Valuable Genetic Material Research Articles

Comparison of two methods of extracting bull epididymal spermatozoa

Extraction of epididymal spermatozoa may be necessary to avoid losing valuable genetic material, for example, from individuals of rare breeds or endangered species, but the resulting sperm samples may be of poor quality. Two methods of extracting bull epididymal spermatozoa from slaughterhouse material were compared. The bulls were 16-23 months of age. Spermatozoa were extracted by making an incision one cm in length in the tail of the epididymis to allow the spermatozoa to flow out (method A), or by flushing the tail of epididymis (method B). The two methods were used for each bull, alternating between right and left epididymis, i.e. if method A was used for the left epididymis in Bull 1, it was used for the right epididymis in bull 2, etc. Sperm concentration in the extracted samples was adjusted to 69 × 106/mL in Andromed; the semen was packed in 0.25 mL straws. After cooling for two h at 5°C, the straws were placed 4 cm above liquid nitrogen for 20 min before transferring them to liquid nitrogen. Sperm motility, viability, reactive oxygen species, membrane integrity and DNA fragmentation were analysed in the fresh samples and again after thawing. The results for all parameters in fresh semen were not different between methods. Although sperm quality was lower in thawed samples than in fresh samples, there was no difference in sperm quality between the two extraction methods in the thawed samples. In conclusion, both methods are useful for the extraction of bull epididymal spermatozoa.

Mitochondrial Genome Assembly and Structural Characteristics Analysis of Gentiana rigescens.

Gentiana rigescens, an alpine plant with significant medicinal value, possesses a complex genetic background. However, comprehensive genomic research on G. rigescens is still lacking, particularly concerning its organelle genome. In this study, G. rigescens was studied to sequence the mitochondrial genome (mitogenome) and ascertain the assembly, informational content, and developmental expression of the mitogenome. The mitogenome of G. rigescens was 393,595 bp in length and comprised four circular chromosomes ranging in size from 6646 bp to 362,358 bp. The GC content was 43.73%. The mitogenome featured 30 distinct protein-coding genes, 26 tRNA genes, and 3 rRNA genes. The mitogenome of G. rigescens also revealed 70 SSRs, which were mostly tetra-nucleotides. In addition, 48 homologous fragments were found between the mitogenome and the chloroplast genome, with the longest measuring 23,330 bp. The documentation of the mitochondrial genome of G. rigescens is instrumental in advancing the understanding of its physiological development. Decoding the G. rigescens mitogenome will offer valuable genetic material for phylogenetic research on Gentianaceae and enhance the use of species germplasm resources.

Artificial Insemination: A New Horizon for Scientific Goat Breeding in Kashmir’s Backyard Farming Sector

Background: Artificial insemination a well-established technique in cattle and buffalo farming, now increasingly adopted in goat farming involves collecting semen from male animals and transferring it to female animals, to disseminate valuable genetic material to bring genetic upgradation, increases milk production, and enhances breeding efficiency of flocks. Methods: Artificial insemination (AI) was performed on 266 goats using fresh and frozen semen from 2021 to 2024 on experimental basis. The data obtained were analyzed on percentage basis. Result: The overall kidding percentage of 30.08% was observed in the present study. Notably, fresh semen yielded a higher kidding percentage (32.69%) compared to frozen semen (29.44%). This study demonstrates the potential of AI in enhancing the genetic quality of backyard goat farming in Kashmir, where quality bucks are often sold at a young age for meat production. By leveraging AI, farmers can improve the genetic makeup of their goat flocks, leading to better productivity and sustainability.

Identification of Drought Tolerant Chickpea Genotypes under Simulated Drought

Chickpea (Cicer arietinum L.) is one of the most important legume crops worldwide, with high protein content. Chickpea productivity is highly threatened by abiotic stresses, of which drought exerts the most crucial role in terms of growth inhibition and yield losses encountered. Since germination is a critical stage that is negatively affected by drought, an experiment was conducted to estimate the genotypic variability among 27 chickpea genotypes and to determine the seed germination and seedling growth ability under water stress conditions. Seeds were subjected to water stress by using polyethylene glycol (PEG-6000) at five stress levels (0, 5, 10, 15 and 20 % PEG). Among the genotype DIBG 205 (94.50%) recorded highest germination percentage compared to other genotypes. Root length recorded was highest in ICCV 4958 (18.48 cm) under 10% PEG -6000 and decreased with increase in osmotic stress. Drought significantly affected germination as well as all other associated traits with the effects of stress being analogous to the stress level applied. Seedling vigour index is a suitable selection criterion for drought tolerance, high seedling vigour index was observed in DIBG 205 and ICCV 4958 showing increased drought tolerance at high stress level, whereas low seedling vigour index was recorded in ICCV 201217 and ICCV 201116 indicating their possible exploitation as valuable genetic material for further breeding programs for drought tolerance.

