Fungal microbiota in seeds, seedlings and mature plants of raspberry (Rubus ideaus L.)

AbstractPresently, there is an intensive search for fungal endophytes to be used in agriculture for the protection and condition improvement of plants and in medicine. We screened for the presence of endophytes in raspberry, which occurs naturally in the Białowieża Forest. The fungal isolates representative of each morphotype were analysed using the molecular markers ITS1 and ITS2. In total, we found 34 taxa of endophytic fungi. The majority were potential pathogens. As many as 27 taxa were found in the leaves of mature plants. No fungi could be isolated from the surface sterilized seeds obtained from these plants. Seedlings were grown from the seeds deposited in the soil seed bank in the Białowieża Geobotanical Station of the University of Warsaw in Białowieża. 8 taxa of endophytic fungi were found in seedlings. It could be due to a possibility of seed infection with these endophytes in soil conditions.

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Isolation and Characterization of Fungal Endophytes Isolated from Medicinal Plant Ephedra pachyclada as Plant Growth-Promoting

Biomolecules ◽

10.3390/biom11020140 ◽

2021 ◽

Vol 11 (2) ◽

pp. 140 ◽

Cited By ~ 2

Author(s):

Ahmed Mohamed Aly Khalil ◽

Saad El-Din Hassan ◽

Sultan M. Alsharif ◽

Ahmed M. Eid ◽

Emad El-Din Ewais ◽

...

Keyword(s):

Medicinal Plants ◽

Plant Growth ◽

Aspergillus Flavus ◽

Endophytic Fungi ◽

Fungal Endophytes ◽

Penicillium Crustosum ◽

Plant Growth Promoting ◽

Penicillium Commune ◽

Maize Plants ◽

Growth Promoting

Endophytic fungi are widely present in internal plant tissues and provide different benefits to their host. Medicinal plants have unexplored diversity of functional fungal association; therefore, this study aimed to isolate endophytic fungi associated with leaves of medicinal plants Ephedra pachyclada and evaluate their plant growth-promoting properties. Fifteen isolated fungal endophytes belonging to Ascomycota, with three different genera, Penicillium, Alternaria, and Aspergillus, were obtained from healthy leaves of E. pachyclada. These fungal endophytes have varied antimicrobial activity against human pathogenic microbes and produce ammonia and indole acetic acid (IAA), in addition to their enzymatic activity. The results showed that Penicillium commune EP-5 had a maximum IAA productivity of 192.1 ± 4.04 µg mL−1 in the presence of 5 µg mL−1 tryptophan. The fungal isolates of Penicillium crustosum EP-2, Penicillium chrysogenum EP-3, and Aspergillus flavus EP-14 exhibited variable efficiency for solubilizing phosphate salts. Five representative fungal endophytes of Penicillium crustosum EP-2, Penicillium commune EP-5, Penicillium caseifulvum EP-11, Alternaria tenuissima EP-13, and Aspergillus flavus EP-14 and their consortium were selected and applied as bioinoculant to maize plants. The results showed that Penicillium commune EP-5 increased root lengths from 15.8 ± 0.8 to 22.1 ± 0.6. Moreover, the vegetative growth features of inoculated maize plants improved more than the uninoculated ones.

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The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr.

Plants ◽

10.3390/plants10071276 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1276

Author(s):

Olga A. Aleynova ◽

Andrey R. Suprun ◽

Nikolay N. Nityagovsky ◽

Alexandra S. Dubrovina ◽

Konstantin V. Kiselev

Keyword(s):

