izvestiiatimacadИзвестия Тимирязевской сельскохозяйственной академииIZVESTIYA OF TIMIRYAZEV AGRICULTURAL ACADEMY0021-342XФГБОУ ВО РГАУ-МСХА имени К.А. Тимирязева10.26897/0021-342X-2023-5-75-87izvestiiatimacad-463Research ArticleГЕНЕТИКА, БИОТЕХНОЛОГИЯ, СЕЛЕКЦИЯ И СЕМЕНОВОДСТВОGENETICS, BIOTECHNOLOGY, BREEDING AND SEED PRODUCTIONАнализ иммунореактивности авенинов сортов овса посевного (Avena sativa L.), возделываемых в Западной СибириAnalysis of the immunoreactivity of avenins in oats (Avena sativa L.) cultivated in Western Siberiahttps://orcid.org/0000-0002-1570-9595ЛюбимоваА. В.LyubimovaA. V.

Любимова Анна Валерьевна, заведующий лабораторией геномных исследований в растениеводстве, канд. биол. наук

625026, Тюменская обл., г. Тюмень, ул. Малыгина, 86

тел.: 8 (952) 34–108–87

Anna V. Lyubimova, CSc (Bio), Head of the Laboratory of Genomic Research in Crop Production

86, Malygina Str., Tyumen, 625026

phone: +7 (952) 34–108–87

ostapenkoav88@yandex.ruhttps://orcid.org/0000-0002-3672-6060ЕрёминД. И.EreminD. I.

Ерёмин Дмитрий Иванович, ведущий научный сотрудник лаборатории геномных исследований в растениеводстве, д-р биол. наук, доцент

625026, Тюменская обл., г. Тюмень, ул. Малыгина, 86

тел.: 89129271386

Dmitriy I. Eremin, DSc (Bio), Leading Research Associate, Laboratory of Genomic Research in Crop Production

86, Malygina Str., Tyumen, 625026

phone: +7 (912) 927–13–86

soil-tyumen@yandex.ruhttps://orcid.org/0000-0003-2923-9448ФоминаМ. Н.FominaM. N.

Фомина Мария Николаевна, ведущий научный сотрудник лаборатории селекции зернофуражных культур, канд. с.-х. наук

625026, Тюменская обл., г. Тюмень, ул. Малыгина, 86

тел.: (3452) 76–40–54

Maria N. Fomina, CSc (Ag), Leading Research Associate, Laboratory of Breeding Grain and Forage Crops

86, Malygina Str., Tyumen, 625026

phone: +7 (3452) 76–40–54

maria_f72@mail.ruФедеральное государственное бюджетное учреждение науки «Федеральный исследовательский центр Тюменский научный центр Сибирского отделения Российской академии наукРоссияFederal Research Centre “The Tyumen Scientific Centre of the Siberian Branch of the Russian Academy of Sciences”Russian Federation202307112023057587Copyright © Любимова А.В., Ерёмин Д.И., Фомина М.Н., 20242024Любимова А.В., Ерёмин Д.И., Фомина М.Н.Lyubimova A.V., Eremin D.I., Fomina M.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://izvestiia.timacad.ru/jour/article/view/463

Целиакия – это аутоиммунное заболевание, возникающее у генетически восприимчивых людей после употребления в пищу проламинов злаков. Единственное средство лечения целиакии – пожизненная безглютеновая диета, но ее соблюдение также может негативно сказаться на состоянии здоровья. Хорошим решением может быть включение в такую диету продуктов из овса. Однако разные сорта овса отличаются по своей иммуногенности, и важнейшее значение имеет изучение иммунореактивности их проламинов для выявления безглютеновых генотипов. Целью исследований стала оценка иммуногенности сортов и перспективных селекционных линий овса посевного, возделываемых на территории Тюменской области в период с 1929 по 2023 гг. для использования в селекции безглютеновых сортов. Определение количества глютена в зерне осуществляли методом иммуноферментного анализа с использованием тест-систем, основанных на применении антител R5 и G12. В ходе исследований было выявлено, что ни один из образцов не проявил иммунореактивности по отношению к антителу R5. При использовании антитела G12 установлено, что все проанализированные сорта относятся к безглютеновым: содержание глютена в них варьирует от 2,14 (Нидар) до 9,55 (ТМ 16–58–4) мг/кг. Особого внимания заслуживает сорт Орёл, содержание глютена в зерне которого – менее 2 мг/кг. Этот генотип является перспективным для включения в селекционный процесс на получение безглютеновых сортов. Среднее содержание глютена в образцах местной селекции составляет 5,23±0,678 мг/кг и достоверно превышает этот показатель у иностранных и инорайоннных сортов, что связано с введением в скрещивания более аллергенных сортов. Иммуногенность сортов связана с конкретными аллелями авенин-кодирующих локусов. Аллели А11, В11 и С2 могут быть перспективны в качестве маркеров безглютеновых сортов. Для успешного развития селекции безглютеновых сортов овса необходимы анализ аллельного состояния авенин-кодирующих локусов исходных генотипов и определение содержания в них глютена с антителом G12.

