, Інститут рибного господарства НААН, м. Київ Мета. Вивчення специфіки генетичної структури, рівня внутрішньо-та міжпопуляційної генетичної мінливості стад українського лускатого коропа різних господарств України за використання ДНК-маркерів (ISSR-PCR). Методика. Для дослідження специфіки генетичної структури на підставі ПЛР використовували ІSSR-PCR-методику з відповідно підібраними праймерами. Результати. На підставі виконаних досліджень за використання трьох мікросателітних локусів ДНК-(AGC) 6 G, (AСC) 6 G, (AGC) 6 С-проведено аналіз генетичної структури українських лускатих коропів. Сумарно за всіма праймерами виявлено 55 ампліконів. При використанні праймеру (AGC) 6 G виявлено 15, (AСC) 6 G-17, (AGC) 6 С-23 амплікони. Молекулярна маса на електрофореграмах була максимальною при використанні праймеру (AСC) 6 G (500 п.н.-3500 п.н. у нивківського коропа). Виявлені специфічні особливості між досліджуваними популяціями лускатих коропів залежать від їх генетичного походження. Варіацій виявлених ампліконів достатньо, щоб відокремлювати особин, або, якщо робота проводиться з групою плідників, підбирати батьківські пари для підвищення генетичного розмаїття. Наукова новизна. За використання ISSR-маркерів встановлено особливості генетичної структури, рівень генетичної мінливості українських лускатих коропів. Вперше отримано нові дані про специфіку генетичної структури за використання методів ПЛР, які сприяють виявленню специфіки механізмів підтримки відносної стабільності генофонду українського лускатого коропа і дозволяють контролювати та зберігати специфічність його генетичної структури. Практична значимість. Запропоновано метод генетичного контролю стад, який дає можливість за використання вказаних праймерів провести аналіз генетичної структури племінних стад і реалізувати генетичну інформацію на різних стадіях селекційного процесу.

ПОСТАНОВКА ПРОБЛЕМИ ТА АНАЛІЗ ОСТАННІХ ДОСЛІДЖЕНЬ І ПУБЛІКАЦІЙВажлива роль у вирішенні проблеми раціонального використання природних ресурсів внутрішніх водойм відводиться рослиноїдним рибам. Для акліматизації вони були обрані за принципом раціонального використання природної кормової бази, яка не повністю використовувалась рибами водойм Європейської частини колишнього СРСР. Природним ареалом рослиноїдних риб є рівнинні ріки Східної Азії -від р. Амур на півночі до Південного Китаю. Нині вони поширені

The rational use of valuable fish species from aquaculture is difficult to implement without knowledge of the state of the genetic structure of local stocks. Different types of DNA markers can be used to achieve the goals of selection and breeding work. The genetic structure of a local stock of rainbow trout (Oncorhynchus mykiss Walbaum, 1792) (Salmoniformes, Salmonidae) farmed in Ukraine was studied using DNA-markers: microsatellite (SSR-markers – simple-sequence repeats-markers) and intermicrosatellite (ISSR – inter-simple sequence repeat). Five fragments of trinucleotide microsatellite motifs with a single anchor nucleotide at the 3'-end were used as a primer for analysis by the ISSR-PCR method. Totally, 85 amplicons were obtained across the five loci, of which 92.9% were polymorphic. The total number of alleles ranged from 10 (marker (ACC)₆G) to 23 (marker (AGC)₆G). The following monomorphic amplicons were determined for the studied local stock of rainbow trout: according to marker (CTC)₆C – 770 and 520 bp bands, for the marker (GAG)₆C – 345, 295 and 260 bp, and for the marker (AGC)₆C – 350 bp. The average number of polymorphic bands per locus was 15.8. The selected ISSR primers had a level of polymorphic information content above the average. The most effective markers for molecular-genetic analysis of rainbow trout were (AGC)₆G and (AGC)₆C according to the percentage of polymorphic bands, marker index, effective multiplex ratio and resolving power. The selected ISSR loci allow the genetic structure of the studied local stock to be characterized using the total and the effective number of alleles per locus (Na and Ne were 1.9 and 1.4, respectively), the Shannon index (average value I was 0.4) and the unbiased expected heterozygosity (mean uHe = 0.3). Microsatellite-based analysis showed features of the genetic structure of the local stock of rainbow trout at six microsatellite loci (OMM 1032, OMM 1077, OMM 1088, Str 15, Str 60, Str 73). Allelic diversity was established and alleles with the highest frequency and most typical for the given stock were identified. The Shannon index and unbiased expected heterozygosity were determined using SSR-markers and were 1.42 and 0.79, respectively. This depicts the complexity of the population structure, a high level of genetic diversity and indicates a high level of heterozygosity of local stock. The “gene pool profile” established as a result of ISSR-PCR in the future will help to differentiate local stocks of rainbow trout in aquaculture of Ukraine. Microsatellite markers provide the ability to determine individual features of genetic variation of local populations and to conduct the management of genetic resources on fish farms.