Screening and analysis of candidate genes conferring alkalinity tolerance in rice (Oryza sativa L.) at the bud burst stage based on QTL-seq and RNA-seq

BackgroundSoil salinization is one of the key factors restricting the production of cropland. Once rice is subjected to alkali stress at the bud burst stage, the yield will suffer irreparable serious loss. Compared with salt tolerance, studies on QTL mapping and candidate gene analysis of rice alkali tolerance are limited. ResultsIn this study, we used the F2:3 population derived from the alkali-tolerant cultivar LD21 and the alkali-sensitive cultivar WL138 to construct an alkali-tolerant DNA mixing pool, and the BSA (Bulked Segregation Analysis) method was used for re-sequencing. The main QTL qRSLB9 controlling the relative shoot length of rice under alkali stress was mapped by QTL-seq. The candidate interval was narrowed to 346.5 kb by regional linkage mapping, which containing 6 DEGs screened through transcriptome sequencing. The qRT-PCR and candidate gene sequencing showed that LOC_Os09g24260 was most likely to control relative shoot length (RSL) in rice as a major gene who encodes the WD domain, G-beta repeat domain-containing protein. ConclusionsBased on these results, LOC_Os09g24260 was the candidate gene of qRSLB9 conferring alkalinity tolerance to rice at the bud burst stage. Our study provides valuable genetic information and materials for breeding new rice varieties with alkalinity tolerance.How to cite: Wang J, Bian J, Liu L, et al. Screening and analysis of candidate genes conferring alkalinity tolerance in rice (Oryza sativa L.) at the bud burst stage based on QTL-seq and RNA-seq. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.07.002.

Modern breeding methods in forestry aimed at preserving genetic diversity

This research is devoted to analysing the impact of modern breeding measures in forestry on the level of genetic diversity of forest tree species. It has been found that the main source of improved seed material for the genetic restoration of forests is base forest seed orchards. Aspects of the influence of determining factors – background pollination and the number of clones – on the indicators of genetic variability of progeny have been analysed in detail. The potential of background pollination in forest seed orchards in the context of a significant decrease in the effectiveness of breeding measures and a parallel increase in the level of genetic variability of progeny has been studied. An analysis of data on the minimum number of clones in seed orchards has been carried out based on the practical experience of other developed countries. The dynamics of clone variability in terms of fertility have been investigated. The potential impact of clonal selection on the genetic diversity of tree species, particularly in reducing it, has been identified. It has been confirmed that integrating the concept of family forestry, which involves the use of vegetative propagation techniques, into the breeding strategy significantly increases the level of genetic variability in progeny. The study has demonstrated that the multiple-population breeding system provides the optimal preconditions for synergising the process of long-term intensive breeding and preserving the gene pool of tree species. It has been determined that there is no negative impact on genetic diversity from implementing a complex of optimally planned breeding programs. At the same time, the potential for intensifying the quality of gene pool conservation in the process of forest ecosystem restoration through the use of improved seed material and clones in artificial orchards has been established. Special attention is given to the maintenance of ex situ – valuable genetic material in forest seed production facilities, including forest seed orchards, trial cultures, and clone archives of plus trees. Research has shown that the implementation of modern innovative solutions and scientific recommendations can minimise the loss of genetic diversity in forest tree species. The results can be applied in contemporary forestry breeding programs

Microsatellite marker-assisted backcross breeding for improvement of wheat salt tolerance using Kharchia 65

