistance, microbiomes, and phytochemistry of a threatened pine species planted in a restoration setting. Correlations of inoculation presence/absence, phytochemistry, spatial location of seedlings, maternal seed sources, and fungal endophytic communities in the foliage of six-year-old whitebark pine (Pinus albicaulis) seedlings were assessed five years after an experimental inoculation of seedlings with foliar endophytic fungi cultured from whitebark pine trees at Crater Lake National Park, including Paramyrothecium roridum. We hypothesized that P. roridum would modify host microbiomes in a manner that combats white pine blister rust disease. Our assessment of seedlings in the field five years after inoculation allowed us to consider whether inoculation stimulated long-lasting changes in microbiome communities and whether effects varied by seedling genetic family. Tests for effects of endophyte inoculation on disease resistance were inconclusive due to current low levels of rust infection observed at the field site. Foliar fungal endophyte richness and Shannon diversity varied with maternal seed sources. Isotopic stoichiometry and phytochemistry did not vary with seedling spatial proximity, inoculation treatment, or maternal seed family. However, endophyte community composition varied with both seedling spatial proximity and maternal seed sources. Endophytic communities did not vary with the inoculation treatment, and the hypothesized biocontrol was not detected in inoculated seedlings. We draw three conclusions from this work: (1) fungal microbiomes of whitebark pine seedlings across our study site did not vary with host phytochemical signatures of ecophysiological status, (2) the inoculation of P. albicaulis seedlings with a mixture of fungal endophytes did not lead to persistent systemic changes in seedling foliar microbiomes, and (3) in correspondence with other studies, our data suggest that maternal seed source and spatial patterns influence fungal endophyte community composition.
... It was also reported as a mycoparasite of powdery mildew (Sullivan and White Jr. 2000). Phoma glomerata (= Didymella glomerata) and P. calorpreferens (= Peyronellaea calorpreferens) were recorded as endophyte fungi from Korean pine (Pinus koraiensis) leaves (Deng et al. 2011). It was also associated with "Pea 'Ascochyta blight complex" in Australia (Tran et al. 2014). ...
This chapter presents an overview of the Phoma-like fungi which can be pathogenic, or weak parasites frequently occurring on papilionaceous plants, the members of Fabaceae family, including Faboideae/Papilionideae/Papilionaceae subfamily; this latest name is generally treated as a family rank in the botany. The members of papilionaceous plants are trees, shrubs or herbaceous plants; the last group includes numerous plant species with agricultural importance. It is critical to note that the identification of the Phoma-like fungi formerly was carried out mainly based on morphological features or only ‘based on the host’. However, such an identification could be incorrect, since it is possible to encounter a similar but widely specialized saprotrophic or weakly pathogenic species as well. The taxonomic status of these phomoid fungi was rather confused, but nowadays it seems reasonable to summarize these fungi by acceptance of the modern ‘One fungus – One name’ concept. These fungi were recently reclassified based on molecular evidence. The fungi were enumerated one by one of the ten hosts that were widely grown geographically as agricultural plants. On the basis of only one valid and confirmed name, all were positioned in their ascomycetous family, indicating the former teleomorph (t) and anamorph (a) stages. The most synonymous names were also listed and notified the morphs of genera: (a) or (t). Probably these will contribute to the better orientation in the proper name usage of these fungi groups.
... Didymella glomerata was the representative of the Didymellaceae family, having been isolated from the water samples in this study. As with most of the Pleosporales, this species has been obtained for a long time from senescent leaves of deciduous trees [76,77]. Three other strains from the same substrate were morphologically identified as Phoma-like due to the presence of pigmented conidia, unlike what occurs with Phoma representatives. ...
