project proposal for public private partnership in …...biotic and abiotic stresses in the future....

Project proposal for Public Private Partnership in pre-breeding:

Combining Knowledge from Field and from Laboratory for Pre-

breeding in Barley II

Project proposal

Summary

The main goal for the project ”Combining Knowledge from Field and from Laboratory for Pre-

breeding in Barley II” is to lay the foundation for effective cereal breeding for disease resistance and

harvest stability in changing climate conditions capable to meet current and future challenges in the

Nordic region. The first part of the project has resulted in the identification of several markers

associated with nematode resistance, powdery mildew resistance, scald resistance, plant height,

earliness, straw length etc. However, three years is too short time for long term pre-breeding

activities. Based on the results of the ongoing PPP project we have obtained a detailed and valuable

knowledge of the genetic pool available in the Nordic material right now. In order to prepare for the

future environmental challenges it is necessary to introduce a larger variability of genes for abiotic

and biotic stresses with stronger emphasis on long term goals. Preparations for the next phase of the

PPP-project have been made in the first PPP project period in ‘WP Preparing for the future’ by

initiating the development of multi-parent advanced generation inter-cross (MAGIC) populations.

The objectives with the next phase of the PPP-project is to develop a new generation of mapping

populations, such as MAGIC that should overcome the limitations of traditional bi-parental and

association mapping populations. Genome-wide association studies (GWAS) will be conducted on

these populations to provide the DNA markers linked to diseases resistance as well as to agronomic

traits. The MAGIC populations will possess combined resistance to different diseases in a single line.

All information regarding the original sources of resistance, DNA markers linked to resistance genes

as well as lines with combined resistance genes will be available for all project partners. The crossing

populations can then be incorporated in their breeding programs and used for e.g. marker assisted

backcrossing (MAB).

PPP-Proposal: Combining knowledge of field and laboratory for PPP in barley II

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Time frame

1st of January 2015 to the 31st of December 2017

Sum requested and total budget

DKK 15.417.814 in total

DKK 7.624.864 requested and DKK 7.792.950 contributed by the breeding institutions

Project coordination (CV attached)

Ahmed Jahoor Nordic Seed Højbygårdvej 14 DK-4960 Holeby Denmark Phone: +45 2913 4757

Email: [email protected]

Scientific output from the first barley PPP including the one year extension period.

Therese Bengtsson et al. Genetic diversity and population structure in Nordic spring barley. In preparation. Therese Bengtsson et al. Identification and mapping of Powdery mildew resistance loci on chromosome 6H. In preparation.

mailto:[email protected]


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Project participants with addresses and contact information

Participant Contact person

Nordic Seed Højbygårdvej 14 DK-4960 Holeby Denmark www.nordicseed.com

Ahmed Jahoor +45 29134757 [email protected]

Sejet Planteforædling I/S Nørremarksvej 67, Sejet DK-8700 Horsens Denmark www.sejet.com

Birger Eriksen +45 75682177 [email protected]

Graminor Breeding AS Hommelstadvegen 60 N-2322 Ridabu Norway www.graminor.no

Lars Reitan +47 48041300 [email protected]

Boreal Plant Breeding Myllytie 10 FI-31600 Jokioinen Finland www.boreal.fi

Outi Manninen +358 407785673 [email protected]

LUKE Natural Resources Institute Finland Tietotie 4 FI-31600 Jokioinen, Finland www.luke.fi

Marja Jalli +358 295317261 [email protected]

Agricultural University of Iceland (LBHI) Faculty of Land and Animal Resources Keldnaholt IS-112 Reykjavik Iceland www.lbhi.is

Áslaug Helgadóttir +354 4335255 [email protected]

Swedish University of Agricultural Sciences (SLU) Department of Plant Breeding Sundsvägen 10 SE-230 53 ALNARP Sweden www.slu.se

Inger Åhman +46 040415240 [email protected]


http://www.luke.fi/


http://www.slu.se/


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Introduction

Barley (Hordeum vulgare L.) is one of the most important crops in the Nordic regions of Europe

(Denmark, Finland, Norway, Iceland and Sweden) where it covered a total area of 1.73 million ha in

2013 excluding Iceland (http://faostat.fao.org). It is mainly used for malting, feed and food.

Nordic barley breeding began more than 100 years ago, starting from utilization of the variation in

the landrace gene pool to cultivation of modern elite varieties. The commercial breeding programs

within the Nordic countries have a long term focus on the current main diseases of barley and the

search for new sources of resistance remains an important factor when breeding for resistance.