BackgroundSalinity is a significant abiotic stress that affects plants from germination through all growth stages. This study was aimed to determine the morpho-physiological and genetic variations in BC1F2, BC2F1 and F3 generations resulting from the cross combination WH1105 × Kharchia 65.ResultsA significant reduction in germination percentage was observed under salt stress in BC1F2 and F3 seeds. Correlation, heritability in the broad sense, phenotypic coefficient of variability (PCV) and genotypic coefficient of variability (GCV) were measured for all traits. The presence of both Nax1 and Nax2 loci was confirmed in twenty-nine plants using the marker-assisted selection technique. Genetic relationships among the populations were assessed using twenty-four polymorphic SSR markers.ConclusionCluster analysis along with two and three-dimensional PCA scaling (Principal Component Analysis) revealed the distinct nature of WH 1105 and Kharchia 65. Six plants closer to the recurrent parent (WH1105) selected through this study can serve as valuable genetic material for salt-tolerant wheat improvement programs.

Comparative analysis of antioxidant activities and chemical compositions in the extracts of different edible parts from Camellia tetracocca Zhang (C. tetracocca) with two distinct color characteristics

The Camellia tetracocca Zhang is a rare and ancient plant, exclusively found in the vicinity of Puan County, Guizhou Province, China. According to leaf color, two distinct variations have been identified: purple C. tetracocca Zhang (PCTZ) and green C. tetracocca (GCTZ). This research was conducted to investigate the antioxidant activities and chemical compositions of different edible parts of PCTZ and GCTZ. Antioxidant activity was evaluated using DPPH, ABTS, HSA, and T-AOC assays, while the content of compounds was determined by HPLC. The findings demonstrated that the antioxidant capacity of PCTZ leaves is significantly superior to that of GCTZ leaves. Notably, theacrine, a rare compound, contains up to 2.075% in PCTZ leaves, indicating potential as a novel natural antidepressant and antioxidant. In conclusion, PCTZ is a distinctive tea beverage and a valuable genetic material for tea tree breeding due to its high theacrine and low caffeine characteristics.

Widening Genetic Diversity Using Embryo Rescue in Cucurbit Crops: A Review.

Embryo rescue is a vital technique in cucurbit breeding and propagation, addressing challenges such as embryo abortion, poor seed viability, and incompatibility barriers. This method involves the excision of immature embryos from seeds followed by their in vitro culture on a nutrient medium, providing an environment conducive to their growth and development. In cucurbits, embryo rescue has been extensively utilized to overcome barriers to hybridization, enabling the production of interspecific and intergeneric hybrids with desired traits. Various factors, including genotype, developmental stage of embryos, and culture conditions, influence the success of embryo rescue in cucurbits. Optimal nutrient formulations, growth regulators, and culture techniques are critical for promoting embryo germination, shoot elongation, and subsequent plantlet establishment. Additionally, embryo rescue facilitates the recovery of valuable genetic material from wild and exotic cucurbit species, expanding genetic diversity and developing novel cultivars with improved traits such as disease resistance, yield, and quality. This review highlights the principles, applications, and advancements in embryo rescue technology in cucurbits, emphasizing its significance in cucurbit breeding programs and crop improvement efforts.

Кoнкурсно изпитване между местни и интродуцирани сортове тютюн Бърлей

During the period 2016-2020, is made in the experimental field of TTРI a comparative complex evaluation of six samples of tobacco from the Burley varietal group, of which three introduced varieties and the same number of indigenous varieties,. The subject of the study are biometric, biological, economic and chemical indicators of the studied variants. As a result of the study, it is found that the researched genotypes presented with favorable Burley tobacco biometric indicators. All studied varieties have a shorter vegetative period than the standard Pliska 2002 variety. The highest yield is presented by Burley 1000 variety, followed by a small but proven difference by Burley 1317 variety. The highest percentage of the first grade is distinguished by the Tennessee 90 variety, which also gives the lowest percentage of the third grade. Tennessee 90 variety also has the most favorable chemical composition in terms of all the investigated chemical indicators.The complex assessment of the biological, economic and chemical indicators shows that it is not possible to distinguish a variety that is superior to the other tested variants. In terms of percentage of grades and chemical composition, the introduced variants are superior to the local varieties, but they are seriously inferior to them in terms of yield. The Tennessee 90 and Tennessee 86 varieties have similar biometric, economic and chemical characteristics with a slight predominance of Tennessee 90. Tennessee 90 variety namely has the highest selection value of the introduced varieties. The studied introduced varieties are valuable genetic material that can be successfully used in the selection of Burley tobacco.

