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

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.

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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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UJI DAYAHAMBAT EUGENOL DARI DAUN CENGKEH HASIL FRAKSINASI TERHADAP PERTUMBUHAN JAMUR PATOGEN Fusarium oxysporum f.sp. Cubense Eugenol Inhibitory Test From Clove Leaf Fractionation Of Fungal Patogen Growth Fusarium oxysporum f.sp. cubense

AgriPeat ◽

10.36873/agp.v20i02.146 ◽

2019 ◽

Vol 20 (02) ◽

pp. 107-113

Author(s):

Admin Journal

Keyword(s):

Fusarium Oxysporum ◽

Vacuum Distillation ◽

Pathogenic Fungus ◽

Lethal Concentration ◽

In Vitro Tests ◽

In Vitro Test ◽

Microsoft Excel ◽

Vitro Test

ABSTRACTThis study aims to determine the inhibition of eugenol derived from fractionation clove leaf essentialoils (CLEO) on the growth of pathogenic fungus Fusarium oxysporum f. sp. cubense (Foc) and LC50(Lethal Concentration 50). This research was in vitro, started with purification of clove leaf essentialoil, fractionation by vacuum distillation and bioassay. In vitro tests include exploration of minimuminhibition and preventability tests. Data were analyzed with Microsoft Excel 2010 program. Theresults of minimum inhibition showed at 218,75 ppm concentration of each level was able to inhibitthe growth of Foc fungi. The minimum inhibition exploration was carried out at 218,75 ppm, 109,38ppm, 54,69 ppm and 27,34 ppm. Exploration results showed that fractionated CLEO has been able toinhibit the growth of Foc fungi at 27,34 ppm in the amount of 15,60%. This concentration is used asthe lowest concentration in the inhibitory test. Furthermore, the inhibitory test was carried out startingat the highest concentration of 218,75 ppm, 109,38 ppm, 54,69 ppm and 27,34 ppm. Observationswere made for 7 days after inoculation (DAI). The results showed the best inhibition was at aconcentration of 218,75 ppm at 90,70% and LC50 at 11.17 µL.Keywords: CLEO, fractionation, Foc, in vitro test and LC50

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Biocontrol potential of Trichoderma harzianum in controlling wilt disease of pistachio caused by Verticillium dahliae

Journal of Plant Protection Research ◽

10.1515/jppr-2017-0025 ◽

2017 ◽

Vol 57 (2) ◽

pp. 185-193 ◽

Cited By ~ 5

Author(s):

Zeinab Fotoohiyan ◽

Saeed Rezaee ◽

Gholam Hosein Shahidi Bonjar ◽

Amir Hossein Mohammadi ◽

Mohammad Moradi

Keyword(s):

Verticillium Dahliae ◽

Trichoderma Harzianum ◽

Pathogenic Fungus ◽

Antagonistic Activity ◽

Wilt Disease ◽

Dual Culture ◽

Volatile Metabolites ◽

Biocontrol Potential

Abstract Verticillium wilt caused by Verticillium dahliae, is one of the most devastating diseases in pistachio orchards in the world including Iran. In search for an effective non-chemical strategy for the management of this disease, we evaluated the biocontrol potential of Trichoderma harzianum isolates obtained from the rhizosphere of healthy pistachio trees in different locations of the Kerman province of Iran against V. dahliae under laboratory and greenhouse conditions. Dual culture tests in the laboratory were conducted in a completely randomized design using 72 T. harzianum isolates. Twenty isolates showed the highest in vitro antagonistic activity. The results indicated that all 20 isolates were capable of inhibiting the mycelial growth of V. dahliae significantly. Among them, isolates Tr8 and Tr19 were the most effective by 88.89% and 85.12% inhibition, respectively. Extracted cell free metabolites of all effective isolates also inhibited the growth of V. dahliae in the culture medium significantly. According to the results, isolates Tr4 and Tr6 inhibited fungal pathogen growth by 94.94% and 88.15% respectively, through production of non-volatile metabolites. In the evaluation of volatile metabolites, isolates Tr5 and Tr4 were the most effective by 26.27% and 24.49% growth inhibition, respectively. Based on the results of the in vitro experiments, the five most effective isolates were selected for evaluation under greenhouse conditions for their biocontrol potential in controlling Verticillium wilt of pistachio. Results of the greenhouse, (in vivo) experiments were positive and indicated that the occurrence of wilt disease in plants treated with the antagonists alone or in combination with pathogenic fungus was lower than in plants inoculated with pathogen alone. The overall results of this study suggest that Trichoderma fungal antagonist may be an effective biocontrol agent for the control of Verticillium wilt of pistachio.