Celiac disease is an autoimmune disease that occurs in genetically susceptible people after eating prolamin-containing grains. The only cure for celiac disease is a lifelong glutenfree diet, but adherence to this diet can also have a negative impact on health. A good solution may be to include oat products in such a diet. However, different oat varieties differ in their immunogenicity, and studying the immunoreactivity of their prolamins to identify gluten-free genotypes is of utmost importance. The aim of the research was to assess the immunogenicity of varieties and promising breeding lines of oats cultivated in the Tyumen region in the period from 1929 to 2023 for use in the selection of gluten-free varieties. The amount of gluten in the grain was determined by enzyme-linked immunosorbent assay using test systems based on the use of R5 and G12 antibodies. It was found that none of the samples showed immunoreactivity to the R5 antibody. Using the G12 antibody, all the varieties analyzed were found to be gluten-free, with gluten content ranging from 2.14 (Nidar) to 9.55 (TM 16–58–4) mg/kg. The variety Orel deserves special attention, as the gluten content of its grain is less than 2 mg/kg. This genotype is promising for inclusion in the breeding process to produce glutenfree varieties. The average gluten content in the samples of local selection is 5.23±0.678 mg/kg and significantly exceeds this index in foreign and non-regional varieties, which is associated with the introduction of more allergenic varieties in crossbreeding. The immunogenicity of varieties is associated with specific alleles of avenin-coding loci. Alleles A11, B11 and C2 may be promising as markers for gluten-free varieties. For the successful development of breeding of gluten-free oat varieties, it is necessary to analyze the allelic state of avenin-coding loci of the original genotypes and to determine their gluten content with the G12 antibody.

