Marker Alleles Research Articles

Genotyping of lines from the VIR sunflower genetic collection using allele-specific markers of the Rf1 locus

Background. The CMS-Rf genetic system based on the PET1-type cytoplasmic male sterility (CMS) is commonly used to create commercial sunflower (Helianthus annuus) hybrids. The Rf1 gene, of key importance for hybrid breeding, is necessary for restoring pollen fertility in F1 plants. The molecular genetic markers tested on various genetic materials are an effective tool for identifying parental line genotypes at the Rf1 locus, controlling homogeneity, and determining the genetic purity of hybrid seed lots. In the present study, the allele-specific markers of the Rf1 candidate genes available from literature were used to genotype lines from the VIR sunflower genetic collection and F2 hybrids. Material and methods. The study concentrated on two sample sets of genotypes, one of which contained 46 lines from the VIR sunflower genetic collection, previously characterized in field experiments for the pollen restoration ability, and the other 80 plants from segregating F2 populations from crosses of the CMS VIR 116A line with fertility restorers VIR 740 and RIL 130, phenotyped for fertility/sterility. The lines differed in respect of the cytoplasm type and the presence of the SCAR marker HRG02 closely linked to the Rf1 locus. The lines have been genotyped using markers specific for the dominant (PPR621.5R, SRF833, 67N04_P_170) and recessive (PPR621.5M, 67N04_P_155) alleles of the Rf1 candidate genes. The PPR621.5M and PPR621.5 R, marker fragments amplified in six genotypes, have been isolated and sequenced. Results. The nucleotide sequences of PPR621.5M and PPR621.5R turned out to be different in four SNPs and completely identical to those presented in the published literature. The PPR621.5M and 67N04_P_155 markers specific for the rf1 allele were identified in CMS lines and the majority of sterility maintainers. Nineteen out of 21 lines characterized by sterile cytoplasm and the presence of the HRG02 marker had three markers specific for the dominant allele; two lines had two allele-specific markers. Four out of seven fertility restorers (sterile cytoplasm, without the HRG02 marker) were found to contain two or three markers specific for the dominant allele, while three lines had only markers for the recessive allele. The F2 genotypes resulting from recombination between the SCAR marker HRG02 and allele specific markers were detected. Conclusion. The study confirmed efficiency of allele-specific markers of the Rf1 locus candidate genes for genotyping sunflower lines, as well as their diagnostic value for selecting target genotypes from segregating hybrid populations.

Genetic Diversity and Drought Stress Tolerance of a Global Collection of Pearl Millet at Germination and Early Seedling Growth Stages.

Pearl millet {Pennisetum glaucum (L.) R. Br} is a C4 panicoid cereal millet crop grown in arid and semi-arid regions in Africa and Asia for food and fodder. This study involves the evaluation of the genetic diversity of 28 worldwide germplasm collection of pearl millet by genetic markers polymorphism and drought tolerance indices. The genetic diversity was expressed by 51 alleles of 9ISSR markers that showed 96.43% totalpolymorphism and 11.76 alleles per marker. Cluster analysis of ISSR markers polymorphism divided the 28 genotypes into four clusters partially in agreement with their origin. The application of drought stress simulated by 20% PEG6000 treatment, retarded the germination percentage, and reduced shoot and root length, seedling fresh and dry weights. Drought tolerance indices (DTIs) were calculated based on the response of the seedling traits under drought stress compared to the control seedlings. ANOVA revealed statistically significant variation among the genotypes (P ≤ 0.05), except for seedling fresh weight (P = 0.17 > 0.05) under control conditions and seedling dry weight (P = 0.99 > 0.05) under drought conditions. Genotypes having higher DTIs for three traits are regarded drought resistant, i.e., those from India, Ethiopia, Pakistan, and Nigeria. The calculated heritability values indicated that seedlings dry weight is the least trait affected by drought stress whereas root length is the most influenced trait. Hierarchical clustering, based on the DTI values, also grouped the genotypes partially concomitant to their origin. The correlation analysis demonstrated a modest positive correlation between shoot length and root length. A low correlation of r ≤ 0.12 was observed between the morphological DTI matrix and the genetic matrix. Nevertheless, high levels of genetic diversity were identified among the examined genotypes that may face genetic erosion by climatic constraints, and a high potential for creating agronomically superior cultivars by crossing widely divergent genotypes.

