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2003). An evaluation of the fungal 'morphotype' concept based on ribosomal DNA sequences. Fungal Diversity 12: 53-66. In studies of fungal endophyte communities, mycelia sterilia are commonly isolated from plant substrates and grouped into morphotypes on the basis of cultural characteristics. In Polygonum multiflorum one hundred and sixty-nine stra...
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... most of these studies anamorphic fungi were commonly isolated and a large number of fungi that did not sporulate in culture were categorized as mycelia sterilia. In order to appreciate the considerable prevalence of these mycelia sterilia, a summary showing the proportions of mycelia sterilia found in previous studies of fungal endophytic communities is given in Table 1. For a given host, mycelia sterilia took up an average of 20% of the population of fungal endophytes. ...
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... for species identi cation, with the ITS region being a signi cant molecular marker known for its high accuracy in fungal identi cation. The ITS region has been widely used for a diverse range of fungi 14,15 . In this study, phylogenetic analysis revealed that the isolates were grouped into distinct clades, and their precise taxonomic placement was determined based on the ITS region analysis. ...
Endophytic fungi are commonly found in the root endosphere and can enhance plant growth through various mechanisms. The aim of this study was to isolate cultivable endophytic fungi associated with the roots of Tamarix ramosissima and to evaluate their plant growth promoting properties. About 35 isolated fungal endophytes belonging to the Ascomycota from four different genera were isolated from the endosphere of T. ramosissima : Alternaria , Aspergillus , Fusarium and Talaromyces . These fungal endophytes showed different abilities to solubilize phosphate and produce indole-3-acetic acid (IAA). The fungal isolates of T. allahabadensis (T3) and A. niger (T4) showed different efficiency in solubilizing phosphate. Almost all fungal isolates were able to produce IAA, and the highest value (0.699 µg/ml) was found in the isolate of F. solani (T11). Inoculation of wheat seeds with endophytic fungi significantly increased the initial growth of wheat roots. The results showed that inoculation with the endophytic fungus A. fumigatus T15 significantly increased root length by 75%. The extensive root system of T. ramosissima may be due to symbiosis with IAA-producing endophytic fungi, which enhance root development and water uptake in dry conditions. These fungi can also boost soil phosphorus levels, promoting plant growth.
... It was molecularly recognized using internal transcribed spacer (ITS-1 and ITS-4) primers and part of the nuclear ribosomal RNA gene large subunit (LSU). A modi ed method was used to extract total gDNA from a fungal isolate (Lacap et al. 2003) and the ITS region was ampli ed (White et al. 1990). The PCR mixture (25 µl) included 12.5 µl of Master mix (2X), 1.25 µl (10 pmol) of each primer, 7.5 µl of the PCR-quality water, and 2.5 µl of the DNA template (~ 50 ng). ...
In a wake of shifting climatic scenarios, plants are frequently forced to undergo a spectrum of abiotic and biotic stresses at various stages of growth, many of which have a detrimental effect on production and survival. Naturally, microbial consortia partner up to boost plant growth and constitute a diversified ecosystem against abiotic stresses. Despite this, little is known pertaining to the interplay between endophytic microbes which release phytohormones and stimulate plant development in stressed environments. In a lab study, we demonstrated that an endophyte isolated from the Kargil region of India, a Fusarium equiseti strain K23-FE, colonizes the maize hybrid MAH 14 − 5, promoting its growth and conferring PEG-induced osmotic stress tolerance. To unravel the molecular mechanism, maize seedlings inoculated with endophyte were subjected to comparative transcriptomic analysis. In response to osmotic stress, genes associated with metabolic, photosynthesis, secondary metabolites, and terpene biosynthesis pathways were highly upregulated in endophyte enriched maize seedlings. Further, in a greenhouse experiment, maize plants inoculated with fungal endophyte showed higher relative leaf water content, chlorophyll content, and antioxidant enzyme activity such as polyphenol oxidase (PPO) and catalase (CAT) under 50% field capacity conditions. Non-enzymatic molecules like proline were higher and malondialdehyde content was reduced in colonized plants. This study set as proof of concept to demonstrate that endophytes adapted to adverse environments can efficiently tweak non-host plant responses to abiotic stresses such as water deficit stress via physiological and molecular pathways, offering a huge opportunity for their deployment in sustainable agriculture.
... A series of species concepts have been applied in fungal taxonomy. Of these, the morphological concept has traditionally been widely used [2]. The polythetic concept has been applied in combination with the characteristic concept [3]. ...
