PPP for pre-breeding in perennial ryegrass (Lolium perenne L.) Petter Marum Graminor AS

Project partners DLF (Niels Christian Nielsen) Lantmännen (Christer Persson) Graminor (Petter Marum) Boreal (Miko Isolathi) Agricultural University of Iceland (Aslaug Hegadottir) Norwegian University of Life Sciences (UMB) (Odd Arne Rognli) Univeristy of Aarhus (Torben Asp) Jõgeva Plant Breeding Institute (Rene Aavola)

Pre-breeding program The aim of this pre-breeding program is to identify and select new plant materials for development of cultivars with a suitable adaption to future climates The pre-breeding program is a continuous and long-term activity

Pre-Breeding In Situ Genebank (Ex Situ) Plant breeding research Pre-breeding Cultivar development Official variety testing DUS Seed Companies Farmers

Perennial ryegrass Lolium perenne L. The main forage grass species in Denmark and south-wards in Europe Important in southern Sweden and parts of southwestern Norway Superior feed quality and productivity under frequent cutting regimes Is expected to expand further north due to milder winters with shorter periods of snow cover

Perennial ryegrass Lolium perenne L. The main weaknesses are: Susceptibility to low-temperature pathogens Inadequate growth cessation in autumn to allow for sufficient cold hardening. Can deharden too early with increased risk of frost injury in spring

Perennial ryegrass Lolium perenne L. Show local adaption to temperature and photoperiod (like all forage grasses) The photoperiod does not change with climate change that cause the temperature to rise. This raises the need for new ideotypes with a different set of physiological traits than the present cultivars.

Perennial ryegrass Lolium perenne L. Important questions are then: Is the available germplasm and active breeding populations sufficiently broad Or should new exotic materials be identified and recombined with existing material to serve as the new genetic resources for development of new cultivars

Project budget 2012-2013 Total cost is DKK 3.143.700 Companies DKK 1.571.850 (50%) NMR DKK 1.571.850 (50%)

Project outline

Work packages WP1 WP2 WP3 WP4 WP5 WP6 Locating and requesting Lolium Perenne L. accessions and cultivars Multiplication of gene bank accessions for further testing Testing the potential winter hardiness of existing perennial ryegrass cultivars Phenotyping of gene bank accessions Establishing a broad breeding population (Base broadening) Genetic diversity analyses and trait/marker associations

Pre-breeding in perennial ryegrass (Lolium perenne L.) WP1 Requesting seed WP2 WP3 WP4 Seed multiplication Cultivar trials Phenotyping Broad breeding population Genotyping, diversity WP5 WP6 2012 N, DK DK N, S, IS FIN, EE N, DK, S, FIN N, DK 2013 2014 WP7 WP10 2015 WP8 WP10 2016 2017 WP10 2018

WP1 Locating and requesting Lolium Perenne L. accessions and cultivars Responsible: DLF, (Graminor) Participating: DLF, Graminor, (NordGen) Timeframe: 2012

WP1 The aim was to acquire 400 accessions for WP2, WP4, WP5 and WP6 Received about 400. 20 commercial cultivars with very good winter hardiness for WP3 Ended up with 22 cultivars

WP2 Multiplication of gene bank accessions for further testing Responsible: DLF Participating: DLF Timeframe: 2012-2013

WP3 Testing the potential winter hardiness of existing perennial ryegrass cultivars Responsible: Agricultural University of Iceland (AUI) Participating: AUI, Lantmännen, Graminor, Boreal, and Jõgeva Timeframe: 2012-2015

Cultivars in WP3 Cultivar Ploidy Country Premium 2n CH Arvicola 2n CH Cavia 2n CH Ivana 2n D Pionero 4n D SW Irene 2n S SW Birger 4n S SW Leia 4n S SW Svea 2n S SW Gunne 2n S Riikka 2n FI Raidi 2n EST Raite 4n EST Faherlin 2n N Spdidola 2n LV Arvella 2n CH Salamandra 4n CH Norlea 2n CAN Falk 4n N Einar 4n N Trygve 4n N Fjaler 4n N

WP4 Phenotyping of gene bank accessions Responsible: Lantmännen Participating: Lantmännen, DLF, Graminor, and Boral Timeframe: 2012-2013

WP4 Observations will be done on 20 plants per accession 5 plants x 4 reps x 400 acc. Important traits Winter survival Spring growth Growth habit Plant height Disease attack Crown rust

WP4 Phenotyping will be divided between participants Crown rust is most frequent in Denmark Winter damage most severe in Norway and Finland Data will be used in WP10 (next phase 2014 -) Establish marker-trait associations

Establishing a broad breeding population (Base broadening) Responsible: Graminor Participating: Graminor and DLF Timeframe: 2012-2013 WP5 WP5

WP5 Two crossing blocks Norway Denmark 10 plants per accession Recombined in two generations Harvest one seed head form each plant and use it to establish one new plant for production of second generation (SSD) Basis for pre-breeding work with a time-frame of 15-20 years or more. Develop site specific populations by natural selection

WP6 Genetic diversity analyses and trait/marker associations Responsible: UMB Participating: UMB and University of Aarhus Timeframe: 2012-2013

WP6 Genotype the accessions using a suitable marker system Preferably SNPs Estimate the genetic distance between the accessions Establish marker-trait associations (WP10 next phase)