AGRIS - 

2024

A. L. Kryazhev | A. S. Novikov

The purpose of the research is isolation, identification, and analysis of ASV (Amplicon Sequence Variant) types of Cryptosporidia spp. in pigs in the Vologda Region of the Russian Federation.Materials and methods. The research has been conducted in the Russian Federation for the first time. The research was conducted on pig farms in the Vologda Region of the Northwestern Federal District of the Russian Federation from January to October 2023. Feces were taken from piglets of various age groups, as well as milking sows. The samples were studied using the equipment of the resource center “Genomic Technologies, Proteomics and Cell Biology” of ARRIAM. Species of the genus Cryptosporidia were identified in fecal samples using high-throughput sequencing of 18S rRNA gene fragment amplicon libraries as obtained from nested PCR followed by “denoising”, sequence combining, and restoring the original phylotypes (ASV, (Amplicon Sequence Variant)).Results and discussion. Cryptosporidia spp. species were identified in each age group studied. As a result of high-throughput sequencing of the libraries using the Illumina technology, 20 to 100 thousand nucleotide sequences (reads) were obtained for each sample after processing of which a total of 2,372 ASVs were identified. The analysis of the ASV taxonomic affiliation performed with phylogenetic analysis supplemented by an analysis using the blastn algorithm in the GenBank database showed that, in total, 10 ASVs were only present in all studied samples that had high similarity to sequences deposited in the GenBank as 18S rRNA gene fragments of Cryptosporidium scrofarum. Eight ASV types were unique and did not repeat from farm to farm. Probably, these sequences belong to local populations of C. scrofarum subspecies. Of interest is the discovery of a unique Cryptosporidium sequence of ASV8 type which is only 91.47% similar to the closest relative of the genus, which may indicate a rather distant taxonomic relationship. This type of nucleotide sequence can be further described as a new species. All identified unique ASV nucleotide sequences were deposited in GenBank.

Show more [+]

Less [-]

Directory of Open Access Journals - Sweden

2023

T. A. Bazanov | I. V. Ushapovsky | N. N. Loginova | E. V. Smirnova | P. D. Mikhaylova

The genetic diversity of crop varieties should be assessed using modern molecular markers. One of the most promising and simple methods of plant genetic analysis is intermicrosatellite labeling (ISSR). The purpose of this research is to study the polymorphism of varieties of fiber flax and linseed flax included in the State Register of Breeding Achievements of the Russian Federation, as well as local forms of fiber flax using ISSR markers. The objects for the research were 83 samples from the collection of the Federal Research Center for Bast Fiber Crops, among them 24 varieties of linseed flax, 56 varieties of fiber flax of domestic and foreign breeding, and three local forms from the Central Non-Black Earth Region. Genetic analysis was carried out by PCR method using a set of 25 ISSR primers. Detection of amplification products was carried out using electrophoresis in 1.5% agarose gel. Based on intraspecific selection of flax varieties to oilseed and fiber forms, a cluster analysis was performed according to the results of ISSR labeling independently in each of two groups that allowed to increase the reliability of conclusions on the existing dependences. Dendrograms of the genetic similarity of the studied samples of linseed flax and fiber flax were studied independently from each other. As a result of the analysis, linseed flax varieties were divided into five clusters, which made it possible to establish dependencies related to their origin (originator of the variety) and oil content in the seed. The dendrogram which included varieties of fiber flax and its local forms, revealed twelve subclusters forming three uneven groups. Almost every group had its own characteristic feature, not related to the geography of origin. The grouping of varieties occurred mainly on the basis of their various characteristics in terms of flax fiber content, ripening time and resistance to environmental stress factors. The results of the research show the applicability of using the ISSR-markers as a valid and accessible method for developing the elements of marker-associative breeding of bast fiber crops.

