scholarly journals Novel Antifungal Activity of Lolium-Associated Epichloë Endophytes

2020 ◽  
Vol 8 (6) ◽  
pp. 955 ◽  
Author(s):  
Krishni Fernando ◽  
Priyanka Reddy ◽  
Inoka K. Hettiarachchige ◽  
German C. Spangenberg ◽  
Simone J. Rochfort ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Insect Pests ◽  
Fungal Endophytes ◽  
Dual Culture ◽  
The Novel ◽  
In Planta ◽  
Pasture Production ◽  
Fusarium Sp ◽  
Invertebrate Pests

Asexual Epichloë spp. fungal endophytes have been extensively studied for their functional secondary metabolite production. Historically, research mostly focused on understanding toxicity of endophyte-derived compounds on grazing livestock. However, endophyte-derived compounds also provide protection against invertebrate pests, disease, and other environmental stresses, which is important for ensuring yield and persistence of pastures. A preliminary screen of 30 strains using an in vitro dual culture bioassay identified 18 endophyte strains with antifungal activity. The novel strains NEA12, NEA21, and NEA23 were selected for further investigation as they are also known to produce alkaloids associated with protection against insect pests. Antifungal activity of selected endophyte strains was confirmed against three grass pathogens, Ceratobasidium sp., Dreschlera sp., and Fusarium sp., using independent isolates in an in vitro bioassay. NEA21 and NEA23 showed potent activity against Ceratobasidium sp. and NEA12 showed moderate inhibition against all three pathogens. Crude extracts from liquid cultures of NEA12 and NEA23 also inhibited growth of the phytopathogens Ceratobasidium sp. and Fusarium sp. and provided evidence that the compounds of interest are stable, constitutively expressed, and secreted. Comparative analysis of the in vitro and in planta metabolome of NEA12 and NEA23 using LCMS profile data revealed individual metabolites unique to each strain that are present in vitro and in planta. These compounds are the best candidates for the differential bioactivity observed for each strain. Novel endophyte strains show promise for endophyte-mediated control of phytopathogens impacting Lolium spp. pasture production and animal welfare.

scholarly journals Fungal Endophytes Control Fusarium graminearum and Reduce Trichothecenes and Zearalenone in Maize

Toxins ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 493 ◽  
Author(s):  
Mohamed F. Abdallah ◽  
Marthe De Boevre ◽  
Sofie Landschoot ◽  
Sarah De Saeger ◽  
Geert Haesaert ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Fusarium Graminearum ◽  
Fungal Endophytes ◽  
Agricultural Science ◽  
Ear Rot ◽  
Seedling Blight ◽  
In Planta ◽  
Detoxification Mechanisms ◽  
Comparative Trials

Fusarium graminearum can cause Giberella Ear Rot (GER) and seedling blight in maize, resulting in major yield losses. Besides GER, the infected grains are consequently contaminated with multiple mycotoxins of F. graminearum. Zearalenone and trichothecenes, such as deoxynivalenol and its acetylated forms, are among the major mycotoxins associated with F. graminearum infection in maize. In the current work, we explored the effect of the endophytic fungal genera of Epicoccum and Sordaria, to control F. graminearum infection in comparative trials with Piriformospora spp., an elusive endophytic genus. Furthermore, we investigated the effect of these endophytes on zearalenone, deoxynivalenol, and 15-acetyldeoxynivalenol levels using in vitro and in planta assays. As plants are endowed with several detoxification mechanisms comprising e.g., glucosylation of trichothecenes, the effect of the isolated fungal endophytes on the deoxynivalenol-3-glucoside level was also assessed. In general, results showed a considerable variability in the antifungal activity, both among species and among isolates within one species. Additionally, the effect on mycotoxin levels was variable, and not necessarily related to the antifungal activity except for zearalenone levels which were consistently reduced by the endophytes. These results highlight the great potential of certain endophytic fungal strains as new biocontrol agents in agricultural science.

