1. The institute covers wide range of activities on areas of practical plant breeding, seed production, preservation of plant genetic resources, applied and fundamental research, evolvement in postgraduate studies and advisory activities. The main aim of Priekuli PBI is to develop and introduce into production new varieties of barley, winter rye, winter triticale, pea and potato with traits suitable for Latvia growing conditions and acceptable to producers demands. State Priekuli Plant Breeding Institute was founded in 1913, and is the oldest and most experienced agricultural scientific centre in Latvia. Within the performance period 103 crop varieties have been created, including 42 potato, 18 perennial grasses, 11 spring barley and 11 pea varieties. In total breeding of 31 crop spiecies has been caried out. Acknowledgements Implementation of molecular markers has been funded by EU Structural Funds (project: “Development, improvement and implementation of environmentally friendly and sustainable crop breeding technologies”, 2009/0218/1DP/1.1.1.2.0/09/APIA/VIAA/099 ), Latvian Council of Science, Latvian National Programme in Agrobiotechnology Acknowledgements Implementation of molecular markers has been funded by EU Structural Funds (project: “Development, improvement and implementation of environmentally friendly and sustainable crop breeding technologies”, 2009/0218/1DP/1.1.1.2.0/09/APIA/VIAA/099 ), Latvian Council of Science, Latvian National Programme in Agrobiotechnology Use of molecular markers in State Priekuli Plant Breeding Institute Ieva Mežaka 1, Linda Legzdiņa 1, Ilze Skrabule 1, Arta Kronberga 1, Māra Bleidere 2, Dainis Ruņģis 3, Nils Rostoks 4 1 State Priekuļi Plant Breeding Institute, Zinatnes Str 1a, Priekuļi, Latvia 2 State Stende Cereals Breeding Institute, Dižstende, Latvia 3 Genetic Resource Centre, Rigas Str 111, Salaspils, Latvia 4 Faculty of Biology, University of Latvia, 4 Konvalda Blvd., Riga, Latvia References Lee, D., Reeves, J.C., Cooke, R.J. 1996. DNA profiling and plant variety registration: 1. The use of random amplified DNA polymorphisms to discriminate between varieties of oilseed rape. Electrophoresis 17: 261–265. Piffanelli, P., Ramsay, L., Waugh, R., Benabdelmouna, A., D'Hont, A., Hollricher, K., Jørgensen, J. H., Schulze-Lefert, P., Panstruga, R. 2004. A barley cultivation-associated polymorphism conveys resistance to powdery mildew. Nature 430, 887-891 -> mlo11 Kokina, A., Rostoks, N.(2008) Genome-wide and Mla locus-specific characterization of Latvian barley varieties, Proceedings of the Latvian Academy of Sciences, Section B, 62: 103 – 109 References Lee, D., Reeves, J.C., Cooke, R.J. 1996. DNA profiling and plant variety registration: 1. The use of random amplified DNA polymorphisms to discriminate between varieties of oilseed rape. Electrophoresis 17: 261–265. Piffanelli, P., Ramsay, L., Waugh, R., Benabdelmouna, A., D'Hont, A., Hollricher, K., Jørgensen, J. H., Schulze-Lefert, P., Panstruga, R. 2004. A barley cultivation-associated polymorphism conveys resistance to powdery mildew. Nature 430, 887-891 -> mlo11 Kokina, A., Rostoks, N.(2008) Genome-wide and Mla locus-specific characterization of Latvian barley varieties, Proceedings of the Latvian Academy of Sciences, Section B, 62: 103 – 109 A few facts about State Priekuļi Plant Breeding Institute First steps of implementation of MAS … of barley Disease and Pest Resistance Quality and agronomical performance Marker use in seed production Previous studies (Lee et al 1996) show that there is a genetic diversity within varieties despite the fact that they are self-polinated species. In last years we have encountered a problem that some triticale and barley breeding lines are not uniform even after 13 times of selfing and fail DUS test. To select genetically homogeneous individuals from such breeding lines we used highly polymorphic molecular markers – AFLP and retrotransposons and examined varieties on plant-by-plant basis. Morphological characteristics were described for each individual and compared with marker data. Individuals with uncommon alleles or morphological differences were discarded. Selection of homogeneous individuals has been successful: triticale and barley varieties displayed uniformity next year after selection and have entered DUS testing. Marker use in seed production Previous studies (Lee et al 1996) show that there is a genetic diversity within varieties despite the fact that they are self-polinated species. In last years we have encountered a problem that some triticale and barley breeding lines are not uniform even after 13 times of selfing and fail DUS test. To select genetically homogeneous individuals from such breeding lines we used highly polymorphic molecular markers – AFLP and retrotransposons and