Morphological, chemical, and molecular characterization of a new late-leafing and high fruit quality hazelnut (Corylus avellana L.) genotype

Hazelnuts are widely employed in various dietary practices, making them one of the most frequently utilized nuts. This study morphologically, chemically, and molecularly characterized a new hazelnut genotype throughout the 2018 to 2020 growing seasons. The variety burst leaf buds between 10th and 15th April and reached harvest maturity between 5th and 15th September. The genotype bore a mean of 2.33 nuts per cluster, with the majority of clusters consisting of double and triple nuts. The means of the nut weight was 2.38 g, the kernel weight was 1.35 g, the kernel ratio was 56.81%, the shell thickness was 0.96 mm, the nut size was 18.32 mm, the kernel size was 14.77 mm, the good kernel ratio was 91.7%, the protein content was 13.5%, the oil content was 55.8%, the oleic acid ratio was 81.43%, and the linoleic acid ratio was 10.68%. The genetic similarity rate between the new hazelnut genotype and the standard Çakıldak, Palaz, Tombul cultivars, as well as randomly selected Çakıldak hazelnut clones from the region, ranged from 0.58 to 0.98. Additionally, the polymorphism rate varied from 45.5 to 100%. On average, the investigated hazelnut genotype exhibited a genetic distinctiveness of 38% compared to randomly selected Çakıldak clones from the same region, and 47% compared to standard hazelnut cultivars. Consequently, this genotype could serve as valuable genetic material for hazelnut breeding programs through genetic distinctiveness and promising nut quality, and could potentially be registered as a new cultivar.

Gonadal transcriptome sequencing reveals sexual dimorphism in expression profiling of sex-related genes in Asian arowana (Scleropages formosus).

Asia arowana (Scleropages formosus) is an ornamental fish with high economic value, while its sex determination mechanism is still poorly understood. By far, no morphological evidence or molecular marker has been developed for effective distinguishment of genders, which poses a critical challenge to our captive breeding efforts. In this study, we sequenced gonadal transcriptomes of adult Asian arowanas and revealed differential expression profiling of sex-related genes. Based on the comparative transcriptomics analysis of testes (n = 3) and ovaries (n = 3), we identified a total of 8,872 differentially expressed genes (DEGs) and 18,490 differentially expressed transposable elements (TEs) between male and female individuals. Interestingly, the expression of TEs usually has been more significantly testis-biased than related coding genes. As expected, several genes related to females (such as foxl2 and cyp19a1a) are significantly transcribed in the ovary, and some genes related to male gonad development (such as dmrt1, gsdf and amh) are highly expressed in the testis. This sexual dimorphism is valuable for ascertaining the differential expression patterns of sex-related genes and enriching the genetic resources of this economically important species. These valuable genetic materials thereby provide instructive references for gender identification and one-to-one breeding practices so as to expand fish numbers for a rapid elevation of economic value.

Лесная селекция и генетическое разнообразие древесных пород

In scientific literature there are different views on the connection between forest tree breeding and genetic variability of wood species. This article has aimed to review the current Russian and foreign literary sources on the impact of breeding measures on the level of genetic diversity of forest wood species. Maintaining adequate genetic variability is necessary for adaptation to the environmental changes, as well as for long-term breeding. As soon as seed plantations are the source of improved seeds for forest regeneration, the influence of such factors as the number of clones and background pollination on the level of genetic variability of the progeny has been considered in detail. The data on the minimal clone number in seed plantations in different countries has been presented and the variability of the clones based on their fertility has been discussed. The importance of background pollination in seed plantations has been highlighted as soon as, on the one hand, it reduces the effect of breeding, but on the other hand, it increases the level of genetic variability of the progeny. A decline in genetic diversity of wood species can be caused by clone selection (which is the basis for clonal forestry). However the analysis of literary sources has shown that the reduction in genetic diversity is minimal if scientific recommendations are fulfilled. The use of the seeds from controlled cross-breeding of plus trees selected as a result of genetic evaluation (the so-called family forestry with vegetative propagation) in the somatic embryogenesis of the common spruce increases the level of genetic variability of the progeny. It has been shown that the multiple population breeding system allows to combine intensive long-term breeding and genetic conservation of wood species. The conclusion has been drawn that the implementation of optimally planned wood species breeding programs does not lead to significant narrowing of genetic diversity. On the contrary, forest tree breeding contributes to the preservation of a better gene pool, transmitting it in the process of forest regeneration through improved seeds and clones into homogeneous stands. Moreover, the objects of forest seed growing such as the archives of plus tree clones and forest seed plantations, as well as test cultures contain valuable genetic material ex situ.