scholarly journals The Known Antimammalian and Insecticidal Alkaloids Are Not Responsible for the Antifungal Activity of Epichloë Endophytes

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2486
Author(s):  
Krishni Fernando ◽  
Priyanka Reddy ◽  
Simone Vassiliadis ◽  
German C. Spangenberg ◽  
Simone J. Rochfort ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Insect Pests ◽  
Ergot Alkaloids ◽  
In Planta ◽  
Turf Quality ◽  
Growth Area ◽  
Pasture Grasses ◽  
Lolitrem B ◽  
Epichloë Endophytes

Asexual Epichloë sp. endophytes in association with pasture grasses produce agronomically important alkaloids (e.g., lolitrem B, epoxy-janthitrems, ergovaline, peramine, and lolines) that exhibit toxicity to grazing mammals and/or insect pests. Novel strains are primarily characterised for the presence of these compounds to ensure they are beneficial in an agronomical setting. Previous work identified endophyte strains that exhibit enhanced antifungal activity, which have the potential to improve pasture and turf quality as well as animal welfare through phytopathogen disease control. The contribution of endophyte-derived alkaloids to improving pasture and turf grass disease resistance has not been closely examined. To assess antifungal bioactivity, nine Epichloë related compounds, namely peramine hemisulfate, n-formylloline-d3, n-acetylloline hydrochloride, lolitrem B, janthitrem A, paxilline, terpendole E, terpendole C, and ergovaline, and four Claviceps purpurea ergot alkaloids, namely ergotamine, ergocornine, ergocryptine, and ergotaminine, were tested at concentrations higher than observed in planta in glasshouse and field settings using in vitro agar well diffusion assays against three common pasture and turf phytopathogens, namely Ceratobasidium sp., Drechslera sp., and Fusarium sp. Visual characterisation of bioactivity using pathogen growth area, mycelial density, and direction of growth indicated no inhibition of pathogen growth. This was confirmed by statistical analysis. The compounds responsible for antifungal bioactivity of Epichloë endophytes hence remain unknown and require further investigation.

scholarly journals Evaluation of in-vitro methods to select effective streptomycetes against toxigenic fusaria

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6905 ◽  
Author(s):  
Elena Maria Colombo ◽  
Cristina Pizzatti ◽  
Andrea Kunova ◽  
Claudio Gardana ◽  
Marco Saracchi ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Culture Media ◽  
Screening Methods ◽  
Dual Culture ◽  
In Planta ◽  
In Vitro Method ◽  
In Vitro Methods ◽  
Strain Diversity ◽  
Fungal Plant Pathogens

Biocontrol microorganisms are emerging as an effective alternative to pesticides. Ideally, biocontrol agents (BCAs) for the control of fungal plant pathogens should be selected by an in vitro method that is high-throughput and is predictive of in planta efficacy, possibly considering environmental factors, and the natural diversity of the pathogen. The purpose of our study was (1) to assess the effects ofFusariumstrain diversity (N= 5) and culture media (N= 6) on the identification of biological control activity ofStreptomycesstrains (N= 20) againstFusariumpathogens of wheat in vitro and (2) to verify the ability of our in vitro screening methods to simulate the activity in planta. Our results indicate that culture media,Fusariumstrain diversity, and their interactions affect the results of an in vitro selection by dual culture assay. The results obtained on the wheat-based culture media resulted in the highest correlation score (r= 0.5) with the in planta root rot (RR) inhibition, suggesting that this in vitro method was the best predictor of in planta performance of streptomycetes against Fusarium RR of wheat assessed as extension of the necrosis on the root. Contrarily, none of the in vitro plate assays using the media tested could appropriately predict the activity of the streptomycetes against Fusarium foot rot symptoms estimated as the necrosis at the crown level. Considering overall data of correlation, the activity in planta cannot be effectively predicted by dual culture plate studies, therefore improved in vitro methods are needed to better mimic the activity of biocontrol strains in natural conditions. This work contributes to setting up laboratory standards for preliminary screening assays ofStreptomycesBCAs against fungal pathogens.