Toward Marker-Assisted Selection in Breeding for Fusarium Wilt Tropical Race-4 Type Resistant Bananas.

Fusarium wilt is a soil borne fungal disease that has devastated banana production in plantations around the world. Most Cavendish-type bananas are susceptible to strains of Fusarium oxysporum f. sp. cubense (Foc) belonging to the Subtropical Race 4 (STR4) and Tropical Race 4 (TR4). The wild banana diploid Musa acuminata ssp. malaccensis (AA, 2n = 22) carries resistance to Foc TR4. A previous study using segregating populations derived from M. acuminata ssp. malaccensis identified a quantitative trait locus (QTL) (12.9 cM) on the distal part of the long arm of chromosome 3, conferring resistance to both Foc TR4 and STR4. An SNP marker, based on the gene Macma4_03_g32560 of the reference genome 'DH-Pahang' v4, detected the segregation of resistance to Foc STR4 and TR4 at this locus. Using this marker, we assessed putative TR4 resistance sources in 123 accessions from the breeding program in Brazil, which houses one of the largest germplasm collections of Musa spp. in the world. The resistance marker allele was detected in a number of accessions, including improved diploids and commercial cultivars. Sequencing further confirmed the identity of the SNP at this locus. Results from the marker screening will assist in developing strategies for pre-breeding Foc TR4-resistant bananas. This study represents the first-ever report of marker-assisted screening in a comprehensive collection of banana accessions in South America. Accessions carrying the resistance marker allele will be validated in the field to confirm Foc TR4 resistance.

Marker Effects for Fusarium Head Blight Resistance and the Dwarfing Gene Rht‐B1 in Cultivated Emmer Wheat (Triticum turgidum ssp. dicoccum) Revealed by Association Mapping

ABSTRACTFusarium head blight (FHB) is a disease that affects all cereals worldwide. This includes emmer wheat (Triticum turgidum ssp. dicoccum), the ancestor of durum and bread wheat. We screened 143 cultivated emmer genotypes from a breeding program and gene bank collections for FHB severity at 2 locations over 2 years. Due to the high negative correlation between FHB severity and heading date (HD) (r = −0.65, p < 0.001), plot‐level FHB scores were corrected for HD before further analysis (FHBcorr). Genetic variation for FHB severity was high, ranging from 2.15 to 8.33 on a 1–9 scale. Twelve genotypes carried the semi‐dwarfing Rht‐B1b marker allele, which reduced plant height by 32 cm but increased FHB severity by 20%. Genome‐wide association study detected seven quantitative trait nucleotides (QTNs) for FHBcorr and three QTNs for plant height. The most important QTN for both traits was located on chromosome 4B, explaining 50.9% and 15.8% of the phenotypic variation in plant height and FHBcorr, respectively, and was localized near the semi‐dwarfing Rht‐B1 locus. Three other large‐effect loci for FHBcorr were found on chromosomes 5B and 7B. In total, 72.6% of phenotypic variation was explained by all markers. The use of Rht‐B1b in emmer breeding has a high effect on plant height but would entail the introgression of potent FHB‐resistance from either native or exotic sources.

