Development of blast-resistant rice varieties based on application of DNA technologies

Diseases of agricultural crops are the main reason for decreased yield and quality of product. Blast (causative agent: Pyricularia oryzae Cav.) is the most harmful disease on rice fields. Economic damage caused by the pathogen is significant in all areas of the world’s rice cultivation. The most effective, economically justified and environmentally friendly strategy for combating it is development of resistant varieties. The application of DNA markers linked to loci of resistance to blast is relevant in this area. This makes it possible to significantly shorten the breeding process and promptly develop disease-resistant rice forms. In this regard, the aim of the work was to develop source material for breeding as well as highly productive rice varieties and lines with genes of resistance to blast based on the use of molecular marking method. To achieve this goal, we have launched a program since 2007 aimed at introduction of the blast resistance Pi-ta gene, effective for the south of Russia, into the domestic rice cultivar Flagman. After a number of recurrent crosses, the breeding material was obtained, which was studied for economically valuable traits in breeding nurseries. As a result of evaluation and rigorous discarding as well as according to the results of a phytopathological test for blast resistance, several lines were identified that have high indicators of milled rice quality, resistance to blast, yield and economically valuable traits. Rice accession KP-171-14 with the Pi-ta gene, adapted to growing conditions in the south of Russia, resistant to the Krasnodar population of P. oryzae, and having high yield and grain quality, in 2017 was submitted for State Variety Trials (SVT) under the name Alyans. Accessions KP-30 and KP-23 are tested for economically valuable traits and disease resistance in competitive variety trials. The best accession will be submitted to SVT. The introduction and cultivation of such varieties will reduce the use of chemical protection products, obtain environmentally friendly agricultural products and avoid contamination of grain ecosystems

1. Ashkani S, Rafii M, Sariah M, Siti NAA, Rusli I, Harun A, Latif M (2011) Ashkani S, Rafii M, Sariah M, Siti NAA, Rusli I, Harun A, Latif M (2011) Analysis of simple sequence linked with blast disease resistance genes in a segregating repeat markers population of rice (Oryza sativa). Genet. Mol. Biol.; 10: 1345-1355

2. Averyanov AA, Lapikova VP, Petelina GG (1990) Rapid laboratory method for assessing varietal resistance of rice to blast (Laboratornyj ehkspress-metod ocenki sortovoj ustojchivosti risa k pirikulyariozu). Bol’shie Vyazemy: VNIIF: 12 p. [in Russian] (Аверьянов А. А., Лапикова В. П., Петелина Г. Г. Лабораторный экспресс-метод оценки сортовой устойчивости риса к пирикуляриозу. Большие Вяземы : ВНИИФ, 1990. 12 с.).

3. Bonman J, Khush G, Nelson R (1992) Breeding rice for resistance to pest. Annu. Rev. Phytopatol. 30: 507-528.

4. Cho YG, Eun MY, McCouch SR, Chae YA (1994) The semidwarf gene, sd-1, of rice (Oryza sativa L.). II. Molecular mapping and marker-assisted selection. Theor. Appl. Genet.; 89: 54-59.

5. Correa-Victoria FJ, Tharreau D, Martinez C, Vales M (2003) Gene combinations in rice for the development of durable resistance to Pyricularia grisea in Colombia. Proc. 3rd Int. Temperate Rice Conference. Punta del Este.

6. Dospekhov BA (1979) Methods of the Field Experiments (Basis of Statistical Processing of the Research Results) (Metodika polevogo opyta (s osnovami statisticheskoj obrabotki rezul’tatov issledovanij)). M. : Kolos. 416 p. [in Russian] (Доспехов Б. А. Методика полевого опыта (с основами статистической обработки результатов исследований). Изд. 4-е, перераб. и доп. М. : Колос. 1979. 416 с.).

7. Dubina EV, Mukhina ZM, Kharitonov EM, Shilovskiy VN, Kharchenko ES, Esaulova LV, Korkina NN, Maximenko EP, Nikitina IB (2015) Creation of blast disease-resistant rice varieties with modern DNA markers. Russian Journal of Genetics; 51(8): 881-886 [in Russian] (Дубина Е. В., Мухина Ж. М., Харитонов Е. М., Шиловский В. Н., Харченко Е. С., Есаулова Л. В., Коркина Н. Н., Максименко Е. П., Никитина И. Б. Создание устойчивой к пирикуляриозу генплазмы риса с использованием технологий ДНК- маркирования // Генетика. 2015. Т. 51 № 8. С. 881-886).

