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AP 14869300 (superviser Usenbekov B.N.)

Brief information about the project

(2022-2024)

Project title: IRN AP14869300 “Improvement of rice resistance to Magnaporthe oryzae using marker assistant selection (MAS)».

Relevance. One of the most dangerous diseases of rice is pyriculariosis, the causative agent of which is a parasitic fungus Magnaporthe oryzae (synonyms: Magnaporthe grisea or Pyricularia oryzae Cavara). Pyriculariosis has a devastating effect on the production of rice in the whole world, the loss of crops is 15-40%, which leads to a significant reduction in crop yields and threatens the food security of the country.

Effective ways to create varieties resistant to rice pyriculariosis are the introduction and pyramidation of genes. The use of molecular markers allows the screening of varieties for the presence of resistance genes, the introduction and pyramidation of genes and the selection of genotypes that carry gene resistance to Magnaporthe oryzae, which significantly shortens the time needed to create resistant rice lines and varieties.

The aim of the project is to create new rice lines based on the introduction and pyramiding of genes using MAS to increase the resistance of rice to Magnaporthe oryzae.

 Expected results:

  1. Using MAS, varieties-donors of genes for resistance to Magnaporthe oryzae will be selected for the introduction and pyramiding of genes into Kazakh rice varieties.
  2. F1BC1 and F2BC2 hybrid offspring will be obtained based on the introduction and pyramiding of Magnaporthe oryzae resistance genes into highly productive rice varieties zoned in Kazakhstan, and lines carrying Magnaporthe oryzae resistance genes will be selected.
  3. Pre-breeding material will be created from promising prototype lines of domestic rice varieties, characterized by high yield and resistance to the Kazakh population of Magnaporthe oryzae.

The scientific supervisor of the project: Usenbekov Bakdaulet Naubayevich – Head of the Laboratories of Plant Physiology and Biochemistry at the RSE with REM «IPBB» SC MES RK, Ph.D., Associate Professor. He published 90 scientific articles in co-authorship, and including for the period 2017-2023y. 9 articles in peer-reviewed foreign scientific publications indexed in Web of Science and/or Scopus (H-index=2, https://www.scopus.com/authid/detail.uri?authorId=56447130000), 5 articles in journals recommended by the COXON of the Ministry of Education and Science of the Republic of Kazakhstan and 1 patent of the RK.

 Members of research team:  

Amirova Aigul Kuzembaevna – Leading scientist, Ph.D. She has more than 150 scientific publications. For the period 2017-2023 published 2 articles in peer-reviewed international journals indexed in databases Web of Science and/or Scopus (H-index 1, https://www.scopus.com/authid/detail.uri?authorId=57193996757); 5 articles in journals recommended by COXON, author of 1 monograph, 1 patent of the RK and 3 study guides.

Bayseitova Gulnaz Abdumanapovna – Senior Researcher, PhD., for the period 2017-2023 published in co-author 11 articles, including 3 articles in peer-reviewed journals indexed in Web of Science  and/or Scopus (H-index=1, https://www.scopus.com/authid/detail.uri?authorId=57224638496), and 4 articles in journals recommended by COXON.

Berkimbay Khorlan Adeshkyzy – Researcher. For the period 2017-2023 published in co-author 2 articles in peer-reviewed journals indexed in Web of Science and/or Scopus (H-index=1, https://www.scopus.com/authid/detail.uri?

authorId=57193390033), and 4 articles in journals recommended by COXON.

Mynbayeva Dana Omarovna – Junior researcher, PhD doctoral student. Co-authors 11 scientific works, including 1 article in journals recommended by COXON.

List of publications and patents of project of members of research team for the period 2017-2023.