An Inexpensive and Quick Method for Genotyping of HLA Variants Included in the Spanish Pharmacogenomic Portfolio of National Health System

The possibility of using the same genotyping technology (TaqMan) for all the genetic tests included in the new Spanish pharmacogenomics portfolio should enable the application of a multigenotyping platform to obtain a whole pharmacogenomics profile. However, HLA-typing is usually performed with other technologies and needs to be adapted to TaqMan assays. Our aim was to establish a set of TaqMan assays for correct typing of HLA-A*31:01, HLA-B*15:02, HLA-B*57:01, and HLA-B*58:01. Therefore, we searched for and selected SNVs described in different populations as surrogate markers for these HLA alleles, designed TaqMan assays, and tested in a set of samples with known HLA-A and HLA-B. HLA-A*31:01 was correctly typed with a combination of rs1061235 and rs17179220 (PPV 100%, 95% CI 84.6–100-%; NPV 100%, 95% CI 96.5–100.0%), HLA-B*15:02 with rs10484555 (PPV 100%, 95% CI 69.2–100.0%; NPV 100%, 95% CI 96.8–100.0%) and rs144012689 (PPV 100%, 95% CI 69.2–100.0%; NPV 100%, 95% CI 96.8–100.0%), and HLA-B*57:01 with rs2395029 (PPV 99.5%, 95% CI 72.9–99.3%; NPV 99.5%, 95% CI 98.3–100.0%). HLA-B*58:01 was typed using two allele-specific TaqMan probes mixed with a ß-Globin reference and treated as a genotyping assay (PPV 100.0%, 95% CI 81.5–100.0%; NPV 100%, 95% CI 96.8–100.0%). In conclusion, we demonstrated a clinically useful way to type HLA-A and HLA-B alleles included in the Spanish pharmacogenomics portfolio using TaqMan assays.

Doubled Haploid Technology: Generation of Doubled Haploid Maize Lines Using Haploid Inducers.

Doubled haploid (DH) technology allows for the development of completely homozygous lines from heterozygous plants in only two generations. This approach has been widely adopted in maize breeding programs, as it expedites the generation of inbred lines compared to traditional methods. The DH approach is based on the use of maize genotypes that have the ability to induce haploid seeds when used as the pollen parent. The most common method for producing maize haploid plants for the generation of DH lines is in vivo maternal haploid induction. The process involves pollination with a haploid inducer maize line to generate haploid seeds. Then, haploids are screened for and identified (typically via the expression of a particular marker gene), germinated, treated with an exogenous doubling agent to induce genome duplication, and transplanted to the field. Following successful self-pollination, seeds harvested from the ear represent fully homozygous lines. The seed set at this stage, however, is often low, necessitating one or two additional rounds of self-pollination to increase the number of fully homozygous inbred lines. Here, we describe a protocol for the generation of maize DH lines using maternal haploid-inducing maize lines. We outline the steps for setting up the donor material, performing induction crosses, selecting haploids based on two different marker alleles, treating seedlings with colchicine to double the genome, transplanting the treated seedlings to the field, and self-pollinating the treated plants.

Potential approaches to create ultimate genotypes in crops and livestock.

Many thousands and, in some cases, millions of individuals from the major crop and livestock species have been genotyped and phenotyped for the purpose of genomic selection. 'Ultimate genotypes', in which the marker allele haplotypes with the most favorable effects on a target trait or traits in the population are combined together in silico, can be constructed from these datasets. Ultimate genotypes display up to six times the performance of the current best individuals in the population, as demonstrated for net profit in dairy cattle (incorporating a range of economic traits), yield in wheat and 100-seed weight in chickpea. However, current breeding strategies that aim to assemble ultimate genotypes through conventional crossing take many generations. As a hypothetical thought piece, here, we contemplate three future pathways for rapidly achieving ultimate genotypes: accelerated recombination with gene editing, direct editing of whole-genome haplotype sequences and synthetic biology.

Characterization of Bila Tserkva winter common wheat cultivars with respect to marker loci