examined varieties on plant-by-plant basis. Morphological characteristics were described for each individual and compared with marker data. Individuals with uncommon alleles or morphological differences were discarded. Selection of homogeneous individuals has been successful: triticale and barley varieties displayed uniformity next year after selection and have entered DUS testing. The benefits we expect from MAS Use of marker assisted selection in early segregating generations could help to identify which segregants carry the desirable trait and discard the rest, therefore diminishing costs Especially significant for detecting resistance to diseases which traditionally are assessed at field conditions under natural infection, but does not occur every year Evaluation of disease resistance which have long and complicated phenotypic evaluation Drawbacks Most of the published markers are applicable only to certain populations Desirable traits often are inherited in a complex manner We are able to use only PCR- based markers The benefits we expect from MAS Use of marker assisted selection in early segregating generations could help to identify which segregants carry the desirable trait and discard the rest, therefore diminishing costs Especially significant for detecting resistance to diseases which traditionally are assessed at field conditions under natural infection, but does not occur every year Evaluation of disease resistance which have long and complicated phenotypic evaluation Drawbacks Most of the published markers are applicable only to certain populations Desirable traits often are inherited in a complex manner We are able to use only PCR- based markers  Protein content  Starch content  Aminoacid content  Lodging-resistance  Vitamin E content  Beta glucan content  Low phytate content  Resistance to pre-harvest sprouting Loose smut Fusarium head blight Powdery mildew Net blotch Leaf stripe … of potato Disease and Pest Resistance  Nematodes  Late blight  Potato viruses  PVX  PVM  PVS  PVY Seed production Traits used in selection … of triticale … of rye … of pea Types of consumption For serving as a side dish Crisps French fries Baking in oven Processing into starch Barley association mapping Molecular markers employed in breeding favorably should be applicable to various populations. Association mapping offers possibility to find a correlation between SNP polymorphisms and desirable traits, phenotyping a wide material of genotypes. We have initiated barley association mapping. Genotyping has been done using Illumina GoldenGateTM Assay which detects Single Nucleotide Polymorphisms. Following traits have been phenotyped in six environments (3 years, 2 locations): beta –glucan, vitamin E, lysine, starch and protein content in grain. The aim is to use SNP polymorphisms linked to desirable traits to develop CAPS markers. Barley association mapping Molecular markers employed in breeding favorably should be applicable to various populations. Association mapping offers possibility to find a correlation between SNP polymorphisms and desirable traits, phenotyping a wide material of genotypes. We have initiated barley association mapping. Genotyping has been done using Illumina GoldenGateTM Assay which detects Single Nucleotide Polymorphisms. Following traits have been phenotyped in six environments (3 years, 2 locations): beta –glucan, vitamin E, lysine, starch and protein content in grain. The aim is to use SNP polymorphisms linked to desirable traits to develop CAPS markers. Stable yield Protein content Falling number Lodging-resistance Winter hardiness Bread –making properties Water absorbtion -testing by MAS or marker adaptation initiated Conservation of plant genetic resources Principial tasks Research on crop producton management Potato Barley Rye Oats Peas Grasses Clovers Wheat Potato Barley Rye Triticale Peas Breeding Lodging-resistance Winter hardiness Vitamin E content Aminoacid content Powdery mildew Snow mould Yield stability Protein content Falling number Feed Biomass Protein content Types of consumption Traditional grey peas Porridge Soups Pyramiding powdery mildew To develop varieties with durable resistance and to avoid formation of new mildew stains, pyramiding barley powdery mildew resistance genes has been initiated in barley breeding. PCR- based markers linked to resistance genes mlo11 (Piffanelli et al 2004) and Mla18 (Kokina and Rostoks 2008), have been used. The strategy is to test F4 generation lines, discard susceptible ones, and to evaluate in next generations only resistant ones. Screeding for Mla 18 (left) mlo11 (right) gene ir hybrids. D- DNA ladder, H- heterozygous, R- resistant, S – succeptible individuals, N- negative control D N R H SD H R R R S S Mla18mlo11 Seed production -testing by MAS desirable