scholarly journals A Yeast-Based Functional Assay to Study Plant N-Degron – N-Recognin Interactions

2022 ◽  
Vol 12 ◽  
Author(s):  
Aida Kozlic ◽  
Nikola Winter ◽  
Theresia Telser ◽  
Jakob Reimann ◽  
Katrin Rose ◽  
...  
Keyword(s):  
Ubiquitin Proteasome System ◽  
Functional Assay ◽  
The Novel ◽  
In Planta ◽  
Amino Terminal ◽  
Weak Binding ◽  
Ubiquitin Conjugating Enzyme ◽  
Ubiquitin Proteasome ◽  
The Stability

The N-degron pathway is a branch of the ubiquitin-proteasome system where amino-terminal residues serve as degradation signals. In a synthetic biology approach, we expressed ubiquitin ligase PRT6 and ubiquitin conjugating enzyme 2 (AtUBC2) from Arabidopsis thaliana in a Saccharomyces cerevisiae strain with mutation in its endogenous N-degron pathway. The two enzymes re-constitute part of the plant N-degron pathway and were probed by monitoring the stability of co-expressed GFP-linked plant proteins starting with Arginine N-degrons. The novel assay allows for straightforward analysis, whereas in vitro interaction assays often do not allow detection of the weak binding of N-degron recognizing ubiquitin ligases to their substrates, and in planta testing is usually complex and time-consuming.

scholarly journals Inhibitory Effect of Trichoderma asperellum Isolate against Ralstonia solanacearum Causing Brinjal Wilt

2021 ◽  
Vol 24 (2) ◽  
pp. 107-120
Author(s):  
SMN Islam ◽  
SS Siddique ◽  
MZH Chowdhury ◽  
NJ Mishu
Keyword(s):  
Ralstonia Solanacearum ◽  
Disease Incidence ◽  
Its Region ◽  
Inhibitory Effect ◽  
Dual Culture ◽  
Trichoderma Asperellum ◽  
In Planta ◽  
Quantitative Test ◽  
Trichoderma Isolate

A native Trichoderma isolate was collected from the agricultural soil of Gazipur. This isolate was identified as a Trichoderma asperellum through morphology and analysis of internal transcribed spacer (ITS) region of ribosomal RNA gene sequence and reconstruction of the phylogenetic tree. The antagonistic effects of the newly identified T. asperellum isolate were assessed against brinjal bacterial wilt caused by Ralstonia solanacearum both in vitro and in planta. Both qualitative and quantitative bioassays were conducted in vitro. For qualitative tests, dual culture and antibacterial activity were carried out, and pathogen growth was observed visually. The antagonism of T. asperellum cell free culture filtrate on the growth of R. solanacearum was conducted in a quantitative test. Successful antagonism was recorded after both in vitro qualitative tests. In addition, the lowest colony forming unit was recorded in 100% of CFC (2.4±0.51 ×103 cfu/ml) in quantitative test. The T. asperellum inoculated plant showed low disease incidence (13.33%) when seedlings were challenged with R. solanacearum in planta experiment. Disease incidence was 100% for seedlings when treated with only R. solanacearum. The results showed that the isolated and identified T. asperellum isolate suppressed R. solanacearum growth in vitro and protected the seedling from wilting in planta. Therefore, this isolate could be considered as a potential isolate. Ann. Bangladesh Agric. (2020) 24(2) : 107-120

scholarly journals Transcriptional Profiling of Diffusible Lipopeptides and Fungal Virulence Genes During Bacillus amyloliquefaciens EZ1509-Mediated Suppression of Sclerotinia sclerotiorum

2020 ◽  
Vol 110 (2) ◽  
pp. 317-326 ◽  
Author(s):  
Ayaz Farzand ◽  
Anam Moosa ◽  
Muhammad Zubair ◽  
Abdur Rashid Khan ◽  
Muhammad Ayaz ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Oxalic Acid ◽  
Sclerotinia Sclerotiorum ◽  
Bacillus Amyloliquefaciens ◽  
Virulence Genes ◽  
Mass Spectrometry Analysis ◽  
Dual Culture ◽  
Biosynthetic Genes ◽  
In Vitro Interaction

