Acknowledgements

1. Use of molecular markers in State Priekuli Plant Breeding Institute Ieva Mežaka1, Linda Legzdiņa1, Ilze Skrabule1, Arta Kronberga1, Māra Bleidere2, Dainis Ruņģis3, Nils Rostoks4 1State Priekuļi Plant Breeding Institute, Zinatnes Str 1a, Priekuļi, Latvia 2State Stende Cereals Breeding Institute, Dižstende, Latvia 3Genetic Resource Centre, Rigas Str 111, Salaspils, Latvia 4Faculty of Biology, University of Latvia, 4 Konvalda Blvd., Riga, Latvia A few facts about State Priekuļi Plant Breeding Institute • Potato • Barley • Rye • Triticale • Peas Breeding Principial tasks • Potato • Barley • Rye • Oats • Peas • Grasses • Clovers • Wheat Seed production Seed production State Priekuli Plant Breeding Institutewas 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. 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 isto 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. Conservation of plant genetic resources Research on crop producton management Traits used in selection • -testing by MAS or marker adaptation initiated • -testing by MAS desirable Quality and agronomical performance • Protein content • Starch content • Aminoacid content• • Lodging-resistance • Vitamin E content • • Beta glucan content• • Low phytate content • Resistance to pre-harvest sprouting Types of consumption • Loose smut• • Fusarium head blight • • Powdery mildew• • Net blotch • • Leaf stripe • • For serving as a side dish • Crisps • French fries • Baking in oven • Processing into starch … of triticale … of rye • Feed • Biomass • Protein content • Types of consumption • Traditional grey peas • Porridge • Soups • Stable yield • Protein content • Falling number • Lodging-resistance • Winter hardiness • Bread –making properties • Water absorbtion • Lodging-resistance • Winter hardiness • Vitamin E content • Aminoacid content • Powdery mildew• • Snow mould • Yield stability • Protein content • Falling number … of potato … of pea … of barley Disease and Pest Resistance Disease and Pest Resistance First steps of implementation of MAS • Nematodes• • Late blight • • Potato viruses • • PVX • • PVM • • PVS • • PVY • 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. • 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 Pyramiding powdery mildew Mla18 mlo11 D H R R R S S D N R H S Screeding for Mla 18 (left) mlo11 (right) gene ir hybrids. D- DNA ladder, H- heterozygous, R- resistant, S – succeptible individuals, N- negative control 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. 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 IlluminaGoldenGateTM 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. • References • Lee, D., Reeves, J.C. , Cooke, R.J. 1996. DNA profiling andplant variety registration: 1. The use of random amplified DNApolymorphisms 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 ofLatvian barley varieties, Proceedings of the Latvian Academy of Sciences, Section B,62: 103 – 109 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