A functional identification system is the core and basis of fungal taxonomy, which provides sufficient diagnostic characteristics for species delimitation. Phenotype-based identification systems have exhibited significant drawbacks, such as being laborious and time-consuming. Thus, a molecular-based identification system (rDNA, DNA fingerprint, etc.) is proposed for application to fungi that lack reliable morphological characteristics. High Throughput Sequencing also makes great contributions to fungal taxonomy. However, the formal naming of nonculturable fungi from environmental sequencing is a significant challenge. Biochemical profile-based identification systems have outstanding value in fungal taxonomy and can occasionally be indispensable. This method utilizes biomarker metabolites and proteins that are expected to be unequivocal and stable. Of these, Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry has become the method of choice for chemotaxonomy. In sum, these described identification systems cannot solve all problems of species delimitation, and considerable attention to the updating of fungal nomenclature, standardization of techniques, knowledge sharing, and dissemination will be necessary.
... 8,9 Moreover, the traditional identification methods are inadequate for distinguishing fungal isolates that do not produce spores in culture, which are categorized as mycelia sterilia. 10 In contradistinction to this, modern molecular techniques showcase high specificity and sensitivity for identifying microorganisms and can be used for classifying microbial strains across various level of taxonomic hierarchy. 11 Molecular methods are progressively being used to identify various kind of fungi [12][13][14] and provide a quick and precise way of characterizing and identifying organisms because they rely on objective information (molecules) derived from the target organism rather than the substantial subjectivity of classical methods. ...
... Basidiospores ellipsoid to ellongate, pale orange to greyish orange, with a brown eusporium bearing fine, short, and distinct echinulae, overlaid by a hyaline myxosporium. 10 Ganoderma mizoramense ...
Identification of fungi to species level is paramount in both basic and applied applications in scientific research. A diverse group of fungi play a crucial role in ecosystem functioning and significantly contribute to the biodiversity of various ecosystems. In the northeastern state of Mizoram, India, the study of wood-inhabiting fungi holds immense potential for uncovering the region's fungal diversity and ecological interactions. The present study investigated the phylogenetic relationships and taxonomic identities of different fungal species of polypores isolated from diverse parts of Mizoram, India.
Fungal species were collected and identified using a macro and micro-morphological characteristics and molecular approaches.
Nuclear ribosomal DNA sequences, along with the internal transcribed spacers (ITS-1 and ITS-2), and the 5.8S gene area, were used to identified the collected samples. 23 fungal isolates of polypores were selected for molecular phylogenetic analysis. The 23 species were identified to the species level based on the fungal sequences with known identities in GenBank.
The combined approach both morphological and molecular techniques proposed in the study holds the potential to address these challenges and provide a more efficient and accurate way of understanding fungal distribution.
... All fungal colonies that grew in the culture medium were morphologically characterized and grouped into morphological types (morphotypes). Morphotype determination was performed according to the criteria proposed by Lacap et al. (2003) and Ibrahim et al. (2017), which included the colony growth rate, shape, and color. To avoid problems of subjectivity in the determination of colors, photographic records were made, and the obverse and reverse colors were defined using the RGB color scales, an abbreviation for the color system formed by Red (red), Green (green), and Blue (blue), used in color reproduction in electronic devices. ...
Spodoptera frugiperda (Lepidoptera: Noctuidae) is an important insect pest in Brazil and worldwide, and its control is based mainly on chemical methods that are harmful to the environment and human and animal health. We investigated the fungal community associated with the body surface of S. frugiperda and evaluated the potential of members of this fungal community to act as biocontrol agents. The caterpillars were collected in the field, reared in the laboratory on a natural diet, evaluated for association with fungi on their surface and the environment, and subjected to a bioassay for insecticidal activity. Our results suggest that fungi are often associated with the body of S. frugiperda, as it was possible to obtain these microorganisms from all insects sampled, with a higher frequency of filamentous fungi than yeast. Fusarium verticillioides was the most frequent filamentous fungus species (F3 = 42.1%), followed by Aspergillus aureolus (F2 = 36.8%). Candida pseudointermedia was the most frequent yeast species (L1 = 57.1%). Aspergillus aureolus F2 was the only strain that exhibited toxicity against S. frugiperda and can be considered a good candidate to be explored from the perspective of obtaining biotechnological products of interest for agriculture.
... Isolates that grew slowly with little to no pigmentation, few to no macroconidia, and sparse macroconidia were considered most likely to be races 1, 5, or 6 (Haglund and Kraft 1979;Harveson et al. 2020). Fast growing, pigmented isolates with abundant macro-and microconidia were presumed to be race 2. Isolates which did not fall into either category with their combination of morphology and growth characteristics were assigned a morphotype, which is defined as a group of individuals of the same species that are distinguishable from each other based on morphological characteristics (Lacap et al. 2003). ...
