bioselБиотехнология и селекция растенийPlant Biotechnology and Breeding2658-62662658-6258VIR10.30901/2658-6266-2018-1-7-15biosel-3Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL ARTICLEDistinctive metabolites of Alternaria, Fusarium and Microdochium fungi as a tool for assessing their relationship in micobiota of wheat grainГагкаеваТ. Ю.GagkaevaT. Yu.

ш. Подбельского, 3, Санкт-Петербург, Пушкин, 196608

Podbelskogo shosse, 3, St.Petersburg, 196608

t.gagkaeva@mail.ruГавриловаО. П.GavrilovaO. P.

ш. Подбельского, 3, Санкт-Петербург, Пушкин, 196608

Podbelskogo shosse, 3, St.Petersburg, 196608

ОринаА. С.OrinaA. S.

ш. Подбельского, 3, Санкт-Петербург, Пушкин, 196608

Podbelskogo shosse, 3, St.Petersburg, 196608

АбловаИ. Б.AblovaI. B.

центральная усадьба КНИИСХ, Краснодар, 350012

central farmstead KNIISH, Krasnodar, 350012

БеспаловаЛ. А.BespalovaL. A.

центральная усадьба КНИИСХ, Краснодар, 350012

central farmstead KNIISH, Krasnodar, 350012

ФГБНУ «Всероссийский научно-исследовательский институт защиты растений»РоссияAll-Russian Institute of Plant Protection (VIZR)Russian FederationФГБНУ «Национальный центр зерна им. П.П. Лукьяненко»РоссияNational Center of Grain named after P.P. LukyanenkoRussian Federation20180502201911715Copyright © Гагкаева Т.Ю., Гаврилова О.П., Орина А.С., Аблова И.Б., Беспалова Л.А., 20192019Гагкаева Т.Ю., Гаврилова О.П., Орина А.С., Аблова И.Б., Беспалова Л.А.Gagkaeva T.Y., Gavrilova O.P., Orina A.S., Ablova I.B., Bespalova L.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://biosel.elpub.ru/jour/article/view/3

Fungal contamination of 17 winter wheat varieties grown in the Krasnodar region under the natural infections was analyzed. In addition to the traditional mycological analysis of infection with fungi, methods based on the quantitative detection of metabolites inherent in fungi and absent in the host plant were used. For this purpose, the ranges of DNA contents of Alternaria, Fusarium and Microdochium fungi, as well as mycotoxins produced by these fungi were characterized in the grain. Abundant presence of Alternaria fungi was detected without a significant difference between the varieties. A quantitative detection of the Microdochium fungi in grain was carried out at the first time in Russia and it was shown that the content of M. nivale DNA was significantly exceeds the content of M. majus DNA. The dominant of F. graminearumin the complex of Fusarium fungi in comparison to other species and the significant range of the content of the metabolites produced by this pathogen were shown. The high variation of contents of marker metabolites inherent in highly aggressive species of fungi and the stability of these parameters for saprotrophic organisms can be of fundamental importance in the selection of objective for quantitative measurements of plants resistant to pathogens. The maximum amount of F.graminearum metabolites was found in the grains of susceptible Brigada. The minimum content of Fusarium metabolites was established in the grain of Adel and Kurs varieties, which are the relatively resistant to pathogenic Fusarium fungi. The proportion of F.graminearum DNA as a percentage of the DNA of Fusarium group forming trichothecene mycotoxins ranged from 6.1% to 100.9%. This value is an indicator of plant resistance to infection by pathogens. According to our observations, values above 30% are characteristic of susceptible varieties. A positive correlation was found between DNA contents of M.nivale and F. sporotrichioides fungi. Any significant relations between the most abundantly present fungi of Alternaria and the parameters of grain contamination by other analyzed fungi have not been revealed. Diversity of the reactions of wheat genotypes to the invasion of various heterotrophic fungi and the possibility of using objective quantitative assessment of resistance were shown.

