The purpose of this research by the way of investigating the molecular genetic structure of a highly variable fungal phytopathogen Pyricularia oryzae Cav., to determine effective genes for the development of a strategy for immunogenetic protection against rice blast in conditions of epiphytotic development of the disease in the south of Russia, which would combine high efficacy with both environmental friendliness and resource and energy saving, to ensure country's food security. The knowledge of local pathotype diversity of Pyricularia oryzae Cav. and the (a)virulence genes in rice-growing regions of Russia may allow the prediction of new races and its interaction in local agro-ecology. The identification of virulence gene may become an indispensable theoretical basis for the development of genetic sources with long-lasting resistance to rice blast. Based on molecular and genetic approaches, the genetic structure and biodiversity of the phytopathogenic fungus Pyricularia oryzae Cav. in the south of Russia were considered. The monitoring was studied and it isolated 57 strains of the pathogen from the damaged herbal material collected from the fields in eight agro-ecological rice-growing regions of the Krasnodar Region (Russian Federation): Krasnoarmeysky, Kalininsky, Krymsky, Abinsky, Temryuksky, Seversky, Slavyansky districts, Krasnodar, Rostov Region (Russian Federation)-Proletarsky district and the Republic of Adygea (Russian Federation). A multiplex PCR technique was applied on the basis of fragment analysis to identify the virulent fungal isolates. 33 fungal genotypes with unique genetic profiles were identified among the studied races of Pyricularia oryzae Cav. Their DNA profiles were created. The studied isolates of the pathogen of rice blast were classified using morphological and microbiology cultural features. Based on the phytopathological test using differentiation rice varieties, the quantitative and qualitative composition of (a)virulence genes in fungal races was established. Effective genes for pathogen resistance, which are recommended for breeding programs for the development of rice varieties resistant to rice blast, were identified in the south of Russia.
Selection done using molecular markers has spread to all types of agricultural plants and animals. Marker assisted selection (MAS) is safe for ecology and food products consumed. The purpose of the study is to develop Sub1A gene (which determines tolerance to prolonged flooding) in the Russian rice varieties using DNA-markers and Polymerase chain reaction (PCR) analysis. A large volume of prebreeding rice material (F2-F4, BC2F2) was created with tolerance genes for prolonged flooding, to control weed in rice agrophytocenosis. The forms with 120-130-day vegetation period were selected. An analysis of the co-inheritance of simple sequence repeat (SSR) markers Sub1Aq, Sub1C173, Sub1A203 with the Sub1 gene was carried out. It revealed the codominance of the selected microsatellites. They can be used to identify and visualize the allelic state of a given target gene in the breeding material. The recurrent parental rice forms, donors of the gene Sub 1 and the hybrids of F2 and ВС2F1 population were tested on tolerance to prolonged flooding. The gene is effective in ensuring resistance to this stress factor within 2 to 3 weeks and it can be recommended for the breeding programs to develop modern genetic resources of rice tolerant to prolonged and deep flooding for weed control on rice fields.
This article presents the results of the development of a methodology for describing the main morphological and cultural traits of the Pyricularia oryzae Cav. strains widespread in the south of Russia. At the same time, the types of traits are identified and listed, which make it possible to unambiguously determine the uniqueness and variety of the pathogen. The relationships and patterns established using cluster and statistical analysis make it possible to identify the conditions for the development of a pathogen that determine its predominant forms. Thus, research shows that leaf forms of P. oryzae strains isolated from rice plants with leaf form of blast disease have an equally directional growth pattern of a colony with a felt structure, and strains isolated from neck-affected plant form often produce a zone of a colony with a clumpy structure. The classification of cultural traits will make it possible to obtain scientifically grounded and comparable data that can be used in the analysis of the interaction of P. oryzae strains with rice plants on various varieties and in various agro-technological conditions in order to improve and rationalize agricultural activities. The study opens up the possibility of using data in breeding, making it possible to identify forms of a pathogen that infect certain varieties.
The article concentrates on studying tolerance to soil salinization, water flooding, and blast in Russian and Asian rice varieties, as well as hybrids of the second and third generations from their crossing in order to obtain sustainable paddy crops based on domestic varieties using DNA markers. Samples IR 52713-2B-8-2B-1-2, IR 74099-3R-3-3, and NSIC Rc 106 were used as donors of the SalTol tolerance gene. Varieties with the Sub1A locus were used as donors of the flood resistance gene: Br-11, CR-1009, Inbara-3, TDK-1, and Khan Dan. The lines C101-A-51 (Pi-2), C101-Lac (Pi-1, Pi-33), IR-58 (Pi-ta), and Moroberekan (Pi-b) were used to transfer blast resistance genes. Hybridization of the stress-sensitive domestic varieties Novator, Flagman, Virazh, and Boyarin with donor lines of the genes of interest was carried out. As a result of the studies carried out using molecular marking based on PCR in combination with traditional breeding, early-maturing rice lines with genes for resistance to salinity (SalTol) and flooding (Sub1A), suitable for cultivation in southern Russia, were obtained. Introgression and pyramiding of the blast resistance genes Pi-1, Pi-2, Pi-33, Pi-ta, and Pi-b into the genotypes of domestic rice varieties were carried out. DNA marker analysis revealed disease-resistant rice samples carrying 5 target genes in a homozygous state. The created rice varieties that carry the genes for blast resistance (Pentagen, Magnate, Pirouette, Argamac, Kapitan, and Lenaris) were submitted for state variety testing. The introduction of such varieties into production will allow us to avoid epiphytotic development of the disease, preserving the biological productivity of rice and obtaining environmentally friendly agricultural products.
Due to the fact that blast (causative agent – Pyricularia oryzae Cav.) is considered to be one of the harmful diseases of rice around the world, weeds compete with the crop for light, mineral nutrition and space, the accelerated development of resistant genotypes for these stressors is very relevant. The use of modern biotechnological approaches (molecular marking) is promising and especially in demand in breeding rice varieties of a new generation. This article presents the results on the introduction and pyramiding in the same genotype blast resistance genes Pi-1, Pi-2, Pi-33, Pi-ta, Pi-b, Pi-40 and the gene for tolerance to prolonged flooding Sub 1A, as a weed control factor, based on domestic rice varieties Flagman, Snezhinka, Novator, Boyarin, as well as large-grain lines, with a short growing season VNIIR5242, KP-25-14, KP-163 and VNIIR9678. As a result of the volumetric work using marker control of target genes in the genotypes of hybrid plants, 4 modern varietal samples and more than 400 backcross self-pollinated rice lines with introduced and pyramided blast resistance genes, as well as backcross self-pollinated lines with Pi and Sub1A genes, were obtained. These plants are adapted for cultivation in the south of Russia, have a duration of 115-117 days, a height of 87-100 cm, a mass of 1000 grains – 30 or more grams, a yield of 8.5 – 11 t/ha, which is significantly higher than that of the standard variety Flagman.
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