8. Dubina EV, Kostylev PI, Ruban MG, Aniskina YuV, Shilov IA, VelishaevaNS, EsaulovaLV(2017) The study of biodiversity of Pyricularia oryzae Cav. in rice-growing zones of the south of Russia on the basis of the methods of PCR. Grain economy of Russia; 54(6): 29-35 [in Russian] (Дубина Е.В., Костылев П.И., Рубан М.Г., АнискинаЮ. В., Шилов И. А., ВелишаеваН. С., Есаулова Л. В. Изучение биоразнообразия Pyricularia oryzae Cav. в рисосеющих зонах юга России на основе метода ПЦР // Зерновое хозяйство России. 2017. Т. 54, № 6. С. 29-35).

9. Frisch M, Bohn M, Melchinger AE (1999) Minimum sample size and optimal positioning of flanking markers in marker-assisted backcrossing for transfer of a target gene. Crop Science; 39: 967-975.

10. Frolova VS, Kovalenko ED, Naskidashvili ZhG, Silicheva TM et al. (1983) Guidelines for assessing the resistance of rice varieties to blast in an infectious nursery (Metodicheskiye ukazaniya po otsenke ustoychivosti sortoobraztsov risa k pirikulyariozu v infektsionnom pitomnike). M. VASKHNIL. 14 p. [in Russian] (Фролова В. С., Коваленко Е. Д., Наскидашвили Ж. Г., Силичева Т. М. и др. Методические указания по оценке устойчивости сортообразцов риса к пирикуляриозу в инфекционном питомнике. М. : ВАСХНИЛ. 1983. 14 с.).

11. Girish Kumar K, Hittalmani S, Srinivasachary К (2000) Marker assisted backcross gene introgression of major genes for blast resistance in rice. Advances in Rice Blast Research: 43-53.

12. Hittalmani S, Parco A, Mew TV, Zeigler RS, Huang N (2000) Fine mapping and DNA marker-assisted pyramiding of the three major genes for blast resistance in rice. Theor. Appl. Genet.; 100: 1121-1128.

13. Jena KK, Moon HP, Mackill DJ (2003) Marker assisted selection - a new paradigm in plant breeding. Korean J. Breed. ; 35: 133-140.

14. Kharchenko ES, Dubina EV, Garkusha SV, Ruban MG, Esaulova LV, Avakyan ER, Balyasny IV (2017) A study of the biodiversity of the causative agent Pyricularia oryzae Cav. in the south of Russia (Izuchenie bioraznoobraziya populyacii vozbuditelya Pyricularia oryzae Cav. na yuge Rossii). Science of the Kuban; 1: 20-27 [in Russian] (Харченко Е. С., Дубина Е. В., Гаркуша С. В., Рубан М. Г., Есаулова Л. В., Авакян Э. Р., Балясный И. В. Изучение биоразнообразия популяции возбудителя Pyricularia oryzae Cav. на юге России // Наука Кубани. № 1. С. 20-27).

15. Khavkin EE (1997) Molecular markers in plant breeding (Molekulyarnyye markery v rasteniyevodstve). Agricultural Biology; 5: 3-21 [in Russian] (Хавкин Э. Е. Молекулярные маркеры в растениеводстве // Сельскохозяйственная биология. № 5. С. 3-21).

16. Kiyosawa S (1989) Breakdown of blast resistance in rice in relation to general strategies of resistance gene deployment to prolong effectiveness of disease resistance in plants. Pages 251-283 in: Plant Disease Epidemiology. K. J. Leonard and W. E. Fry, eds. McGraw-Hill Publishing Company, New York: 251-283.

17. Kolomiets TM (1990) The selection of the source material of rice for breeding for immunity to blast (Otbor iskhodnogo materiala risa dlya selekcii na immunitet k pirikulyariozu). Avtoref. dis. kand. biol. nauk.-Summary of PhD thesis. Golicyno: 21 c.

18. Los GD (1987) A promising method of rice hybridization. Agricultural Biology; 12: 15-17 [in Russian]. (Лось Г. Д. Перспективный способ гибридизации риса // Сельскохозяйственная биология. 1987. Т. 12. С. 15-17).