micromycetesgrain infectionDNAquantitative PCRmycotoxinsELISAReferencesAndersen B, Frisvad JC (2004) Natural occurrence of fungi and fungal metabolites in moldy tomatoes. J Agric Food Chem 52(25): 7507–7513. doi:10.1021/jf048727kAndersen B, Frisvad JC (2004) Natural occurrence of fungi and fungal metabolites in moldy tomatoes. J Agric Food Chem 52(25): 7507–7513. doi:10.1021/jf048727kAudenaert K, Vanheule A, Höfte M, Haesaert G (2013) Deoxynivalenol: a major player in the multifaceted response of Fusariumto its environment. Toxins (Basel) 6: 1–19. doi:10.3390/toxins6010001Audenaert K, Vanheule A, Höfte M, Haesaert G (2013) Deoxynivalenol: a major player in the multifaceted response of Fusariumto its environment. Toxins (Basel) 6: 1–19. doi:10.3390/toxins6010001Burkin AA, Kononenko GP (2011) Enzyme immunassay of alternariol for the assessment of risk of agricultural products contamination. Appl Biochem Microbiol 47(1): 72-76. doi:10.1134/S0003683811010030Burkin AA, Kononenko GP (2011) Enzyme immunassay of alternariol for the assessment of risk of agricultural products contamination. Appl Biochem Microbiol 47(1): 72-76. doi:10.1134/S0003683811010030Burkin AA, Kononenko GP, Gavrilova OP, Gagkaeva TYu (2015) Mycotoxicological inspection of oat grain and its processing products (Mikotoksikologicheskoe obsledovanie zerna ovsa i produktov ego pererabotki). Sovremennaya mikologiya v Rossii – Current mycology in Russia 5: 221–223 [in Russian] (Буркин А. А., Кононенко Г. П., Гаврилова О. П., Гагкаева Т. Ю. Микотоксикологическое обследование зерна овса и продуктов его переработки // Современная микология в России. 2015. T.5. C. 221–223).Burkin AA, Kononenko GP, Gavrilova OP, Gagkaeva TYu (2015) Mycotoxicological inspection of oat grain and its processing products (Mikotoksikologicheskoe obsledovanie zerna ovsa i produktov ego pererabotki). Sovremennaya mikologiya v Rossii – Current mycology in Russia 5: 221–223 [in Russian] (Буркин А. А., Кононенко Г. П., Гаврилова О. П., Гагкаева Т. Ю. Микотоксикологическое обследование зерна овса и продуктов его переработки // Современная микология в России. 2015. T.5. C. 221–223).Ellis MB (1971) Dematiaceous Hyphomycetes. Kew, Surrey: Commonwealth Mycological Institute.Ellis MB (1971) Dematiaceous Hyphomycetes. Kew, Surrey: Commonwealth Mycological Institute.Gagkaeva TYu, Gavrilova OP, Orina AS, Blinova EV, Loskutov IG (2017) Response of wild Avena species to fungal infection of grain. The Crop Journal 5(6): 499–508. doi: 10.1016/j.cj.2017.04.005Gagkaeva TYu, Gavrilova OP, Orina AS, Blinova EV, Loskutov IG (2017) Response of wild Avena species to fungal infection of grain. The Crop Journal 5(6): 499–508. doi: 10.1016/j.cj.2017.04.005Gagkaeva TYu, Orina AS, Gavrilova OP, Ablova IB, Bespalova LA (2018) Characterization of resistance of winter wheat varieties to Fusarium head blight (Harakteristika sortov ozimoj pshenicy po ustojchivosti k fuzariozu zerna). Vavilovskij zhurnal genetiki i selekcii – Vavilov Journal of Genetics and Breeding 22 (6): 685– 692 [in Russian] (Гагкаева Т. Ю., Орина А. С., Гаврилова О. П., Аблова И. Б., Беспалова Л. А. Характеристика сортов озимой пшеницы по устойчивости к фузариозу зерна // Вавиловский журнал генетики и селекции. 2018. T. 22. № 6. С. 685– 692). doi:10.18699/VJ18.411Gagkaeva TYu, Orina AS, Gavrilova OP, Ablova IB, Bespalova LA (2018) Characterization of resistance of winter wheat varieties to Fusarium head blight (Harakteristika sortov ozimoj pshenicy po ustojchivosti k fuzariozu zerna). Vavilovskij zhurnal genetiki i selekcii – Vavilov Journal of Genetics and Breeding 22 (6): 685– 692 [in Russian] (Гагкаева Т. Ю., Орина А. С., Гаврилова О. П., Аблова И. Б., Беспалова Л. А. Характеристика сортов озимой пшеницы по устойчивости к фузариозу зерна // Вавиловский журнал генетики и селекции. 