... Among the 20 North Dakota Fop isolates classified into previously defined wilt races based on bioassays, 11 isolates displayed morphological characteristics consistent with that race. The 14 North Dakota isolates which did not correspond to the reference isolates as described in previous literature were divided into four morphological groups (morphotypes) (Lacap et al. 2003). Morphotype I was comprised of six isolates, one of an unclassified wilt race and five race 2, with purple colony pigmentation, a slow growth rate, and abundant conidia. ...
... Summary of colony pigmentation, growth rate, conidia production, and race classification of North Dakota Fusarium oxysporum f. sp. pisi (Fop) isolates and reference Fop isolates race 1 (ATCC 26043), race 2 (ATCC 26087), race 5 (NRLL 37621), and race 6 (NRLL 37610) using pea differential lines (wilt race) and morphological characteristics on potato dextrose agar (PDA) (presumptive race) Morphotype IV a Morphotype = a group of different types of individuals of the same species that are distinguishable from each other on the base of morphological characteristics (Lacap et al. 2003). b As determined by the wilt score ratings on pea differentials. ...
Fusarium root rot is an important disease of field pea (Pisum sativum var. sativum L.) that occurs everywhere pea is grown, causing yield loss of up to 75%. Fusarium root rot is caused by a complex of Fusarium species, most notably Fusarium solani in the Pacific Northwest of the United States and F. avenaceum in the northern Great Plains of the United States and Canada. F. oxysporum f. sp. pisi (Fop) was frequently isolated from peas exhibiting root rot symptoms in North Dakota during recent surveys. Fop causes wilt (races 1, 5, and 6) and near wilt (race 2) on pea. However, its contribution to pea root rot remains unclear. Fop race was determined for isolates from North Dakota pea root rot surveys. ND Fop isolates were evaluated for root rot pathogenicity and aggressiveness at standard and elevated temperatures. Results from greenhouse wilt assays indicated that all Fop races exist in North Dakota, with race 2 most prevalent among the 25 North Dakota isolates evaluated. Root rot evaluations conducted at 21/18°C and 25/19°C day/night temperatures demonstrated that most Fop isolates were as aggressive or more aggressive than F. solani and F. avenaceum under both temperature regimes. Aggressiveness of Fop isolates tended to increase at elevated assay temperatures. Results from these experiments indicate that Fop may be an important contributor to the root rot complex of field pea in North Dakota and should be considered in integrated pest management strategies, including pea breeding efforts to improve resistance to Fusarium root rot.
[Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .
... Purification consisted of raising each isolate separately until confirmation that the isolate represented an individual with unique morphological characteristics (until there was a single morphotype). The use of the concept of morphotype has been useful in estimating the number of fungi since the species is conventionally the basic unit in biodiversity studies [96]. ...
Invasive plants affect ecosystems across various scales. In particular, they affect the quality and quantity of litter, which influences the composition of decomposing (lignocellulolytic) fungal communities. However, the relationship among the quality of invasive litter, lignocellulolytic cultivated fungal community composition, and litter decomposition rates under invasive conditions is still unknown. We evaluated whether the invasive herbaceous Tradescantia zebrina affects the litter decomposition in the Atlantic Forest and the lignocellulolytic cultivated fungal community composition. We placed litter bags with litter from the invader and native plants in invaded and non-invaded areas, as well as under controlled conditions. We evaluated the lignocellulolytic fungal communities by culture method and molecular identification. Litter from T. zebrina decomposed faster than litter from native species. However, the invasion of T. zebrina did not alter decomposition rates of either litter type. Although the lignocellulolytic fungal community composition changed over decomposition time, neither the invasion of T. zebrina nor litter type influenced lignocellulolytic fungal communities. We believe that the high plant richness in the Atlantic Forest enables a highly diversified and stable decomposing biota formed in conditions of high plant diversity. This diversified fungal community is capable of interacting with different litter types under different environmental conditions.
... Only a fraction of fungal endophytes can be captured using the culture-based approach [39] , but the morphotaxa-based characterization of endophytic communities does not reveal the detailed taxonomic diversity of this fraction [40]. However, the morphotaxa provide a starting point to explore the differences in endophyte diversity in vaccinated and unvaccinated elms. ...