Aim. We studied special features of the sample of Bila Tserkva winter common wheat cultivars with respect to storage protein loci and some disease resistance genes. Methods. To identify alleles at gliadin and high-molecular-weight glutenin loci, acid polyacrylamide gel electrophoresis and SDS-electrophoresis of grain proteins were carried out. Alleles of markers for the disease resistance genes Lr34, Tsn1, and TDF_076_2D were identified using PCR. Results. The composition and frequencies of alleles at the Gli-1, Glu-1, and Gli-A3 loci were studied in samples of Bila Tserkva cultivars released before 2011 and after 2010. For the latter group of cultivars, alleles of the disease resistance gene markers were identified. We observed a significant increase in the frequency of the allele Gli-A1x(9) and a decrease in the frequency of Gli-A1c, as well as a tendency for an increase in the frequency of Gli-A3c. In the total sample of Bila Tserkva cultivars, nonrandom associations of pairs of certain storage protein alleles were revealed. Conclusions. The group of Bila Tserkva cultivars is similar to the previously studied groups of the Central Forest Steppe of Ukraine with respect to allele set and frequencies of the main prolamin loci, but differs in the frequencies of the disease resistance genes. A special feature of the group of Bila Tserkva cultivars is a high frequency of the allele c at the minor locus Gli-A3, as well as the allele association Gli-A1x Gli-B1l Gli-A3c.

ОЦЕНКА КОРОВ-ПЕРВОТЁЛОК ЧЁРНО-ПЁСТРОЙ ПОРОДЫ ПО ДНК-МАРКЕРАМ, АССОЦИИРОВАННЫХ С ХОЗЯЙСТВЕННО-ПОЛЕЗНЫМИ ПРИЗНАКАМИ

At present time are widely used genetic markers as means in the selection of highly productive animals. Since the genes: CSN3 (kappa-casein), PIT-1 (pituitary-specific transcription factor), PRL (prolactin), GH (somatotropin) are polymorphic and encode various proteins, the study of their influence on the productive traits of cattle is relevant. In this regard, the goal of the work is to determine the influence of complex genotypes of the studied genes on the milk productivity of first-calf cows of the Black-and-White breed bred in the collective farm-breeding plant «Kazminsky» in the Kochubeevsky district of the Stavropol Territory. The place of research is the Department of Genetics and Biotechnology, VNIIOK, a branch of the North Caucasian Federal National Research Center. This article presents the results of DNA genotyping of allelic polymorphism for the CSN3, PIT-1, PRL, GH genes using PCR-RFLP methods in first-calf cows of the Black-and-White breed. It was established that these genes in the studied sample are polymorphic. The frequency of occurrence of the desired CSN3B allele in the studied sample was 0.23. The presence of alleles PIT-1A and PIT-1B of the pituitary-specific transcription factor gene was noted with a frequency of occurrence of 0.52 and 0.48. A feature of the PRL gene polymorphism was the fairly high frequency of occurrence of the PRLA allele (0.60). In this study sample of dairy cattle, the frequency of occurrence of the desired GHL allele of the somatotropin gene was 0.62. All variants of complex genotypes were also identified and their relationship with milk productivity in black-and-white cows was noted. The study sample was dominated by animals carrying complex desirable genotypes, consisting of four and three marker alleles of three and two genes (CSN3AB/PIT-1BB/GHLV; CSN3AB/PIT1AB/GHLL or PIT-1AB/PRLAB/GHLV; PIT-1AA/GHLL ) - 46.7 %. Milk yield per lactation between groups of black-and-white first-calf cows with different complex genotypes ranged from 5839.14 to 8611.33 kg. The mass fraction of fat in milk ranged from 3.93 to 4.04 %, and protein from 2.99 to 3.14 %.

Association of Prehypertension with Renin Angiotensin System: A Cross-Sectional Study on Tribal Population of Northern India

