Comparative Genomics Reveals Insights into the Divergent Evolution of Astigmatic Mites and Household Pest Adaptations

Highly diversified astigmatic mites comprise many medically important human household pests such as house dust mites causing roughly 1–2% of the allergic diseases globally; however, their evolutionary origin, diverse lifestyles including reversible parasitism and quick adaptation to rather new human household environments have not been illustrated at genomic level, which hamper the allergy prevention and our exploration of these household pests. Using six high-quality assembled and annotated genomes, this comparative genomics study not only refuted the monophyly of mites and ticks, but also thoroughly explored the divergence of Acariformes and the divergent evolution of astigmatic mites. In the monophyletic Acariformes, Prostigmata known as notorious plant pests first evolved, then rapidly evolving Astigmata diverged from soil oribatid mites. Within astigmatic mites, a wide range of gene families rapidly expanded via tandem gene duplications, including ionotropic glutamate receptors, triacylglycerol lipases, serine proteases and UDP glucuronosyltransferases (UGTs), which enriched their capacities of adapting to rapidly changing household environments. The gene diversification after tandem duplications provided plenty of genetic resources for their adaptations of sensing environmental signals, digestion, and detoxification. Whilst many gene decay events only occurred in the skin-burrowing parasitic mite Sarcoptes scabiei. Throughout the evolution of Acariformes, massive horizontal gene transfer events occurred in gene families such as UGTs and several important fungal cell wall lytic enzymes, which enable the detoxification and associated digestive functions and provide perfect drug targets for pest control. Our comparative study sheds light on the rapid divergent evolution of astigmatic mites from the divergence of Acariformes to their diversification and provides novel insights into the genetic adaptations and even control of human household pests.

Biological Control of Fusarium culmorum, Fusarium graminearum and Fusarium poae by Antagonistic Yeasts

The genus Fusarium is considered to be one of the most pathogenic, phytotoxic and toxin-producing group of microorganisms in the world. Plants infected by these fungi are characterized by a reduced consumer and commercial value, mainly due to the contamination of crops with mycotoxins. Therefore, effective methods of reducing fungi of the genus Fusarium must be implemented already in the field before harvesting, especially with alternative methods to pesticides such as biocontrol. In this study we identified yeasts that inhibit the growth of the pathogenic fungi Fusarium culmorum, F. graminearum and F. poae. Tested yeasts came from different culture collections, or were obtained from organic and conventional cereals. The greater number of yeast isolates from organic cereals showed antagonistic activity against fungi of the genus Fusarium compared to isolates from the conventional cultivation system. Cryptococcus carnescens (E22) isolated from organic wheat was the only isolate that limited the mycelial growth of all three tested fungi and was the best antagonist against F. poae. Selected yeasts showed various mechanisms of action against fungi, including competition for nutrients and space, production of volatile metabolites, reduction of spore germination, production of siderophores or production of extracellular lytic enzymes: chitinase and β-1,3-glucanase. Of all the investigated mechanisms of yeast antagonism against Fusarium, competition for nutrients and the ability to inhibit spore germination prevailed.

THE COMPOST, A SOURCE OF PLANT BENEFICIAL BACTERIA WITH BIOCONTROL POTENTIAL

Recycling the sewage sludge from treatment plants is a common activity. The resulting compost is usually rich in plant nutrients and beneficial microorganisms. However, compost properties greatly differ depending on the nature of the fermented biomass and fermentation processes. The aim of this study was to analyze the microbial load of three different composts, in order to detect new bacterial strains with plant protection properties. Isolated bacteria were microbiologically characterized and evaluated for their potential to reduce soil-borne phytopathogenic fungi. Results showed a microbial load of approximately 106 CFU/g of compost. In the analyzed samples it was revealed that as bacterial load increases, the fungal amount decreases. Analyzing some newly isolated bacteria obtained from these composts, a good biocontrol potential against soil-borne pathogenic fungi was revealed. Some of the isolated bacterial strains revealed antifungal activity against Rhizoctonia solani and Sclerotinia sclerotiorum. These bacteria showed good colonization capacity and lytic enzymes production, correlated to antimicrobial activity. These compost-originated bacteria reveal high potential in pathogens inhibition. Therefore, the analyzed composts are recommended not only as soil fertility improvers, but also as potential suppressors of soil-borne pathogens. Results revealed these composts as source of plant beneficial bacteria with biological control potential.

Impact of Pseudomonas spp. on Plant Growth, Lytic Enzymes and Secondary Metabolites Production

Seven strains of Pseudomonas spp. were isolated from the south Gujarat region of India. Antifungal and bacterial activities of bacterial strains were evaluated against important plant pathogens in vitro, among them, PaRS was found most effective. The indole acetic acid production was recorded in all isolated Pseudomonas spp. Seed treatment with PaRS at 6 g/kg was significantly superior over other treatments for plant growth promotion (germination, shoot and root length, shoot and root weight, vigor index, and both shoot and root colonization) under standard roll towel method and pot conditions. The maximum siderophore production was observed in PaRS and medium production in PfRB, PaNS, and PfNC. PaNS and PaRS strains recorded strong HCN production but moderate production recorded in PaWP, PaWS, and PfNC. The maximum phosphate solubalization zone (22 mm) was found in PaRS. PaRS recorded maximum chitinase, ß-1,3-glucanase activity, hydrogen cyanide, and salicylic acid production as compared to other strains.