овес (Avena sativa L.)целиакиябезглютеновая диетаглютенавениниммуноферментный анализантитело R5 и G12авенин-кодирующий локусoats (Avena sativa L.)celiac diseasegluten-free dietglutenaveninenzyme-linked immunosorbent assayR5 and G12 antibodiesavenin-coding locusРабота выполнена по госзаданию №122011300103-0 и при поддержке Западно-Сибирского межрегионального научно-образовательного центра мирового уровня.ReferencesЛюбимова А.В., Еремин Д.И., Мамаева В.С. и др. Каталог биохимических паспортов сортов овса посевного сибирской селекции // Вестник КрасГАУ. – 2022. – № 5(182). – С. 73-83. – DOI 10.36718/1819-4036-2022-5-73-83.Lyubimova A.V., Eremin D.I., Mamaeva V.S. i dr. Siberian oat varieties' biochemical passports catalog // Bulletin of KrasSAU. 2022; 5(182): 73-83. (In Rus.) DOI 10.36718/1819-4036-2022-5-73-83.Методы определения глютена в продовольственном сырье и пищевых продуктах: Методические указания. – М.: Федеральный центр гигиены и эпидемиологии Роспотребнадзора, 2011. – 32 с.Metody opredeleniya glyutena v prodovol'stvennom syr'e i pishchevyh produktah: Metodicheskie ukazaniya [Methods for determining gluten in food raw materials and food products: Guidelines]. M.: Federal'nyj centr gigieny i epidemiologii Rospotrebnadzora, 2011. 32 p. (In Rus.)Amador M., Iehisa J., Martín C., Losada F. Functional genomic characterization of immunogenic gluten proteins from oat cultivars that differ in toxicity for celiac disease // Proceedings of the Nutrition Society. – 2020. – 79(OCE2). – E550. DOI 10.1017/S0029665120004991Amador M., Iehisa J., Martín C., Losada F. Functional genomic characterization of immunogenic gluten proteins from oat cultivars that differ in toxicity for celiac disease. Proceedings of the Nutrition Society. 2020; 79(OCE2): E550. DOI 10.1017/S0029665120004991Cardo A., Churruca I., Lasa A. et al. Nutritional Imbalances in Adult Celiac Patients Following a Gluten-Free Diet // Nutrients. – 2021. – №13. – Pp. 2877. DOI 10.3390/nu13082877Cardo A., Churruca I., Lasa A. et. al. Nutritional Imbalances in Adult Celiac Patients Following a Gluten-Free Diet. Nutrients. 2021; 13: 2877. DOI 10.3390/nu13082877CODEX STAN WHO/FAO (2008) Standard for foods for special dietary use for persons intolerant to gluten. CODEX STAN 118–1979. Adopted in 1979. Amendment: 1983 and 2015. URL: https://www.fao.org/fao-who-codexalimentarius/sh-proxy/en/?lnk=1&url=https%253A%252F%252Fworkspace.fao.org%252Fsites%252Fcodex%252FStandards%252FCXS%2B118-1979%252FCXS_118e_2015.pdf (Дата обращения: 10.09.2023)CODEX STAN WHO/FAO (2008) Standard for foods for special dietary use for persons intolerant to gluten. CODEX STAN 118–1979. Adopted in 1979. Amendment: 1983 and 2015. URL: https://www.fao.org/fao-who-codexalimentarius/sh-proxy/en/?lnk=1&url=https%253A%252F%252Fworkspace.fao.org%252Fsites%252Fcodex%252FStandards%252FCXS%2B118-1979%252FCXS_118e_2015.pdf (Date of the application: 10.09.2023)Cohen I.S., Day A.S., Shaoul R. To Be Oats or Not to Be? An Update on the Ongoing Debate on Oats for Patients with Celiac Disease // Front. Pediatr. –2019. – №7. – Pp. 384. DOI 10.3389/fped.2019.00384Cohen I.S., Day A.S., Shaoul R. To Be Oats or Not to Be? An Update on the Ongoing Debate on Oats for Patients with Celiac Disease. Front. Pediatr. 2019; 7: 384. DOI 10.3389/fped.2019.00384Col Hoffmanová I., Sánchez D., Szczepanková A. et al. The Pros and Cons of Using Oat in a Gluten-Free Diet for Celiac Patients // Nutrients. – 2019. – №11. – Pp. 2345. DOI 10.3390/nu11102345Col Hoffmanová I., Sánchez D., Szczepanková A. et. al. The Pros and Cons of Using Oat in a Gluten-Free Diet for Celiac Patients. Nutrients. 2019; 11: 2345. DOI 10.3390/nu11102345Colombo F., Di Lorenzo C., Biella S., et al. Ancient and Modern Cereals as Ingredients of the Gluten-Free Diet: Are They Safe Enough for Celiac Consumers? // Foods. – 2021. – №10(4). – Pp. 906. DOI 10.3390/foods10040906.Colombo F., Di Lorenzo C., Biella S., et al. Ancient and Modern Cereals as Ingredients of the Gluten-Free Diet: Are They Safe Enough for Celiac Consumers? Foods. 