Analysis of sequencing chromatograms for the identification of ash fungal pathogens in the territory of Voronezh

  • Inna Yu. Burakova Voronezh State University of Engineering Technologies, Voronezh, Russian Federation
  • Artem P. Gureev Voronezh State University of Engineering Technologies, Voronezh, Russian Federation
  • Elena Yu. Amineva All-Russian Research Institute of Forest Genetics, Breeding, and Biotechnology, Voronezh, Russian Federation
  • Stanislav G. Rzhevsky All-Russian Research Institute of Forest Genetics, Breeding, and Biotechnology, Voronezh, Russian Federation
  • Vasily N. Vasily N. Voronezh State University, Voronezh, Russian Federation
Keywords: Fraxinus excelsior L.; Hymenoscyphus fraxineus; fungal pathogen; sequencing; chromatogram analysis

Abstract

Within the territory of the city of Voronezh, as in many other regions of Russia, intensive die-back of ash is observed. However, to date, the study of pathogens in the city of Voronezh have not been performed. For the identification of the species of ash-infecting fungi, we cultivated them on a nutrient medium, isolated and purified the DNA using sorption spin columns, and amplified and sequenced the ITS regions of DNA. Analysis of the chromatograms obtained as a result of sequencing made it possible to determine the species of fungi that infect ash trees. It should be noted that the analysis of chromatograms obtained as a result of DNA sequencing of the studied fungi did not reveal Hymenoscyphus fraxineus, a fungal pathogen that was the most obvious candidate for the main cause of ash die-back, since this fungus greatly reduced the population of ash in Central and Northern Europe, and was also found in the Voronezh region on the territory of the Tellerman experimental forestry. On the contrary, in one of the trees Fusarium avenaceum was identified, which is considered an endophyte of the ash tree and, according to some reports, has antagonistic activity against H. fraxineus. However, the DNA of Erysiphe salmonii, a powdery mildew fungus that previously affected ash trees in Eastern Europe, was determined on two trees. In turn, the chromatograms revealed peaks that corresponded to the nucleotide sequences of DNA of Aspergillus niger, Coprinellus micaceus, and C. ovatus. The results of this study are applicable in the economic and environmental sector of forestry in the Voronezh region for designing a strategy for targeted combating the epidemic of ash die-back in the city of Voronezh.

Downloads

Download data is not yet available.

Author Biographies

Inna Yu. Burakova, Voronezh State University of Engineering Technologies, Voronezh, Russian Federation

Postgraduate student, Department of Biochemistry and Biotechnology, VSUET, Voronezh, Russian Federation, e-mail: vitkalovai@inbox.ru

Artem P. Gureev, Voronezh State University of Engineering Technologies, Voronezh, Russian Federation

Senior Lecturer, Department of Genetics, Cytology and Bioengineering, Faculty of Medicine and Biology, VSU, Voronezh; Junior Researcher, Laboratory of Metagenomics and Food Biotechnologies, Voronezh, Russian Federation, e-mail: gureev@bio.vsu.ru

Elena Yu. Amineva, All-Russian Research Institute of Forest Genetics, Breeding, and Biotechnology, Voronezh, Russian Federation

researcher, Department of Forest Genetics and Biotechnology, All-Russian Research Institute of Forest Genetics, Breeding and Biotechnology (In Russian: VNIILGISbiotech), Voronezh, Russian Federation, e-mail: elena.pardaeva@mail.ru

Stanislav G. Rzhevsky, All-Russian Research Institute of Forest Genetics, Breeding, and Biotechnology, Voronezh, Russian Federation

Junior Researcher, Department of Forest Genetics and Biotechnology, All-Russian Research Institute of Forest Genetics, Breeding and Biotechnology (In Russian: VNIILGISbiotech), Voronezh, Russian Federation, e-mail: slavaosin@yandex.ru

Vasily N. Vasily N. , Voronezh State University, Voronezh, Russian Federation

Doctor of Biological Sciences, Professor, Head of the Department of Genetics, Cytology and Bioengineering, Faculty of Medicine and Biology, VSU, Voronezh; Rector of VSUET, Voronezh, Russian Federation, e-mail: pvn@bio.vsu.ru

References

Fedorova A.I., Mikheeva M.A. Tree plants of Voronezh (biodiversity and sus-tainability). Voronezh, Publishing and Print-ing Center of Voronezh State University. 2008. 100 p. (In Russ.).