2018. T. 22. № 6. С. 685– 692). doi:10.18699/VJ18.411Gannibal PhB (2018) Factors affecting Alternaria appearance in grains in European Russia (Izuchenie faktorov, vliyayushchih na razvitie al'ternarioza zerna u zlakov, vozdelyvaemyh v evropejskoj chasti Rossii). Sel’skokhozyaistvennaya biologiya – Agricultural Biology 53(3): 605–615 [in Russian] (Ганнибал Ф. Б. Изучение факторов, влияющих на развитие альтернариоза зерна у злаков, возделываемых в европейской части России // Сельскохозяйственная биология. 2018. Т. 53. № 3. С. 605–615). doi: 10.15389/agrobiology.2018.3.605rusGannibal PhB (2018) Factors affecting Alternaria appearance in grains in European Russia (Izuchenie faktorov, vliyayushchih na razvitie al'ternarioza zerna u zlakov, vozdelyvaemyh v evropejskoj chasti Rossii). Sel’skokhozyaistvennaya biologiya – Agricultural Biology 53(3): 605–615 [in Russian] (Ганнибал Ф. Б. Изучение факторов, влияющих на развитие альтернариоза зерна у злаков, возделываемых в европейской части России // Сельскохозяйственная биология. 2018. Т. 53. № 3. С. 605–615). doi: 10.15389/agrobiology.2018.3.605rusGerlach W, Nirenberg HI (1982) The genus Fusarium — a pictorial atlas. Berlin: Dahlem.Gerlach W, Nirenberg HI (1982) The genus Fusarium — a pictorial atlas. Berlin: Dahlem.Halstensen AS, Nordby KC, Eduard W, Klemsdal SS (2006) Real-time PCR detection of toxigenic Fusarium in airborne and settled grain dust and associations with trichothecene mycotoxins. J Environ. Monit 8: 1235–1241. doi: 10.1039/b609840aHalstensen AS, Nordby KC, Eduard W, Klemsdal SS (2006) Real-time PCR detection of toxigenic Fusarium in airborne and settled grain dust and associations with trichothecene mycotoxins. J Environ. Monit 8: 1235–1241. doi: 10.1039/b609840aKahl SM, Ulrich A, Kirichenko AA, Müller ME (2015) Phenotypic and phylogenetic segregation of Alternaria infectoria from smallspored Alternaria species isolated from wheat in Germany and Russia. J Appl Microbiol 119(6): 1637–1650. doi:10.1111/jam.12951Kahl SM, Ulrich A, Kirichenko AA, Müller ME (2015) Phenotypic and phylogenetic segregation of Alternaria infectoria from smallspored Alternaria species isolated from wheat in Germany and Russia. J Appl Microbiol 119(6): 1637–1650. doi:10.1111/jam.12951Lawrence DP, Rotondo F, Gannibal PhB (2016) Biodiversity and taxonomy of the pleomorphic genus Alternaria. Mycol Prog 15:3. doi:10.1007/s11557-015-1144-xLawrence DP, Rotondo F, Gannibal PhB (2016) Biodiversity and taxonomy of the pleomorphic genus Alternaria. Mycol Prog 15:3. doi:10.1007/s11557-015-1144-xMesterhάzy A, Bartόk T, Kάszonyi G, Varga M, Tόth B, Varga J (2005) Common resistance to different Fusarium spp. causing Fusarium head blight in wheat. Eur J Plant Pathol 112: 267–281. doi:10.1007/s10658-005-2853-9Mesterhάzy A, Bartόk T, Kάszonyi G, Varga M, Tόth B, Varga J (2005) Common resistance to different Fusarium spp. causing Fusarium head blight in wheat. Eur J Plant Pathol 112: 267–281. doi:10.1007/s10658-005-2853-9Nicolaisen M, Justesen AF, Knorr K, Wang J, Pinnschmidt HO (2014) Fungal communities in wheat grain show significant co-existence patterns among species. Fungal Ecol 11: 145–153. doi: 10.1016/j.funeco.2014.06.002Nicolaisen M, Justesen AF, Knorr K, Wang J, Pinnschmidt HO (2014) Fungal communities in wheat grain show significant co-existence patterns among species. Fungal Ecol 11: 145–153. doi: 10.1016/j.funeco.2014.06.002Nicolaisen M, Suproniene S, Nielsen LK, Lazzaro I, Spliid NH, Justesen AF (2009) Real-time PCR