Microvolume DNA extraction methods for microscale amplicon and metagenomic studies

Investigating the composition and metabolic capacity of aquatic microbial assemblages usually requires the filtration of multi-litre samples, which are up to 1 million-fold larger than the microenvironments within which microbes are predicted to be spatially organised. To determine if community profiles can be reliably generated from microlitre volumes, we sampled seawater at a coastal and an oceanic site, filtered and homogenised them, and extracted DNA from bulk samples (2 L) and microvolumes (100, 10 and 1 μL) using two new approaches. These microvolume DNA extraction methods involve either physical or chemical lysis (through pH/thermal shock and lytic enzymes/surfactants, respectively), directly followed by the capture of DNA on magnetic beads. Downstream analysis of extracted DNA using both amplicon sequencing and metagenomics, revealed strong correlation with standard large volume approaches, demonstrating the fidelity of taxonomic and functional profiles of microbial communities in as little as 1 μL of seawater. This volume is six orders of magnitude smaller than most standard operating procedures for marine metagenomics, which will allow precise sampling of the heterogenous landscape that microbes inhabit.

Exploring biofertilizer potential of plant growth-promoting rhizobacteria Bacillus clausii strain B8 (MT305787) on Brassica napus and Medicago sativa

Plant Growth Promoting Rhizobacteria (PGPR) are soil bacteria that can stimulate plant growth by supplying substances that are usually in limited quantities in the soil especially phosphorous, nitrogen and growth hormone such as indole acetic acid (AIA). These bacteria can also slow the growth of plant pathogens through the production of several antimicrobial metabolites. To investigate the role of rhizobacteria as a biostimulant agent a novel bacterium B8, isolated from the rhizospheric soil of medlar (Mespilus germanica L.- Family Rosaceae), was evaluated on Brassica napus and Medicago sativa. In addition to the classical methods of identification (physiological and biochemical tests), B8 was identified by 16S rRNA gene sequencing as Bacillus clausii. The ability of the strain to produce lytic enzymes such as cellulases, chitinases, pectinases, and phospholipases was studied. Furthermore, the strain B8 was tested for the capability to produce plant growth metabolites like phosphatases and phytases in order to solubilize inorganic phosphate and production of siderophores, cyanhydric acid (HCN) and indole-3-acetic acid. The strain was able to produce lytic enzymes, with an intense production of siderophores and to solubilize inorganic phosphate. Result of in vivo experiments indicated that the application of B8 at 107 CFU/mL, improved markedly the germination rate of rapeseed, whereas alfalfa seeds treated with the same strain showed a lower germination rate than the controls. The vegetative growth parameters; Roots length, lateral roots number, stem length, number of leaves, diameters of stems and plant weight were significantly improved. We also noted capacity of bacteria to colonize root systems of both plants B. napus and M. sativa in one week of inoculation. The overall results of this study showed that B clausii B8 has a great potential to be commercialized as a biostimulant agent and provide promising new option for sustainable agriculture.

Discovering the Potentials of Four Phage Endolysins to Combat Gram-Negative Infections

Endolysin-based therapeutics are promising antibacterial agents and can successfully supplement the existing antibacterial drugs array. It is specifically important in the case of Gram-negative pathogens, e.g., ESKAPE group bacteria, which includes Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species, and are highly inclined to gain multiple antibiotic resistance. Despite numerous works devoted to the screening of new lytic enzymes and investigations of their biochemical properties, there are significant breaches in some aspects of their operating characteristics, including safety issues of endolysin use. Here, we provide a comprehensive study of the antimicrobial efficacy aspects of four Gram-negative bacteria-targeting endolysins LysAm24, LysAp22, LysECD7, and LysSi3, their in vitro and in vivo activity, and their biological safety. These endolysins possess a wide spectrum of action, are active against planktonic bacteria and bacterial biofilms, and are effective in wound and burn skin infection animal models. In terms of safety, these enzymes do not contribute to the development of short-term resistance, are not cytotoxic, and do not significantly affect the normal intestinal microflora in vivo. Our results provide a confident base for the development of effective and safe candidate dosage forms for the treatment of local and systemic infections caused by Gram-negative bacterial species.

Identification and Functional Analysis of Cytokine-Like Protein CLEC-47 in Caenorhabditis elegans

A variety of effector proteins contribute to host defense in the nematode Caenorhabditis elegans . However, little is known about the exact function of these infection-related effectors beyond lytic enzymes and antimicrobial peptides and proteins.

Lytic Enzymes of Aspergillus piperis as a Tool for Attacking Some Phytopathogenic Fungi In vitro with Special Reference to its Cytotoxicity

The antagonistic activity of Aspergillus piperis against Fusarium oxysporum f. sp. fabae (FOF) and Sclerotinia sclerotiorum were examined and showed multiple signs of hyphal interactions. Microscopic examination of contact regions among A. piperis and each pathogen revealed distinct enzymatic lysis of pathogenic hyphal cell walls. Therefore, it is important to estimate the lytic enzyme activity of A. piperis. Extracellular lytic enzymes are important offensive forces for A. piperis as a biological control agent. Chitinase, phospholipase, and protease recorded relatively high activity from a culture age of 10 days (82.3, 42.4, and 6.2 U/ml, respectively). Enzymatic persistence was measured at room temperature, recording relatively long periods, saving 54%, 46%, and 21% of their activity, respectively. The cytotoxicity of the crude culture filtrate of A. piperis was examined in MCF7 and WI38 human cell lines. The cell viability (IC50) value of the fungal filtrate was estimated after 24 h and 48 h. The results revealed that IC50 values against the MCF7 cell line were inoperative after 24 h and were recorded 80 μg/ml after 48 h. In contrast, IC50 values against the WI38 cell line were 85.69 and 69.8 μg/ml after 24 and 48 h, respectively.