  1. Batayeva D.S., Usenbekov B.N., A.B. Rysbekova, Mukhina Zh.M., Kazkeyev D.T., Zhanbyrbayev Ye.A., Sartbayeva I.A., Garkusha S.V., Volkova S.A. Estimation and selection of parental forms for breeding kazakhstan salt tolerant rice varietIES // Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2017, Vol. 52, № 3, p. 544-552. doi: 10.15389/agrobiology.2017.3.544eng; Citation Index -1, percentile -35 (Scopus)
  2. Batayeva, D., Labaco, B., Ye, C., Li, X., Usenbekov, B., Rysbekova, A., Dyuskalieva, G., Vergara, G., Reinke, R., Leung, H. Genome-wide association study of seedling stage salinity tolerance in temperate japonica rice germplasm // BMC Genetics, 2018. — Volume 19, Issue 1. 19:2. – P. 1-11. DOI: 10.1186/s12863-017-0590-7 (IF-2.777); Citation Index -20, -WOS – Q3 (percentile -57 (Scopus).
  3. Sartbayeva I.A., Usenbekov B., Rysbekova A., Kazkeev D., Zhanbyrbayev E., Batayeva D., Berkimbay H., Zhambakin K., Matsuba Sh., Umemoto T. Haplotype Analysis of Wx and Alk Genes and Amylopectin Chain-Length Distribution among Kazakhstan Glutinous Rice Lines //Japan Agricultural research Quaterly (JARQ). – Vol.51, №2. – 2017. – P. 147-153. https://www.jircas.go.jp DOI: 10.6090/jarq.51.147 (IF-0.567); Citation Index -0, WOS – Q4 (percentile -35 (Scopus).
  4. A. B. Rysbekova, D. T. Kazkeyev, B. N. Usenbekov, Zh. M. Mukhina, E. A. Zhanbyrbaev, I.A. Sartbaeva, K.Zh. Zhambakin, Kh.A. Berkimbay, D.S. Batayeva. Prebreeding selection of rice with colored pericarp based on genotyping Rc and Pb genes //Russian Journal of Genetics 2017 volume 53, pages 49–58. DOI: 10.1134/S1022795416110119; Citation Index -2, WOS – Q4 (IF-0.567), (percentile -7 (Scopus).
  5. I.A. Sartbaeva, B.N. Usenbekov, A.B. Rysbekova, Zh.M. Mukhina, D.T. Kazkeev, K.Zh. Zhambakin, E.A. Zhanbyrbaev, Kh.A. Berkimbai, D.Sh. Akhmetova, and A.A. Meldebekova. Obtaining of Doubled Haploid Lines for Selection of Glutinous Rice //Biotechnology. Vol 34(2018) N 2 p. 26-36; DOI 10.21519/0234-2758-2018-34-2-26-36; Citation Index — 0, percentile — 9 (Scopus).
  6. Zhanbyrbayev, Y.A., Rysbekova, A.B., Usenbekov, B.N., Kazkeyev, D.T., Sarsenbayev, B.A., Zelensky, G.L.Physiologic al and molecular-genetic evaluation of rice breeding resources on cold tolerance for creating new varieties for the northern zone of rice cultivation // OnLine Journal of Biological Sciences, 2017, 17(3), стр. 240–250. DOI: https://doi.org/10.3844/ojbsci.2017.240.250 Citation Index -0, (percentile -27 (Scopus).
  7. D.T. Kazkeev, A.B. Rysbekova, B.N. Usenbekov, E.A. Zhanbyrbaev. Research of initial forms according to technological and biochemical parameters for selection of rice varieties with colored pericarp // Bulletin of the Shakarim State University of Semey. No. 2 (86), 2019, pp. 349-353.
  8. Berkimbay Kh., Zhanbirbayev E.A., Ussenbekov B.N. Molecular-genetic analysis of rice with painted pericarp for cold resistance // Research, results KazNAU, 2020. №2 (86), P. 236-242.
  9. Bishimbayeva Nazira, Yertayeva Bakhyt, Amirova Aigul and Rakhimbayev Izbasar Morphogenesis in Tissue Culture of Local Kazakh Cotton Varieties // OnLine Journal of Biological Sciences, 2017. ISSN: 1608-4217. DOI: 10.3844/ojbsci.2017. URL: http://thescipub.com/abstract /10.3844/ofsp.11335 (WOS – Q3, Scopus percentile 27, IF 0,14)
  10. Dossymbetova S.A., Amirova A.K., Kurbangaliyeva T.A., Abdresh Kh.Zh., Kurtibayeva G.R. Germination of seeds of medicinal plants Origanum vulgare L. and Salvia officinalis L. in vitro // Bulletin of Karaganda University. Series Biology. Medicine. Geography, 2021. №2 (102)7. P.23-29.
  11. Dossymbetova S.A., Amirova A.K., Kurbangaliyeva T.A. The effect of phytohormones on callusogenesis of the medicinal plant Origanum vulgare/ Bulletin of the North Kazakhstan University named after Manash Kozybayev, 2021. № 1 (50). P. 84-90.
  12. Amirova, A.; Dossymbetova, S.; Rysbayeva, Y.; Usenbekov, B.; Tolegen, A.; Ydyrys, A. Multiple Plant Regeneration from Embryogenic Calli of Paulownia tomentosa (Thunb.) Steud. Plants 2022, 11 (8), 1020. Citation Index -0, WOS – Q1 (IF-3.935), (percentile -71 (Scopus) https://doi.org/10.3390/plants11081020.
  13. Mynbayeva D.O., Usenbekov B.N., Amirova A.K., Zhunusbayeva Zh.K. Rice blast and control methods // Bulletin of Karaganda University. Series Biology. Medicine, 2023. № 1 (109). P. 98-109. DOI 10.31489/2023BMG1/98-109
  14. Usenbekov B.N., Amirova A.K., Berkimbay Kh.A., Zhanbyrbayev Ye.A., Kazkeev D.T., Zhaisankulova U.B., Sartbayeva I.A. Evaluation of initial material for the obtaining of high-amylose rice // Bulletin of Karaganda University. Series Biology. Medicine, 2022. № 2(106). P. 118-126. DOI 10.31489/2022BMG2/118-12
  15. Berkimbay1 Kh.A., Ussenbekov B.N., Amirova A.K. Screening of genotypes of rice with colored pericarp for amylose content // BULLETIN of ENU named after L.N. Gumilyov. Series Biological Sciences, 2022. № 1(138). P. 38-46. DOI: 10.32523/2616-7034-2022-138-1-38-46
  16. Usenbekov B.N., Amirova A.K., Zhanbyrbayev Y.A., Berkimbay Kh.A., Sartbayeva I.A., Kazkeev D.T. // Method of haploid biotechnology for obtaining of green regenerated plants in anther culture of rice with colored pericarp // Patent RK №5962 // Bulletin № 13 from 02.04.2021.

