Mycobiota in Mycobiota in the grain of the oat breeding lines produced in 2019 in competitive variety trials on the fields of Nemchinovka Federal Research Center, Moscow Provincepetitive variety trials on the fields of Nemchinovka Federal Research Center, Moscow Province

Background. The requirement for high-quality oat grain for food production leads to the need for studying the resistance of this cereal crop to harmful diseases and for the development of new productive cultivars.

Materials and methods. A comparative characterization was performed among the breeding lines of naked oats from Nemchinovka Federal Research Center according to the parameters of microbiological purity of grain. The mycological method was applied for the analysis of infection in the oat grain, and fungal species composition was identified. The DNA amount of toxin-producing species from genus Fusarium Link in the grain was detected using the real-time PCR, and the amount of mycotoxins was measured by the enzyme immunoassay.

Results. The fungi of Alternaria Nees, Fusarium, Cladosporium Link, Cochliobolus Drechsler, Epicoccum Link were associated with oat grain. Uniquely high infection of the grain by F. langsethiae Torp & Nirenberg (14%) was found under the natural conditions of the Central Region of Russia. The differences revealed among the analyzed breeding lines and cultivars in the DNA amount of F. langsethiae and F. sporotrichioides Sherb. were from 1.3×10^-4 to 7.2×10^-3 pg/ng in the total DNA, and from 5 to 1230 μg/kg in the content of T-2/ HT-2 toxins.

Conclusion. All naked oat breeding lines were relatively resistant to Fusarium infection and proved the best as regards the mycotoxin contamination of the grain against the reference hulled cultivar ‘Yakov’. In the competitive variety trials, three lines – 66h2618, 54h2476 and 70h2613 – were characterized as relatively resistant to Fusarium disease, compared with the naked oat reference ‘Vyatsky golozerny’.

1. Andersen B., Krøger E., Roberts R.G. Chemical and morphological segregation of Alternaria arborescens, A. infectoria and A. tenuissima species-groups. Mycological Research. 2002;106(2):170-182. DOI: 10.1017/S0953756201005263

2. Batalova G.A. Perspectives and results of naked oats breeding. Legumes and Groat Crops. 2014;2(10):64-69. [in Russian]

3. Bensch K., Braun U., Groenewald J.Z., Crous P.W. The genus Cladosporium. Studies in Mycology. 2012;72(1):1-401. DOI: 10.3114/sim0003

4. Condon B.J., Leng Y., Wu D., Bushley K.E., Ohm R.A., Otillar R. et al. Comparative genome structure, secondary metabolite, and effector coding capacity across Cochliobolus pathogens. PLOS Genetics. 2013;9(1):e1003233. DOI: 10.1371/journal.pgen.1003233

5. Divon H.H., Bøe L., Tveit M.M.N., Klemsdal S.S. Infection pathways and penetration modes of Fusarium langsethiae. European Journal of Plant Pathology. 2019;154(2):259-271. DOI: 10.1007/s10658-018-01653-3

6. Ellis M.B. Dematiaceous Hyphomycetes. Kew, Surrey: Commonwealth Mycological Institute; 1971.

7. Gagkaeva T.Yu., Gavrilova O.P. Contamination of oats grain by fungi and pollution by mycotoxins in the northwest of Russia in 2014. Zashchita i karantin rasteniy = Plant Protection and Quarantine. 2015;(8):10-12. [in Russian]

8. Gagkaeva T.Yu., Gavrilova O.P., Levitin M.M. Biodiversity and distribution of the main toxigenic Fusarium fungi. Biosfera = Biosphere. 2014;6(1):36-45. [in Russian]

9. Gagkaeva T.Yu., Gavrilova O.P. Orina A.S., Blinova E.B., Loskutov I.G. Diversity of the species of genus Avena revealed by morphological characters and resistance to Fusarium infection of grain. Ecological Genetics. 2017:15(1):20-29. [in Russian] DOI: 10.17816/ecogen15120-29

10. Gagkaeva T.Yu., Gavrilova O.P., Yli-Mattila T., Loskutov I.G. Sources of resistance to Fusarium head blight in VIR oat collection. Euphytica. 2013;191(3):355-364. DOI: 10.1007/s10681-013-0865-7

11. Gannibal F.B. Alternaria disease of grain – modern view of the problem. Zashchita i karantin rasteniy = Plant Protection and Quarantine. 2014;(6):11-15. [in Russian]

