Analysis of Structure of Septoria Nodorum Berk., Septoria Tritici Rob. Et Desm Populations in Kazakhstan and Neighbouring Territories and Creation of Tolerant to Septoriosis Wheat Base Lines Using Cell Selection Methods
Tech Area / Field
- AGR-PPR/Plant Protection/Agriculture
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
3 Approved without Funding
Kazakh Research Institute of Plant Protection, Kazakstan, Almaty reg., Rakhat
- National Biotechnology Center of Kazakstan / Institute of Plant Biology and Biotechnology, Kazakstan, Almaty\nThe Republican Government Enterprise on the basic of economic control rights “Research Institute for Biological Safety Problems” , Kazakstan, Gvardeiski
Project summaryThe aim of the project is analysis of the structure of Septoria nodorum Berk., Septoria tritici Rob. Et Desm populations in Kazakhstan and neighboring territories and creation of tolerant to Septoriosis wheat base lines using cell selection methods.
Septoriosis is a wide spread and very harmful wheat disease in almost all countries. In epiphytotic years crop yield losses due to septoriosis may be up to 20-30% of the total. It also worsens grain quality. Agricultural measures are ineffective whereas chemical measures are expensive. Besides, chemical pesticides affect the environment. Therefore, cultivation of tolerant to septoriosis wheat varieties is the most effective way to solve the problem both from economic and ecological points of view. However, traditional selection methods are rather expensive and time consuming (10-15 years). During this period of time new races of pathogens able to affect the new sorts may appear.
Biotechnological methods accelerate selection and increase its efficiency. The most perspective method is cell selection. It let to obtain quickly resistant to septoriosis lines on selective media, containing toxins, cultural filtrates and pure fungi strains. These lines may be used as the initial material for selection of resistant to septoriosis wheat sorts.
Investigation of mycotoxin action mechanisms and development of cell selection technology for culturing of tolerant to fungi infection sorts meet up to date scientific level of researches targeted to investigation of interaction in “parasite –host” system and usage of biotechnological methods in selection. The main idea of the project was reported by Karabaev M.K. at the conference organized in Kazakhstan under the aegis of CIMMYT (International Maize and Wheat Improvement Center) in 1997. Usage of biotechnological methods in selection was approved by N.E.Borlaug, world famous geneticist and selectioner, the Nobel Prize winner, during his visit to Almaty in 1998.
· 1986-1999. We investigated Septoria nodorum and Septoria tritici distribution in Kazakhstan and neighboring West Siberia, South Urals and Kyrghyzstan. We possess many year information on the rate of harm of the disease and on its seasonal dynamics in different soil and climate zones of Kazakhstan. We elaborated protection measures for septoriosis of spring and winter wheat and established the optimum terms for fungicides use taking into account the rate of disease development and genotype features of wheat sorts. We determined predictors of disease development depending on weather conditions. We assessed septoriosis strike rate of wheat sorts cultivated in Kazakhstan as well as newly selected wheat samples. We found out that only few genotypes out of 1685 tested germoplasma variants possess tolerance to septoriosis (strike rate up to 20%).
· We elaborated nutrition media, light and temperature parameters for cultivation of Septoria nodorum and Septoria tritici and regimes for long term storage of inoculums (up to 6 years). We elaborated original methods for assessment of wheat tolerance to septoriosis using fungi isolates with different virulence as inoculums. We isolated FT-2 and FT-5 toxins of Septoria nodorum.
· We got 4 type of wheat callus tissues with different morphogenetic potential. We isolated embryogenic calluses that keep cell totipotency for a long time (36 months). From embryogenic calluses we obtained suspended cultures of wheat cells.
The project will be carried out by qualified experts of three research institutes:
· Kazakh Research Institute of Plant Protection
Koishibaev M., Professor (Agriculture), head of Phytopathology Department
Alipbekova S.A., Ph.D. (Agriculture), senior researcher of the same department.
Alipbekov O.A., Ph.D. (Agriculture), senior researcher of Pesticide Application Laboratory.
Ismailova E.T., Ph.D.. (Agriculture), senior researcher of Phytopathology Department.