Environmental pollution with alkylbenzene hydrocarbons such as toluene is a recurring phenomenon. Their toxicity and harmful effect on people and the environment drive the search for sustainable removal techniques such as bioremediation, which is based on the microbial metabolism of xenobiotic compounds. Melanized fungi present extremophilic characteristics, which allow their survival in inhospitable habitats such as those contaminated with hydrocarbons. Screening methodologies for testing the microbial assimilation of volatile organic compounds (VOC) are scarce despite their importance for the bioremediation of hydrocarbon associated areas. In this study, 200 strains of melanized fungi were isolated from four different hydrocarbon-related environments by using selective methods, and their biodiversity was assessed by molecular and ecological analyses. Seventeen genera and 27 species from three main orders, namely Chaetothyriales, Cladosporiales, and Pleosporales, were identified. The ecological analysis showed a particular species distribution according to their original substrate. The isolated strains were also screened for their toluene assimilation potential using a simple and inexpensive methodology based on miniaturized incubations under controlled atmospheres. The biomass produced by the 200 strains with toluene as the sole carbon source was compared against positive and negative controls, with glucose and with only mineral medium, respectively. Nineteen strains were selected as the most promising for further investigation on the biodegradation of alkylbenzenes.
... Interestingly, P. herbarum has been described as potentially beneficial for plant growth (Muhammad et al. 2009), including in Scots pine (Sanz-Ros et al. 2015), and has also been used as a biological control agent against Taraxacum officinale (Neumann Brebaum 1998). P. glomerata has been described as having mycoparasitic properties (Sullivan, White 2000), as an endophyte (Deng et al. 2011), as a pathogen causing boxwood tip blight (Horst 2001), cankers of peach trees (Thomidis et al. 2011) and, according to the American Phytopathological Society, phoma canker in elm (https:// www.apsnet.org/publications/commonnames/Pages/Elm. aspx). ...
Phoma blight is a disease affecting Norway spruce, Scots pine and other conifer seedlings in many forest tree nurseries throughout the world. Members of the Phoma genus, the causatives of this disease, are difficult to distinguish morphologically and genetically. In this study the use of a retrotransposon-based polymerase chain reaction approach using iPBS amplification for intra-species genetic discrimination between Phoma samples is described. Eight retrotransposon-based iPBS primers were used to genotype DNA from pure cultures of several Phoma species. The utilised markers were able to discriminate between Phoma species, but not all of them were able to differentiate all Phoma sp. isolates investigated. Belarusian samples were found to be distinct from the Latvian Phoma isolates. The Belorussian isolates were very similar to each other. A combination of three iPBS markers (2001, 2076 and 2242) enabled partial differentiation of the investigated Belarusian Phoma isolates.
... The strain QLP12 was further identified by ITS rDNA sequence analysis. Genomic DNA was extracted as described in earlier reports [11]. Universal primers ITS1-F and ITS4-R were used for amplification of the internal transcribed spacer (ITS) region [12]. ...
A fungus with broad spectrum antifungal activity was isolated from the soil in Qinling Mountain, Shaanxi Province, in China. The fungus was identified as Purpureocillium lilacinum based on ITS rDNA gene analysis. The strain, coded as QLP12, showed high inhibition activity on fungal mycelium growth in vitro , especially to Mucor piriformis , Trichothecium roseum , Rhizoctonia solani, and Verticillium dahliae , and its potential for biocontrol efficacy of eggplant. Verticillium wilt disease caused by Verticillium dahliae among 10 fungal species tested was explored. In greenhouse experiments, QLP12 showed an excellent growth-promoting effect on eggplant seed germination (76.7%), bud growth (79.4%), chlorophyll content (47.83%), root activity (182.02%), and so on. QLP12 can colonize the eggplant interior and also develop in rhizosphere soil. In greenhouse, the incidence of Verticillium wilt decreased by 83.82% with pretreated QLP12 fermentation broth in the soil. In the field, QLP12 showed prominent biocontrol effects on Verticillium wilt by reducing the disease index over the whole growth period, a decline of 40.1%. This study showed that the strain QLP12 is not only an effective biocontrol agent for controlling Verticillium wilt of eggplant, but also a plant growth-promoting fungus that deserves to be further developed.
... Genomic DNA was extracted from mycelium using the method described by Deng et al. [21]. Amplification of the internal transcribed spacer (ITS) region was performed using the ITS5 and ITS4 primers, after which the PCR products were purified using a Wizard PCR prep kit (Promega, Madison, WI, USA). ...