Antipov V.G. Resistance of woody plants to industrial gases. Minsk. Science and Technology. 1979. 215 p. (In Russ.).

Boyd I.L., Freer-Smith P.H., Gilli-gan C.A., Godfray H.C.J., The consequence of tree pests and diseases for ecosystem services. Science. 2013; 342(6160): 1235773-1235773. https://doi.org/10.1126/science.1235773

Coker T.L.R., Rozsypálek J., Ed-wards A., Harwood T.P., Butfoy L., Buggs, R.J.A., Estimating mortality rates of Euro-pean ash (Fraxinus excelsior) under the ash dieback (Hymenoscyphus fraxineus) epi-demic. PLANTS, PEOPLE, PLANET. 2018; 1(1): 48-58. https://doi.org/10.1002/ppp3.11

Orlova-Bienkowskaja M.J., Drog-valenko A.N., Zabaluev I.A., Sazhnev A.S., Peregudova E.Y., Mazurov S.G., Komarov E.V., Struchaev V.V., Martynov V.V., Nikulina T.V., Bieńkowski A.O. Current range of Agrilus planipennis Fairmaire, an alien pest of ash trees, in European Russia and Ukraine. Annals of Forest Science. 2020; 77(2): 1-14. https://doi.org/10.1007/s13595-020-0930-z

Musolin D.L., Selikhovkin A.V., Shabunin D.A., Zviagintsev V.B., Baranchikov Yu.V. Between Ash Dieback and Emerald Ash Borer: Two Asian In-vaders in Russia and the Future of Ash in Europe. Baltic Forestry. 2017; 23(1): 316-333.

Herms D.A., McCullough D.G., Emerald Ash Borer Invasion of North America: History, Biology, Ecology, Im-pacts, and Management. Annual Review of Entomology. 2014; 59(1): 13-30. https://doi.org/10.1146/annurev-ento-011613-162051

Forest plan of the Voronezh region. Decree of the Governor of the Voronezh Region of August 15. 2011 N 303-u (as amended on October 17, 2017). Access mode: https://www.govvrn.ru/1462 (date of access: 12.11.2021) (In Russ.).The first tim-ber industry portal. Forestry news. Access mode: http://www.wood.ru/ru/lonewsid-75837.html. wood.ru (date of treatment 11/12/2021)

Orlova-Benkovskaya M.Ya. “Ash emerald narrow-bodied goldfish (Agrilus planipennis) settled in nine regions of Euro-pean Russia: from Yaroslavl to Voronezh”, Materials of the international conference, St. Petersburg, November 25-27, 2013. VII Readings in memory of O.A. Kataeva. Pests and diseases of woody plants in Rus-sia. Ed. A.V. Selikhovkin and D.L. Musoli-na. SPb. SPbGLTU, 2013: 65-66. (In Russ.).

Kolganikhina GB, Panteleev SV. “First report of the harmful phytopathogen-ic fungus Hymenoscyphus fraxineus in Tellermanovsky forest (south forest-steppe of the European part of Russia)” Proceed-ings of the 2d International Conference “Biology, systematic and ecology of fungi and lichen in natural and agricultural eco-systems”. Minsk. Kolorgrad. 2016: 115-118. (In Russ.).

Kolganikhin G.B., Pathogenic and saprotroph fungi on ash in plantings of the tellerman experimental forest area. Forestry Bulletin. 2018; 22(6): 40-48. https://doi.org/10.18698/2542-1468-2018-6-40-48(In Russ.).

Fantin Yu., Favorov A., Neverov A., Mironov A.A., Chulanov V. Analysis of complex chromatograms of population se-quencing. Access mode: http://itas2011.iitp.ru/pdf/1569459319.pdf (date of access 12.11.2021) (In Russ.).

Kowalski T., Łukomska A., Badania nad zamieraniem jesionu (Fraxinus excelsior L.) w drzewostanach Nadleśnictwa Włoszczowa [The studies on ash dying (Fraxinus excelsior L.) in the Włoszczowa Forest Unit stands]. Acta Agrobotanica. 2005; 58(2): 429-440. https://doi.org/10.5586/aa.2005.068 (In Russ.).

