STUDY OF BIODIVERSITY OF PYRICULARIA ORYZAE CAV. BY PCR-METHOD IN THE RICE-GROWING AREAS IN THE SOUTH OF RUSSIA

E. V. Dubina^1, M. G. Ruban¹, U. V. Aniskina³, I. A. Shilov³, N. S. Velishaeva³, P. I. Kostiylev², Yu. A. Makukha¹, D. A. Pishenko^1
1All-Russian Rice Research Institute, Krasnodar, p. Belozerny, 3, 350921, Russian Federation
²Agrarian Scientific Center «Donskoy», Zernograd, Nauchny gorodok, 3, 347740, Russian Federation
³All-Russian Research Institute of Agricultural Biotechmology, ul. Timiryazevskaya, 42, Moskva, 127550, Russian Federation

Abstract. Genetic structures and biodiversity of fungal phytopathogen Pyricularia oryzae Cav. were studied on the basis of molecular and genetic approaches with the use of fragment analysis as well as classic microbiological methods in the south of Russia. The investigations were carried out in 2015–2017 in the All-Russian Research Institute of Rice Breeding in collaboration with the All-Russian Research Institute of Agricultural Biotechnology and Agrarian Scientific Center “Donskoj”. It was selected 57 strains of the pathogen from the damaged herbarium material collected in the fields of eight ecological rice-growing zones of Krasnodar Krai (Krasnoarmejsky, Kalininsky, Abinsky, Temryuksky, Seversky, Slavyansky districts, the city of Krasnodar), Rostov region (Proletarsky district) and the Republic of Adygeya. Pathogens were cultivated on carrot-sugar medium at the humidity of 100% and the temperature of 27±1 C. DNA was isolated from a mixture of mycelium of monoconidial culture by CTAB-method with the use of hexadecyl trimethyl ammonium bromide as a detergent in the lysis buffer. Twenty-two genotypes of Pyricularia oryzae Cav. were revealed on the basis of multiplex PCR developed earlier. Every one of them is characterized by a unique genetic profile. Their DNA-passports were composed. The isolates were divided into 11 morphotypes according to morphological and cultural characteristics. The discrepancy between the results of DNA analysis and analysis by morphological and cultural features is due to the fact that in the second case some strains were characterized by heterogeneity of the colony (by sectors). This knowledge makes it possible to predict the appearance of new races and pathotypes of Pyricularia oryzae Cav., and also to determine genes of (a)virulence in the rice growing regions of Russia. The latter serves as a necessary theoretical basis for the creation of genetic sources with a long-lasting resistance to rice blast.

Keywords: rice; blast; Pyricularia oryzae Cav.; isolates; PCR; micro-satellite markers; DNA-passports.

Acknowldgement. Research was conducted with support of grant RFFR № 16-44-230178 «Study of genetic structure of population of Pyricularia oryzae Cav. agent and scientific ground of immune and genetic protection of rice crop».

Author Details: E.V. Dubina¹, D.Sc (Biol.), head of the laboratory (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); M.G. Ruban¹, junior researcher; Yu.V. Aniskina², D.Sc (Biol.), Senior Researcher; I.A. Shilov², D.Sc (Biol.), head of the laboratory; N.S.Velishaeva², D.Sc (Biol.), Senior Researcher; P.I. Kostylev³, D.Sc (Agr.), head of the laboratory; Yu.A. Makukha¹, post graduate student; D.A. Pishenko¹, head of the laboratory.

For citation: Dubina E. V., Ruban M. G., Aniskina U. V., Shilov I. A., Velishaeva N. S., Kostiylev P. I., Makukha Yu. A., Pishenko D. A. Study of Biodiversity of Pyricularia oryzae Cav. by PCR-method in the Rice-Growing Areas in the South of Russia. Dostizheniya nauki i tekhniki APK. 2018. Vol. 32. No. 10. Pp. 19–23 (in Russ.). DOI: 10.24411/0235-2451-2018-11004.