Volume 2 (2018): Issue 2 (April 2018)

The paper presents main results and recommendations of the recent European Workshop dedicated to the evaluation of current state and prospects for Biotechnology with the attention to Central and European countries. The contribution of Biotechnology to the RIS3 strategy is also briefly presented. It is believed that there is still insufficient integration of research and innovation, especially in Central and Eastern European countries. Another problem is the weak interest and lack of mechanisms for the entry of potential investors into biotechnology capital–intensive areas. Series of proposals and recommendations coming from the Workshop on how to increase the regional cooperation in the field of Biotechnology is briefly described together with the potential role of international institutions (JRC-EC, CEI, ICGEB, EBTNA) in such cooperation.

Next Generation Sequencing (NGS) or deep sequencing technology enables parallel reading of multiple individual DNA fragments, thereby enabling the identification of millions of base pairs in several hours. Recent research has clearly shown that machine learning technologies can efficiently analyse large sets of genomic data and help to identify novel gene functions and regulation regions. A deep artificial neural network consists of a group of artificial neurons that mimic the properties of living neurons. These mathematical models, termed Artificial Neural Networks (ANN), can be used to solve artificial intelligence engineering problems in several different technological fields (e.g., biology, genomics, proteomics, and metabolomics). In practical terms, neural networks are non-linear statistical structures that are organized as modelling tools and are used to simulate complex genomic relationships between inputs and outputs. To date, Convolutional Neural Networks (CNNs) and Recurrent Neural Networks (RNN) have been demonstrated to be the best tools for improving performance in problem solving tasks within the genomic field.

Polyhydroxyalkanoates (PHA), the only group of “bioplastics” sensu stricto, are accumulated by various prokaryotes as intracellular “carbonosomes”. When exposed to exogenous stress or starvation, presence of these microbial polyoxoesters of hydroxyalkanoates assists microbes to survive.

“Bioplastics” such as PHA must be competitive with petrochemically manufactured plastics both in terms of material quality and manufacturing economics. Cost-effectiveness calculations clearly show that PHA production costs, in addition to bioreactor equipment and downstream technology, are mainly due to raw material costs. The reason for this is PHA production on an industrial scale currently relying on expensive, nutritionally relevant “1^st-generation feedstocks”, such as like glucose, starch or edible oils. As a way out, carbon-rich industrial waste streams (“2^nd-generation feedstocks”) can be used that are not in competition with the supply of food; this strategy not only reduces PHA production costs, but can also make a significant contribution to safeguarding food supplies in various disadvantaged parts of the world. This approach increases the economics of PHA production, improves the sustainability of the entire lifecycle of these materials, and makes them unassailable from an ethical perspective.

In this context, our EU-funded projects ANIMPOL and WHEYPOL, carried out by collaborative consortia of academic and industrial partners, successfully developed PHA production processes, which resort to waste streams amply available in Europe. As real 2^nd-generation feedstocks”, waste lipids and crude glycerol from animal-processing and biodiesel industry, and surplus whey from dairy and cheese making industry were used in these processes. Cost estimations made by our project partners determine PHA production prices below 3 € (WHEYPOL) and even less than 2 € (ANIMPOL), respectively, per kg; these values already reach the benchmark of economic feasibility.

The presented studies clearly show that the use of selected high-carbon waste streams of (agro)industrial origin contributes significantly to the cost-effectiveness and sustainability of PHA biopolyester production on an industrial scale.

Drought and soil salinity are at present the major factors responsible for the global reduction of crop yields, and the problem will become more severe in the coming decades because of climate change effects. The most promising strategy to achieve the increased agricultural production that will be required to meet food demands worldwide will be based on the enhancement of crop stress tolerance, by both, traditional breeding and genetic engineering. This, in turn, requires a deep understanding of the mechanisms of tolerance which, although based on a conserved set of basic responses, vary widely among plant species. Therefore, the use of different plant models to investigate these mechanisms appears to be a sensible approach. The genus Portulaca could be a suitable model to carry out these studies, as some of its taxa have been described as tolerant to drought and/or salinity. Information on relevant mechanisms of tolerance to salt and water stress can be obtained by correlating the activation of specific defence pathways with the relative stress resistance of the investigated species. Also, species of the genus could be economically attractive as ‘new’ crops for ‘saline’ and ‘arid’, sustainable agriculture, as medicinal plants, highly nutritious vegetable crops and ornamentals.

