Eurasian Journal of Applied Biotechnology

SSR-BASED ASSESSMENT OF GENETIC DIVERSITY IN TOMATO CULTIVARS AND LINES GROWN IN KAZAKHSTAN

Yu. Genievskaya, S. Jantasov, E. Nurbayeva , Ye. Turuspekov — Tue, 25 Jun 2024 00:00:00 +0500

Tomato (Solanum lycopersicum L.) is a versatile crop known for its nutritional value and health benefits, thriving in diverse climates worldwide. Nevertheless, regional variations persist in tomato yield, with Kazakhstan serving as an example of lower productivity in contrast to global averages. Closing this disparity requires comprehensive genetic studies and breeding efforts. This study is focused on the genetic diversity of 49 tomato cultivars and hybrids sourced from Kazakh Research Institute of Fruit and Vegetable Growing (Almaty), employing 10 SSR markers associated with important agronomic traits. SSR genotyping unveiled polymorphisms across 6 markers with variable allele numbers. Genetic diversity metrics highlighted significant genetic diversity within both outdoor and greenhouse tomato cultivars and lines. Bayesian clustering, Neighbor-joining (NJ) clustering, and principal coordinate analysis (PCoA) delineated genetic differentiation between outdoor and greenhouse tomatoes with small admixture, indicating distinct breeding directions for these two types. Highly polymorphic SSRs (PIC > 0.5) associated with essential fruit traits emerge as promising targets for marker-assisted selection (MAS) that can be used to enhance tomato breeding efficiency in Kazakhstan. According to 8 SSRs, 22 out of 30 outdoor accessions and 8 greenhouse tomato accessions were genetically uniform. This study offers comprehensive insights into the genetic diversity and population structure of tomato cultivars and lines in Kazakhstan, laying the foundation for informed breeding endeavors aimed at bolstering yield and resilience in tomato crops.

MONOCLONAL ANTIBODIES AGAINST THE BABESIA BOVIS RAP1 ANTIGEN: OBTAINING AND CHARACTERIZATION

K. Mukantayev , K. Tursunov, Zh. Adish , D. Kanayev, L. Tokhtarova, B. Abirbekov — Tue, 25 Jun 2024 00:00:00 +0500

Bovine babesiosis is a disease caused by parasites of the genus Babesia. One of the main Babesia genera causing disease in cattle, according to the International Office of Epizootics (OIE), is Babesia bovis. Recovered cattle become permanent latent carriers and can be a source of tick infestation and spread of infection. Despite susceptibility and specificity, diagnostic methods such as enzyme-linked immunosorbent assays (ELISA) and indirect fluorescent antibody assays (IFA) have some disadvantages. These methods require a long analysis time, the use of specialized equipment and trained specialists. At the same time, immunochromatographic tests (ICA) are widely used in veterinary practice today. ICA is characterized by a short analysis time, ease of performance and interpretation of results. The main components of the ICA test system are monoclonal antibodies. As a result of the studies, three mAbs against the recombinant Rap1 antigen of B. bovis were obtained. The resulting mAbs have high diagnostic characteristics. A conjugate of colloidal gold with mAbs is capable of detecting the Rap1 antigen of the causative agent of babesiosis in cattle.