Results achieved in 2022:

35 rice varieties were taken for research from the IPBB rice collection, 31 of which are introgressive lines (IL) provided by the All-Russian Research Institute of Rice (Krasnodar, RF) and the FGBNU All-Russian Research Institute of Rice Research. I.G. Kalinenko (Zernograd, Rostov-on-Don, Russian Federation) in accordance with the agreement on the exchange and provision of rice germplasm dated May 12, 2015.

Screening of 35 rice genotypes of domestic and foreign breeding for the presence of blast resistance genes was carried out using six SSR markers. As a result, genetic polymorphism of the studied varieties was revealed using the molecular markers RM224, MSM1, Pita, MSM6, Z56592 and TRS26 associated with the blast resistance genes Pi-1, Pi-2, Pi-ta, Pi-40, Piz-t and Pi-54.

The genetic variability of studied samples by the genes of Pi complex. Electrophoresis of PCR products showed that 3 samples from 35 studied samples did not contain the amplicon with size 157 bp corresponding to the Pi-1 gene using SSR marker RM224 localized on chromosome 11. The frequency of occurrence of the Pi-1 gene in the studied genotypes is 91.4%.

It was found that the 175 bp amplicon is absent in most of the studied samples using the SSR marker MSM1 located on chromosome 6 and associated with Pi-2 gene of rice resistance to blast, the frequency of this gene is low — 14.28%.

In the case of the using of molecular marker Pita located on chromosome 12 associated with the Pi-ta resistance gene, the amplicon 500 bp is present in many genotypes (94.29%) and only in 2 samples is absent.

The use of the molecular marker MSM6 located on chromosome 6 allow to identify the presence of the Pi-40 gene by PCR product with a size 256 bp, the frequency of the gene – 80.0%.

The Piz-t gene was detected using the SSR marker Z56592 localized on chromosome 6 by the presence of 292 bp amplicon, the frequency of this gene was 62.86%.

The molecular marker TRS26 located on chromosome 11 and associated with the Pi-54 resistance gene was determined by the presence of an amplicon of size 266 bp, the frequency of the gene was 82.86%. In the studied Kazakh varieties, this gene is absent.

It is known that according to the strategy for increasing the resistance of rice varieties to blast, the presence of not one gene, but 5 or more genes is desirable in order to ensure stable and long-term resistance of varieties to changing races of the fungus Magnaporthe oryzae. Presence (1) and absence (0) of genes Pi-1 Pi-ta, Pi-2, Pi-40, Piz-t and Pi-54 in the studied samples were evaluated by visualization of amplicons about 157 bp, 500 bp, 175 bp, 256 bp, 292 bp and 266 bp, respectively. We have clarified the presence of resistance genes to Magnaporthe oryzae in introgressive lines and varieties of foreign and domestic breeding from the IPBB rice collection.

Most samples of foreign selection contain from 1 to 5 resistance genes, while domestic varieties – from 0 to 4 blast resistance genes.

As a result of the work pyramidal lines containing several fungus resistance genes will be used as donors of the gene for increasing the resistance of domestic rice varieties to phytopathogen.

Conclusions: Rice samples of domestic and foreign breeding were screened for the presence of resistance genes to Magnaporthe oryzae based on the molecular genetic approach (molecular labeling).

Rice samples with genes controlling resistance to Magnaporthe oryzae were selected from the IPBB rice working collection for hybridization of domestic rice varieties with donors of Pi-1 Pi-ta, Pi-2, Pi-40, Piz-t and Pi-54 genes, and breeding blast resistant rice lines.