Gross A., Holdenrieder O., Pautasso M., Queloz V., Sieber T. N. Hymenoscy-phus pseudoalbidus, the causal agent of Eu-ropean ash dieback. Molecular Plant Pa-thology. 2014; 15(1): 5-21. https://doi.org/10.1111/mpp.12073

Baral H.O., Bemmann M., Hymen-oscyphus fraxineus vs. Hymenoscyphus al-bidus – A comparative light microscopic study on the causal agent of European ash dieback and related foliicolous, stroma-forming species. Mycology. 2014; 5(4): 228-290. https://doi.org/10.1080/21501203.2014.963720

Zviagintsev V.B., Baranov O.Yu., Panteleev S.V. “Distribution of ash dieback disease caused by invasive mycopathogen Hymenoscyphus fraxineus Baral et al., in Moscow region and along the route M1”. Problems of forest phytopathology and my-cology. Materials of the IX international conference. Belarussian State Technological Institute. Minsk. 2015: 87-89.

Erfandoust R., Habibipour R., Soltani J. Antifungal activity of endophytic fungi from Cupressaceae against human pathogenic Aspergillus fumigatus and As-pergillus niger. Journal de Mycologie Médi-cale. 2020; 100987. https://doi.org/10.1016/j.mycmed.2020.100987

Martínez J., Nevado A., Suñén E., Gabriel M., Vélez-Del-Burgo A., Sánchez P., Postigo I. The Aspergillus niger Major Allergen (Asp n 3) DNA-Specific Sequence Is a Reliable Marker to Identify Early Fun-gal Contamination and Postharvest Damage in Mangifera indica Fruit. Front Microbiol. 2021; 12: 663323. https://doi.org/10.3389/fmicb.2021.663323

Dijksterhuis J. Fungal spores: Highly variable and stress-resistant vehicles for dis-tribution and spoilage. Food Microbiology. 2018; 81: 2-11. https://doi.org/10.1016/j.fm.2018.11.006

Braun U., Cook R.T.A., Taxonomic Manual of the Erysiphales (powdery mil-dews). CBS Biodiversity Series no 11. Utrecht: CBS-KNAW Fungal Biodiversity Centre. 2012; 707.

Heluta V.P., Takamatsu S., Siahaan, S.A.S. Erysiphe salmonii (Erysiphales, As-comycota), another East Asian powdery mildew fungus introduced to Ukraine. Ukranian Botanical Journal. 2017; 74: 212-219. https://doi.org/10.15407/ukrbotj74.03.212

Yamaguchi Y., Meeboon J., Heluta V.P., Liu S-Y., Feng J., Takamatsu S. Phy-logeny and taxonomy of Erysiphe species (powdery mildew: Erysiphaceae) occurring on the ash trees (Fraxinus spp.). Mycosci-ence. 2021; 62(2): 205-211. https://doi.org/10.47371/mycosci.2020.11.009

Kowalski T., Bilański P., Kraj W. Pathogenicity of fungi associated with ash dieback towards Fraxinus excelsior. Plant Pathology. 2017; 66(8): 1228-1238. https://doi.org/10.1111/ppa.12667

Kosawang C., Amby D.B., Bussa-ban B., McKinney L.V., Xu J., Kjær E.D., Collinge, D.B., Nielsen L.R. Fungal com-munities associated with species of Fraxinus tolerant to ash dieback, and their potential for biological control. Fungal Biology. 2018; 122(2-3): 110-120. https://doi.org/10.1016/j.funbio.2017.11.002

Wang S., Zhang J., He Y., Chang M., Meng J. Characterization of the com-plete mitochondrial genome of Coprinellus micaceus, a wild saprobic mushroom in China. Mitochondrial DNA Part B. 2021; 6(7): 1979-1981. https://doi.org/10.1080/23802359.2021.1938717

Papic S., Longauer R., Milenković I., Rozsypálek J. Genetic predispositions of common ash to the ash dieback caused by ash dieback fungus. Genetika. 2018; 50(1): 221-229. https://doi.org/10.2298/GENSR1801221P

Published
2022-05-19
How to Cite
Burakova, I. Y., Gureev, A. P., Amineva, E. Y., Rzhevsky, S. G., & Vasily N. , V. N. (2022). Analysis of sequencing chromatograms for the identification of ash fungal pathogens in the territory of Voronezh. Sorbtsionnye I Khromatograficheskie Protsessy, 22(2), 214-223. https://doi.org/10.17308/sorpchrom.2022.22/9226