Dutch elm disease (DED) is causing extensive mortality of ecologically and culturally valuable elm trees (Ulmus spp.). Treatment of elms with the biological vaccine Dutch Trig® has been found to provide effective protection against DED by stimulating the defensive mechanisms of the trees. We hypothesized that the same mechanisms could also affect non-target organisms associated with elms. We explored the possible effects of vaccination on epiphytes (mainly lichens) and fungal endophytes living in the bark and young xylem of treated elms. Epiphyte cover percentage was assessed visually using a grid placed on the trunks, and a culture-based approach was used to study endophytes. Epiphyte cover was lower on the trunks of vaccinated trees as compared with unvaccinated trees, but the difference was not statistically significant. The presence of slow-growing and uncommon endophytes seemed to be reduced in continuously vaccinated elms; however, the highest endophyte diversity was found in elms four years after cessation of the vaccination treatments. Our findings suggest that although vaccination may shape epiphyte and endophyte communities in elms, its impacts are not straightforward. More detailed studies are, therefore, needed to inform the sustainable application of the vaccine as a part of the integrated management of DED.
... The culture and growth characteristics of the 437 isolates are summarized in Table 1. These isolates were grouped into ten morphotypes based on the similarity of their cultural characters (Lacap et al. 2003;Samson et al. 2014;Visagie et al. 2014a,b;Vesth et al. 2018;Frisvad et al. 2019;Kowalski and Cramer 2020). ...
Arifah F, Aini LQ, Muhibuddin A. 2023. Molecular and morphological characterization of fungi isolated from nutmeg (Myristica fragrans) in North Sulawesi, Indonesia. Biodiversitas 24: 441-453. Nutmeg (Myristica fragrans Houtt) is one of the predominant commodities from Indonesia exported to the EU market. However, it is frequently rejected due to mycotoxin (especially aflatoxin) content exceeding the aflatoxin standard set by the EU. The high levels of aflatoxin were hypothesized to be produced by a toxigenic fungus infecting nutmeg during and after harvest. The study aims to determine and identify fungal contaminants in nutmeg kernels and provide information on the types of mycotoxins produced. Ten fungal morphotypes were isolated from nutmeg kernels collected from North Minahasa, North Sulawesi, Indonesia, and were grouped into four genera (Aspergillus, Lasiodiplodia, Rhizopus, and Penicillium) based on morphological characters. Ten fungal morphotypes were identified, i.e., Aspergillus flavus (Groups I and II), A. tamarii (Groups I and II), A. niger, A. aculeatus, A. ochraceus, Lasiodiplodia sp., Rhizopus delemar, and Penicillium sp. Isolates of A. flavus and A. tamarii were divided into two groups based on slight differences in their colony appearances. Molecular identification using homology of 18S rDNA sequences and ITS regions supports morphological identification except for isolates of A. flavus (Groups I and II), Lasiodiplodia sp., and Penicillium sp. These isolates were identified as A. nomiae and A. aflatoxiformans (A. autswikcii), L. theobromae, and P. citrinum. The maximum likelihood phylogenetic tree showed that isolates from the same species were grouped in the same clade. The L. theobromae was the most dominant type of fungus in the nutmeg kernels (55.61%), followed by A. niger (23.8%) and A. tamarii (10.53%). It is the first report of A. aflatoxiformans or A. autswikcii in nutmeg kernels, and no incidence of L. theobromae in nutmeg kernels from Indonesia previously.
... Fungi usually grow in conditions where suitable nutrition is provided with proper aeration. However, higher oxygen level encourages them to sporulate [43] . Here, enhanced aeration (shaking at 120 rpm) and reduced nutrient levels showed less correlation with the sporulation of tested endophytic fungi. ...
To date, many plants explored for their endospheric microbiomes have revealed a substantial amount of sterile mycelia, a group of fungi with pseudo-taxonomic positions. While unravelling the endophytic fungal assemblages of a plant Ephedra gerardiana Wall. ex Stapf. via culture-dependent methods, 73.2% of the total isolates failed to sporulate on the commonly used artificial media. Therefore, the current study was designed to investigate the diverse methods, either individually or in combination to check their impact in inducing fungal sporulation. The maximum number of isolates (Nmax) sporulated after individual treatments with 12 h alternate light and dark cycles and cold shocks at 4−6 °C were 22.91% and 18%, respectively. However, the combination of cold treatment with alternate light and dark cycles showed the Nmax of 27.84% and proved to be the best method of sporulation induction out of the methods investigated. To the best of our knowledge, this involves the pioneering attempt of using cold treatment as a triggering factor for sporulating endophytic fungi. Accentuating better methods for endophytic fungal sporulation would not only solve the taxonomic position of these mycosymbionts, but also lessen the time to make them sporulate along with unveiling their eco-physiological behavior