Sclerotinia sclerotiorum is a devastating necrotrophic pathogen that infects multiple crops, and its control is an unremitting challenge. In this work, we attempted to gain insights into the pivotal role of lipopeptides (LPs) in the antifungal activity of Bacillus amyloliquefaciens EZ1509. In a comparative study involving five Bacillus strains, B. amyloliquefaciens EZ1509 harboring four LPs biosynthetic genes (viz. surfactin, iturin, fengycin, and bacilysin) exhibited promising antifungal activity against S. sclerotiorum in a dual-culture assay. Our data demonstrated a remarkable upsurge in LPs biosynthetic gene expression through quantitative reverse transcription PCR during in vitro interaction assay with S. sclerotiorum. Maximum upregulation in LPs biosynthetic genes was observed on the second and third days of in vitro interaction, with iturin and fengycin being the highly expressed genes. Subsequently, Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry analysis confirmed the presence of LPs in the inhibition zone. Scanning electron microscope analysis showed disintegration, shrinkage, plasmolysis, and breakdown of fungal hyphae. During in planta evaluation, S. sclerotiorum previously challenged with EZ1509 showed significant suppression in pathogenicity on detached leaves of tobacco and rapeseed. The oxalic acid synthesis was also significantly reduced in S. sclerotiorum previously confronted with antagonistic bacterium. The expression of major virulence genes of S. sclerotiorum, including endopolygalacturonase-3, oxalic acid hydrolase, and endopolygalacturonase-6, was significantly downregulated during in vitro confrontation with EZ1509.

scholarly journals Antifungal Activity of Silver Ions and Nanoparticles on Phytopathogenic Fungi

Plant Disease ◽  
2009 ◽  
Vol 93 (10) ◽  
pp. 1037-1043 ◽  
Author(s):  
Young-Ki Jo ◽  
Byung H. Kim ◽  
Geunhwa Jung
Keyword(s):  
Antifungal Activity ◽  
Colony Formation ◽  
Magnaporthe Grisea ◽  
Silver Ions ◽  
Pathogenic Fungi ◽  
Chloride Ions ◽  
Plant Diseases ◽  
Plant Pathogenic Fungi ◽  
In Planta

Silver in ionic or nanoparticle forms has a high antimicrobial activity and is therefore widely used for various sterilization purposes including materials of medical devices and water sanitization. There have been relatively few studies on the applicability of silver to control plant diseases. Various forms of silver ions and nanoparticles were tested in the current study to examine the antifungal activity on two plant-pathogenic fungi, Bipolaris sorokiniana and Magnaporthe grisea. In vitro petri dish assays indicated that silver ions and nanoparticles had a significant effect on the colony formation of these two pathogens. Effective concentrations of the silver compounds inhibiting colony formation by 50% (EC50) were higher for B. sorokiniana than for M. grisea. The inhibitory effect on colony formation significantly diminished after silver cations were neutralized with chloride ions. Growth chamber inoculation assays further confirmed that both ionic and nanoparticle silver significantly reduced these two fungal diseases on perennial ryegrass (Lolium perenne). Particularly, silver ions and nanoparticles effectively reduced disease severity with an application at 3 h before spore inoculation, but their efficacy significantly diminished when applied at 24 h after inoculation. The in vitro and in planta evaluations of silver indicated that both silver ions and nanoparticles influence colony formation of spores and disease progress of plant-pathogenic fungi. In planta efficacy of silver ions and nanoparticles is much greater with preventative application, which may promote the direct contact of silver with spores and germ tubes, and inhibit their viability.