12. Gannibal Ph.B. Factors affected Alternaria appearance in grains in European Russia. Agricultural Biology. 2018:53(3):605-615. [in Russian] DOI: 10.15389/agrobiology.2018.3.605rus

13. Gavrilova O.P., Gagkaeva T.Yu., Burkin A.A., Kononenko G.P. Mycological infection by Fusarium strains and mycotoxins contamination of oats and barley in the north of Nonchernozem’e. Agricultural Biology. 2009a;6:89-93. [in Russian]

14. Gavrilova O.P., Gagkaeva T.Yu., Loskutov I.G. Evaluation of oat accessions from the VIR collection for fusarial head blight resistance Bulletin of Applied Botany, Genetics and Plant Breeding. 2009b;165:182-186. [in Russian]

15. Gavrilova O.P., Gannibal Ph.B., Gagkaeva T.Yu. Fusarium and Alternaria fungi in grain of oats grown in the North-Western Russia regarding cultivar specificity. Agricultural Biology. 2016;51(1):111-118. [in Russian] DOI: 10.15389/agrobiology.2016.1.111rus

16. Gavrilova O.P., Gribchenko E.S., Loskutov I.G., Gagkaeva T. Yu. Influence of a flower film on the mycobiota of oat genotypes (Vlianiye tsvetkovoy plenki na mikobiotu genotipov ovsa). In: Yu.T. Dyakov (ed.). Current Mycology in Russia. Vol. 5. Proceedings of the III International Mycological Forum; April 14–15, 2015; Moscow, Russia. Moscow: National Academy of Mycology; 2015. p.44- 46. [in Russian] URL: http://www.mycology.ru/files/cmr2015vol5.pdf [Аccessed on: 05.03.2020].

17. Gavrilova O.P., Skritnika A., Gagkaeva T.Yu. Identification and characterization of spontaneous auxotrophic mutants in Fusarium langsethiae. Microorganisms. 2017;5(2):E14. DOI: 10.3390/microorganisms5020014

18. Gerlach W., Nirenberg H.I. The genus Fusarium – a pictorial atlas. Mitteilungen der Biologischen Bundesanstalt für Land- und Forstwirtschaft. 1982;209:1-406.

19. Methodology of state crop variety trials. Second issue. Cereals, groats, legumes, maize, and fodder crops (Metodika gosudarstvennogo sortoispytaniya selskokhozyaystvennykh kultur. Vypusk vtoroy. Zernovye, krupyanye, zernobobovye, kukuruza i kormovye kultury). Moscow: Gosagroprom SSSR; 1989. [in Russian] URL: https://docplayer.ru/28203913-Metodikagosudarstvennogo-sortoispytaniya-selskohozyaystvennyh-kultur.html [Аccessed on: 05.03.2020].

20. Goncharenko A.A. Moscow Breeding Center for Cereals: history, achievements, problems and prospects (Moskovskiy selektsionny tsentr po zernovym kulturam: istoria, dostizheniya, problemy i perspektivy). Informatsionny vestnik VOGiS = Bulletin of the Vavilov Society of Geneticists and Plant Breeders. 2005;9(3):324-332. [in Russian]

21. Imathiu S.M., Ray R.V., Back M., Hare M., Edwards S.G. In vitro growth characteristics of Fusarium langsethiae isolates recovered from oats and wheat grain in the UK. Acta Phytopathologica et Entomologica Hungarica. 2016;51(2):159-170. DOI: 10.1556/038.51.2016.2.1

22. Imathiu S.M., Ray R.V., Back M.I., Hare M.C., Edwards S.G. A survey investigating the infection of Fusarium langsethiae and production of HT-2 and T-2 mycotoxins in UK oat fields. Journal of Phytopathology. 2013;161(7-8):553-561. DOI: 10.1111/jph.12105

23. Isachkova O.A., Ganichev B.L. Sources of economically valuable signs and their combination for breeding naked oat in Western Siberia. Mezhdunarodny nauchno-issledovatelskiy zhurnal = International Research Journal. 2013;4-1(11):127-131. [in Russian]

24. Ivanova Y.S., Fomina M.N. Technological evaluation of grain in collection samples of naked oat in the conditions of Northern forest-steppe of the Tyumen region. Agricultural Science Euro-North-East. 2017;5(60):4-9. [in Russian]

25. Jensen B.D., Knorr K., Nicolaisen M. In vitro competition between Fusarium graminearum and Epicoccum nigrum on media and wheat grains. European Journal of Plant Pathology. 2016;146(3):657-670. DOI: 10.1007/s10658-016-0950-6