Plant pathogens are a constant threat in cereal production in Northern Europe. By reducing crop

quality and yield they not only threaten food safety but cause economic losses as well as an

environmental burden. Today the disease control in barley mainly relies on moderately resistant to

resistant cultivars and the use of fungicides. The implementation of the EU legislation that will

restrict the use of fungicides and the wish to increase organic production, further highlights the

importance of finding new sources of resistance for disease control. Disease resistance based on

genetic resistance is an environmentally sound and economical alternative for plant disease control.

The Nordic region exhibits a large variation in climate and soil, where the eastern part of Finland and

inland Sweden has an inland climate, the southern part has a mild maritime climate and coastal

Norway and Iceland has a cool maritime climate. The climate change is predicted to cause more

extreme weather conditions such as prolonged periods of drought and severe cloud bursts in the

Nordic countries (http://www.dmi.dk/dmi/index/klima/). Diseases such as Spot blotch (Bipolaris

sorokiniana), Ramularia leaf spot (Ramularia colly-cygni) and Fusarium head blight (Fusarium

graminearum) are examples of new and emerging barley diseases related to climate change in the

Nordic region. In addition to emerging diseases an anticipated increased temperature with retained

photoperiod is expected to alter the growth habit (Bragason, 1985). In the first phase of the project

a variation was seen between locations for maturity due to differences in the length of growth

season and temperature. Therefore, it is important to increase the variability and diversify the

source of desired genes/traits that has a low genotype by environment interaction to sustain both

biotic and abiotic stresses in the future.

Understanding which genes are affecting important traits such as yield, quality, disease resistance

and environmental adaptation is a fundamental part in modern plant breeding. Most plant traits

show complex genetic inheritance, with phenotypic expression under polygenic control at

quantitative trait loci (QTL). Although the identification of simply inherited traits has been successful,

the discovery of genes for more complex traits has been restricted by the absence of proper genetic

resources, both in terms of genotyping methods and in terms of breeder-relevant germplasm

(Cavanagh et al. 2008). However, thanks to recent advances in genotyping capabilities, genetic

marker density no longer restricts QTL discovery in plants. Rather the concern in QTL discovery now

is the limitations of the type of mapping population used and the bottle neck of phenotyping.

Typically F2, backcross (BC), double haploid or recombinant inbred line (RIL) populations derived

from two parents have been used for QTL mapping. The problem with using bi-parental populations

is that only two alleles are analyzed (in diploid species) and that the resolution for QTL detection is

restricted due to limited gene recombination (Li et al. 2010; Rakshit et al. 2012).

http://faostat.fao.org/

http://www.dmi.dk/dmi/index/klima/


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Barley has been a model species for the development of QTL mapping based on bi-parental

populations but due to the development of high density genotyping platforms, there has been a shift

from traditional QTL mapping to genome-wide association studies (GWAS) in barley (Waugh et al.

2009). Association mapping relies on linkage disequilibrium (LD) to exploit the correlation between

phenotype and genotype in populations of unrelated individuals, hence it has the advantage of many

more generations of recombination resulting in high-resolution mapping (Myles et al. 2009).

However, one concern in association mapping is the false positive associations caused by a strong

population structure (Pritchard et al. 2000). In cultivated barley strong population structure is often

associated with spike row number and the growth habit (e.g. spring vs. winter barley) (Hamblin et al.

2010), thus GWAS need to correct for the population structure stratification. For controlling

population structure and relatedness within GWAS, mixed linear models (MLM) have been useful,

but they can still be computationally challenging for large datasets (Zhang et al. 2010). Recent

development of statistical methods such as the unified mixed model, efficient mixed model

association (EMMA), the compressed MLM, and population parameters previously determined

(P3D), have reduced computing time and improved the statistical power by clustering individuals

into groups (Zhang et al. 2010). Association mapping was usefully employed in the first phase of

project and several DNA markers were identified and validated in the breeding material by each

company.

To overcome these limitations caused by bi-parental populations and association mapping

populations due to presence of rare alleles, next-generation mapping populations have been

developed and started to be utilized (Morrell et al. 2012). Some of the identified markers from the

first PPP project, e.g., for earliness and straw length, were associated with rare alleles in the Nordic

barley population and in such cases GWAS is an unsuitable method. Hence, a way to overcome this

problem is to develop segregating populations that will increase the frequency of the rare alleles

thus enable GWAS. The multi-parent advanced generation inter-cross (MAGIC) is one example of

these new mapping resources, in this case created by several generations of inter-crossing among

multiple founder lines. The MAGIC population contributes to a higher allelic diversity than a bi-

parental population and gives greater opportunities for recombination and, hence greater precision

in QTL location (Cavanagh et al. 2008). In addition, MAGIC populations allow the use of both linkage

and association analysis without the restrictions encountered with highly structured populations

(Rakshit et al. 2012). So far, MAGIC and MAGIC-like populations have been developed in a few crop

species including rice (Bandillo et al. 2013), barley (Sannemann, 2013), spring wheat (Huang et al.