19. Matsumura H, Nirasawa S, Kiba A, Urasaki N, Saitoh H, Ito M, Kawai- Yamada M, Uchimiya H, Terauchi R (2003) Overexpression of Bax inhibitor suppresses the fungal elicitor-induced cell death in rice (Oryza sativa L.) cells. Plant J. ; 33: 425-434.

20. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research; 8(10): 4321-4325.

21. Myagkih YuA (2009) The use of molecular marking to improve the efficiency of rice selection (Primenenie molekulyarnogo markirovaniya dlya povysheniya ehffektivnosti selekcii risa). Dissertaciya na soiskanie uchenoj stepeni kandidata biologicheskih nauk - Thesis for the degree of candidate of biological sciences. Krasnodar, 2009: 100 p. [in Russian] (Мягких Ю. А. Применение молекулярного маркирования для повышения эффективности селекции риса. Диссертация на соискание ученой степени кандидата биологических наук. Краснодар, 2009: 100 с.).

22. Openshau SJ, Jarboe SJ, Bears WD (1994) Marker assisted selection in backross breeding. In: Analysis of molecular marker data. Joint Plant Breed. Symp. Ser., Corvallis, Oregon, USA: 41-43.

23. Osterman LA (1981) Methods for studying nucleic acids (Metody issledovaniya nukleinovyh kislot). М: Nauka Publ. 288 p. [in Russian] (Остерман Л.А. Методы исследования нуклеиновых кислот. М. : Наука, 1981. 288 с.).

24. Practical Guide to Intensive Rice Cultivation Technology in the Krasnodar Territory (reference edition) (Prakticheskoe rukovodstvo po intensivnoj tekhnologii vozdelyvaniya risa v Krasnodarskom krae (spravochno-metodicheskoe izdanie). Krasnodar. 1986: 38 p. [in Russian] (Практическое руководство по интенсивной технологии возделывания риса в Краснодарском крае (справочно-методическое издание). Краснодар, 1986. 38 с.).

25. Smetanin AP, Dzyuba VA, Aprod AL (1972) Methods of experimental work on breeding, genetics, seed production, seed science and quality control of rice seeds (Metodiki opytnykh rabot po selektsii, genetike, semenovodstvu, semenovedeniyu i kontrolyu za kachestvom semyan risa). Krasnodar: 156 p.[in Russian] (Сметанин А. П., Дзюба В. А., Апрод А. И. Методики опытных работ по селекции, генетике, семеноводству, семеноведению и контролю за качеством семян риса. Краснодар, 1972. 156 с.).

26. Shilov LA, Kolobova OS, Aniskina YuV, Shalaeva TV, Velishaeva NS, Kostylev PN, Dubina EV (2016) Improvement of the identification method of the genes Pi-ta, Pi-b for resistance to rice blast (Usovershenstvovaniye metoda identifikatsii genov ustoychivosti k pirikulyariozu risa Pi-ta, Pi-b). Achievements of science and technology of agroindustrial complex; 30(8): 45-48 [in Russian] (ШиловИ. А., Колобова О. С., Анискина Ю. В., Шалаева Т. В., Велишаева Н. С., Костылев П. Н., Дубина Е. В. Усовершенствование метода идентификации генов устойчивости к пирикуляриозу риса Pi-ta, Pi-b // Достижения Науки и Техники АПК. № 8. С. 45-48).

27. Sasaki R (1922) Inheritance of rice blast resistance. Japan. J. Genet.; 1: 81-85.

28. Tabien RE, Li Z, Paterson AH, Marchetti MA, Stansel JW, Pinson SRM (2000) Mapping of four major blast resistance genes from “Lemont” and “Teqing” and evaluation of their combination effect for field resistance. Theor. Appl. Genet.; 101(8): 1215-1225.

29. Witcombe JR, Hash CT (2000) Resistance gene deployment strategies in cereal hybrids using marker-assisted selection: gene pyramiding, three-way hybrids, and synthetic parent population. Euphytica; 112(2): 175-186.

30. Yu ZH, Mackill DJ, Bonman JM, McCouch SR, Guiderdoni E (1996) Molecular mapping of genes for resistance to rice blast. Theor. Appl. Genet.; 93: 859-863.