for quantification of eleven individual Fusarium species in cereals. J Microbiol Methods 76: 234–240. doi: 10.1016/j.mimet.2008.10.016Nicolaisen M, Suproniene S, Nielsen LK, Lazzaro I, Spliid NH, Justesen AF (2009) Real-time PCR for quantification of eleven individual Fusarium species in cereals. J Microbiol Methods 76: 234–240. doi: 10.1016/j.mimet.2008.10.016Nielsen LK, Justesen AF, Jensen JD, Jørgensen LN (2013) Microdochium nivale and Microdochium majus in seed samples of Danish small grain cereals. Crop Prot 43: 192–200. doi: 10.1016/j.cropro.2012.09.002Nielsen LK, Justesen AF, Jensen JD, Jørgensen LN (2013) Microdochium nivale and Microdochium majus in seed samples of Danish small grain cereals. Crop Prot 43: 192–200. doi: 10.1016/j.cropro.2012.09.002Orina AS, Gavrilova OP, Gagkaeva TYu, Loskutov IG (2017) Symbiotic relationships between aggressive Fusarium and Alternaria fungi colonizing oat grain (Simbioticheskie vzaimootnosheniya gribov roda Fusarium i Alternaria, koloniziruyushchih zerno ovsa). Sel’skokhozyaistvennaya biologiya – Agricultural Biology 52(5): 986–994 [in Russian] (Орина А. С., Гаврилова О. П., Гагкаева Т. Ю., Лоскутов И. Г. Симбиотические взаимоотношения грибов рода Fusarium и Alternaria, колонизирующих зерно овса // Сельскохозяйственная биология. 2017. Т. 52. № 5. С. 986–994). doi: 10.15389/agrobiology.2017.5.986rusOrina AS, Gavrilova OP, Gagkaeva TYu, Loskutov IG (2017) Symbiotic relationships between aggressive Fusarium and Alternaria fungi colonizing oat grain (Simbioticheskie vzaimootnosheniya gribov roda Fusarium i Alternaria, koloniziruyushchih zerno ovsa). Sel’skokhozyaistvennaya biologiya – Agricultural Biology 52(5): 986–994 [in Russian] (Орина А. С., Гаврилова О. П., Гагкаева Т. Ю., Лоскутов И. Г. Симбиотические взаимоотношения грибов рода Fusarium и Alternaria, колонизирующих зерно овса // Сельскохозяйственная биология. 2017. Т. 52. № 5. С. 986–994). doi: 10.15389/agrobiology.2017.5.986rusPavón MÁ, González I, Martín R, García Lacarra T (2012) ITS-based detection and quantification of Alternaria spp. in raw and processed vegetables by real-time quantitative PCR. Food Microbiol 32: 165–171. doi:10.1016/j.fm.2012.05.006Pavón MÁ, González I, Martín R, García Lacarra T (2012) ITS-based detection and quantification of Alternaria spp. in raw and processed vegetables by real-time quantitative PCR. Food Microbiol 32: 165–171. doi:10.1016/j.fm.2012.05.006Rotem J (1994) The genus Alternaria. Biology, epidemiology and pathogenicity. St. Paul, Minnesota: APS Press.Rotem J (1994) The genus Alternaria. Biology, epidemiology and pathogenicity. St. Paul, Minnesota: APS Press.Samson RA, Hoekstra ES, Frisvad JC, Filtenborg O (2002) Introduction to food- and airborne fungi. Sixth Ed. Utrecht: Centraalbureau Voor Schimmelcultures.Samson RA, Hoekstra ES, Frisvad JC, Filtenborg O (2002) Introduction to food- and airborne fungi. Sixth Ed. Utrecht: Centraalbureau Voor Schimmelcultures.Schiro G, Verch G, Grimm V, Müller MEH (2018) Alternaria and Fusarium fungi: differences in distribution and spore deposition in a topographically heterogeneous wheat field. J Fungi (Basel) 4: 63. doi:10.3390/jof4020063Schiro G, Verch G, Grimm V, Müller MEH (2018) Alternaria and Fusarium fungi: differences in distribution and spore deposition in a topographically heterogeneous wheat field. J Fungi (Basel) 4: 63. doi:10.3390/jof4020063Siou D, Gélisse S, Laval V, Suffert F, Lannou C (2015) Mutual exclusion between fungal species of the Fusarium head blight complex in a wheat spike. Appl Environ Microbiol 81(14): 4682–4689. doi:10.1128/AEM.00525-15Siou D, Gélisse S, Laval V, Suffert F, Lannou C (2015) Mutual