Biotechnology of virus eradication and plant vaccination in phytobiome context

A plant’s associated biota plays an integral role in its metabolism, nutrient uptake, stress tolerance, pathogen resistance and other physiological processes. Although a virome is an integral part of the phytobiome, a major contradiction exists between the holobiont approach and the practical need to eradicate pathogens from agricultural crops. In this review, we discuss grapevine virus control, but the issue is also relevant for numerous other crops, including potato, cassava, citrus, cacao and other species. Grapevine diseases, especially viral infections, cause main crop losses. Methods have been developed to eliminate viruses and other microorganisms from plant material, but elimination of viruses from plant material does not guarantee protection from future reinfection. Elimination of viral particles in plant material could create genetic drift, leading in turn to an increase in the occurrence of pathogenic strains of viruses. A possible solution may be a combination of virus elimination and plant propagation in tissue culture with in vitro vaccination. In this context, possible strategies to control viral infections include application of plant resistance inducers, cross protection and vaccination using siRNA, dsRNA and viral replicons during plant ‘cleaning’ and in vitro propagation. The experience and knowledge accumulated in human immunization can help plant scientists to develop and employ new methods of protection, leading to more sustainable and healthier crop production.     

Proof of transovarial transmission of Lymantria dispar nucleopolyhedrovirus (fam. Baculoviridae) with the RAPD-PCR method

Transovarial transmission of Lymantria dispar Nucleopolyhedrovirus is shown with the RAPD-PCR method. Genetic markers of virus DNA found with the primer OPA-08 are recommended for diagnostics of Lymantria dispar nucleopolyhedrosis, in particular at the egg stage. Genetic markers of virus DNA found with the use of OPA-08 primer are helpful for estimation of genetic diversity of virus, level of the virus presence among the eggs, and for a decision about forestry protective measures.     

Prediction of the three-dimensional structure of aflatoxin of Aspergillus flavus by homology modelling

Aflatoxins are polyketide-derived secondary metabolites produced by Aspergillus spp. The toxic effects of aflatoxins have adverse consequences for human health and agricultural economics. The aflR gene, a regulatory gene for aflatoxin biosynthesis, encodes a protein containing a zinc-finger DNA-binding motif. AFLR-Protein three-dimensional model was generated using Robetta server. The modeled AFLR-Protein was further optimization and validation using Rampage. In the simulations, we monitored the backbone atoms and the C-α-helix of the modeled protein. The low RMSD and the simulation time indicate that, as expected, the 3D structural model of AFLR-protein represents a stable folding conformation. This study paves the way for generating computer molecular models for proteins whose crystal structures are not available and which would aid in detailed molecular mechanism of inhibition of aflatoxin.     

Genetical distances between nominative and paratypical forms of Colias crocea Fourc. and C. erate Esp. (lepidoptera, Pieridae) according to RAPD-PCR analysis

Genetic polymorphism of the two Palaearctic sulphur butterflies Colias crocea Fourc. and Colias erate Esp. was examined in serial samples using RAPD-PCR analysis of total cellular DNA. The primers OPA-01, OPA-03, OPA-04, OPA-08 were used. Taxon-specific molecular markers have been found for each species (markers OPA-08(250), OPA-08(380) were associated with rounded valve margin; marker OPA-04(400) was associated with angled valve margin). The genetic diversity of C. erate is considerably less than that of C. crocea. Genetic evidence for interspecific hybridization between C. crocea and C. erate is presented. Interspecific hybrids in question (paratypical forms such as f. chlorodona and f. pseudochrysodona) are situated at intermediate position with regard to the parental species on dendrograms (UPGMA) and maximal spanning trees (MST). Completely additional RAPD-spectra were not obtained though taxonomic status of these paratypical forms stay still vague. Only f. eratoides has DNA markers that are absent in spectra of other specimens tested. Probably it is a cryptic species unrevealed by common morphological and biochemical methods and its investigation by the methods of molecular genetics (sequence of cloned DNA) is very desirable.     

Mapping the amino acid properties of constituent nucleoporins onto the yeast nuclear pore complex

Visualization of molecular structures aids in the understanding of structural and functional roles of biological macromolecules. Macromolecular transport between the cell nucleus and cytoplasm is facilitated by the nuclear pore complex (NPC). The ring structure of the NPC is large and contains several distinct proteins (nucleoporins) which function as a selective gate for the passage of certain molecules into and out of the nucleus. In this note we demonstrate the utility of a python code that allows direct mapping of the physiochemical properties of the constituent nucleoporins on the scaffold of the yeast NPC׳s cytoplasmic view. We expect this tool to be useful for researchers to visualize the NPC based on their physiochemical properties and how it alters when specific mutations are introduced in one or more of the nucleoporins. The code developed using Python is available freely from the authors.     

The treatment of isolated genera

Problems presented by genera, or small groups of genera, which have been given family rank are reviewed, and the genera are divided into a number of categories according to the closeness of their affinity to other genera or families. Satellite genera that stand in close relation to families should be united with them. Binary families, that have been divided into two (or more) related families, should be re–united. Families connected by linking genera, should, logically, be united but practical considerations usually prevent this. Clusters of diverse but more or less distantly related genera present unusual problems, being treated either as several, often monogeneric families or as a loosely structured family. Truly isolated genera must be given family and often ordinal rank.     

Effect of ethacrynic acid on the course of cytotoxic glomerulonephritis]

Ethacrynic acid (10 mg/kg subcutaneously 15 days) significantly facilitated the experimental glomerulonephritis in rats. The efficacy of diuretic was confirmed by the improvement of the functional characteristics (the degree of proteinuria, the plasma contents of protein and urea, the creatinine excretion), and by the results of the histological investigation, especially at the second phase of the pathological process.