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Penapisan Cendawan Trichoderma spp. untuk Pengendalian Phytophthora infestans secara in vitro

Jurnal Fitopatologi Indonesia ◽

10.14692/jfi.12.3.96 ◽

2016 ◽

Vol 12 (3) ◽

pp. 96

Author(s):

Susiana Purwantisari ◽

Achmadi Priyatmojo ◽

Retno Peni Sancayaningsih ◽

Rina Sri Kasiamdari

Keyword(s):

Growth Inhibition ◽

Phytophthora Infestans ◽

Pathogenic Fungus ◽

Culture Method ◽

Inhibition Zone ◽

Dual Culture ◽

Trichoderma Spp ◽

Suppressive Soil ◽

Blight Disease

Late blight disease on potato caused by a plant pathogenic fungus (Phytophthora infestans) is the most important disease in Indonesia. The use of antagonist fungi Trichoderma is an environmentally friendly technology to control the potato disease. The purpose of this study was to determine the ability of Trichoderma spp. to control P. infestans in vitro. Trichoderma spp. have been isolated from suppressive soil at central potato areas in district of Lembang, Bandung, West Java Province. To determine the inhibition ability of Trichoderma spp against P. infestans, a dual culture method was performed. Variable observed was inhibition zone of Trichoderma spp. against P. infestans. The results showed that 9 isolates of Trichoderma were successfuly isolated from suppressive soil, i.e. T. viride (2 isolates), T. atroviride (1 isolate), T. harzianum (1 isolate) and T. aureoviride (5 isolates). All the Trichoderma isolates revealed growth inhibition abality against P. infestans and. The highest growth inhibition (68.6%) was observed by T. viride isolate ( Ti 9).

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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

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Antibacterial Activity of Endophytic Fungi from Sembukan (Paederia foetida L.) Leaves

Science & Technology Indonesia ◽

10.26554/sti.2021.6.3.189-195 ◽

2021 ◽

Vol 6 (3) ◽

pp. 189-195

Author(s):

Hary Widjajanti ◽

Christina Vivid Handayani ◽

Elisa Nurnawati

Keyword(s):

Antibacterial Activity ◽

Secondary Metabolites ◽

Endophytic Fungi ◽

Antimicrobial Agents ◽

Antibacterial Properties ◽

Antibacterial Activities ◽

In Vitro Tests ◽

E Coli ◽

Paederia Foetida

The antibiotic resistance of phatogenic bacteria has become a serious health problem and has encouraged the search for novel and effective antimicrobial metabolites. Meanwhile, endophytic fungi have great potential as a natural source for antimicrobial agents. The endophytic fungi that live in plant tissue produces secondary metabolites which potentially act as an antibacterial compound. The isolation of fungi for antibacterial sources reduces the large amount of plant as a source of antibacterial agents. Hence, this study aims to obtain endophytic fungi isolates from Paederia foetida L. that are capable of producing secondary metabolites as antibacterial, carry out in vitro tests to verify the antibacterial properties of secondary metabolites of the Paederia foetida L. endophytic fungi, and identify the potential of Paederia foetida L. endophytic fungi in producing antibacterial compounds. The antibacterial activity was tested against Escherichia coli ATCC8739 and Staphylococcus aureus ATCC6538 while seven isolates of endophytic fungi that potentially produced antibacterial were obtained from Sembukan (P. foetida L.). The results showed that antibacterial activities of SL1, SL4 and SL6 secondary metabolites against S. aureus ATCC6538 and E. coli ATCC8739 were moderate to strong activities. Furthermore, the Minimum Inhibition Concentration (MIC) of secondary metabolites extract of SL1 against S. aureus ATCC6538 value was 250 ????g/mL while the values of MIC extract of SL4 against S. aureus ATCC6538 and E. coli ATCC8739 were 125 ????g/mL and 250 ????g/mL respectively and MIC extract of SL6 against E. coli ATCC8739 value was 125 ????g/mL. The secondary metabolites extract of SL1 isolate were alkaloid and tannin, SL4 were phenolic and alkaloid while SL6 isolate were alkaloid and terpenoid. Hence, endophytic fungi SL1 isolate was identified as Fusarium sp., SL4 as Dematophora sp., and SL6 isolate as Acremonium sp.

