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.

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.

Download Full-text

Identification and quantitative determination of pinoresinol in Taxus ×media Rehder needles, cell suspension and shoot cultures

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2014.038 ◽

2015 ◽

Vol 84 (1) ◽

pp. 125-132 ◽

Cited By ~ 2

Author(s):

Paulina Mistrzak ◽

Hanna Celejewska-Marciniak ◽

Wojciech J. Szypuła ◽

Olga Olszowska ◽

Anna K. Kiss

Keyword(s):

Cell Culture ◽

Suspension Culture ◽

Quantitative Determination ◽

Cell Suspension ◽

Dry Weight ◽

Shoot Cultures ◽

Taxus Media ◽

In Vitro Shoot Cultures

The aim of our study was to investigate the presence and quantitative contents of lignans in the tissues of Taxus ×media. The presence of the lignans: pinoresinol, matairesinol and secoisolariciresinol was assessed in needles, shoots cultures and suspension culture. Pinoresinol was the only lignan found in the tissue of T. ×media. The total pinoresinol content in the needles and in the shoots was 1.24 mg/g dry weight (dw) and 0.69 mg/g dw, respectively. Most of the pinoresinol identified was appeared glycosidically bound. In needles, the amount of glycosidically bound pinoresinol (0.81 mg/g dw) was about twice as high as that of free pinoresinol (0.43 mg/g dw). The content of free and glycosidically bound pinoresinol showed the level of 0.18 mg/g dw and 0.51 mg/g dw, respectively in the in vitro shoot cultures. In the cell culture, no pinoresinol was found.

Download Full-text

Evaluation of endophytic fungal fractions of Andrographis paniculata (Burm.f.) Wall. Nees leaves for in vitro free radical scavenging and hepatoprotective activity

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v9i1.1150 ◽

2018 ◽

Vol 9 (1) ◽

pp. 1

Author(s):

Smita Kishor Puri ◽

Prasanna Vasantrao Habbu ◽

Preeti Venkatrao Kulkarni ◽

Venkatrao Hanumanthrao Kulkarni

Keyword(s):

Free Radical ◽

Secondary Metabolites ◽

Endophytic Fungi ◽

Fungal Endophytes ◽

Radical Scavenging ◽

Hepatoprotective Activity ◽

Free Radical Scavenging ◽

Andrographis Paniculata ◽

Rdna Sequencing

Fungal endophytes are the microbes residing in internal tissues of the plant forming symbiotic, mutualistic, communalistic and trophobiotic relationship. Endophytes from medicinal plants are considered as essential source of secondary metabolites accompanied by interesting biological/pharmacological activities. In this study, an effort was made to isolate, characterize endophytic fungi from leaves of Andrographis paniculata and to screen the fungal fractions for in-vitro antioxidant and hepatoprotective activity. Two fungal endophytes (APLF-1 and APLF-2) from Andrographis paniculata were isolated and fermented to get chloroform (A1C, A2C), ethyl acetate (A1EA, A2EA) and n butanol (A1nB, A2nB) extracts. All the endophytic fractions of APLF-1 and APLF-2 were assayed for free radical scavenging properties against 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical and reducing power. Fractions of APLF-2 showed good scavenging activity compared to APLF-1 extracts. Further, A2EA and A2nB were evaluated for hepatoprotective activity against CCl4 induced hepatotoxicity. A2EA (50 mg/kg &100 mg/kg) and A2nB (50 mg/kg &100 mg/kg) reversed the elevated biochemical parameters with respect to CCl4 treated group (p<0.001). The LPO, SOD and CAT levels were also restored by A2EA and A2nB (100 mg/kg p.o). APLF-1 and APLF-2 were studied for rDNA sequencing by PCR technique. The endophytic fungi, APLF-1 and APLF-2 were identified as Diaporthe sp. A25 and Preussia sp. PPV3.6 respectively based on their morphology and molecular characterization. The presence of polyvalent secondary metabolites in A2EA and A2nB were confirmed by HPTLC analysis. Keyword: Endophytic fungi; Hepatoprotective; Antioxidant; Andrographis paniculata; Diaporthe sp. A25; Preussia sp. PPV3.6

