Morphological and Molecular Diversity of Ginger (Zingiber officinale Roscoe) Pathogenic Fungi in Chilga District, North Gondar, Ethiopia

Ginger diseases caused by fungal pathogens have become one of the most serious problems causing reduced production around the world. It has also caused a major problem among farmers in different parts of Ethiopia resulting in a huge decline in rhizome yield. However, the exact causative agents of this disease have not been identified in the state. Although there are few studies related to pathogenic fungus identification, molecular level identification of fungal pathogen was not done in the area. Therefore, this study was undertaken to isolate and characterized the fungal causative agent of ginger disease from the diseased plant and the soil samples collected around the diseased plant from Chilga district, Gondar, Ethiopia. Samples from infected ginger plants and the soil around the infected plant were collected. Culturing and purification of isolates were made using Potato Dextrose Agar supplemented with antibacterial agent chloramphenicol. The morphological characterization was done by structural identification of the isolates under the microscope using lactophenol cotton blue stains. Isolated fungi were cultured and molecular identification was done using an internal transcribed spacer (ITS) of ribosomal DNA (rDNA). A total of 15 fungal morphotypes including 11 Aspergillus spp. (73.3%), 2 Penicillium spp. (13.3%), and single uncultured fungus clone S23 were isolated from the samples representing all the plant organs and the soil. Aspergillus spp. (73.3%) was the most common and seems to be the major causative agent. To the best of our knowledge, this is the first report of ginger pathogenic fungi in Ethiopia identified using ITS rDNA molecular techniques. This study will lay foundation for the development of management strategies for fungal diseases infecting ginger.

Epichloë Endophyte-Promoted Seed Pathogen Increases Host Grass Resistance Against Insect Herbivory

Plants host taxonomically and functionally complex communities of microbes. However, ecological studies on plant–microbe interactions rarely address the role of multiple co-occurring plant-associated microbes. Here, we contend that plant-associated microbes interact with each other and can have joint consequences for higher trophic levels. In this study we recorded the occurrence of the plant seed pathogenic fungus Claviceps purpurea and aphids (Sitobion sp.) on an established field experiment with red fescue (Festuca rubra) plants symbiotic to a seed transmitted endophytic fungus Epichloë festucae (E+) or non-symbiotic (E–). Both fungi are known to produce animal-toxic alkaloids. The study was conducted in a semi-natural setting, where E+ and E– plants from different origins (Spain and Northern Finland) were planted in a randomized design in a fenced common garden at Kevo Subarctic Research Station in Northern Finland. The results reveal that 45% of E+ plants were infected with Claviceps compared to 31% of E– plants. Uninfected plants had 4.5 times more aphids than Claviceps infected plants. By contrast, aphid infestation was unaffected by Epichloë symbiosis. Claviceps alkaloid concentrations correlated with a decrease in aphid numbers, which indicates their insect deterring features. These results show that plant mutualistic fungi can increase the infection probability of a pathogenic fungus, which then becomes beneficial to the plant by controlling herbivorous insects. Our study highlights the complexity and context dependency of species–species and multi-trophic interactions, thus challenging the labeling of species as plant mutualists or pathogens.

Toxin-producing endosymbionts shield pathogenic fungus against micropredators

The phytopathogenic fungus Rhizopus microsporus harbours a bacterial endosymbiont (Mycetohabitans rhizoxinica) for the production of the toxin rhizoxin, the causative agent of rice seedling blight. This toxinogenic bacterial-fungal alliance is, however, not restricted to the plant disease, but has been detected in numerous environmental isolates from geographically distinct sites covering all five continents. Yet, the ecological role of rhizoxin beyond rice seedling blight has been unknown. Here we show that rhizoxin serves the fungal host in fending off protozoan and metazoan predators. Fluorescence microscopy and co-culture experiments with the fungivorous amoeba Protostelium aurantium revealed that ingestion of R. microsporus spores is toxic to P. aurantium. This amoebicidal effect is caused by the bacterial rhizoxin congener rhizoxin S2, which is also lethal towards the model nematode Caenorhabditis elegans. By combining stereomicroscopy, automated image analyses, and quantification of nematode movement we show that the fungivorous nematode Aphelenchus avenae actively feeds on R. microsporus that is lacking endosymbionts, while worms co-incubated with symbiotic R. microsporus are significantly less lively. This work uncovers an unexpected ecological role of rhizoxin as shield against micropredators. This finding suggests that predators may function an evolutionary driving force to maintain toxin-producing endosymbionts in non-pathogenic fungi.