Abstract Introduction: Hypertension (HTN), the third risk factor driving death and disability as per Global Burden of Disease 2019, is a major public health concern. Prehypertension, the intermediate stage between HTN and normal blood pressure (BP), is associated with subclinical atherosclerosis and target-organ damage. Evaluating the prevalence of prehypertension is imperative to prevent disease progression. Prehypertension, alike HTN, is a complex disorder that manifests when genes are triggered by the environment. The current study attempts to evaluate the prevalence of prehypertension in a tribal population of Northern India. Further, the association with extensively studied renin–angiotensin system (RAS) pathway genes with prehypertension has been explored. Methods: A cross-sectional study was conducted on adults of 20–60 years old belonging to a tribal community, Rang Bhotia of Northern India. Information on demographic, lifestyle, and anthropometric risk factors was collected. The blood sample was collected for lipid and genetic analysis. DNA extraction and genotyping of five genetic markers of RAS-related pathway were performed. Chi-square and t-tests were used for assessing significant differences between the groups. Logistic regression was used to examine the association of genes with prehypertension and HTN. Results: Among 254 adults participating in the study, 39% and 42% had prehypertension and HTN, respectively. T allele of M235T genetic marker was found to be posing two-fold risk for prehypertension (odds ratio [OR] = 2.9; 95% confidence interval [CI] =1.10–7.72; P = 0.03 in dominant model and OR = 1.83; 95% CI = 1.09-3.09; P = 0.02 in additive model) after adjusting for age and sex. On the other hand, G allele of A6G polymorphism was found to be providing protection toward prehypertension (OR = 0.32; 95% CI = 0.11–0.92; P = 0.03 in recessive model and OR = 0.55; 95% CI = 0.32–0.94; P = 0.03 in additive model). Conclusion: More than one-third of individuals exhibited prehypertension, suggesting an impending burden of HTN and comorbidities among the tribal population in the near future. Moreover, increased genetic susceptibility toward elevated BP commands immediate attention. Effective management strategies including interventions on lifestyle, diet modification, and weight management need to be introduced to combat increasing HTN among the tribal population.

Assessing the carotenoid profiles and allelic diversity of yellow maize inbred lines adapted to mid-altitude subhumid maize agroecology in Ethiopia.

Biofortification of provitamin A in maize is an attractive and sustainable remedy to the problem of vitamin A deficiency in developing countries. The utilization of molecular markers represents a promising avenue to facilitate the development of provitamin A (PVA)-enriched maize varieties. We screened 752 diverse tropical yellow/orange maize lines using kompetitive allele-specific PCR (KASP) makers to validate the use of KASP markers in PVA maize breeding. To this end, a total of 161 yellow/orange inbred lines, selected from among the 752 lines, were evaluated for their endosperm PVA and other carotenoid compounds levels in two separate trials composed of 63 and 98 inbred lines in 2020 and 2021, respectively. Significant differences (p < 0.001) were observed among the yellow maize inbred lines studied for all carotenoid profiles. An inbred line TZMI1017, introduced by the International Institute of Tropical Agriculture (IITA) showed the highest level of PVA (12.99 µg/g) and β-carotene (12.08 µg/g). The molecular screening showed 43 yellow maize inbred lines carrying at least three of the favorable alleles of the KASP markers. TZMI1017 inbred line also carried the favorable alleles of almost all markers. In addition, nine locally developed inbred lines had medium to high PVA concentrations varying from 5.11 µg/g to 10.76 µg/g and harbored the favorable alleles of all the KASP PVA markers. Association analysis between molecular markers and PVA content variation in the yellow/orange maize inbred lines did not reveal a significant, predictable correlation. Further investigation is warranted to elucidate the underlying genetic architecture of the PVA content in this germplasm. However, we recommend strategic utilization of the maize-inbred lines with higher PVA content to enhance the PVA profile of the breeding program's germplasm.

Striatal GDNF Neurons Chemoattract RET-Positive Dopamine Axons at Seven Times Farther Distance Than Medium Spiny Neurons.

Glial cell line-derived neurotrophic factor (GDNF) is among the strongest dopamine neuron function- and survival-promoting factors known. Due to this reason, it has clinical relevance in dopamine disorders such as Parkinson's disease and schizophrenia. In the striatum, GDNF is exclusively expressed in interneurons, which make up only about 0.6% of striatal cells. Despite clinical significance, histological analysis of striatal GDNF system arborization and relevance to incoming dopamine axons, which bear its receptor RET, has remained enigmatic. This is mainly due to the lack of antibodies able to visualize GDNF- and RET-positive cellular processes; here, we overcome this problem by using knock-in marker alleles. We find that GDNF neurons chemoattract RET+ axons at least seven times farther in distance than medium spiny neurons (MSNs), which make up 95% of striatal neurons. Furthermore, we provide evidence that tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, is enriched towards GDNF neurons in the dopamine axons. Finally, we find that GDNF neuron arborizations occupy approximately only twelve times less striatal volume than 135 times more abundant MSNs. Collectively, our results improve our understanding of how endogenous GDNF affects striatal dopamine system function.