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Isolation of Endophytic Fungi from Vitex trifolia L and Antagonism Test against Sclerotium rolfsii and pathogenic bacteria

JURNAL BIOLOGI TROPIS ◽

10.29303/jbt.v21i1.2340 ◽

2021 ◽

Vol 21 (1) ◽

pp. 72

Author(s):

Muhammad Hasan Basri ◽

Lalu Zulkifli ◽

Abdul Syukur

Keyword(s):

Biological Control ◽

Endophytic Fungi ◽

Pathogenic Bacteria ◽

Pathogenic Fungus ◽

Sclerotium Rolfsii ◽

Pathogenic Fungi ◽

Promising Alternative ◽

Environmental Friendly ◽

Low Activity

Plant damage by pathogenic fungi is often found in plants, one of which is caused by Sclerotium rolfsii. Biological control strategy offers a promising alternative for managing disease in plants because they are environmental friendly compared to pesticides application. One of the biological control offered is by using endophytic fungi isolated from Vitex trivolia L. The aim of the study was to isolate, to identify macroscopic and microscopic endophytic fungi from Vitex trifolia L and to test their antagonism potency against the pathogenic fungus Sclerotium rolfsii in vitro. The isolation obtained 7 endophytic fungi isolates identified based on their genus characteristics, nsmely Periconia sp, Aspergillussp, Dendrophoma sp, Geotrichum sp, Ampulliferina sp, Chalara sp, dan Bispora sp and 2 isolates have not been identified. The Antibacterial test of the fungi isolate on the 4 tested bacteria showed that of all the fungi isolate have low activity. The antagonism test using the direct opposition method with the PIRG formula, showed that the 3 isolates had high percentage of growth inhibition, in which ALJ1, BLJ5, and ALJ3 isolate has 85%, 90%, and 100% respectively. This potency could be used as biological agents on the pathogenic fungus Sclerotium rolfsii.

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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.

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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.

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Isolation and characterization of Mycoplasma meleagridis

Canadian Journal of Microbiology ◽

10.1139/m72-022 ◽

1972 ◽

Vol 18 (2) ◽

pp. 127-135 ◽

Cited By ~ 1

Author(s):

M. S. Ogra

Keyword(s):

Inhibitory Effect ◽

Metabolic Inhibition ◽

In Vitro Tests ◽

Chronic Type ◽

Tetrazolium Chloride ◽

Productive Life ◽

Isolation And Characterization ◽

The Status

The ability of Mycoplasma meleagridis to metabolize L-argininehydrochloride (HCl) and its inability to ferment dextrose can be used in identifying and differentiating this serotype from other Mycoplasma known to infect turkeys. Mycoplasma meleagridis may be the only species of turkey origin which lacks the ability to reduce tetrazolium chloride. A selective inhibitory effect of erythromycin and oleandomycin on M. gallisepticum can be used to differentiate it from M. meleagridis in in vitro tests. The metabolic inhibition test can be applied successfully to identification of M. meleagridis and its antibody. In birds carrying a chronic type of infection, Mycoplasma may be isolated from the vagina throughout much of their productive life. Since the infection is not readily transmitted, the maintenance of flocks free from M. meleagridis may not be difficult once the status is achieved.

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