Biscogniauxia Charcoal Canker—A New Potential Threat for Mid-European Forests as an Effect of Climate Change

Biscogniauxia nummularia (Bull.) Kuntze is a fungus which induces strip-cankers on beech, commonly referred to as charcoal canker. The symptoms of infection are visible on the host tree’s bark as elongated, blackish bark lesions on the trunk and branches. Recent years have shown that, due to climate change causing local epidemics, the species is increasing its economic impact in Mediterranean regions. Until recently, B. nummularia was considered rare and uncommon in central Europe. However, in the last few years it has been noticed more often, mostly in coniferous trees, which are out of B. nummularia’s host range. A similar situation has been observed with the closely related species Biscogniauxia mediterranea (De Not.) Kuntze, which prior to 2017 had not been observed in central Europe at all. This study shows the genetic diversity of mid-European strains of Biscogniauxia spp. (based on the ITS, TEF1, TUB2 and ACT regions) and, as the first in Europe, presents a molecular investigation of this species isolated from coniferous trees. It is also the first attempt at estimating the potential impact of this pathogenic fungus on European forestry management in the close future.

Molecular characterization and vegetative growth of pathogenic seed-borne fungus, Curvularia lunata of tomato and its in vitro control measures

Present studies were conducted to isolate and identify the seed-borne pathogenic fungus from the selected tomato variety through morphological and molecular techniques based on the sequencing of internal transcribed spacer (ITS) region of 18S rDNA. According to the colony and conidial features, the fungus was identified as Curvularia sp. The obtained ITS sequencing showed above 99% similarity with Curvularia lunata in the NCBI database. The sequence of the fungus was deposited in NCBI GenBank under the accession number: ITS, MH382879.1. Besides, the phylogenetic tree further confirmed the taxonomic position of the studied fungus. Growth characteristics of the fungus on nine different fungal culture media were evaluated, in which Honey peptone agar, Carrot agar, Potato sucrose agar, and Kauffman’s agar were found the most suitable. The maximum vegetative growth of the fungus was recorded at 30°C temperature and pH conditions. The bio-control potential of five different antagonists against the studied fungus was assessed, in which Trichoderma harzianum showed the better performance to restrict mycelial growth. Three ethanolic plant extracts were also evaluated, in which Lowsonia inermis L. exhibited above 60% mycelial growth inhibition of the fungus. Among three tested fungicides, Tilt 250 EC was found as an excellent fungicide to inhibit mycelial growth of C. lunata under in vitro conditions. Int. J. Agril. Res. Innov. Tech. 11(2): 124-132, Dec 2021

Screening of Bacillus velezensis E2 and the Inhibitory Effect of Its Antifungal Substances on Aspergillus flavus

Aspergilus flavus is the main pathogenic fungus that causes food mold. Effective control of A. flavus contamination is essential to ensure food safety. The lipopeptides (LPs) produced by Bacillus strains have been shown to have an obvious antifungal effect on molds. In this study, an antagonist strain of Bacillus velezensis with obvious antifungal activity against A. flavus was isolated from the surface of healthy rice. Using HPLC-MS analysis, the main components of LPs produced by strain E2 were identified as fengycin and iturins. Further investigations showed that LPs could inhibit the spore germination, and even cause abnormal expansion of hyphae and cell rupture. Transcriptomic analyses showed that some genes, involved in ribosome biogenesis in eukaryotes (NOG1, KRE33) and aflatoxin biosynthesis (aflK, aflR, veA, omtA) pathways in A. flavus were significantly down-regulated by LPs. In conclusion, this study provides novel insights into the cellular and molecular antifungal mechanisms of LPs against grain A. flavus contamination.

Vishaghn Dhoop, Nano-Scale Particles with Detoxifying Medicinal Fume, Exhibits Robust Anti-Microbial Activities: Implications of Disinfection Potentials of a Traditional Ayurvedic Air Sterilization Technique

The rapidly increasing global burden of healthcare associated infections (HAI) is resulting in proportionate increase in chemical disinfection in healthcare settings, adding an extra burden of environmental toxicity. Therefore, alternative disinfection techniques with less or no adverse side-effects need to be explored. In this regard, ayurvedic ‘ dhoopan’ technique involving slow combustion of medicinal herbs, minerals and animal products hold great promise. In this study, dhoopan of a traditionally defined ayurvedic medicinal mix, ‘Vishaghn Dhoop’ (VD) has been assessed for its anti-microbial potentials against both Gram-positive and negative pathogenic bacteria, Mycobacterium and pathogenic fungus, Candida albicans. Fume generated from slow combustion of VD was subjected to physico-chemical characterization and was assessed for anti-microbial effects. VD fume contained particles of 354 ± 84 nm size, laden with anti-microbial metabolites. On agar plates, VD fumigation reduced bacterial growth by 13 - 38%. Liquid culture aeration with VD fume inhibited bacterial growth by 50 - 85%, and fungal growth by 80%. In real life settings (in vivo), un-sanitized rooms fumigated with VD fumes for 30 min reduced the environmental microbial loads by 10 folds. In addition, the safety of VD fumigation was evaluated through in vitro cytotoxicity assay on human lung epithelial (A549) cells. Cells exposed to media-collected VD fumes for 24 h exhibited normal cyto-safety profile. Collectively, these observations provide scientific evidence in support of a traditional technique of disinfection, which can be fine-tuned to have implications in clinical, healthcare and food industry where, disinfection is a prime requirement.