26. Kabashov A.D., Kolupaeva A.S., Leybovich Y.G., Ra zu movskaya L.G., Filonenko Z.V. Preliminary results of hullless oats breeding. Selektsiya, semenovodstvo i genetika = Breeding, Seed Production and Genetics. 2018;4(4):20-24. [in Russian] DOI: 10.24411/2413-4112-2018-10003

27. Kabashov A.D., Mamedov R.Z., Leybovich Ya.G. The actual directions in oats selection at the present stage. The Bulletin of Izhevsk State Agricultural Academy. 2013;1(34):25-28. [in Russian]

28. Kosiak B, Torp M, Skjerve E, Andersen B. Alternaria and Fusarium in Norwegian grains of reduced quality – a matched pair sample study. International Journal of Food Microbiology. 2004;93(1):51-62. DOI: 10.1016/j.ijfoodmicro.2003.10.006

29. Lee H.B., Patriarca A., Magan N. Alternaria in food: ecophysiology, mycotoxin production and toxicology. Mycobiology. 2015;43(2):93-106. DOI: 10.5941/MYCO.2015.43.2.93

30. Li Y., Wang Z., Beier R.C., Shen J., De Smet D., De Saeger S. et al. T-2 toxin, a trichothecene mycotoxin: review of toxicity, metabolism, and analytical methods. Journal of Agricultural and Food Chemistry. 2011;59(8):3441-3453. DOI: 10.1021/jf200767q

31. Loskutov I.G., Blinova E.B., Gavrilova O.P., Gagkaeva T. Yu. The valuable characteristics of oats genotypes and resistance to Fusarium disease. Vavilov Journal of Genetics and Breeding. 2016;20(3):286-294. [in Russian]

32. Martin C., Schöneberg T., Vogelgsang S., Mendes Ferreira C.S., Morisoli R., Bertossa M. et al. Responses of oat grains to Fusarium poae and F. langsethiae infections and mycotoxin contaminations. Toxins. 2018;10(1):E47. DOI: 10.3390/toxins10010047

33. Mazzara M., Paoletti C., Puumalaainen J., Rasulo D., Van Den Eede G. Event-Specific Method for the Quantitation of Maize Line NK603 Using Real-Time PCR –Validation Report and Protocol. EUR 21825. JRC32103. Ispra: European Commission; 2005.

34. Nikolaev Yu.N., Androsova O.V. Seed production and nurseries in the Russian Federation: analytical statistics (Semenovodstvo i pitomnikovodstvo RF: analiticheskaya statistika). Vestnik Rosselhozcentra = Bulletin of the Russian Agricultural Center. 2018;(1):13-16. [in Russian]

35. Oviedo M.S., Sturm M.E., Reynoso M.M., Chulze S.N., Ramirez M.L. Toxigenic profile and AFLP variability of Alternaria alternata and Alternaria infectoria occurring on wheat. Brazilian Journal of Microbiology. 2013;44(2):447-455. DOI: 10.1590/S1517-83822013000200017

36. Samson R.A., Hoekstra E.S., Frisvad J.C., Filtenborg O. (eds). Introduction to Food- and Airborne Fungi (revised 6th edition). Utrecht: Centraalbureau voor Schimmelcultures; 2002.

37. SanPiN 2.3.2.1078-01. Hygienic requirements for safety and nutritional value of food products. Chief Sanitary Inspector of the Russian Federation. Resolution of November 14, 2001, No. 36 “Concerning the introduction of sanitary regulations” (with amendments of July 6, 2011. [in Russian] URL: http://docs.cntd.ru/document/901806306 [Аccessed on: 12.02.2020].

38. Schöneberg T., Kibler K., Wettstein F.E., Bucheli T.D., Forrer H.R., Musa T. et al. Influence of temperature, humidity duration and growth stage on the infection and mycotoxin production by Fusarium langsethiae and Fusarium poae in oats. Plant Pathology. 2019;68(1):173-184. DOI: 10.1111/ppa.12922

39. Schuhmacher-Wolz U., Heine K., Schneider K. Report on toxi city data on trichothecene mycotoxins HT-2 and T-2 toxins. EFSA Supporting Publications. 2010;7(7):65E. DOI: 10.2903/sp.efsa.2010.EN-65