2021; 10(4): 906. DOI 10.3390/foods10040906.Comino I., Real A., de Lorenzo L. et al. Diversity in oat potential immunogenicity: basis for the selection of oat varieties with no toxicity in coeliac disease // Gut. – 2011. – №60(7). – Pp. 915-22. DOI: 10.1136/gut.2010.225268.Comino I., Real A., de Lorenzo L. et al. Diversity in oat potential immunogenicity: basis for the selection of oat varieties with no toxicity in coeliac disease. Gut. 2011; 60(7): 915-22. DOI: 10.1136/gut.2010.225268.Comino I., Torres M. I., Ruiz-Carnicer A. et al. Comparative reactivity of avenins from different pure oat varieties to gluten R5 and G12 ELISA immunomethods // Proceeding of the 32 Meeting, working group on prolamin analysis and toxicity: edited by Peter Koehler. United Kingdom, 27-29 september 2018. – United Kingdom, 2019. – Pp. 39-43.Comino I., Torres M. I., Ruiz-Carnicer A. et al. Comparative reactivity of avenins from different pure oat varieties to gluten R5 and G12 ELISA immunomethods. Proceeding of the 32 Meeting, working group on prolamin analysis and toxicity: edited by Peter Koehler. United Kingdom, 27-29 september 2018. United Kingdom, 2019: 39-43.Daly M., Bromilow S. N., Nitride C. et al. Mapping Coeliac Toxic Motifs in the Prolamin Seed Storage Proteins of Barley, Rye, and Oats Using a Curated Sequence Database // Frontiers in Nutrition. – 2020. – №7. DOI 10.3389/fnut.2020.00087Daly M., Bromilow S. N., Nitride C. et al. Mapping Coeliac Toxic Motifs in the Prolamin Seed Storage Proteins of Barley, Rye, and Oats Using a Curated Sequence Database. Frontiers in Nutrition. 2020; 7. DOI 10.3389/fnut.2020.00087Dvořáček V., Kotrbová-Kozak A., Kozová-Doležalová J. et al. Identification of Perspective Oat Cultivars with a Minimum Content of Gluten Homologous Peptides // Preprints. – 2020. – Pp. 2020100596. DOI 10.20944/preprints202010.0596.v1Dvořáček V., Kotrbová-Kozak A., Kozová-Doležalová J. et al. Identification of Perspective Oat Cultivars with a Minimum Content of Gluten Homologous Peptides. Preprints. 2020; :2020100596. DOI 10.20944/preprints202010.0596.v1Gell G., Bugyi Z., Florides C. G. et al. Investigation of Protein and Epitope Characteristics of Oats and Its Implications for Celiac Disease // Frontiers in Nutrition. – 2021. – №8. DOI 10.3389/fnut.2021.702352Gell G., Bugyi Z., Florides C. G. et al. Investigation of Protein and Epitope Characteristics of Oats and Its Implications for Celiac Disease. Frontiers in Nutrition. 2021; 8. DOI 10.3389/fnut.2021.702352Kamal N., Tsardakas Renhuldt N., Bentzer J. et al. The mosaic oat genome gives insights into a uniquely healthy cereal crop // Nature. – 2022. – №606. – Pp. 113–119. DOI 10.1038/s41586-022-04732-yKamal N., Tsardakas Renhuldt N., Bentzer J. et al. The mosaic oat genome gives insights into a uniquely healthy cereal crop. Nature. 2022; 606: 113–119. DOI 10.1038/s41586-022-04732-yKosová K, Leišová-Svobodová L, Dvořáček V. Oats as a Safe Alternative to Triticeae Cereals for People Suffering from Celiac Disease? A Review // Plant Foods Hum Nutr. – 2020 Jun. – V.75(2). – Pp.131-141. DOI 10.1007/s11130-020-00800-8.Kosová K, Leišová-Svobodová L, Dvořáček V. Oats as a Safe Alternative to Triticeae Cereals for People Suffering from Celiac Disease? A Review. Plant Foods Hum Nutr. 2020; 75(2): 131-141. DOI 10.1007/s11130-020-00800-8.Lacorn M. Determination of Gliadin as a Measure of Gluten in Food by R5 Sandwich ELISA RIDASCREEN® Gliadin Matrix Extension: Collaborative Study 2012.01 // Journal of AOAC INTERNATIONAL. – 2022. – V. 105. – I. 2. – Pp. 442–455. DOI 10.1093/jaoacint/qsab148Lacorn M. Determination of Gliadin as a Measure of Gluten in Food by R5 Sandwich ELISA RIDASCREEN® Gliadin Matrix Extension: Collaborative Study 2012.01. Journal of AOAC INTERNATIONAL. 