DArTseq genotyping facilitates the transfer of "exotic" chromatin from a Secale cereale × S. strictum hybrid into wheat.

Cultivated and wild species of the genus rye (Secale) are important but underexploited gene sources for increasing the genetic diversity of bread wheat. Gene transfer is possible via bridge genetic materials derived from intergeneric hybrids. During this process, it is essential to precisely identify the rye chromatin in the wheat genetic background. In the present study, backcross generation BC2F8 from a cross between Triticum aestivum (Mv9kr1) and S. cereanum ('Kriszta,' a cultivar from the artificial hybrid of S. cereale and S. strictum) was screened using in-situ hybridization (GISH and FISH) and analyzed by DArTseq genotyping in order to select potentially agronomically useful genotypes for prebreeding purposes. Of the 329,267 high-quality short sequence reads generated, 27,822 SilicoDArT and 8,842 SNP markers specific to S. cereanum 1R-7R chromosomes were identified. Heatmaps of the marker densities along the 'Lo7' rye reference pseudomolecules revealed subtle differences between the FISH- and DArTseq-based results. This study demonstrates that the "exotic" rye chromatin of S. cereanum introgressed into wheat can be reliably identified by high-throughput DArTseq genotyping. The Mv9kr1-'Kriszta' addition and translocation lines presented here may serve as valuable prebreeding genetic materials for the development of stress-tolerant or disease-resistant wheat varieties.

Obtaining a culture of somatic cells using tissue material from the ear of dead sheep/snow sheep hybrid

Production and cryopreservation of somatic cells (SCs) from valuable and endangered animals allows a preservation of genetic diversity and ensuring their future reproduction. The aim of present work was to isolate SCs from the ear of unique hybrid sheep (Ovis aries) and snow sheep (Ovis nivicola borealis) post-mortem. In this purpose, enzymatic and mechanical methods of tissue preparation were compared.Materials and Methods. Ears from deceased animal were brought to the laboratory 12 hours after the death in a pasture, and biological material was thoroughly washed under running water. The hairs were removed from the part of the ear shell by the blade. Skin fragments were treated with 70% ethyl alcohol, washed three times in a saline solution with antibiotics and ground up to small pieces. The ear pieces were washed several times in phosphate buffer saline and divided into two parts. One part of the explants started in vitro culture without enzymatic treatment (group 1), whereas another part was pre-treated with a 0.25% trypsin/EDTA solution. After trypsinization, either tissue fragments (group 2), or cell complexes separated from cell suspension fraction (group 3) were taken for in vitro culture for 9 days. Monitoring of cell colony formation and growth was carried out daily. Results. In the group 3, cell colonies were formed on the second day of in vitro culture. In groups 1 and 2, cell growth was observed from tissue fragments after five days regardless of the treatment. On the 9th day, all the groups produced the primary cultures, represented by two types of SCs. In general, single cell complexes from the group 3 formed cell growth zones more quickly than tissue explants from the groups 1 and 2, however, final cultures of SCs and their morphological aspects were no different between the groups. Conclusion. Methodological protocols were proposed and successfully used to obtain in vitro cultures of SCs from the ear of dead sheep/snow sheep hybrid animal, 12 hours post-mortem that may allow further storage of valuable genetic material.