Application of bioluminescence assay to assess PCR carryover contamination in forensic DNA laboratories

Application of bioluminescence assay to assess PCR carryover contamination in forensic DNA laboratories

Molecular Markers Help with Breeding for Agronomic Traits of Spring Wheat in Kazakhstan and Siberia.

The Kazakhstan-Siberia Network for Spring Wheat Improvement (KASIB) was established in 2000, forming a collaboration between breeding and research programs through biannual yield trials. A core set of 142 genotypes from 15 breeding programs was selected, genotyped for 81 DNA functional markers and phenotyped for 10 agronomic traits at three sites in Kazakhstan (Karabalyk, Shortandy and Shagalaly) and one site in Russia (Omsk) in 2020-2022. The study aim was to identify markers demonstrating significant effects on agronomic traits. The average grain yield of individual trials varied from 118 to 569 g/m2. Grain yield was positively associated with the number of days to heading, plant height, number of grains per spike and 1000-kernel weight. Eight DNA markers demonstrated significant effects. The spring-type allele of the Vrn-A1 gene accelerated heading by two days (5.6%) and was present in 80% of the germplasm. The winter allele of the Vrn-A1 gene significantly increased grain yield by 2.7%. The late allele of the earliness marker per se, TaMOT1-D1, delayed development by 1.9% and increased yield by 4.5%. Translocation of 1B.1R was present in 21.8% of the material, which resulted in a 6.2% yield advantage compared to 1B.1B germplasm and a reduction in stem rust severity from 27.6 to 6.6%. The favorable allele of TaGS-D1 increased both kernel weight and yield by 2-3%. Four markers identified in ICARDA germplasm, ISBW2-GY (Kukri_c3243_1065, 3B), ISBW3-BM (TA004946-0577, 1B), ISBW10-SM2 (BS00076246_51, 5A), ISBW11-GY (wsnp_Ex_c12812_20324622, 4A), showed an improved yield in this study of 3-4%. The study recommends simultaneous validation and use of selected markers in KASIB's network.

Identification of Sr67, a new gene for stem rust resistance in KU168-2 located close to the Sr13 locus in wheat

Key messageSr67 is a new stem rust resistance gene that represents a new resource for breeding stem rust resistant wheat cultivarsRe-appearance of stem rust disease, caused by the fungal pathogen Puccinia graminis f. sp. tritici (Pgt), in different parts of Europe emphasized the need to develop wheat varieties with effective resistance to local Pgt populations and exotic threats. A Kyoto University wheat (Triticum aestivum L.) accession KU168-2 was reported to carry good resistance to leaf and stem rust. To identify the genomic region associated with the KU168-2 stem rust resistance, a genetic study was conducted using a doubled haploid (DH) population from the cross RL6071 × KU168-2. The DH population was phenotyped with three Pgt races (TTKSK, TPMKC, and QTHSF) and genotyped using the Illumina 90 K wheat SNP array. Linkage mapping showed the resistance to all three Pgt races was conferred by a single stem rust resistance (Sr) gene on chromosome arm 6AL, associated with Sr13. Presently, four Sr13 resistance alleles have been reported. Sr13 allele-specific KASP and STARP markers, and sequencing markers all showed null alleles in KU168-2. KU168-2 showed a unique combination of seedling infection types for five Pgt races (TTKSK, QTHSF, RCRSF, TMRTF, and TPMKC) compared to Sr13 alleles. The phenotypic uniqueness of the stem rust resistance gene in KU168-2 and null alleles for Sr13 allele-specific markers showed the resistance was conferred by a new gene, designated Sr67. Since Sr13 is less effective in hexaploid background, Sr67 will be a good source of stem rust resistance in bread wheat breeding programs.