Two new species of endophytic fungi were encountered during a diversity study of healthy tissues of chili pepper plants in Korea. The species were identified as Paecilomyces inflatus and Bionectria ochroleuca based on molecular and morphological analyses. Morphological descriptions of these endophytic isolates matched well with their molecular analysis. In the present study, detailed descriptions of internal transcribed spacer regions and morphological observations of these two fungi are presented.
... in a laminar air flow chamber. Five sterile samples with the point of rupture downside were placed on potato dextrose agar (PDA; Difco, Detroit, MI, USA). To inhibit bacterial growth, the PDA media were supplemented with the antibiotic streptomycin sulfate. Five replications of the assay using either weakly infected or healthy fruit were performed. [18] was used for isolation of genomic DNA from mycelium. Primers ITS5 and ITS4 were used for amplification of the internal transcribed spacer (ITS) region [19]. In addition, primers Bt2a and Bt2b were used for amplification of the -tubulin (BT2) gene of the Penicillium isolate CNU 111554 [20] and primers EF-728F and EF2 were used for amplif ...
Two species, Penicillium adametzioides and Purpureocillium lilacinum, were isolated from decayed grapes (cv. Cheongsoo) in Korea. Each species was initially identified by phylogenetic analysis of a combined dataset of two genes. Internal transcribed spacer (ITS) and β-tubulin (BT2) genes were used for identification of Penicillium adametzioides, and ITS and partial translation elongation factor 1-α (TEF) genes were used for identification of Purpureocillium lilacinum. Morphologically, they were found to be identical to previous descriptions. The two species presented here have not been previously reported in Korea.
Endophytic fungi are symptomless internal inhabits of plant tissues. They are implicated in the production of antibiotic and other compounds of therapeutic importance. Ecologically they provide several benefits to plants, including protection from plant pathogens. There have been numerous studies on the biodiversity and ecology of endophytic fungi. Some taxa dominate and occur frequently when compared to others due to adaptations or capabilities to produce different primary and secondary metabolites. It is therefore of interest to examine different fungal species and major taxonomic groups to which these fungi belong for bioactive compound production. In the present paper a list of endophytes based on the available literature is reported. More than 800 genera have been reported worldwide. Dominant genera are Alternaria, Aspergillus, Colletotrichum, Fusarium, Penicillium, and Phoma. Most endophyte studies have been on angiosperms followed by gymnosperms. Among the different substrates, leaf endophytes have been studied and analyzed in more detail when compared to other parts. Most investigations are from Asian countries such as China, India, European countries such as Germany, Spain and the UK in addition to major contributions from Brazil and the USA.
Molecular morphometrics is an emerging third dimensional aspect of fungal species delimitation. They have been demonstrated to be more informative than conventional barcoding methods. Hence in this study, foliar endophytic fungal (FEF) assemblages in three Magnoliopsida plants were delimited using nuclear ribosomal internal transcribed spacer 2 (ITS2) sequence—secondary structural features based phylogenetic analysis, also known as molecular morphometrics. A total of 392 FEF isolates were obtained from the Aglaia elaeagnoidea, Flacourtia inermis, and Premna serratifolia leaves and grouped into 98 morphotypes. Among these host plants, P. serratifolia showed the maximum percentage of colonization frequency. Representatives of each morphotype was sequenced and subjected to further molecular characterization. The results revealed that morphotypes were belonged to the phylum of Ascomycota, distributed over two classes (Sordariomycetes (68.59%) and Dothideomycetes (31.41%)), 6 orders and 19 genera. Based on compensatory base changes (CBC) analysis and absolute identity of ITS2 structure, 21, 20 and 23 species were recognized from A. elaeagnoidea, F. inermis, and P. serratifolia respectively. Diversity indices were higher in A. elaeagnoidea, despite it accounted for a modest 16.8% of total isolates recorded in this study. The genus Colletotrichum was predominant in A. elaeagnoidea (39%) and P. serratifolia (48%). Similarly, Diaporthe (43%) was dominant in F. inermis. Several host-specific species were also observed. This study concludes that these plants host diverse species of Ascomycota. To the best of our knowledge, this is the first detailed report on FEF diversity from these plants. Also, the inclusion of ITS2 secondary structure information along with the sequence provides a further dimension to resolve the inherent problems in identification of fungal species.
