Accelerating the understanding of Aspergillus terreus: Epidemiology, physiology, immunology and advances.

Aspergillus species encompass a variety of infections, ranging from invasive aspergillosis to allergic conditions, contingent upon the immune status of the host. In this spectrum, Aspergillus terreus stands out due to its emergence as a notable pathogen and its intrinsic resistance to amphotericin-B. The significance of Aspergillus-associated infections has witnessed a marked increase in the past few decades, particularly with the increasing number of immunocompromised individuals. The exploration of epidemiology, morphological transitions, immunopathology, and novel treatment approaches such as new antifungal drugs (PC945, olorofim) and combinational therapy using antifungal drugs and phytochemicals (Phytochemicals: quercetin, shikonin, artemisinin), also using immunotherapies to modulate immune response has resulted in better outcomes. Furthermore, in the context COVID-19 era and its aftermath, fungal infections have emerged as a substantial challenge for both immunocompromised and immunocompetent individuals. This is attributed to the use of immune-suppressing therapies during COVID-19 infections and the increase in transplant cases. Consequently, this review aims to provide an updated overview encompassing the epidemiology, germination events, immunopathology, and novel drug treatment strategies against Aspergillus terreus-associated infections.

Two Novel Plant-Growth-Promoting Lelliottia amnigena Isolates from Euphorbia prostrata Aiton Enhance the Overall Productivity of Wheat and Tomato.

Euphorbiaceae is a highly diverse family of plants ranging from trees to ground-dwelling minute plants. Many of these have multi-faceted attributes like ornamental, medicinal, industrial, and food-relevant values. In addition, they have been regarded as keystone resources for investigating plant-specific resilience mechanisms that grant them the dexterity to withstand harsh climates. In the present study, we isolated two co-culturable bacterial endophytes, EP1-AS and EP1-BM, from the stem internodal segments of the prostate spurge, Euphorbia prostrata, a plant member of the succulent family Euphorbiaceae. We characterized them using morphological, biochemical, and molecular techniques which revealed them as novel strains of Enterobacteriaceae, Lelliotia amnigena. Both the isolates significantly were qualified during the assaying of their plant growth promotion potentials. BM formed fast-growing swarms while AS showed growth as rounded colonies over nutrient agar. We validated the PGP effects of AS and BM isolates through in vitro and ex vitro seed-priming treatments with wheat and tomato, both of which resulted in significantly enhanced seed germination and morphometric and physiological plant growth profiles. In extended field trials, both AS and BM could remarkably also exhibit productive yields in wheat grain and tomato fruit harvests. This is probably the first-ever study in the context of PGPB endophytes in Euphorbia prostrata. We discuss our results in the context of promising agribiotechnology translations of the endophyte community associated with the otherwise neglected ground-dwelling spurges of Euphorbiaceae.

Microbiome based approaches for the degradation of polycyclic aromatic hydrocarbons (PAHs): A current perception.

Globally, polycyclic aromatic hydrocarbons (PAHs) pollution is primarily driven by their release into the air through various combustion processes, including burning fossil fuels such as coal, oil, and gas in motor vehicles, power plants, and industries, as well as burning organic matter like wood, tobacco, and food in fireplaces, cigarettes, and grills. Apart from anthropogenic pollution sources, PAHs also occur naturally in crude oil, and their potential release during oil extraction, refining processes, and combustion further contributes to contamination and pollution concerns. PAHs are resistant and persistent in the environment because of their inherent features, viz., heterocyclic aromatic ring configurations, hydrophobicity, and thermostability. A wide range of microorganisms have been found to be effective degraders of these recalcitrant contaminants. The presence of hydrocarbons as a result of numerous anthropogenic activities is one of the primary environmental concerns. PAHs are found in soil, water, and the air, making them ubiquitous in nature. The presence of PAHs in the environment creates a problem, as their presence has a detrimental effect on humans and animals. For a variety of life forms, PAH pollutants are reported to be toxic, carcinogenic, mutation-inducing, teratogenic, and immune toxicogenics. Degradation of PAHs via biological activity is an extensively used approach in which diverse microorganisms (fungal, algal, clitellate, and protozoan) and plant species and their derived composites are utilized as biocatalysts and biosurfactants. Some microbes have the ability to transform and degrade these PAHs, allowing them to be removed from the environment. The goal of this review is to provide a critical overview of the existing understanding of PAH biodegradation. It also examines current advances in diverse methodologies for PAH degradation in order to shed light on fundamental challenges and future potential.

Establishment of Agrobacterium tumefaciens - mediated genetic transformation of apple pathogen Marssonina coronaria using marker genes under the control of CaMV 35S promoter.