· Institute of Plant Physiology, Genetics and Bioengineering
Rakhimbaev I., Professor (Biology), Director of the Institute and Head of Laboratory of Biotechnology
Bishimbaeva N.K., Ph.D. (Biology), senior researcher of Cell Engineering Laboratory
Kushnarenko S.V., Ph.D. (Biology), senior researcher of Laboratory of Biotechnology.
· Research Agricultural Institute
Rsaliev S.S., Ph.D., (Biology), head of Laboratory of Plant Immunity
Zaitsev V.L. Ph.D., (Biology), head of Laboratory of Molecular Biology and Toxicology
Savenkov A.F., Ph.D. (Technology), head of Laboratory of Radiobiology and Ecology.
Scientific Consultant: Karabaev M.K., Professor (Biology), representative of CIMMYT in Kazakhstan.
· Information on Septoria nodorum and Septoria tritici population structure depending on their virulence.
· Replenishment of the fungi collection with new clones with morphologically and physically stable features.
· Information on cell mechanisms of mycotoxin action for development of the knowledge on “parasite- host” interaction.
· Technology for cell selection of tolerant to mycotoxins wheat.
· Express methods for screening of wheat tolerance to septoriosis in vivo and in vitro.
· Tolerant to septoriosis wheat lines, obtained via cell selection.
Application of the results:
· Obtained wheat lines will be used in selection to produce new highly productive and tolerant to septoriosis sorts.
Meeting ISTC Goals and Objectives:
· The project gives the possibility for scientists related to development of biological weapon to route their activity to agricultural problems, in particular, to wheat selection.
· Integration of Kazakhstan scientists into international science community.
· The project will promote solution of national and global problems increasing of wheat productivity and developing genetic basis for wheat selection.
Scope of activities. The following tasks will be accomplished:
· Isolation of pure Septoria nodorum and Septoria tritici cultures from different geographical areas of wheat cultivation, comparative assessment of their virulence, creation of fungi collection, selection of highly virulent strains.
· Isolation of mycotoxins and their chemical characterization. Investigation of cell mechanisms of mycotoxin action.
· Creation of experimental systems for cell selection in vitro (culture of tissues, culture of cells).
· Obtaining of tolerant to septoriosis diploid and haploid wheat cell lines. Regeneration of diploid and di-haploid plants and testing of their tolerance to septoriosis in laboratory conditions.
· Estimation of the rate of tolerance to septoriosis in field conditions with infection background. Selection of tolerant to septoriosis wheat lines.
Role of foreign collaborator:
· Identification of the gene providing wheat tolerance to septoriosis.
· Usage of tolerant to septoriosis wheat base lines in selection.
· Assessment of the technical reports.
· Participation in technical audit of the project carried out by ISTC personnel.
· Joint workshops and information exchange.
Technical approach and methodology:
· Route investigation of wheat crops and collection of affected plants in different soil and climate zones of Kazakhstan and neighboring countries in order to analysis of Septoria nodorum and Septoria tritici population structure. Isolation of the pathogens and obtaining of the pure fungi cultures will be carried out using conventional research methods of mycology and phytopathology. Virulence of the fungi isolates will be analyzed using local wheat sorts and germoplasma samples from the CIMMYT collection with different rate of tolerance to septoriosis.
· Cell selection for tolerance to septoriosis will be carried out in callus tissues and suspension cell cultures of winter and spring wheat. We plan to use toxins, cultural filtrates and pure Septoria nodorum and Septoria tritici cultures as selection agents. To increase the level of genetic cell variability we plan to use chemical mutagene, N-nitrosomethylurea. Plants will be regenerated from somatic embryos formed from tolerant to selection agents- cell and tissue cultures. Tolerance of plant-regenerates to septoriosis will be assessed in laboratory conditions. Tolerance to septoriosis of wheat lines selected by cell selection method will be determined in field conditions with infection background. Infection will be carried out in stooling and earing phases through treatment of the plants with conidium suspension of highly virulent fungi strains, cultivated in artificial or natural substrates.