exclusion between fungal species of the Fusarium head blight complex in a wheat spike. Appl Environ Microbiol 81(14): 4682–4689. doi:10.1128/AEM.00525-15Smirnova OG, Shumnyj VK, Kochetov AV (2018) The gene network and the database of gene of resistance of wheat to pathogenic fungi (Gennaya set' i baza dannyh genov ustojchivosti pshenicy k patogennym gribam). Fiziologiya rastenij – Russian Journal of Plant Physiology 65(3): 181–195 [in Rissian] (Смирнова О. Г., Шумный В. К., Кочетов А. В. Генная сеть и база данных генов устойчивости пшеницы к патогенным грибам // Физиология растений. 2018. Т. 65. № 3. С. 181–195). doi:10.7868/S0015330318030028Smirnova OG, Shumnyj VK, Kochetov AV (2018) The gene network and the database of gene of resistance of wheat to pathogenic fungi (Gennaya set' i baza dannyh genov ustojchivosti pshenicy k patogennym gribam). Fiziologiya rastenij – Russian Journal of Plant Physiology 65(3): 181–195 [in Rissian] (Смирнова О. Г., Шумный В. К., Кочетов А. В. Генная сеть и база данных генов устойчивости пшеницы к патогенным грибам // Физиология растений. 2018. Т. 65. № 3. С. 181–195). doi:10.7868/S0015330318030028Tralamazza SM, Piacentini KC, Iwase CHT, de Oliveira Rocha L (2018) Toxigenic Alternaria species: impact in cereals worldwide. Curr Opin Food Sci 23: 57. doi: 10.1016/j.cofs.2018.05.002Tralamazza SM, Piacentini KC, Iwase CHT, de Oliveira Rocha L (2018) Toxigenic Alternaria species: impact in cereals worldwide. Curr Opin Food Sci 23: 57. doi: 10.1016/j.cofs.2018.05.002Vilkova NA (2000) Plant immunity against pest organisms and its role in stabilizing agroecosystems and plant growing (Immunitet rastenij k vrednym organizmam i ego biocenoticheskoe znachenie v stabilizacii agroehkosistem i povyshenii ustojchivosti rastenievodstva). Vestnik zashchity rastenij – Plant Protection News (2): 3–15. [in Russian] (Вилкова Н. А. Иммунитет растений к вредным организмам и его биоценотическое значение в стабилизации агроэкосистем и повышении устойчивости растениеводства // Вестник защиты растений. 2000. № 2. С. 3–15).Vilkova NA (2000) Plant immunity against pest organisms and its role in stabilizing agroecosystems and plant growing (Immunitet rastenij k vrednym organizmam i ego biocenoticheskoe znachenie v stabilizacii agroehkosistem i povyshenii ustojchivosti rastenievodstva). Vestnik zashchity rastenij – Plant Protection News (2): 3–15. [in Russian] (Вилкова Н. А. Иммунитет растений к вредным организмам и его биоценотическое значение в стабилизации агроэкосистем и повышении устойчивости растениеводства // Вестник защиты растений. 2000. № 2. С. 3–15).Yli-Mattila T, Paavanen-Huhtala S, Jestoi M, Parikka P, Hietaniemi V, Gagkaeva T, Sarlin T, Haikara A, Laaksonen S, Rizzo A (2008) Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia. Arch Phytopathology Plant Protect 41: 243–260. doi:10.1080/03235400600680659Yli-Mattila T, Paavanen-Huhtala S, Jestoi M, Parikka P, Hietaniemi V, Gagkaeva T, Sarlin T, Haikara A, Laaksonen S, Rizzo A (2008) Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia. Arch Phytopathology Plant Protect 41: 243–260. doi:10.1080/03235400600680659Zwickel Th, Kahl SM, Rychlik M, Müller MEH (2018) Chemotaxonomy of mycotoxigenic small-spored Alternaria fungi – do multitoxin mixtures act as an indicator for species differentiation? Front Microbiol 9: 1368. doi:10.3389/fmicb.2018.01368Zwickel Th, Kahl SM, Rychlik M, Müller MEH (2018) Chemotaxonomy of mycotoxigenic small-spored Alternaria fungi – do multitoxin mixtures act as an indicator for species differentiation? Front Microbiol 9: 1368. doi:10.3389/fmicb.2018.01368

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