Paddlefish (Polyodon spathula (WALBAUM, 1792)) is reared in aquaculture on Ukrainian farms as an alternative source of gourmet meat and caviar to compensate for the lack of this valuable sturgeon product, which was caused by a drastic reduction of sturgeon stocks in Ukraine's natural waters. However these stocks require analysis of the genetic structure of breeding material cultivated in the aquaculture of Ukraine for further control and maintainance of their polymorphism and heterogeneity. Genetic control will allow monitoring the condition of stocks, selection of the most acceptable source of breeding stock and optimizing the breeding of this object. The aim of the research was to determine the level of efficiency of the ISSR-PCR method for studying polymorphism and identifying specific features of the genetic structure of the paddlefish reared at fish farms in Ukraine. Materials and methods of research. The analysis was performed using two age groups of paddlefish (younger – age-4 and older – age-13–15), which were reared at the state institution "Production and Experimental Dnieper Sturgeon Breeding Plant named after Academician S.T. Artushchyk". Molecular genetic analysis was performed using three ISSR markers (B – (GAG)₆C; C – (AGC)₆G and E – (AGC)₆C) followed by separation of amplification products by agarose gel electrophoresis. The informativeness of ISSR primers was determined by such indicators as the effective multiplex ratio (EMR), marker index (MI), resolution (Rp), calculated according to the methods for dominant markers. Research results. The specifics of the genetic structure of the paddlefish of the Kherson stock were described using ISSR markers. The study showed that a total of 32 amplicons were obtained for the three primers, of which 87.5% were polymorphic. The largest number of amplicons was obtained using primer E – (AGC)₆C – 14 specimens, while primers B – (GAG)₆C and C – (AGC)₆G were observed 9 bands. The information polymorphism index (PIC) ranged from 0.227 (marker C) to 0.314 (marker E), with an average value of 0.256. The informativeness of the selected ISSR primers was assessed during the study of the paddlefish (mean values were: PIC = 0.256; PPB = 85.2%; EMR = 8.4; MI = 2.3; Rp = 9.7). These indicators demonstrated the level of informativeness and efficiency of selected intermicrosatellite loci for the analysis of polymorphisms of the genetic structure of the paddlefish. Marker E – (AGC)₆C had the highest values of PPB, PIC, EMR, MI and Rp compared to other selected ISSR-markers, which indicated the greatest effectiveness of this primer in population genetic studies of paddlefish. Ranges of amplicons at selected intermicrosatellite loci were determined: for marker B – (GAG)₆C from 215 to 1275 bp; for markers C – (AGC)₆G and E – (AGC)₆C in the range of 125–1265 bp and 320–1470 bp, respectively. Following monomorphic amplicons for all studied specimens of both age groups were determined: for marker C – allele 125, 245 and 395 bp; for marker B – (GAG)₆C – 1085 bp. Identified monomorphic bands in the future can be used for interspecific comparisons of different species and to establish the origin of samples. Differences between the studied different age groups of paddlefish were characterized, which consisted in: 1) the specificity of the frequency distribution of alleles encountered with high frequency (over 40%), 2) the absence of larger amplicons for markers B – (GAG)₆C and C – (AGC)₆G in older paddlefish groups and 3) the presence of different monomorphic alleles for marker E – (AGC)₆C (allele 380 bp in specimens of the younger and allele 320 bp in the older group). In the younger age group, a higher total number of amplified fragments (NTB) and the number of polymorphic fragments (NPB) were observed at the three loci studied compared to the older group. The biodiversity level according to the Shannon index I (0.394 ± 0.043 in the younger and 0.321 ± 0.043 in the older age group) and the level of heterozygosity (objective expected heterozygosity – 0.286 ± 0.034 and 0.231 ± 0.035, respectively) of the studied groups of paddlefish were determined. Conclusions. The obtained results demonstrated the effectiveness of the use of selected ISSR-markers to assess the polymorphism of the genetic structure of the otter, the state of biodiversity and the heterozygosity of stocks. The analysis performed by the ISSR-PCR method allowed characterizing the specifics of the genetic structure of the paddlefish and showing age differences. The obtained gene pool profile can be used as a basis for the certification of paddlefish stocks.

The purpose. To study genetic structure of breed herd of Amur carp selected in «Karpatskyi vodohrai» Ltd. (Lviv oblast) with the use of different types of markers and to determine the level of somatic mutagenesis by the microkernel test. Methods. Laboratory researches, computer statistical analysis. Results. Genetic structure of Amur carp on separate types of molecular-genetic markers (DNA-markers, genetic-biochemical systems) is studied. Species-specific features of genetic structure on probed loci are determined. The level of actual and expected heterozygosity is calculated. Increased allele and genotypic variety of genetic structure can be caused by a little heightened intensity of the carried out selection work. The detected excess of heterozygotes on separate loci testifies to presence of stabilization processes of genetic structure. Research of the level of somatic mutagenesis is also carried out at use of micronucleus test. Conclusions. At use of different types of molecular-genetic markers the information on genetic structure of a carp and its variety on genome level is gained. These results testify to the following: genome preserved resistant gene complexes, despite of significant selection effort. The applied genetically-statistical approaches can be used for monitoring genetic structure of groups of a carp of Ukraine, determination of the level of consolidation and phylogenetic links between them with their subsequent genetic certification. Results of researches of the level of somatic mutagenesis at use of micronucleus test shown that the probed group of fishes was characterized by average level of erythrocytes with micronucleus MNE (4,7±0,3‰), low level of lymphocytes with micronucleus and binucleate lymphocytes which total has made 3,5±0,3‰ that in its turn testifies to satisfactory conditions of selection.