Premature leaf fall of apple caused by Marssonina coronaria is economically very important apple disease and all the commercially available apple cultivars are susceptible to this disease. The non-availability of an efficient transformation system for this fungus hinders the functional genomics research. Herein, we report for the first time, the successful Agrobacterium-mediated transformation in apple leaf blotch fungus M. coronaria by transferring T-DNA harbouring the genes for hygromycin phosphotransferase (hpt), ß-glucuronidase (uidA) and green fluorescent protein (gfp) under the control of CaMV 35S promoter. The key factors that affect the transformation efficiency including type of recipient fungal material, acetosyringone concentration, the conditions for co-cultivation, Agrobacterium concentration, Agrobacterium strains and membrane types as support were investigated. The present results have recommended that 250 µM concentration of acetosyringone, 24 °C temperature and 48 h time, 0.5 OD600 of A. tumefaciens, EHA105 Agrobacterium strain and Whatman filter paper were the optimal co-cultivation conditions for the transformation of M. coronaria by using fragmented mycelia suspension and mycelial plugs. We observed that conidia were tedious to transform as compared to the fragmented mycelia and mycelial plugs of this slow growing fungus. These optimized parameters yielded 54 and 70 average transformants per 60 mycelial plugs and 104 fragmented mycelia, respectively. Fungal transformants were analysed for T-DNA integration, gus gene expression and gfp gene expression. Strong gus histochemical staining and green fluorescence expression indicated that the CaMV 35S promoter can drive gene expression in M. croronaria. Some mutants showed difference in the morphology of the colony as compared to the wild type control. This report will be very useful to inspect molecular basis of apple-M. coronaria interactions by deciphering the functional roles of various genes in this pathogenic fungus.

Comparison of various RNA extraction methods, cDNA preparation and isolation of calmodulin gene from a highly melanized isolate of apple leaf blotch fungus Marssonina coronaria.

Marssonina coronaria causes apple blotch disease resulting in severe premature defoliation, and is distributed in many leading apple-growing areas in the world. Effective, reliable and high-quality RNA extraction is an indispensable procedure in any molecular biology study. No method currently exists for RNA extraction from M. coronaria that produces a high quantity of melanin-free RNA. Therefore, we evaluated eight RNA extraction methods including manual and commercial kits, to yield a sufficient quantity of high-quality and melanin-free RNA. Manual methods used here resulted in low quality and black colored RNA pellets showing the presence of melanin, despite all the modifications employed to original procedures. However, these methods when coupled with clean up resulted in melanin-free RNA. On the other hand, all commercial kits used were able to yield high-quality melanin-free RNA having variable yields. TRIzol™ Reagent + RNA Clean & Concentrator™-5 and Ambion-PureLink® RNA Mini Kit were found to be the best methods as the RNA extracted with these methods from 15 day old fungal culture grown on solid medium were free of melanin with good yield. RNA extracted by this improved methodology was applied for RT-PCR, subsequent PCR amplification, and isolation of calmodulin gene sequences from M. coronaria and infected apple leaf pieces. These methods are more time effective than traditional methods and take only an hour to complete. To our knowledge, this is the first report on the method of isolation of high-quality RNA for cDNA synthesis as well as isolation of the calmodulin gene sequence from this fungus.

Application of response surface methodology in optimization of lactic acid fermentation of radish: effect of addition of salt, additives and growth stimulators.

Lactic acid fermentation of radish was conducted using various additive and growth stimulators such as salt (2 %-3 %), lactose, MgSO4 + MnSO4 and Mustard (1 %, 1.5 % and 2 %) to optimize the process. Response surface methodology (Design expert, Trial version 8.0.5.2) was applied to the experimental data for the optimization of process variables in lactic acid fermentation of radish. Out of various treatments studied, only the treatments having ground mustard had an appreciable effect on lactic acid fermentation. Both linear and quadratic terms of the variables studied had a significant effect on the responses studied. The interactions between the variables were found to contribute to the response at a significant level. The best results were obtained in the treatment with 2.5 % salt, 1.5 % lactose, 1.5 % (MgSO4 + MnSO4) and 1.5 % mustard. These optimized concentrations increased titrable acidity and LAB count, but lowered pH. The second-order polynomial regression model determined that the highest titrable acidity (1.69), lowest pH (2.49) and maximum LAB count (10 × 10(8) cfu/ml) would be obtained at these concentrations of additives. Among 30 runs conducted, run 2 has got the optimum concentration of salt- 2.5 %, lactose- 1.5 %, MgSO4 + MnSO4- 1.5 % and mustard- 1.5 % for lactic acid fermentation of radish. The values for different additives and growth stimulators optimized in this study could successfully be employed for the lactic acid fermentation of radish as a postharvest reduction tool and for product development.