Show more [+]

Less [-]

Directory of Open Access Journals - Sweden

2023

A. L. Kryazhev | A. S. Novikov

The purpose of the research is to determine infection rate and oocyst isolation degree followed by identification of taxa of Cryptosporidium species in piglets of different age groups using the latest molecular genetic methods, on private farms in the Vologda Region of the North-Western Federal District of the Russian Federation.Materials and methods. These studies were performed in the Russian Federation for the first time. The research was performed on private pig farms located in the Vologda Region of the North-Western Federal District of the Russian Federation from January to September 2022. Feces were taken from piglets of different age groups, namely, sucklings under the age of 1 month, weaners (1–3 months), feeder pigs (4 months and older), as well as from milking sows. Age groups were formed taking into consideration technological parameters of keeping animals on farms. Using microscopic research methods, “positive” samples were detected in which Cryptosporidium oocysts were found, and the number of oocysts was determined. Subsequently, the samples were studied using the equipment of the resource center «Genomic Technologies, Proteomics and Cell Biology» of ARRIAM. Cryptosporidium species were identified in samples of animal feces using high-throughput sequencing of 18S rRNA gene fragment amplicon libraries as obtained by nested PCR.Results and discussion. Cryptosporidium species were identified in each studied age group both in the animal’s presenting indigestion and the animals without any clinical sign of the disease. The average cryptosporidium infection rate was 32.4% in the animals on private farms. The most infected with cryptosporidium oocysts were feeder pigs aged 4–6 months (72%). As a result of sequencing of 18S rRNA gene fragment amplicon libraries obtained using selected primers and subsequent taxonomic analysis of the resulting nucleotide sequences, it was shown that only representatives of the Cryptosporidium scrofarum species were present in all the studied samples.

Show more [+]

Less [-]

Directory of Open Access Journals - Sweden

2023

Belan, I.A. | Rоssеeva, L.P. | Mukhordova, M.E. | Ignat'eva, E.Yu. | Fedorenko, E.N.

The research is aimed at studying economically valuable traits and genetic control of resistance to leaf diseases, photoperiodic response and short stature of soft spring wheat variety 'Semenovna', created by scientists of the Omsk Agrarian Scientific Center (Russia) and the North Kazakhstan Agricultural Experimental Station (Kazakhstan). Using methods of state variety testing, molecular genetic and in vitro methods, morphological features of the new variety, features of its development have been described, the yield level at different ecological test points for 3 years (2015-2017) has been analyzed. The studies conducted at two ecological points made it possible to select a mid ripening promising line of soft spring wheat ('Lutescens 354/04-6'), which was transferred to the State Registration Service of the Republic of Kazakhstan and after two years of testing was included into the State Register of the Republic of Kazakhstan under the name 'Semenovna' (patent No. 1023). In terms of grain quality, it was at the level of valuable wheat, exceeded the standard in a protein content by 1-2% and raw gluten by 3-4%. The new med-ripening 'Semenovna' variety combines a higher yield (2.73-4.40 t/ha) with resistance to drought (resistance index Ir=0.57), brown and stem rust (Ir=0.00-0.23). The genotype of the variety contains wheat-rye translocation 1RS.1BL (with a cluster of Lr26/Sr31/Pm8/Yr9 genes). The medium-stem variety carries the Rht8b allele in its genotype (174 bp) and is photosensitive to the day length (allele 414 bp). The parameters of ecological plasticity of the new variety are determined: linearity regression coefficient (bi=1.08), stability index (sigma d2=0.27). | Исследование направлено на изучение хозяйственно ценных признаков и генетического контроля устойчивости к листостебельным заболеваниям, фотопериодической реакции и короткостебельности сорта пшеницы мягкой яровой Семеновна, созданного учеными Омского аграрного научного центра (Россия) и Северо-Казахстанской с.-х. опытной станции (Казахстан). С использованием методов государственного сортоиспытания, молекулярно-генетических и in vitro описаны морфологические признаки нового сорта, особенности его развития, проанализирован уровень урожайности в разных экологических точках испытаний за 3 года (2015-2017). Исследования, проведенные в двух экологических точках, позволили отобрать среднеспелую перспективную линию пшеницы мягкой яровой (Lutescens 354/04-6), которая передана в Государственную регистрационную службу Республики Казахстан и после двухлетнего тестирования была включена в Государственный реестр Республики Казахстан под названием Семеновна (патент № 1023). По показателям качества зерна она была на уровне ценной пшеницы, превосходила стандарт по содержанию белка на 1-2% и сырой клейковины на 3-4%. Новый среднеспелый сорт Семеновна сочетает повышенную урожайность (2,73-4,40 т/га) с устойчивостью к засухе (индекс устойчивости Ir=0,57), бурой и стеблевой ржавчине (ИУ=0,00-0,23). Генотип сорта содержит пшенично-ржаную транслокацию 1RS.1BL (с кластером генов Lr26/Sr31/Pm8/Yr9). Среднестебельный сорт несет в своем генотипе аллель Rht8b (174 п.н.) и светочувствителен к продолжительности дня (аллель 414 п.н.). Определены параметры экологической пластичности нового сорта: коэффициент линейной регрессии (bi=1,08), показатель стабильности (сигмаd2=0,27).