Cell Culture ◽

Cell Suspension ◽

Endophytic Fungi ◽

Endophytic Bacteria ◽

Fungal Endophytes ◽

Biomass Accumulation ◽

Vitis Amurensis ◽

Grape Cell ◽

Stilbene Biosynthesis

Plant endophytes are known to alter the profile of secondary metabolites in plant hosts. In this study, we identified the main bacterial and fungal representatives of the wild grape Vitis amurensis Rupr. microbiome and investigated a cocultivation effect of the 14 endophytes and the V. amurensis cell suspension on biomass accumulation and stilbene biosynthesis. The cocultivation of the V. amurensis cell culture with the bacteria Agrobacterium sp., Bacillus sp., and Curtobacterium sp. for 2 weeks did not significantly affect the accumulation of cell culture fresh biomass. However, it was significantly inhibited by the bacteria Erwinia sp., Pantoea sp., Pseudomonas sp., and Xanthomonas sp. and fungi Alternaria sp., Biscogniauxia sp., Cladosporium sp., Didymella sp. 2, and Fusarium sp. Cocultivation of the grapevine cell suspension with the fungi Didymella sp. 1 and Trichoderma sp. resulted in cell death. The addition of endophytic bacteria increased the total stilbene content by 2.2–5.3 times, while the addition of endophytic fungi was more effective in inducing stilbene accumulation by 2.6–16.3 times. The highest content of stilbenes in the grapevine cells cocultured with endophytic fungi was 13.63 and 13.76 mg/g of the cell dry weight (DW) after cultivation with Biscogniauxia sp. and Didymella sp. 2, respectively. The highest content of stilbenes in the grapevine cells cocultured with endophytic bacteria was 4.49 mg/g DW after cultivation with Xanthomonas sp. The increase in stilbene production was due to a significant activation of phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) gene expression. We also analyzed the sensitivity of the selected endophytes to eight antibiotics, fluconazole, and trans-resveratrol. The endophytic bacteria were sensitive to gentamicin and kanamycin, while all selected fungal strains were resistant to fluconazole with the exception of Cladosporium sp. All endophytes were tolerant of trans-resveratrol. This study showed that grape endophytes stimulate the production of stilbenes in grape cell suspension, which could further contribute to the generation of a new stimulator of stilbene biosynthesis in grapevine or grape cell cultures.

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Revealing the endophytic mycoflora in tea (Camellia sinensis) leaves in Sri Lanka: the first comprehensive study

Phytotaxa ◽

10.11646/phytotaxa.514.3.5 ◽

2021 ◽

Vol 514 (3) ◽

pp. 247-260

Author(s):

KASUN THAMBUGALA ◽

DINUSHANI DARANAGAMA ◽

SAGARIKA KANNANGARA ◽

THENUKA KODITUWAKKU

Keyword(s):

Sri Lanka ◽

Camellia Sinensis ◽

Endophytic Fungi ◽

Sequence Data ◽

Fungal Endophytes ◽

Fungal Endophyte ◽

Tea Leaves ◽

Molecular Sequence Data ◽

Molecular Sequence ◽

Rdna Sequence Data

Endophytic fungi are a diverse group of microorganisms that live asymptomatically in healthy tissues of host and they have been reported from all kinds of plant tissues such as leaves, stems, roots, flowers, and fruits. In this study, fungal endophytes associated with tea leaves (Camellia sinensis) were collected from Kandy, Kegalle, and Nuwara Eliya districts in Sri Lanka and were isolated, characterized, and identified. A total of twenty endophytic fungal isolates belonging to five genera were recovered and ITS-rDNA sequence data were used to identify them. All isolated endophytic fungal strains belong to the phylum Ascomycota and the majority of these isolates were identified as Colletotrichum species. Phyllosticta capitalensis was the most commonly found fungal endophyte in tea leaves and was recorded in all three districts where the samples were collected. This is the very first investigation on fungal endophytes associated with C. sinensis in Sri Lanka based on molecular sequence data. In addition, a comprehensive account of known endophytic fungi reported worldwide on Camellia sinensis is provided.