Download Full-text

Molecular Identification of Endophytic Fungi and Their Pathogenicity Evaluation Against Dendrobium nobile and Dendrobium officinale

International Journal of Molecular Sciences ◽

10.3390/ijms21010316 ◽

2020 ◽

Vol 21 (1) ◽

pp. 316 ◽

Cited By ~ 7

Author(s):

Surendra Sarsaiya ◽

Archana Jain ◽

Qi Jia ◽

Xiaokuan Fan ◽

Fuxing Shu ◽

...

Keyword(s):

Molecular Identification ◽

Endophytic Fungi ◽

Fungal Endophytes ◽

Dendrobium Officinale ◽

Plant Host ◽

Efficient System ◽

Dendrobium Nobile ◽

Living Tissues ◽

Pathogenic Effects

Dendrobium are tropical orchid plants that host diverse endophytic fungi. The role of these fungi is not currently well understood in Dendrobium plants. We morphologically and molecularly identified these fungal endophytes, and created an efficient system for evaluating the pathogenicity and symptoms of endophytic fungi on Dendrobium nobile and Dendrobium officinale though in vitro co-culturing. ReThe colony morphological traits of Dendrobium myco-endophytes (DMEs) were recorded for their identification. Molecular identification revealed the presence of Colletotrichum tropicicola, Fusarium keratoplasticum, Fusarium oxysporum, Fusarium solani, and Trichoderma longibrachiatum. The pathogenicity results revealed that T. longibrachiatum produced the least pathogenic effects against D. nobile protocorms. In seedlings, T. longibrachiatum showed the least pathogenic effects against D. officinale seedlings after seven days. C. tropicicola produced highly pathogenic effects against both Dendrobium seedlings. The results of histological examination of infected tissues revealed that F. keratoplasticum and T. longibrachiatum fulfill Koch’s postulates for the existence of endophytes inside the living tissues. The DMEs are cross-transmitted inside the host plant cells, playing an important role in plant host development, resistance, and alkaloids stimulation.

Download Full-text

Endophytic fungi as a promising biocontrol agent to protect wheat from Fusarium graminearum head blight

Plant Disease ◽

10.1094/pdis-06-21-1253-re ◽

2021 ◽

Author(s):

Zachary Albert Noel ◽

Ludmilla Roze ◽

Mikaela Breunig ◽

Frances Trail

Keyword(s):

Microbial Community ◽

Fusarium Graminearum ◽

Endophytic Fungi ◽

Seed Weight ◽

Biocontrol Agent ◽

Fungal Endophytes ◽

In Planta ◽

Head Blight ◽

Species Definitions

The search for beneficial endophytes that can be part of a constructed microbial community has increased in recent years. We characterized three endophytic fungi previously isolated from wheat for their in vitro and in planta antagonism toward the Fusarium head blight pathogen, Fusarium graminearum. The endophytes were phylogenetically characterized and shown to be Alternaria destruens, Fusarium commune, and Fusarium oxysporum. Individual fungal endophytes significantly increased seed weight and lowered the accumulation of the mycotoxin deoxynivalenol compared to F. graminearum infected wheat heads without endophyte pretreatment. Investigation into the mechanism of competition in vitro showed that endophytes competitively excluded F. graminearum by pre-emptive colonization and possible inhibition over a distance. Investigations on the use of these endophytes in the field are in progress. Identification of these three endophytes highlights a common quandary in searching for beneficial microbes to use in agriculture: species definitions often do not separate individual isolates’ lifestyles. A greater understanding of the risks in using intraspecies variants for biocontrol is needed and should be examined in the context of the ecology of the individuals being investigated.