Show more [+]

Less [-]

Central Scientific Agricultural Library - Russian Federation

2023

Semenova, O.V. | Koteneva, S.V. | Nefedchenko, A.V. | Sudorgina, T.E. | Glotova, T.I. | Glotov, A.G.

The research was carried out in 2020-2022 at a farm specialized in milk production. The total number of the herd is 1,750 heads. An outbreak of infection caused by Pestivirus H (bovine viral diarrhea virus, BVDV-3, that is agent of mucosal disease of the third kind) is described. The disease was accompanied with high morbidity and lethality among animals of different ages. In some sick animals, a full complex of pronounced symptoms characteristic of "classical" bovine mucosal disease was registered: erosions and ulcers on the nasal mirror and tongue, foaming from the mouth, serous nasal discharge, hemorrhagic inflammation and pronounced longitudinal erosions on the mucosa of the esophagus, rennet stomach and intestine. Cows miscarried at different stages of pregnancy. The coefficient of effective insemination decreased to 20%. The course of the disease was complicated by the involvement of the bovine herpes virus type 4, bacteria of the family Pasteurellaceae and Clostridium spp. in the infectious process. The BVDV-3 genome was found in a wide range of internal organs of aborted fetuses, calves, and adult animals. According to sequencing data, the pathogen was classified as subtype 3a. Phylogenetic analysis of the 5'-untranslated region of the virus genome (5'-UTR) showed its close relationship to the strains isolated in Italy and Brazil, most of which were previously identified as contaminants of fetal bovine serum and live vaccines against viral infections of cattle. No specific prophylaxis against BVDV-3 infection has been developed at this time, therefore, it is necessary to update and improve diagnostic methods, optimize control measures to prevent the spread of virulent pathogen strains, and to control safety of the vaccines used. | Исследования проводили в 2020-2022 гг. в хозяйстве, специализирующемся на производстве молока. Общая численность стада – 1750 гол. Описана вспышка инфекции, вызванной Pestivirus H (вирус вирусной диареи – болезни слизистых оболочек третьего вида, BVDV-3), которая сопровождалась высокой заболеваемостью и летальностью среди животных разных возрастов. У части больных животных зарегистрировали полный комплекс ярко выраженных симптомов, характерных для "классической" болезни слизистых оболочек крупного рогатого скота: эрозии и язвы на носовом зеркальце и языке, выделение пены из ротовой полости, серозные выделения из носа, геморрагическое воспаление и выраженные продольные эрозии на слизистой пищевода, сычуга и кишечника. Коровы абортировали на разных стадиях стельности. Коэффициент плодотворного осеменения снизился до 20%. Течение болезни осложнилось вовлечением в инфекционный процесс вируса герпеса крупного рогатого скота 4-го типа, бактерий семейства Pasteurellaceae и Clostridium spp. Геном BVDV-3 обнаружили в широком спектре внутренних органов абортированных плодов, телят и взрослых животных. По данным секвенирования возбудитель отнесли к субтипу 3а. Филогенетический анализ участка 5'-нетранслируемой области генома вируса (5'-UTR) показал близкое его родство со штаммами, выделенными в Италии и Бразилии, большинство из которых ранее идентифицированы как контаминанты эмбриональной сыворотки и живых вакцин против вирусных инфекций крупного рогатого скота. В настоящее время средства специфической профилактики против инфекции, вызванной BVDV-3, не разработаны, поэтому необходимы обновление и совершенствование методов диагностики, оптимизация противоэпизоотических мероприятий для недопущения распространения вирулентных штаммов возбудителя, контроль безопасности используемых вакцин.

Show more [+]

Less [-]

Central Scientific Agricultural Library - Russian Federation

2023

Il'nitskaya, E.T. | Makarkina, M.V. | Kozina, T.D. | Kozhevnikov, E.A. | Petrov, V.S.

