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Ilyushko M.V., Romashova M.V., Guchenko S.S. Study of Oryza sativa L. samples at a collection nursery in Primorsky krai and screening for of blast resistance genes. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2025, Vol. 60, № 5, p. 810-826.
(how to cite this article)

doi: 10.15389/agrobiology.2025.5.810eng

UDC: 633.18:631.522/.524:577.2

 

STUDY OF Oryza sativa L. SAMPLES AT A COLLECTION NURSERY IN PRIMORSKY KRAI AND SCREENING FOR OF BLAST RESISTANCE GENES

M.V. Ilyushko✉, M.V. Romashova, S.S. Guchenko

Chaika Federal Research Center of Agricultural Biotechnology of the Far East, 30, ul. Volozhenina, pos. Timityazevskii, Ussuryisk, Primorskii Krai, 692539 Russia, e-mail ilyushkoiris@mail.ru (✉ corresponding author), romashova_1969@mail.ru, lana_svet8@mail.ru

ORCID:
Ilyushko M.V. orcid.org/0000-0001-7042-8641
Romashova M.V. orcid.org/0000-0002-7426-8523
Guchenko S.S. orcid.org/0000-0003-3492-8934

Final revision received December 20, 2024
Accepted March 22, 2025

 In the south of the Far East, Oryza sativa L. rice is grown at the northern limit of its range, which is characterized by recurrent cold weather during the initial periods of rice growth and development. Soil and climatic conditions allow obtaining grain yields of up to 6.0 t/ha using early maturing varieties. The region has always been characterized by significant phytopathological stress. A wide variety of Pyricularia oryzae Cav. races is noted here, since the monsoon climate creates favorable humid conditions for their development. In this study, productive accessions were isolated for the first time from 159 accessions of the O. sativa rice collection in the conditions of Primorsky Territory; collection accessions with resistance alleles of the Pi genes were highlighted; polymorphic, heterozygous accessions and accessions with a pyramid of the Pi genes were identified. The aim of the study was to select the most productive and early maturing forms in an updated rice collection under the conditions of Primorsky Territory  and determine the allelic state of seven blast resistance genes (Pi-ta, Pi-1, Pi-2, Pi-b, Pi-zt, Pi-ta2, Pi-9) to P. oryzae using molecular markers. A total of 159 collection accessions of O. sativa rice from 22 countries were examined. Seeds were collected in 2018-2021 and stored in a refrigerator at 4 °C. The studies were conducted at the vegetation site of the A.K. Chaika Federal Scientific Center of Agrobiotechnology of the Far East (Primorsky Territory, Ussuriysk, Timiryazevsky settelment) in 2022-2023. Rice was sown at the recommended dates: May 25, 2022 and May 19, 2023. The experiment was conducted in 460 l pots filled with soil typical of rice fields – meadow-brown with a heavy mechanical composition. The irrigation regime was shortened flooding. Weather conditions during the growing season met the biological requirements of the crop, exceeding the long-term regional average by month by 0.2-0.8 °C in 2022 and by 1.4-2.4 °C in 2023. Plants were harvested before September 30. Late-ripening samples in 2022 were transplanted into plastic containers and placed in a greenhouse at a temperature of 18-20 °C for ripening. In 2023, all plants ripened before harvesting in the growing site. Manual tending of crops was carried out. The date of germination and ripening (at least three plants per replicate at the required phase) was recorded, averaging for each sample. The following biometric parameters were measured: tillering (pcs.), plant height (cm), panicle length (cm), number of grains in the main panicle (pcs.), empty grain content (%), weight of grains in the main panicle and plant (g), the weight of 1000 grains was obtained by recounting (g). A total of 4804 plants were analyzed in the work. A comparative assessment of the collection samples with the control was performed using LDS-test. To isolate DNA, we used the leaves of three to four plants of the same sample; 598 plants were analyzed. If sample polymorphism was detected, an additional 26-30 plants were grown to determine the proportion of resistance alleles. The PCR reaction was carried out in 25 μl of the reaction mixture according to the generally accepted technique. As a result of the work, variety samples (Tavrichesky, (4764) and UkrNIS 3455) exceeding the control variety Primorsky 29 in productivity were isolated. Fefteen samples with the resistance allele of the Pi-ta gene, 9 with Pi-b, 2 with Pi-zt, 12 with Pi-ta2, 8 with Pi-1, 52 with Pi-2 were isolated. Samples with the Pi-9 resistance allele were not identified. The obtained results can be included in the databases characterizing O. sativa rice collections and used in breeding for resistance to blast. Polymorphism for the genes of resistance to P. oryzae was revealed among 15 variety samples. The proportion of samples with the susceptibility allele varied from 3.6 to 96.4 %. Samples Long Jing 15, Sui Jing 4, Mu 07-1233 and Dubrava turned out to be polymorphic for two genes; Lon-do-6 for three genes. Sample LD-122 was polymorphic for four genes. Heterozygous plants were detected in three samples, reaching 21.4 %. In the sample LD-122, heterozygotes were detected for three genes Pi-b, Pi-2 and Pi-1. The samples can be used in breeding for heterosis in the northern part of the range of the cultivated species O. sativa, since they are capable of allogamy under such conditions. A pyramid of two or three most valuable genes of rice resistance to P. oryzae for Primorsky Territory was determined in five samples: LD-122, X-н-20-09, Long Jing 19, Oxy 2x, Long Jing 14. All of them are inferior in productivity and early maturity to the control variety Primorsky 29 and are suitable for introgression of target genes a complex in selection for resistance to P. oryzae with molecular genetic support.

Keywords: Oryza sativa, varieties, collection, productivity, early maturity, blast resistance genes Pi.

 

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