ИЗУЧЕНИЕ СОРТА ВИНОГРАДА ВАРЮШКИН В РЕГИОНЕ ПРОИСХОЖДЕНИЯ

The article presents the results of a 17-year study of autochthonous Don grapevine variety Varyushkin. The studies were carried out in 2006−2022 at Don ampelographic collection named after Ya.I. Potapenko (Novocherkassk, Rostov region). The variety is cultivated in a grafted (rootstock Kober 5BB) covering cul-ture, the scheme of planting bushes is 3.0 × 1.5 m. The study was carried out using methods generally accepted in viticulture and State Standarts. An ampelographic description of the variety (crowns of a young shoot, leaf, bunch, berry) and an analysis of meteorological condi-tions of the research are given. According to the results of long-term observations, the Don autochthonous grapevine variety Varyushkin has a high agrobiological potential in the climat-ic conditions of the Lower Don region (71 % of blossoming buds, 58 % of fruitful shoots, fruit-ing coefficient 0.8, average bunch weight 253 g, yield 109 c/ha). Medium ripening variety (141 days). When reaching technological ma-turity, it accumulates sugars of 22.3 g/100 cm3 with a titratable acidity of 8.2 g/dm3. It has the following characteristics: bunch length 17.2 cm, bunch width − 9.2 cm, berry length − 14.7 mm, berry width − 13.9 mm. The average weight of 1 berry was 2.1 g. Dry red wine from Varyush-kin variety has a harmonious taste and has a tasting score at the level of wine from the clas-sic Pinot noir variety (8.6 points). Therefore, the variety can be used to expand the raw material resource in order to produce high-quality wines that help meet the needs of the population, as well as a valuable genetic material for breeding.

Parallel Lives

This article investigates cow caretaking practices in a Finnish prison and living gene bank. Building upon work in Feminist Science and Technology Studies, Critical Animal Studies, and Critical Heritage Studies, we seek to understand how human and nonhuman animal lives are partially folded together, while also discussing the worlds cultivated in a space of conservation and incarceration. Empirically, the article draws upon multispecies ethnography undertaken during two separate visits to Pelso Prison in central Finland. We conclude that cows emerge as not simply working animals. They are viewed as valuable genetic material vital to the Finnish nation state as well as given names and granted personalities. Meanwhile, in the company of an endangered cattle breed, inmates gain new value as care-workers and conservationists “saving” the breed. In this space of exception, precarious interspecies lives are interwoven.

Discovering useful genetic variation in the seed parent gene pool for sorghum improvement.

Multi-parent populations contain valuable genetic material for dissecting complex, quantitative traits and provide a unique opportunity to capture multi-allelic variation compared to the biparental populations. A multi-parent advanced generation inter-cross (MAGIC) B-line (MBL) population composed of 708 F6 recombinant inbred lines (RILs), was recently developed from four diverse founders. These selected founders strategically represented the four most prevalent botanical races (kafir, guinea, durra, and caudatum) to capture a significant source of genetic variation to study the quantitative traits in grain sorghum [Sorghum bicolor (L.) Moench]. MBL was phenotyped at two field locations for seven yield-influencing traits: panicle type (PT), days to anthesis (DTA), plant height (PH), grain yield (GY), 1000-grain weight (TGW), tiller number per meter (TN) and yield per panicle (YPP). High phenotypic variation was observed for all the quantitative traits, with broad-sense heritabilities ranging from 0.34 (TN) to 0.84 (PH). The entire population was genotyped using Diversity Arrays Technology (DArTseq), and 8,800 single nucleotide polymorphisms (SNPs) were generated. A set of polymorphic, quality-filtered markers (3,751 SNPs) and phenotypic data were used for genome-wide association studies (GWAS). We identified 52 marker-trait associations (MTAs) for the seven traits using BLUPs generated from replicated plots in two locations. We also identified desirable allelic combinations based on the plant height loci (Dw1, Dw2, and Dw3), which influences yield related traits. Additionally, two novel MTAs were identified each on Chr1 and Chr7 for yield traits independent of dwarfing genes. We further performed a multi-variate adaptive shrinkage analysis and 15 MTAs with pleiotropic effect were identified. The five best performing MBL progenies were selected carrying desirable allelic combinations. Since the MBL population was designed to capture significant diversity for maintainer line (B-line) accessions, these progenies can serve as valuable resources to develop superior sorghum hybrids after validation of their general combining abilities via crossing with elite pollinators. Further, newly identified desirable allelic combinations can be used to enrich the maintainer germplasm lines through marker-assisted backcross breeding.