Currently, when implementing breeding programs to produce pathogen-resistant grape genotypes, the DNA marker assessment method is used both directly in breeding-oriented selection and in evaluation of the initial gene pool. The 'Talisman' grape cultivar, with its fine consumer characteristics, resistance to diseases and frost, and a functionally female type of flower, is promising for breeding. We performed a DNA marker assessment of the 'Talisman' genotype and cultivars as well as new hybrids of grapes developed with the participation of cv. 'Talisman' for the presence of the Rpv3 and Rpv12 downy mildew resistance loci. It is known that the presence of these two genes in the same grape genotype has an additive effect. According to the pedigree of cv. 'Talisman' (Frumoasa Albe x Vostorg), the presence of these genes can be assumed in the studied sample. The study was based on PCR with an analysis of the results on an automatic genetic analyzer. DNA was isolated from young shoots of the analyzed plants by the CTAB method. There were used closely linked microsatellite markers recommended for DNA identification of the allelic status of the Rpv3 (UDV305, UDV737) and Rpv12 (UDV343, UDV360) genes. The simultaneous presence of Rpv3 and Rpv12 was detected only in the cv. 'Talisman' genotype. When analyzing the progeny genotypes of cv. 'Talisman', the presence of the Rpv3 downy mildew resistance gene was found in the grape hybrids "Agat Dubovskiy", "Pestry", and Rpv12 gene in the 'Viktor' and 'Preobrazheniye' genotypes. DNA-marker analysis confirmed the prospects of the cv. 'Talisman' genotype for breeding of table cultivars as a donor of the downy mildew resistance genes Rpv3 and Rpv12. | В настоящее время при реализации селекционных программ по созданию устойчивых к патогенам генотипов винограда используют методику ДНК-маркерной оценки как непосредственно в селекционном отборе, так и при оценке исходного генофонда. Сорт винограда 'Талисман', обладая высокими потребительскими характеристиками, устойчивостью к болезням и морозу и функционально женским типом цветка, является привлекательным для селекции. Выполнена ДНК-маркерная оценка генотипа 'Талисман' и сортов, а также новых гибридных форм винограда, созданных с участием сорта 'Талисман' на наличие локусов устойчивости к милдью Rpv3 и Rpv12. Известно, что наличие этих двух генов в одном генотипе винограда имеет аддитивный эффект. Согласно родословной сорта 'Талисман' (Фрумоаса Албэ x Восторг), можно предполагать наличие данных генов в исследуемой выборке. Исследование проведено методом ПЦР с анализом результатов на автоматическом генетическом анализаторе. ДНК выделяли из молодых побегов анализируемых растений методом ЦTAБ. В работе использованы тесно сцепленные микросателлитные маркеры, рекомендованные для ДНК-идентификации аллельного состояния генов Rpv3 (UDV305, UDV737) и Rpv12 (UDV343, UDV360). Одновременное присутствие Rpv3 и Rpv12 определено только в генотипе сорта 'Талисман'. При анализе генотипов – потомков этого сорта наличие гена устойчивости к милдью Rpv3 определено в гибридах винограда под наименованиями "Агат дубовский", "Пёстрый" и гена Rpv12 – в генотипах 'Виктор', 'Преображение'. ДНК-маркерный анализ подтверждает перспективность генотипа 'Талисман' для селекции столовых сортов как донора генов устойчивости к милдью Rpv3 и Rpv12.

Show more [+]

Less [-]

Central Scientific Agricultural Library - Russian Federation

2023

Tarasova, E.V. | Кapitova, I.A. | Evdokimenko, S.N. | Nemtsova, E.V.