Download Full-text

Fungal endophytes of some medicinal plant growing in northwestern coast of Egypt: Isolation, identification and two-way clustering analysis of its antimicrobial activity

10.21203/rs.3.rs-828514/v1 ◽

2021 ◽

Author(s):

Mohammad Magdy El-Metwally ◽

Atia Mohamed Eisa ◽

Amal AI Mekawey ◽

Samy F. Mahmoud ◽

Yasser El Halmouch

Keyword(s):

Antimicrobial Activity ◽

Medicinal Plants ◽

Endophytic Fungi ◽

Fungal Endophytes ◽

Diversity Indices ◽

Antimicrobial Activities ◽

Penicillium Expansum ◽

Nicotiana Glauca ◽

Northwestern Coast

Abstract One of hidden mine of antibiotics is endophytic fungi especially that inhibited medicinal plants. In this regard, leaves, stems, fruits and bulbs of some commonly medicinal plants growing in Northwestern coast of Egypt were subjected for isolation of endophytic fungi with screening study of its antimicrobial activity. Practically, more than one hundred (101) endophytic fungal species isolated from Scorpiurus muricatus, Mellilotus indicus, Lotus polyphyllos, Ononis vaginalis, Nicotiana glauca, Lycium europaeum, Asphodelus aestivus, Echium angustifolium,Fagonia cretica, Pancratium maritimum,and Carduus getulus were tested in vitro for their antimicrobial activities against E. coli, Pseudomonas argenosa, Staphylococcus aureus, Bacillus subtilus, Candida albicanus, Candida glabrata, Penicillium expansum, Aspergillus flavus. The values of applied diversity indices revealed significant differences in presence, absence and abundance among endophytic fungal isolates. To the best of our knowledge, the present study is the first to report of Alternaria pluriseptata as endophytic species with most dominant and most active in its anitimicrobial activity among the isolated species. Seven distinctive groups were revealed from the two-way cluster analysis showing the intensity of antimicrobial activity against tested pathogens: Twenty-five percent of the isolates (26 strains assembled together in group V) exhibited no antimicrobial activity against all tested pathogens while six percent (6 isolates) assembled in group VII revealed high antimicrobial activity against five pathogens.

Download Full-text

Biodiversity, Ecology, and Secondary Metabolites Production of Endophytic Fungi Associated with Amaryllidaceae Crops

Agriculture ◽

10.3390/agriculture10110533 ◽

2020 ◽

Vol 10 (11) ◽

pp. 533 ◽

Cited By ~ 1

Author(s):

Gianluca Caruso ◽

Nadezhda Golubkina ◽

Alessio Tallarita ◽

Magdi T. Abdelhamid ◽

Agnieszka Sekara

Keyword(s):

Endophytic Fungi ◽

Fungal Endophytes ◽

Market Potential ◽

Plant Diseases ◽

In Vitro Synthesis ◽

Pharmaceutical Industries ◽

Fungal Microbiome ◽

Antagonistic Potential ◽

Metabolites Production

Amaryllidaceae family comprises many crops of high market potential for the food and pharmaceutical industries. Nowadays, the utilization of plants as a source of bioactive compounds requires the plant/endophytic microbiome interactions, which affect all aspects of crop’s quantity and quality. This review highlights the taxonomy, ecology, and bioactive chemicals synthesized by endophytic fungi isolated from plants of the Amaryllidaceae family with a focus on the detection of pharmaceutically valuable plant and fungi constituents. The fungal microbiome of Amaryllidaceae is species- and tissue-dependent, although dominating endophytes are ubiquitous and isolated worldwide from taxonomically different hosts. Root sections showed higher colonization as compared to bulbs and leaves through the adaptation of endophytic fungi to particular morphological and physiological conditions of the plant tissues. Fungal endophytes associated with Amaryllidaceae plants are a natural source of ecofriendly bioagents of unique activities, with special regard to those associated with Amarylloidae subfamily. The latter may be exploited as stimuli of alkaloids production in host tissues or can be used as a source of these compounds through in vitro synthesis. Endophytes also showed antagonistic potential against fungal, bacterial, and viral plant diseases and may find an application as alternatives to synthetic pesticides. Although Amaryllidaceae crops are cultivated worldwide and have great economic importance, the knowledge on their endophytic fungal communities and their biochemical potential has been neglected so far.