2012) and winter wheat (Mackay et al. 2014). In crop plants MAGIC population can provide not only

sources of novel trait QTL combinations for breeding but also can provide training population for

genomic selection.

Educating the next generation of plant breeders

There is a need for strengthened research education in plant breeding in the Nordic region,

combining more traditional methods and quantitative genetics with different aspects of plant

biotechnology (Nilsson and von Bothmer, 2010). Only few candidates have this general

agricultural/horticultural profile including necessary aspects on practical breeding and genetics.

Candidates with a background in plant biotech normally lack these skills in their training. In the first

phase of the Barley-PPP (2012 – 2014) one PhD student and two Post-Docs was assigned to the


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project. By continuing the educational aspect in phase two, the next generation of plant breeders

will be trained.

Underlying idea with the second phase

In the first phase of the Barley-PPP project (2012-2014) we have obtained a detailed and valuable

knowledge of the genetic pool available in the Nordic material right now. Several markers have been

identified such as markers associated with nematode resistance, powdery mildew resistance, scald

resistance, plant height, earliness, straw length etc. In some cases easy-to-use-markers have been

developed for the companies to use in their screening of breeding materials. However, three years is

too short time for long term pre-breeding activities. One of the objectives of the second phase is to

introduce a larger variability of genes for abiotic and biotic stresses with emphasis on various

climatic conditions and on long term goals. This project will:

a) Develop pre-competitive multi-parent advanced generation intercross (MAGIC) populations

for mapping of novel disease and agronomic traits (The development of MAGIC populations

was already initiated in spring 2014 and a second round of crosses was conducted in autumn

2014). WP1

b) Produce homozygous segregating plant lines from each MAGIC population that will be

screened for a broad selection of diseases and agronomic traits at different locations in the

Nordic countries. WP2

c) Provide the preliminary molecular knowledge and tools for current and future

implementation of high-throughput marker system in Nordic barley cultivar

development.WP3, WP6

d) Provide knowledge of disease resistance genetics and screening tools for current and future

important disease evaluation. WP3, WP4, WP6

e) Provide physiological knowledge and screening tools for harvest stability traits that are

linked to climate.WP5, WP6

f) Incorporate the new sources in breeding programs. WP7

A GANTT diagram at the end of this application, shows the workload in different WP over the project

period. Shaded in gray are the activities which should continue beyond the project period to get the

full benefit of the work done in this project (Combining Knowledge from Field and from Laboratory

for Pre-breeding in Barley II).


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References

Bandillo et al. 2013. Multi-parent advanced generation inter-cross (MAGIC) populations in

rice: progress and potential for genetics research and breeding. Rice 6:11.

Bragason Á. 1985. Sammenligning af vårbygpopulationer i Danmark og Island. Den kgl.

Veterinær- og Landbohøjskole, Afdelingen for Landbrugets Plantekultur, Copenhagen,

Denmark.

Cavanagh et al. 2008. From mutations to MAGIC: resources for gene discovery, validation

and delivery in crop plants. Curr. Opin. Plant Biol 11:215–221.

Hamblin et al. 2010. Population Structure and Linkage Disequilibrium in U.S. Barley

Germplasm: Implications for Association Mapping. Crop Sci 50:556-566.

Huang et al. 2012. A multiparent advanced generation inter-cross population for genetic

analysis in wheat. Plant Biotechnol. J 10:826–839.

Li et al. 2010. Statistical properties of QTL linkage mapping in biparental genetic populations.

Heredity 105:257-267.

Lipka et al. 2012. GAPIT: Genome Association and Prediction Integrated Tool. Bioinformatics,

2012, doi:10.1093/bioinformatics/bts444.

Mackay et al. 2014. An Eight-Parent Multiparent Advanced Generation Inter-Cross

Population for Winter-Sown Wheat: Creation, Properties and Validation. G3 4:1603-1609.

Morrell et al. 2012. Crop genomics: advances and applications. Nature Rev Genet 13:85–96.

Myles et al. 2009. Association Mapping: Critical Considerations Shift from Genotyping to

Experimental Design. Plant Cell 21:2194-2202.

Nilsson and von Bothmer. 2010. Measures to promote Nordic plant breeding. TemaNord

2010:518. Nordic Council of Ministers, Copenhagen.