Antifungal Activity of the Novel Fungicide Cyazofamid against Phytophthora infestans and Other Plant Pathogenic Fungi in Vitro

2001 ◽  
Vol 70 (2) ◽  
pp. 92-99 ◽  
Author(s):  
Shigeru Mitani ◽  
Satoshi Araki ◽  
Tomona Yamaguchi ◽  
Yasuko Takii ◽  
Takeshi Ohshima ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Phytophthora Infestans ◽  
Pathogenic Fungi ◽  
Plant Pathogenic Fungi ◽  
The Novel

scholarly journals Characterisation of a novel endophyte NRPS gene and its role in endophyte-grass symbioses

2007 ◽  
Vol 13 ◽  
pp. 491-493
Author(s):  
H. Harzer ◽  
R.D. Johnson ◽  
S. Rasmussen ◽  
C.R. Voisey ◽  
L.J. Johnson
Keyword(s):  
Fungal Endophytes ◽  
Gene Clusters ◽  
Gene Replacement ◽  
In Planta ◽  
Dna Library ◽  
Peptide Synthetase ◽  
Genomic Dna Library ◽  
Targeted Gene Replacement ◽  
Fungal Secondary Metabolite

Symbiotic grass associations with fungal endophytes (genera Neotyphodium and Epichloë) display enhanced fitness as well as prolonged field persistence over their endophyte free equivalents. Perennial ryegrass, an important agronomic grass, is typically associated with the N. lolii endophyte. The endophyte lives within the intercellular spaces without inducing any symptoms in the plant. The aim of this study is to elucidate the biosynthetic function of fungal secondary metabolite gene clusters. Non-ribosomal peptide synthetase genes (NRPSs) of unknown function were targeted, as these genes are commonly associated with the production of bioactive peptides some of which are ecologically important. Some novel endophyte NRPS genes have been identified using a degenerate PCR screen; one of these, NRPS5 will be discussed here. Clones were obtained by screening a fosmid Epichloë festucae genomic DNA library and we are currently determining gene function by using targeted gene replacement followed by an assessment in vitro and in planta using metabolomics and appropriate bioassay screens. Keywords: endophyte, NRPS, secondary metabolism

scholarly journals MEKANISME PARASITISME TRICHODERMA HARZIANUM TERHADAP FUSARIUM OXYSPORUM PADA SEMAI ACACIA MANGIUM

2015 ◽  
Vol 15 (1) ◽  
pp. 72
Author(s):  
Susanti Tasik ◽  
Siti Muslimah Widyastuti ◽  
Harjono .
Keyword(s):  
Fusarium Oxysporum ◽  
Trichoderma Harzianum ◽  
Acacia Mangium ◽  
Culture Method ◽  
Inhibition Mechanism ◽  
Dual Culture ◽  
In Planta ◽  
Know How ◽  
Soil Borne Fungi

Mechanism of parasitism of Trichoderma harzianum on Fusarium oxysporum on Acacia mangium seedlings. Fusarium oxysporum is one of the most important soil-borne fungi the causal agent of damping-off disease. Detailed information it needed to know how the pathogen can be inhibited by Trichoderma harzianum. The objective of this research was to investigate the inhibition mechanism of T. harzianum on F. oxysporum in vitro and in planta. Green Flourescent Protein (GFP) T. harzianum was used as biocontrol agent of F. oxysporum. An in vitro inhibition test of T. harzianum was performed using dual culture method. In the in planta inhibition tests, seedlings of A. mangium were applied with GFP T. harzianum two days before inoculation of F. oxysporum; GFP T. harzianum was simultaneously applied with F. oxysporum and GFP T. harzianum was applied two days after inoculation of F. oxysporum. The inhibition effect of T. harzianum GFP was observed at seven days incubation, indicated by attachment of T. harzianum to F. oxysporum hyphae. GFP T. harzianum hyphae covered the colonies of F. oxysporum at 12 days after incubation. The highest life percentage of A. mangium seedlings was found on the treatment of GFP T. harzianum two days before inoculation of F. oxysporum (82.22%), whereas the lowest life percentage was found on seedling applied with GFP T. harzianum two days after inoculation of F. oxysporum (64.44%).

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