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Isolation and antimicrobial effects of endophytic fungi from Edgeworthia chrysantha

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v10i3.23575 ◽

2015 ◽

Vol 10 (3) ◽

pp. 529 ◽

Cited By ~ 4

Author(s):

Huawei Zhang ◽

Chuanfeng Ruan ◽

Xuelian Bai

Keyword(s):

Ethyl Acetate ◽

Endophytic Fungi ◽

Fermentation Broth ◽

Fungal Endophytes ◽

Antimicrobial Effect ◽

Antibacterial Activities ◽

Antimicrobial Activities ◽

Human Pathogens ◽

Antimicrobial Effects ◽

Staphyloccocus Aureus

Ten fungal strains isolated from Edgeworthia chrysantha, one of traditional medicinal plants in China, were evaluated their antimicrobial activities against three human pathogens, Escherichia coli, Staphyloccocus aureus and Candida albicans, and two phytopathogens, Rhizoctonia cerealis and Colletotrichum gloeosporioides. The results indicated that most ethyl acetate extracts of fermentation broth of these fungal endophytes had stronger antimicrobial activities than their fermentation broth. Among these endophytic strains, both fermentation broth and the ethyl acetate extract of strain D showed the strongest inhibitory effects on all pathogens. Strains 5-19 and BZ also exhibited potent antibacterial activities. However, other strains had weak or no antimicrobial effect. This was the first report on the isolation and antimicrobial effects of endophytic fungi from E. chrysantha.

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A Note on the Application of an AgLeader® Cotton Yield Monitor for Measuring Peanut Yield: An Investigation in Two US states.

Peanut Science ◽

10.3146/ps19-16.1 ◽

2020 ◽

Vol 47 (2) ◽

pp. 115-122

Author(s):

W.M. Porter ◽

J. Ward ◽

R.K. Taylor ◽

C.B. Godsey

Keyword(s):

Soil Conditions ◽

State University ◽

Yield Estimation ◽

Yield Monitor ◽

University Of Georgia ◽

High Data ◽

Data Validity ◽

Deflector Plate ◽

The University ◽

Multiple Field

ABSTRACT Previous researchers demonstrated the ability to adapt an AgLeader® Cotton Monitor to a peanut combine. It was demonstrated that the field weight could be accurately predicted with average errors of less than 10% across all trials when at least five calibration loads are applied. This project focused on expanding previous work performed at the University of Georgia and other peanut optical yield monitor work by incorporating a protective deflector plate for the sensors, obtaining multiple field weights, and using the peanut sale sheets to correlate yield monitor yield to sale weight. This study was a two-university, two-state effort, including Oklahoma State University (Oklahoma), and Mississippi State University (Mississippi). Data collected during this study included multiple loads which included yield monitor weight, field weight, field moisture content, and all the information presented on the standard USDA peanut grade sheet, when available. The multi-state effort allowed for the incorporation of the two major peanut types and for the incorporation of different soil types. The goal of this study was to develop guidelines for using, calibrating, and adapting the AgLeader® Cotton Monitor for peanut harvest. Five calibration loads referenced to buy-point net weight were typically needed to bring error within acceptable limits. Results indicated that multiple local calibrations were needed to ensure high data validity and yield estimation across multiple harvest environments. The data showed that peanut type (virginia, runner and spanish) and variable soil conditions impacted yield estimation.

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Production of eugenol from fungal endophytesNeopestalotiopsissp. andDiaporthesp. isolated fromCinnamomum loureiroileaves

PeerJ ◽

10.7717/peerj.6427 ◽

2019 ◽

Vol 7 ◽

pp. e6427 ◽

Cited By ~ 5

Author(s):

Chutima Tanapichatsakul ◽

Sarunpron Khruengsai ◽

Sakon Monggoot ◽

Patcharee Pripdeevech

Keyword(s):