2022; 05; 2: 442–455. DOI 10.1093/jaoacint/qsab148Leišová-Svobodová, L., Sovová, T. & Dvořáček, V. Analysis of oat seed transcriptome with regards to proteins involved in celiac disease // Sci Rep. – 2022. – №12. – Pp. 8660. DOI 10.1038/s41598-022-12711-6Leišová-Svobodová, L., Sovová, T. & Dvořáček, V. Analysis of oat seed transcriptome with regards to proteins involved in celiac disease. Sci Rep. 2022; №12: 8660. DOI 10.1038/s41598-022-12711-6Lyubimova A.V., Tobolova G.V., Eremin D.I. et al. Dynamics of the genetic diversity of oat varieties in the Tyumen region at avenin-coding loci // Vavilov Journal of Genetics and Breeding. – 2020. – Vol. 24, No. 2. – Pp. 123-130. DOI 10.18699/VJ20.607.Lyubimova A.V., Tobolova G.V., Eremin D.I. et al. Dynamics of the genetic diversity of oat varieties in the Tyumen region at avenin-coding loci. Vavilov Journal of Genetics and Breeding. 2020; 24(2): 123-130. DOI 10.18699/VJ20.607.Machado M.V. New Developments in Celiac Disease Treatment // Int J Mol Sci. – 2023. – №24(2). – Pp. 945. DOI 10.3390/ijms24020945.Machado M.V. New Developments in Celiac Disease Treatment. Int J Mol Sci. 2023; 24(2): 945. DOI 10.3390/ijms24020945.Marasco G., Cirota G.G., Rossini B. et al. Probiotics, Prebiotics and Other Dietary Supplements for Gut Microbiota Modulation in Celiac Disease Patients // Nutrients. – 2020. – №12. – Pp. 2674. DOI 10.3390/nu12092674Marasco G., Cirota G.G., Rossini B. et al. Probiotics, Prebiotics and Other Dietary Supplements for Gut Microbiota Modulation in Celiac Disease Patients. Nutrients. 2020; 12: 2674. DOI 10.3390/nu12092674Marín-Sanz M., Giménez M.J., Barro F. et al. Prolamin content and grain weight in RNAi silenced wheat lines under different conditions of temperature and nitrogen availability // Frontiers in plant science. – 2020. – V.11. – №314. – Pp. 1–11.Marín-Sanz M., Giménez M.J., Barro F. et al. Prolamin content and grain weight in RNAi silenced wheat lines under different conditions of temperature and nitrogen availability. Frontiers in plant science. 2020; 11(314): 1–11.Orjiekwe O.A. Nutritional Management of Celiac Disease // J Clin Exp Immunol. – 2023. – V. 8(2). – Pp. 561-572Orjiekwe O.A. Nutritional Management of Celiac Disease. J Clin Exp Immunol. 2023; 8(2): 561-572Paudel D., Dhungana B., Caffe M. et al. A Review of Health-Beneficial Properties of Oats // Foods. – 2021. – № 10. – Pp. 2591. DOI 10.3390/foods10112591Paudel D., Dhungana B., Caffe M. et al. A Review of Health-Beneficial Properties of Oats. Foods. 2021; 10: 2591. DOI 10.3390/foods10112591Roeckendorf N., Meckelein B., Scherf K. et al. Identification of novel antibodyreactive detection sites for comprehensive gluten monitoring // PLoS One. – 2017. – V.12(7). – Pp. e0181566/1-e0181566/17.Roeckendorf N., Meckelein B., Scherf K. et al. Identification of novel antibodyreactive detection sites for comprehensive gluten monitoring. PLoS One. 2017; 12(7): e0181566/1-e0181566/17.Silano M., Di Benedetto R., Maialetti F. et al. Avenins from different cultivars of oats elicit response by coeliac peripheral lymphocytes // Scand J Gastroenterol. – 2007. – №42. – Pp.1302-1305 DOI 10.1080/00365520701420750Silano M., Di Benedetto R., Maialetti F. et al. Avenins from different cultivars of oats elicit response by coeliac peripheral lymphocytes. Scand J Gastroenterol. 2007; 42: 1302-1305 DOI 10.1080/00365520701420750Wieser H., Segura V., Ruiz-Carnicer Á. et al. Food Safety and Cross-Contamination of Gluten-Free Products: A Narrative Review // Nutrients. – 2021. – №13. – Pp.2244. DOI 10.3390/nu13072244Wieser H., Segura V., Ruiz-Carnicer Á. et al. Food Safety and Cross-Contamination of Gluten-Free Products: A Narrative Review. Nutrients. 2021; 13: 2244. DOI 10.3390/nu13072244Yu J.M., Lee J.H., Park J.-D. et al. Analyzing Gluten Content in Various Food Products Using Different Types of ELISA Test Kits // Foods. – 2021. – №10. – Pp.108. DOI 10.3390/foods10010108Yu J.M., Lee J.H., Park J.-D. et al. Analyzing Gluten Content in Various Food Products Using Different Types of ELISA Test Kits. Foods. 2021; 10: 108. DOI 10.3390/foods10010108

The authors declare that there are no conflicts of interest present.