The morphology, morphometry and functionality of fresh and cryopreserved wisent (Bison bonasus) epididymal spermatozoa

Epididymal spermatozoa obtained post mortem are considered a valuable source of genetic material which is often irrevocably lost. This makes these gametes constitute a key element in protection and restitution programs. The wisent (Bison bonasus, Linnaeus 1758) is a species that survived in zoos after extinction from its natural habitat. This resulted in a narrowing of the genetic pool of the whole population, which is at present derived from only 12 ancestors. Currently, wisent protection programs are aimed at preserving the genetic diversity by establishing a germplasm bank. The objective of this study was to comprehensively characterize the morphology, morphometry and functionality of wisent epididymal spermatozoa and evaluate the effectiveness of their cryopreservation in extender based on Tris buffer and chicken egg yolk. The median total number of spermatozoa obtained from one individual was 1985.0 × 106 (62.5 × 106–7452.0 × 106). These gametes were characterized by median: 40.0% (0.5–70.0%) subjective motility, 69.8% (32.5–90.0%) viability and 54.3% (10.5–83.3%) normal morphology. The sperm head had a median size of 5.0 μm (3.5–6.7 μm) width, 8.5 μm (6.4–11.3 μm) length and 36.9 μm2 (23.7–48.6 μm2) surface area. The viable population of the obtained gametes was characterized by median values 53.2% (4.5–80.3%) of intact sperm membrane, 50.8 (26.0–76.6%) of intact acrosome, 0.4% (0–98.7%) of fragmented chromatin, 5.9% (0.0–88.8%) of cells with high mitochondrial potential and 42.1% (8.3–63.7%) without lipid peroxidation. The viable population of the frozen/thawed gametes was characterized by median values: 18.4% (2.4–57.9%) of intact sperm membrane, 35.1 (11.9–56.7%) of intact acrosome, 0.07% (0–89.2%) of fragmented chromatin, 12.8% (0.0–49.7%) of cells with high mitochondrial potential and 16.3% (2.2–53.6%) without lipid peroxidation. Due to the material originating from a relatively large number of wild individuals, the research presented here contributed to the description of certain species standards for the assessment of wisent epididymal spermatozoa. The presented effect of cryopreservation on these gametes justifies the use of an extender based on Tris buffer with the addition of chicken egg yolk. The obtained effects are satisfactory from the point of view of preserving valuable genetic material and their use in ART.

ИЗУЧЕНИЕ АВТОХТОННОГО ДОНСКОГО СОРТА ВИНОГРАДА ЦИМЛЯНСКИЙ БЕЛЫЙ В УСЛОВИЯХ НИЖНЕГО ПРИДОНЬЯ

The study was carried out in order to assess the agrobiological and uvological indicators of the rare autochthonous Don grape variety Tsimlyansky Belyy, growing in the conditions of the Lower Don, and can be used to expand the raw material resource in the production of quality wines and as a valuable genetic material for grape breeding. The experiments were performed on the ampelographic collection (Rostov region) in 2015-2021, according to generally accepted methods and GOST. The bushes are grafted onto the rootstock Kober 5BB. Varieties are cultivated in a covering culture. It has been established that under these growing conditions, Tsimlyansky Belyy variety has an early ripening period (117 days), and the control variety Sibir'kovyy has an early-middle period ripening of berries (128 days). According to the results of agrobiological surveys, good overwintering and fruitfulness of plants were noted, the percentage of fruiting buds in Tsimlyansky Belyy variety was at the level of 72.7%, and in Sibir'kovyy variety - 66.2%. In terms of the percentage of fruitful shoots and the fruitfulness ratio, the differences were not significant and amounted to 5.8% and 0.1, respectively. The yield of the studied variety Tsimlyansky Belyy was 91 centners/ha, in the control variety - 86 centners/ha. The characteristics of grape were practically at the same level (medium sugar content - 20.6 g/100 cm3 for Tsimlyansky Belyy variety and 20.2 g/100 cm3 for the Sibir'kovyy variety; and the acidity of the varieties was low: Tsimlyansky Belyy - 6.4 g/dm3, Sibir'kovyy - 6.3 g/dm3). The wine from Tsimlyansky Belyy variety has a pale straw color, with a shine, has delicate floral and honey notes in the aroma, turning into the taste. The taste is full and harmonious. Tasting score 8.6 points. Wine from the control variety Sibir'kovyy received a tasting score of 8.7 points. In 2022, Tsimlyansky Belyy grape variety is included in the State Register of Russian varieties approved for use, has a high agrobiological potential in the climatic conditions of the Lower Don region, and can be used to expand the raw material resource in the production of quality wines, and as a valuable genetic material for grape breeding.