Antimicrobial Activity ◽

Antioxidant Activity ◽

Endophytic Fungi ◽

Antioxidant Activities ◽

Chemical Constituents ◽

Fungal Endophytes ◽

Biologically Active ◽

Crude Extracts ◽

Antioxidant Agents ◽

Fungal Extracts

Endophytic fungi, which colonize within a host plant without causing any apparent diseases, have been considered as an important source of bioactive secondary metabolites containing antimicrobial and antioxidant activities. The aim of this research was to isolate the endophytic fungi ofCinnamomum loureiroiand then to screen their antimicrobial and antioxidant activities. A total of 11 fungal endophytes were isolated from healthy leaves ofCinnamomum loureiroibelonging to six genera:Botryosphaeria,Colletotrichum,Diaporthe,Fusarium,Neopestalotiopsis, andPestalotiopsis. All isolated strains were cultured and further extracted with ethyl acetate solvent. Antimicrobial activity of all crude endophytic fungal extracts was analyzed using disc diffusion assay against six bacterial and two fungal pathogens. Crude extracts of strains MFLUCC15-1130 and MFLUCC15-1131 showed broad-spectrum antimicrobial activity against all tested pathogens. Activity againstBacillus cereusandStaphylococcus epidermidiswas notable, showing the lowest minimum inhibitory concentration at 3.91 μg/mL. Antioxidant activity of all crude endophytic fungal extracts was also evaluated based on 2,2-diphenyl-1-picrylhydrazyl assay. Significant antioxidant activity was detected in the crude extracts of fungus MFLUCC15-1130 and MFLUCC15-1131 with IC50of 22.92 ± 0.67 and 37.61 ± 0.49 μg/mL, respectively. Using molecular identification, MFLUCC15-1130 and MFLUCC15-1131 were identified asNeopestalotiopsissp. andDiaporthesp., respectively. The major chemical constituents produced by both crude extracts were identified by gas chromatography-mass spectrometry. Eugenol, myristaldehyde, lauric acid, and caprylic acid were the primary antimicrobial and antioxidant compounds in both crude extracts. This is the first report of eugenol being a biologically active compound ofNeopestalotiopsissp. andDiaporthesp. fungal endophytes. Eugenol has been reported as antimicrobial and antioxidant agents with agronomic applications. Thus the two newly-isolated endophytes may be used for eugenol production, which in turn can be used in a variety of applications.

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Chemical and Functional Diversity of Natural Products from Plant Associated Endophytic Fungi

Natural Product Communications ◽

10.1177/1934578x0900401114 ◽

2009 ◽

Vol 4 (11) ◽

pp. 1934578X0900401 ◽

Cited By ~ 20

Author(s):

Vijay C. Verma ◽

Ravindra N. Kharwar ◽

Gary A. Strobel

Keyword(s):

Natural Products ◽

Biological Activity ◽

Functional Diversity ◽

Bioactive Compounds ◽

Endophytic Fungi ◽

Host Plants ◽

Fungal Endophytes ◽

Sources Identification ◽

Naturally Occurring

This review describes examples of naturally occurring bioactive compounds obtained from fungal endophytes from various host plants. The main topics addressed are sources, identification, biological activity, biosynthesis, and ecological and chemosystematic significance of those bioactive compounds whose sources were well defined.

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Diverse Bacteria Inhabit Living Hyphae of Phylogenetically Diverse Fungal Endophytes

Applied and Environmental Microbiology ◽

10.1128/aem.02928-09 ◽

2010 ◽

Vol 76 (12) ◽

pp. 4063-4075 ◽

Cited By ~ 147

Author(s):

Michele T. Hoffman ◽

A. Elizabeth Arnold

Keyword(s):

Endophytic Fungi ◽

Tree Species ◽

Host Plants ◽

Abiotic Factors ◽

Fungal Endophytes ◽

Molecular Techniques ◽

Biogeographic Provinces ◽

Viable Bacteria ◽

Bacterial Endosymbionts ◽

First Time

ABSTRACT Both the establishment and outcomes of plant-fungus symbioses can be influenced by abiotic factors, the interplay of fungal and plant genotypes, and additional microbes associated with fungal mycelia. Recently bacterial endosymbionts were documented in soilborne Glomeromycota and Mucoromycotina and in at least one species each of mycorrhizal Basidiomycota and Ascomycota. Here we show for the first time that phylogenetically diverse endohyphal bacteria occur in living hyphae of diverse foliar endophytes, including representatives of four classes of Ascomycota. We examined 414 isolates of endophytic fungi, isolated from photosynthetic tissues of six species of cupressaceous trees in five biogeographic provinces, for endohyphal bacteria using microscopy and molecular techniques. Viable bacteria were observed within living hyphae of endophytic Pezizomycetes, Dothideomycetes, Eurotiomycetes, and Sordariomycetes from all tree species and biotic regions surveyed. A focus on 29 fungus/bacterium associations revealed that bacterial and fungal phylogenies were incongruent with each other and with taxonomic relationships of host plants. Overall, eight families and 15 distinct genotypes of endohyphal bacteria were recovered; most were members of the Proteobacteria, but a small number of Bacillaceae also were found, including one that appears to occur as an endophyte of plants. Frequent loss of bacteria following subculturing suggests a facultative association. Our study recovered distinct lineages of endohyphal bacteria relative to previous studies, is the first to document their occurrence in foliar endophytes representing four of the most species-rich classes of fungi, and highlights for the first time their diversity and phylogenetic relationships with regard both to the endophytes they inhabit and the plants in which these endophyte-bacterium symbiota occur.