OPTIMIZATION OF THE MODE OF DEPOSITION OF ASEPTIC CULTURES IN VITRO OF A RARE AND ENDEMIC SPECIES ALLIUM IVASCZENKOAE

D. Tagimanova, O. Raizer, G. Nagmetova , O. Khapilina — Tue, 25 Jun 2024 00:00:00 +0500

Optimization of the method of medium-term storage of the rare and endangered plant species Allium ivasczenkoae under in vitro conditions using modern approaches of biotechnology is an urgent task. Aseptic microplants Allium ivasczenkoae were used as starting material. The cultivation of microplants was carried out on nutrient media supplemented in various combinations with an increased concentration of sucrose, mannitol, chlorocholine chloride, ABA and BAP at various temperature and light conditions of deposition. The survival dynamics of A. ivasczenkoae varied depending on cultivation regimes. After 9 months of deposition, the survival of test-tube plants in mode 1 (control) was 20%, mode 2 - 60-80%, mode 3 - 30-50%. Under experimental conditions, the best results were obtained by depositing aseptic cultures for 9 months at a low positive temperature and low light intensity (mode 2) on nutrient media with the addition of CCC 120 mg/l and a sucrose concentration of 60 g/l and (B-1) and MS medium containing 0.5 mg/l BAP and 3 g/l mannitol (MSВМ). This mode of deposition made it possible to lengthen the period between transplants, as well as to maintain the viability of A. ivasczenkoae microplants from 60 to 80%. The data obtained make it possible to optimize the mode of medium-term storage of Allium ivasczenkoae under in vitro conditions for the mass production of rare species of bulbous plants in order to restore, reproduce and preserve the number of valuable gene pool and natural populations, create gene banks in vitro, reintroduce and green building.

Development of a real-time PCR test system for the identification of Francisella tularensis

Tue, 25 Jun 2024 00:00:00 +0500

Tularaemia is caused by the Gram-negative bacterium Francisella tularensis, which has three subspecies: holarctica, mediacia and tularensis. Due to the high virulence of the pathogen, the wide range of susceptible animals and the presence of numerous vectors and natural reservoirs, the development of sensitive diagnostic methods for the epidemiological surveillance of tularemia is crucial. New opportunities for understanding the epidemiological situation of tularemia are opening up with the use of new molecular analysis technologies for typing F. tularensis. The development and use of molecular methods for the diagnosis of tularemia is important in this context. The present study developed a protocol for detection of F. tularensis by real-time polymerase chain reaction (qPCR). The selection of primers and a fluorescent probe was based on the aligned sequences of ISFtu1 (multi-copy insertion element). qPCR conditions were optimised, including the determination of the optimal annealing and extension temperatures for the primers. Sensitivity was tested using successive 4-fold dilutions of DNA derived from F. tularensis subsp. mediasiatica, indicating a minimum sensitivity threshold of 15 genomic equivalents per reaction. Two subspecies of F. tularensis (subsp. mediasiatica and subsp. holarctica), 27 non-target bacterial species and 3 eukaryotic organisms were used to assess specificity.

ASSESSMENT OF BIOTECHNOLOGICAL POTENTIAL OF MICROORGANISMS CULTURES PROMISSING FOR SUPRESSING ANTIBIOTIC-RESISTANT STRAINS

Wed, 26 Jun 2024 00:00:00 +0500

One of the problems in global practical medicine is antibiotic-resistant strains. The problem is that pathogens are resistant to a number of antibacterial drugs and this makes treatment difficult, which can be long and/or ineffective. One solution is to develop drugs based on bacterial lysates. In this article, the objects of study are producers of extended spectrum β-lactamases - Enterobacteriaceae, Acinetobacter, Pseudomonas, Staphylococcus, Haemophilus, Streptococcus. We assessed the survival, antibiotic resistance and biological compatibility of 25 microbial cultures. Cultures have acceptable viability numbers reguired for industrially valuable strains: more than 10⁶ CFU/ml (10⁹– 10¹⁰ CFU/ml). Antibiotic resistance to various groups of is antibiotics was studied: carbapenems, fluoroguinolones, cephalosporins, macrolides, aminoglycosides, penicillins, chloramphenicols. The choise of antibacterial drugs is based on the freguent use of these antibiotics. The spectrum of antibiotic resistance of each strain is different, from one antibiotic (Ps. aeruginosa 13) to ten antibiotics (S. pneumoniae 5). Cultures are resistant to amoxicillin, ampicillin, benzylpenicillin, erythromycin and cephalosporins. The strains are mostly biocompatible with each other and have antibiotic resistance to a number of antibacterial drugs.