Registration of WGC002 spring wheat containing wild grass‐derived Fusarium head blight resistance gene Fhb7The2

AbstractThe USDA‐ARS and North Dakota State University Agricultural Experiment Station jointly released the Fusarium head blight (FHB)‐resistant spring wheat (Triticum aestivum L.) germplasm WGC002 (Reg. no. GP‐1089, PI 702949) in May 2023. WGC002 is a wheat‐Thinopyrum elongatum 7B‐7E translocation line, designated 7BS·7BL‐7EL, with the wheat chromosome 7BL terminal region replaced by the homoeologous counterpart of the Th. elongatum chromosome 7EL that contains the novel FHB resistance gene Fhb7The2. WGC002 was developed from the Chinese Spring (CS) wheat‐Th. elongatum disomic substitution line DS 7E(7B) using our genomics‐enabled chromosome engineering pipeline. The pedigree of WGC002 is DS 7E(7B)/2*CS ph1b mutant//DS 7E(7B). WGC002 has consistently exhibited resistance to FHB in inoculations of greenhouse grown plants. WGC002 does not contain the yellow flour pigment gene in the Fhb7The2 haplotype present on the terminal 7EL segment of the 7BS·7BL‐7EL translocation and has not exhibited any obvious linkage drag on the 7EL segment. Therefore, WGC002 is ready for immediate use in wheat breeding. One sequence‐tagged site (STS) and two polymerase chain reaction allelic competitive extension markers were developed specifically for Fhb7The2 and validated in different wheat genotypes. They are highly diagnostic for Fhb7The2 and extremely useful in marker‐assisted introgression of Fhb7The2 in wheat breeding. In summary, WGC002 is a new wild grass‐derived FHB‐resistant spring wheat germplasm with diagnostic DNA markers available to conduct marker‐assisted selection of Fhb7The2, that will enhance and diversify FHB resistance of wheat.

GENETIC DIVERSITY OF IRAQI LOCAL COWS AND THEIR COMPARISON WITH IMPORTED COWS USING MICROSATELLITE MARKERS

This study was conducted to identify the genetic diversity of microsatellite markers CSSM66, ETH10 and ILSTS34 using 100 cows of several breeds distributed as follows: 30 local Iraqi cows (equally divided into Restaki, Janoubi and Sharabi) as well as 50 Holstein cows (German) and 20 Brahman bulls after amplification in PCR reaction, 16 alleles of ETH10, CSSM66 and ILSTS34 markers were detected in a sample of studied cattle and they were 7, 6 and 5 alleles, respectively. The total Shannon (I) was 1.5400, and it was found that the average value of Obs_Hom is smaller than the average value of Exp_Het, being 0.2600 and 0.7544, respectively. This indicates the presence of high genetic heterogeneity or diversity among cows. The average values ​​of FIS, FIT and FST and genetic migration Nm -0.6257, -0.0162, 0.3749 and 0.4168, respectively. These results showed through the UPGMA tree a common ancestry between domestic cattle and Brahman.

ОСОБЕННОСТИ СИСТЕМНОГО ОТВЕТА НА АДЪЮВАНТНУЮ ЛУЧЕВУЮ ТЕРАПИЮ У НОСИТЕЛЕЙ ПОЛИМОРФИЗМА -308(G/A)TNF БОЛЬНЫХ РАКОМ МОЛОЧНОЙ ЖЕЛЕЗЫ