Download Full-text

Fish Chromosome Preparation: Air-Dried Displays of Cultured Ovarian Cells in Two Killifishes (Fundulus)

Journal of the Fisheries Research Board of Canada ◽

10.1139/f70-016 ◽

1970 ◽

Vol 27 (1) ◽

pp. 158-161 ◽

Cited By ~ 18

Author(s):

T. R. Chen

Keyword(s):

Cell Culture ◽

Cell Suspension ◽

Ovarian Tissue ◽

Single Cell Suspension ◽

High Quality ◽

Karyotypic Analysis ◽

Ovarian Cells ◽

Short Period ◽

General Similarity

A relatively simple and reliable in vitro methodology for fish chromosome study is described. A single cell suspension obtained by trypsinizing ovarian tissue was cultured for 5–7 days at 31 C. The cells were treated with colchicine and hypotonic solutions then fixed in acetic-alcohol fixative. Air-dried preparations were made from these cells and were stained with aceto-orcein.Numerous metaphase plates unmarred by overlapping chromosomes were recovered and high quality karyograms were available for detailed karyotypic analysis. The general similarity of squash and air-dried karyotypes suggests that short-period cell culture preserves natural karyotypes.

Download Full-text

Isolation, Characterization and Targeted Metabolic Evaluation of Endophytic Fungi Harbored in 14 Seed-Derived Hypericum Species

Planta Medica ◽

10.1055/a-1130-4703 ◽

2020 ◽

Vol 86 (13/14) ◽

pp. 997-1008 ◽

Cited By ~ 1

Author(s):

Jana Henzelyová ◽

Michaela Antalová ◽

Katarína Nigutová ◽

Mariia Logoida ◽

Andrea Schreiberová ◽

...

Keyword(s):

Endophytic Fungi ◽

Extracellular Enzymes ◽

Fungal Endophytes ◽

Ecological Niches ◽

Metabolic Evaluation ◽

Protein Coding ◽

Emodin Anthrone ◽

First Time

AbstractMedicinal plants of the genus Hypericum are rich sources of bioactive naphthodianthrones, which are unique in the plant kingdom, but quite common in fungal endophytes. Cultivable endophytic fungi were isolated from 14 different Hypericum spp. originating from seeds grown under in vitro conditions and further acclimated to outdoor conditions. Among 37 fungal isolates yielded from the aerial and underground plant organs, 25 were identified at the species level by the fungal barcode marker internal transcribed spacer rDNA and protein-coding gene region of tef1α. Ten of them were isolated from Hypericum spp. for the first time. The axenic cultures of the isolated endophytes were screened for the production of extracellular enzymes, as well as bioactive naphthodianthrones and their putative precursors by Bornträgerʼs test and HPLC-HRMS. Traces of naphthodianthrones and their intermediates, emodin, emodin anthrone, skyrin, or pseudohypericin, were detected in the fungal mycelia of Acremonium sclerotigenum and Plectosphaerella cucumerina isolated from Hypericum perforatum and Hypericum maculatum, respectively. Traces of emodin, hypericin, and pseudohypericin were released in the broth by Scedosporium apiospermum, P. cucumerina, and Fusarium oxysporum during submerged fermentation. These endophytes were isolated from several hypericin-producing Hypericum spp. Taken together, our results reveal the biosynthetic potential of cultivable endophytic fungi harbored in Hypericum plants as well as evidence of the existence of remarkable plant-endophyte relationships in selected non-native ecological niches. A possible role of the extracellular enzymes in plant secondary metabolism is discussed.