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Isolation of fungal endophytes from black spruce (Picea mariana) dormant buds and needles from New Brunswick, Canada

Canadian Journal of Botany ◽

10.1139/b92-217 ◽

1992 ◽

Vol 70 (9) ◽

pp. 1754-1757 ◽

Cited By ~ 43

Author(s):

J. A. Johnson ◽

N. J. Whitney

Keyword(s):

New Brunswick ◽

Key Words ◽

Endophytic Fungi ◽

Picea Mariana ◽

Black Spruce ◽

Fungal Endophytes ◽

The Other ◽

Age Classes ◽

Needle Age

Endophytic fungi were isolated from black spruce (Picea mariana (Mill.) BSP) dormant buds and needles of four age-classes. Only one isolate was obtained from 400 buds. A total of 914 isolates were obtained from the needles. With increasing needle age the rate of colonization increased between current-year and 3-year-old needles from 4 to 90%, respectively. The needle segment attached to the twig was colonized more often (p < 0.05) than other segments overall and for 6 of the 11 taxa isolated. The first endophyte from current-year needles was isolated on July 14, 1988, but endophytes were obtained from needles of the other age-classes (1, 2, and 3 years) on all sampling dates between June 10 and September 16, 1988. Key words: endophyte, Picea mariana, dormand buds, needles.

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DNA-Dependent Detection of the Grapevine Fungal Endophytes Aureobasidium pullulans and Epicoccum nigrum

Plant Disease ◽

10.1094/pdis-93-10-0993 ◽

2009 ◽

Vol 93 (10) ◽

pp. 993-998 ◽

Cited By ~ 55

Author(s):

M. Martini ◽

R. Musetti ◽

S. Grisan ◽

R. Polizzotto ◽

S. Borselli ◽

...

Keyword(s):

Aureobasidium Pullulans ◽

Potential Role ◽

Fungal Endophytes ◽

Specific Primers ◽

Variable Regions ◽

Epicoccum Nigrum ◽

Polymerase Chain ◽

Species Specific ◽

Reference Strains ◽

Its1 And Its2

Aureobasidium pullulans and Epicoccum nigrum are frequently reported as endophytes of various crops, including grapevine (Vitis vinifera). Because of their potential role as biological control agents against grapevine pathogens, we examined the occurrence of A. pullulans and E. nigrum in two grapevine varieties (Merlot and Prosecco) in Italian vineyards where spontaneous recovery from phytoplasma disease is recurrent. Species-specific primers for A. pullulans and two genetically distinct strains of E. nigrum were designed in variable regions of ITS1 and ITS2. Primer specificity was confirmed by polymerase chain reaction using purified DNA from other fungal endophytes that are usually encountered during isolation attempts from grapevine tissues and from several other strains of A. pullulans and E. nigrum isolated from other sources. In order to determine the occurrence of the two endophytes in grapevine plants, DNA was extracted from shoots of 44 grapevines collected in six vineyards from different localities of northeast Italy. Both endophytes were detected and their identity was confirmed by restriction fragment length polymorphism (RFLP) patterns obtained from reference strains. RFLP analyses confirmed the presence of two E. nigrum strains belonging to different RFLP groups in grapevine. The molecular methods described allowed a sensitive, specific, and reliable identification of the two endophytes in grapevine.

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