Red yeast Cystofilobasidium capitatum autofluorescence was studied by means of confocal laser scanning microscopy (CLSM) to reveal distribution of carotenoids inside the cells. Yeasts were cultivated in 2L fermentor on glucose medium at permanent light exposure and aeration. Samples were collected at different times for CLSM, gravimetric determination of biomass and HPLC determination of pigments. To compare FLIM (Fluorescence Lifetime Imaging Microscopy) images and coupled data (obtained by CLSM) with model systems, FLIM analysis was performed on micelles of SDS:ergosterol and SDS:coenzyme Q with different content of ergosterol and coenzyme Q, respectively, and with constant addition of beta-carotene. Liposomes lecithin:ergosterol:beta-carotene were investigated too. Two different intracellular forms of carotenoids were observed during most of cultivations, with third form appeared at the beginning of stationary phase. Observed behavior is probably due to formation of some kind of carotenoid protective system in membranes of different compartments of yeast cell, especially cytoplasmic membrane.

Agmatine has been functionally characterized as an important hormone and co-neurotransmitter in mammals. Given its ability in binding Imidazoline sites, a regolatory site of monoaminoxydase, it has been suggested to be involved in many neurological aspects. However, its inhibitory effect on this enzyme still remains an unanswered question. This present study is aimed to asses whether different experimental conditions could affect the agmatine action on monoaminoxydase activity. We demonstrate that the monoaminoxydase inhibition by agmatine is obtained under alkaline conditions and a long time of incubation. No inhibitiory action was found for shorter times of reaction at elevated pH, or at neutral condition and long time of incubation. No inhibition was also detected by substituting the monoamineoxydase substrate tyramine with kynuramine, however, while in these conditions a remarkable inhibition was shown by two aminoxydase inhibitors tranylcypromine and idazoxan. Herein, we discuss a mechanism model and the functional consequences of agmatine action on monoaminoxydase.

To develop tomato hybrids and varieties with a high fruit quality, we gradually solved the following tasks: development of DNA-marking methods for long shelf life genes; the genes modifying the biosynthesis of carotenoids and their composition; testing of the elaborated methods on the developed breeding material; the selection of samples with different allelic composition of fruit quality genes; the development of F₁ hybrids using the method of successive crosses and their study; the selection of tomato forms by DNA-typing methods with target genes in F₂ populations to develop valuable breeding samples; the study of carotenoids’ accumulation peculiarities and their inheritance.

We used DNA-identification methods for fruit quality genes: nor, rin, nor^A (long shelf life), B, og^c, hp2^dg, gf-3 (carotenoid content). The tomato hybrids, combining two pigment content genes and one long shelf life gene and the model forms with different combinations of fruit quality genes (B/rin/gf-3, B/rin/hp2^dg; B /nor/gf-3, B /nor/hp2^dg; оg^c /rin/gf-3, оg^c /rin/hp2^dg; оg^c /nor/gf-3, оg^c /nor/hp2^dg) in a homozygous state were developed.

Use of the developed accessions with carotenoid content genes (og^c/hp2^dg, B/hp2^dg) as maternal forms and the accessions with complex fruit quality genes (og^c/hp2^dg/nor, og^c/hp2^dg/rin, B/hp2^dg/nor, B/hp2^dg/rin, og^c/gf-3/nor, og^c/gf-3/rin, B/gf-3/rin, B/gf-3/nor) as paternal forms for hybridization contributes to high accumulation of carotenoids and a lond period of fruit storability.