Download Full-text

In vitro Antagonistic Activity of Endophytic Fungi Isolated from Shirazi Thyme (Zataria multiflora Boiss.) against Monosporascus cannonballus

Polish Journal of Microbiology ◽

10.33073/pjm-2020-029 ◽

2020 ◽

Vol 69 (3) ◽

pp. 379-383

Author(s):

RAHIL SAID AL-BADI ◽

THAMODINI GAYA KARUNASINGHE ◽

ABDULLAH MOHAMMED AL-SADI ◽

ISSA HASHIL AL-MAHMOOLI ◽

RETHINASAMY VELAZHAHAN

Keyword(s):

Endophytic Fungi ◽

Pathogenic Fungus ◽

Fungal Endophytes ◽

Inhibition Zone ◽

Antagonistic Activity ◽

In Vitro Tests ◽

Zataria Multiflora ◽

Monosporascus Cannonballus ◽

Isolation And Characterization

Endophytic fungi viz., Nigrospora sphaerica (E1 and E6), Subramaniula cristata (E7), and Polycephalomyces sinensis (E8 and E10) were isolated from the medicinal plant, Shirazi thyme (Zataria multiflora). In in vitro tests, these endophytes inhibited the mycelial growth of Monosporascus cannonballus, a plant pathogenic fungus. Morphological abnormalities in the hyphae of M. cannonballus at the edge of the inhibition zone in dual cultures with N. sphaerica were observed. The culture filtrates of these endophytes caused leakage of electrolytes from the mycelium of M. cannonballus. To our knowledge, this is the first report on the isolation and characterization of fungal endophytes from Z. multiflora as well as their antifungal effect on M. cannonballus.

Download Full-text

Bioactivity Screening and Identification of Secondary Metabolites from Fungal Endophytes of Carica papaya L. Leaves

Journal of Natural Remedies ◽

10.18311/jnr/2020/24110 ◽

2021 ◽

Vol 20 (4) ◽

pp. 217

Author(s):

Israt Farha Lini ◽

Rabita Zinnurine ◽

Md. Habibur Rahman ◽

Mst. Nadira Begum ◽

Farhana Afroz ◽

...

Keyword(s):

Secondary Metabolites ◽

Endophytic Fungi ◽

Large Scale ◽

Carica Papaya ◽

Fungal Endophytes ◽

Diffusion Method ◽

Cytotoxic Activities ◽

Alternariol Monomethyl Ether ◽

Chemical Screening

Objectives: This work describes the chemical and in-vitro bioactivity studies on endophytic fungi derived from the leaves of Carica papaya L. Methods: Column Chromatography was employed for isolation of fungal metabolites. Nuclear Magnetic Resonance (NMR) spectral data was utilized to elucidate the structure of the isolated compounds and physicochemical properties of them were also examined. As a part of in-vitro bioactivity screening disc diffusion method, 1, 1-diphenyl-2- picryl-hydrazyl (DPPH) scavenging assay and brine shrimp lethality bioassay were conducted to evaluate antimicrobial, antioxidant and cytotoxic activities, respectively. Results: A total of four fungal isolates were identified as endophytic fungi and purified from the young leaves of Carica papaya. These fungi, encoded as CPLE-1, CPLE-2, CPLE-3 and CPLE-4, were identified up to the genus level on the basis of their macroscopic and microscopic characteristics as Colletotrichum sp., Carvularia sp., and next two as Alternaria sp., respectively. Chemical profiling and preliminary bioactivity screening of the fungal endophytes signified strain CPLE-3 with highest potentiality to produce bioactive compounds which directs its large scale cultivation. Ergosterol and Alternariol monomethyl ether were isolated from the ethyl acetate extract of the fungus, CPLE-3. Conclusion: Bioactivity and chemical screening of the endophytic fungal extracts of Carica papaya leaf along with the isolation of fungal secondary metabolites suggested these endophytic fungi to be possible source of bioactive leads for developing new and improved medicines.

Download Full-text