40. Šliková S., Šrobárová A., Šudyová V., Polišenská I., Gregová E., Mihálik D. Response of oat cultivars to Fusarium infection with a view to their suitability for food use. Biologia. 2010;65(4):609-614. DOI: 10.2478/s11756-010-0055-1

41. Stenglein S.A. Fusarium poae: a pathogen that needs more attention. Journal of Plant Pathology. 2009;91(1):25-36. DOI: 10.4454/jpp.v91i1.621

42. Tekauz A.B., Mitchell Fetch J.W., Rossnagel B.G., Savard M.E. Progress in assessing the impact of Fusarium head blight on oat in western Canada and screening of Avena germplasm for resistance. Cereal Research Communications. 2008;36:49-56. DOI: 10.1556/CRC.36.2008.Suppl.B.8

43. Tekle S., Lillemo M., Skinnes H., Reitan L., Buraas T., Bjørnstad Å. Screening of oat accessions for Fusarium head blight resistance using spawn-inoculated field experiments. Crop Science. 2018;58(1):143-151. DOI: 10.2135/cropsci2017.04.0264

44. Tihonovich I.A., Provorov N.A. Agricultural microbiology as the basis of ecologically sustainable agriculture: fundamental and applied aspects. Agricultural Biology. 2011;46(3):3-9. [in Russian]

45. Torp M., Nirenberg H.I. Fusarium langsethiae sp. nov. on cereals in Europe. International Journal of Food Microbiology. 2004;95(3):247-256. DOI: 10.1016/j.ijfoodmicro.2003.12.014

46. TR CU 015/2011. Technical Regulations of the Customs Union “On grain safety” with the changes of September 15, 2017. [in Russian] URL: http://docs.cntd.ru/document/902320395 [Аccessed on: 12.02.2020].

47. TR CU 021/2011. Council of the Eurasian Economic Commission. Resolution of August 8, 2019, No. 115. Amendments to the Technical Regulations of the Customs Union “On food safety” URL: http://docs.cntd.ru/document/564134628 [Аccessed on: 12.02.2020].

48. Ueno Y. Toxicological features of T-2 toxin and related trichothecenes. Fundamental and Applied Toxicology. 1984;4(2, Pt 2):S124-S132. DOI: 10.1016/0272-0590(84)90144-1

49. Vargach J.I., Golovin S.E., Loskutov I.G. Research on micromycetes in oats (Avena sativa L.) under the conditions of Stupino District, Moscow Province. Proceedings on Applied Botany, Genetics and Breeding. 2019;180(3):96-105. [in Russian] DOI: 10.30901/2227-8834-2019-3-96-105

50. Vogelgsang S., Sulyok M., Hecker A., Jenny E., Krska R., Schuhmacher R. et al. Toxigenicity and pathogenicity of Fusarium poae and Fusarium avenaceum on wheat. European Journal of Plant Pathology. 2008;122(2):265-276. DOI: 10.1007/s10658-008-9279-0

51. Yan W., Fregeau-Reid J., Rioux S., Pageau D., Xue A., Martin R. et al. Response of oat genotypes to Fusarium head blight in Eastern Canada. Crop Science. 2010;50(1):134-142. DOI: 10.2135/cropsci2009.03.0129

52. Yli-Mattila T., Paavanen-Huhtala S., Jestoi M., Parikka P., Hietaniemi V., Gagkaeva T. et al. Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia. Archives of Phytopathology and Plant Protection. 2008;41(4):243-260. DOI: 10.1080/03235400600680659

53. Zhuikova O.A., Batalova G.A. Analysis of the development of main diseases in naked oat varieties in the filmed nursery of the competitive trials at the North-Eastern FASC (Analiz proyavleniya osnovnykh bolezney sortov ovsa plenchatogo pitomnika konkursnogo ispytaniya FANTs SeveroVostoka). In: G.A. Batalova, E.M. Lisitsyn, T.K. Sheshegova, I.N. Shchennikova, E.I. Utkina (eds). Proceedings of the V Scientific and Practical Conference “Methods and Technologies in Plant Breeding and Plant Growing” (Materialy V Mezhdunarodnoy nauchno-prakticheskoy konferentsii “Metody i tekhnologii v selektsii rasteniy i rasteniyevodstve”). Kirov: North-Eastern FASC; 2019. p.21-24. [in Russian] URL: http://fanc-sv.ru/uploads/docs/2019/Konferenciya-Batalova-2019.pdf [Аccessed on: 05.03.2020].