The article presents the results of DNA testing for 21 raspberry cultivars using the ISSR-PCR method to study genetic polymorphism and identify genotypes potentially resistant to the raspberry bushy dwarf virus (RBDV). It was revealed that promising hybrids of everbearing raspberry (Rubus idaeus L.), including 9-155-1, 11-165-10 and 9-163-1, exhibited significant genetic relations with RBDV-resistant cultivars. Hybrids 9-163-1 and 11-165-10, both having the hybrid 13-118-1 as one of the parent forms, were grouped with the RBDV-resistant cultivar 'Poklon Kazakovu'. These forms showed similarities with the resistant cultivars 'Meteor' and 'Atlant', as well as the sensitive cultivars 'Evrasia' and 'Podarok Kashinu'. The hybrid 9-155-1 was characterized by genetic proximity with the sensitive cultivars 'Podarok Kashinu' and 'Evrasia', as well as the resistant cultivars 'Meteor', 'Atlant', 'Zhar-Ptitsa' and 'Heritage'. The 'Heritage' variety is known to be resistant to RBDV, attributed to the Bu gene. This gene is responsible for the resistance to the virus, including the most dangerous isolate R15. The analysis of the constructed dendrites suggested the potential resistance to RBDV in the hybrid 9-155-1. | Проведено молекулярно-генетическое исследование 21 сорта малины методом ISSR-PCR. Целью работы было изучение генетического полиморфизма и поиск генотипов, потенциально устойчивых к вирусу кустистой карликовости малины (RBDV - Raspberry bushy dwarf virus). В процессе исследования выявлено, что перспективные гибриды ремонтантной малины (Rubus idaeus L.) 9-155-1, 11-165-10 и 9-163-1 обладали наибольшим генетическим родством с устойчивыми к RBDV сортами. Родственные гибриды 9-163-1 и 11-165-10, одной из родительских форм которых являлся гибрид 13-118-1, группировались с устойчивым к RBDV сортом Поклон Казакову. Данные формы имели сходство с устойчивыми сортами Метеор, Атлант, а также с чувствительными сортами Евразия и Подарок Кашину. Гибрид 9-155-1 характеризовался генетической близостью с чувствительными сортами Подарок Кашину и Евразия, а также устойчивыми сортами Метеор, Атлант, Жар-Птица и Heritage. Сорт Heritage известен как устойчивый к RBDV за счёт наличия гена Bu. Этот ген обуславливает устойчивость к вирусу, в том числе к наиболее опасному изоляту R15. Анализ построенных дендритов позволил предположить наличие устойчивости к RBDV у гибрида 9-155-1.

Show more [+]

Less [-]

Central Scientific Agricultural Library - Russian Federation

2023

ХАУСТОВ, Евгений

Studies have been carried out to study the process of the spread of the phytoplasmic disease of Bois noir on grape plantations of the Republic of Moldova. It has been established that the necessary factors for the spread of the disease are: the presence of plants-reservers of the disease, leafhoppers-vectors and the appropriate climatic conditions that contribute to the premature drying of the grass cover, which forces the leafhopper vectors to change to other food sources, including grapevine.

Show more [+]

Less [-]

Scientific-Practical Institute of Horticulture and Food Technologies - Republic of Moldova

2023

ГАШЕНКО, О.А | КУХАРЧИК, Н.В. | ПРОДАНЮК, Л.Н. | ЧЕРНЕЦ, A.M.

Field and micropropagated plants were tested for the presence of RBDV by DAS-ELISA and PCR. For in vitro therapy, plants with a proven absence of ArMV, ApMV, RpRSV, TBRV, TomRSV, SLRV, CMV, TRSV and infected with Raspberry bushy dwarf virus (RBDV) were selected.Virazole and salicylic acid at concentrations of 30, 50 and 70 mg/l were used as antiviral preparation.Low phytotoxicity of salicylic acid and high phytotoxicity of virazole were established. Long-term use of virazole, at concentrations of 50 and 70 mg/l, by the end of the 3rd passage leads to the complete death of microshoots,The studies were carried out within the framework of the project „Development of biotechnological methods (in vitro and ex vitro) obtaining virus-free material from the raspberry bushy dwarf virus” in order to increase the productivity of industrial raspberry plantations.

Show more [+]

Less [-]

Scientific-Practical Institute of Horticulture and Food Technologies - Republic of Moldova

2023

A. L. Kryazhev | A. S. Novikov

The purpose of the research is to determine taxa of the genus Cryptosporidium species in pigs using molecular genetic methods in the north-west of the Russian Federation.Materials and methods. Fecal samples were taken from pigs of different age groups on farms of different types of incorporation that differ in climatic and geographical zones and animal keeping technologies in the Vologda Region. Microscopic research methods identified “positive” samples in which Cryptosporidium species were present; they were sorted out and deep-frozen. Then the samples were examined using the equipment of the resource center «Genomic Technologies, Proteomics and Cell Biology» of ARRIAM. The Cryptosporidium species in the fecal samples from farm animals were identified using high-throughput sequencing of 18S rRNA gene amplicon libraries obtained by a nested polymerase chain reaction (PCR) assay.Results and discussion. A primer system was designed for the nested PCR to amplify a potentially species-specific 393 bp fragment of the 18S rRNA gene. The sequence of the ILL_R2_ Zheng primer was modified with included degenerated positions to make the primer more versatile. As a result of sequencing of the libraries of 18S rRNA gene fragments obtained with the selected primers and subsequent taxonomic analysis of the nucleotide sequences, it was shown that all the studied samples included representatives of only one species, Cryptosporidium scrofarum.

Show more [+]

Less [-]

Directory of Open Access Journals - Sweden