Background: Adjuvant radiation therapy (ART), an integral part of locoregional breast cance (BC) therapy, acting not only locally, but also systemically and leads to a shift in homeostasis, which is reflected in routine general clinical tests. Tumor necrosis factor (TNF) is a proinflammatory cytokine, the production of which can be influenced by the single-nucleotide substitution -308(G/A)TNF. The minor allele -308A can be included in the stable inherited haplotype AH8.1 of the HLA gene complex. At the same time, the carriage of the -308A without the AH8.1 haplotype is associated with a poor prognosis in patients with BC. The aim of the study was to evaluate the features of the systemic response to the course of ART in carriers of TNF-associated genotypes with BC. Material and methods: The sample is represented by 147 BC patients who underwent a course of ART (2 Gy in 25 fractions). Clinical and morphological characteristics and data of general clinical blood analysis were obtained from medical histories. Venous blood samples for the study were obtained at the beginning and at the end of the ART course. Alleles -308(G/A)TNF and marker alleles of haplotype AH8.1 (HLA-A×01, HLA-B×08 and HLA-DRB1×03) were determined by allele-specific PCR. sTNF concentrations were determined by the ELISA in 102 blood plasma samples. Results: TNF-associated comparison groups were identified based on genotyping: (1) 114 carriers -308GG of the TNF gene, regardless of the AH8.1 haplotype (77,6 %); (2) 23 carrier -308A(AH8.1pos) had at least one AH8.1 marker allele (15.6 %); (3) 10 carriers -308A(AH8.1neg) did not have any AH8.1 marker allele (6.8 %). In the -308A(AH8.1neg) group the average concentration of sTNF both at the beginning and at the end of ART was significantly higher and, unlike other comparison groups, did not significantly decrease at the end of the ART course. A significant decrease in absolute values was revealed during ART in a number of cases for leukocytes, platelets and lymphocytes, however within the reference values. In the group -308A(AH8.1neg) correlation analysis revealed a high strength of positive connections between sTNF and leukocytes (r=0.71; p=0.027), platelets (r=0.67; p=0.04), neutrophils (r=0.70; p=0.027) only at the end of ART, whereas at the beginning ART these correlations were weak (r≤0.3) and statistically unreliable. For other genetic groups, the revealed correlations were not strong enough. Conclusion: The revealed features of the systemic response to ART for carriers of a prognostically unfavorable genotype -308A(AH8.1neg) – a high concentration of sTNF and a positive correlation with the content of leukocytes (probably due to neutrophils) and platelets – can be considered as targets of individualized therapy.

The Favorable Allele of CAPN1-316 Genetic Marker is Absent in Bali and Sumbawa Cattle

The micromolar calcium-activated neutral protease 1 (CAPN1) gene encodes the u-calpain enzyme, which plays a crucial role in meat tenderisation. Genetic diversity within the CAPN1 gene, specifically a nucleotide substitution from G to C in exon nine resulting in a change from glycine to alanine at position 316 (CAPN1-316 marker), is known to significantly affect meat tenderness. This study aimed to assess the polymorphism of the CAPN1-316 locus in Bali and Sumbawa cattle. A total of 293 blood samples, 193 from Bali cattle and 100 from Sumbawa cattle were extracted and genotyped using PCR-RFLP with BtgI restriction enzyme (recognition sequence: 5'-C*CRYGG-3') applied to 706 bp PCR products. The results showed the presence of only one genotype (GG genotype) and one allele (G allele) in all DNA samples obtained from the Bali and Sumbawa cattle populations studied. In conclusion, the CAPN1-316 genetic marker showed a lack of diversity or monomorphism in Bali and Sumbawa cattle, making it unsuitable for further association studies in these breeds. Consequently, the CG/AG haplotype identified in Sumbawa cattle warrants further investigation and could serve as an alternative genetic marker, especially due to its monomorphism at the CAPN1-316 locus.

Features of Distribution of the Allelic Variant of the OAS1 Gene Associated with Severe Form of the Coronavirus Infection in the Russian and Global Populations.

The study of the geographic distribution of the allelic variant of the OAS1 gene associated with severe form of the infections caused by RNA viruses was carried out using the rs10774671 polymorphic locus. The mutant allele encoding the p42 protein isoform was most prevalent in the Russian populations. A comparative analysis of the prevalence of the mutant allele in world populations showed that its frequency is 0.9 among the inhabitants of Northern Eurasia, while the allele encoding the p46 protein isoform is widespread among the population of West Central Africa. A cartographic analysis of the relationship between the population-frequency characteristics of the marker alleles and the geographical remoteness of the populations showed that the mutant allele is most often observed in the indigenous populations of the Far East, which suggests its East Asian origin.