Pritchard et al. 2000. Inference of population structure using multilocus genotype data.

Genetics 155: 945–959.

Rakshit et al. 2012. Multiparent intercross populations in analysis of quantitative traits. J

Genet 91:111-117.

Sannemann, 2013. Marker-trait-sensor association in a multi-parent advanced generation

intercross (MAGIC) population in barley (Hordeum vulgare ssp. vulgare). Doctoral

thesis."Institut für Nutzpflanzenwissenschaften und Ressourcenschutz Professur für

Pflanzenzüchtung Prof. Dr. J. Léon."

Waugh et al. 2009. The emergence of whole genome association scans in barley. Curr Opin

Plant Biol 12:218-222.


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Work package 1: Development of Multiparent Advanced Generation

InterCross (MAGIC) populations

Leader: Jens Due Jensen; Nordic Seed – Denmark

1.1 INTRODUCTION

The main goal of WP1 is the development of pre-competitive Multi-parent Advanced Generation

InterCross (MAGIC) populations for mapping of novel disease resistance and agronomic traits. From

these populations, breeders will be able to incorporate the mapped novel traits in their elite

material using marker assisted backcrossing (MAB). The development of a new generation of

mapping populations, such as MAGIC, should overcome the limitations of traditional bi-parental and

association mapping populations. Some of the advantages of these new populations, including

MAGIC populations, are the following: 1) increased rate of effective recombination per generation,

2) higher level of genetic variability, 3) higher resolution, 4) effective sampling of rare alleles, 5)

elimination of population structure issues, 6) good estimations of allelic effects.

Figure 1.1: Schematic on 8 parent Multiparent Advanced Generation InterCross (MAGIC) population

For the development of MAGIC populations, multiple parents, in our case 8 parents, are used per

populations (Figure 1). The parents in such population consists of 4 elite lines from the Nordic region

combined with 4 lines either coming from intensive screening of the 180 lines from the first barley

PPP project (2012-2014) or from the MAGIC donor screening, conducted in the extension year

(2014) of the first barley PPP project. The development of the MAGIC populations was already

initiated in spring 2014 and a second round of crosses was conducted in autumn 2014. We are

aiming at getting 5 populations, four in which we focus on disease resistance and one where the

focus is earliness. The elite lines and donors have been selected based on their resistance to Leaf


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rust (rph7, rph16 and MBR1012), Scald (new resistance), Fusarium head blight, Bipolaris spot blotch,

Net blotch (net and spot type) and earliness. This was done in the extension of the first barley PPP

project in order to be prepared for a future PPP project but also because all Nordic breeding

companies saw the development of pre-competitive MAGIC populations as a crucial tool in a very

competitive market where the relative small breeding companies in Nordic region are under press

from big multi-national companies. New MAGIC populations will also be developed in the new

project. The principle for them will be the same as those that are already being developed. New

donors will come from the intensive screening of donors done in WP3, described later in this

application.

1.2 TASKS

Task 1.1 Last rounds of crosses in the development of the first set of MAGIC population:

Description: Coordination of the third (last) round of crosses for the already initiated MAGIC

populations during spring 2015, five crosses in total.

Time: Month 2-4.

Deliverables: 20 F1 seed per cross (population) for development of homozygous lines in WP2.

Task 1.2 First rounds of crosses in the second set of MAGIC population:

Description: Initiation and coordination of the development of five new MAGIC populations. Four

populations for mapping of diseases resistance and one for agronomic traits, 20 crosses in the first

round, 10 crosses in the second round and 5 in last round.

Time: Month 14-16 first round of crosses, month 22-24 second round crosses and month 26-28 third

round of crosses.

Deliverables: 20 F1 seed per cross (population) for development of homozygous lines in WP2.

1.3 PARTNERS

Main crossing sites will be Sejet and Nordic seed in Denmark. The coordination and accomplishment

of the crosses is more successful when there is a small physical distance between the two companies

which allows easy exchange of seed and plant material. In special cases other project partners will

be included in carrying out the crosses.

Each partner will have a member in the WP1 MAGIC working group, which will discuss and decide

which parental lines and donors should be crossed.


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Work package 2: Development of homozygous lines from Magic populations

Leader: Lene Krusell; Sejet Planteforædling – Denmark

2.1 INTRODUCTION

One of the main goals of the second PPP project is to introduce a larger variability of genes involved

in disease resistance and important agronomical traits with emphasis on various climatic conditions.

A collection of new sources for resistances and agronomic traits has been identified during the

ongoing PPP project and by comprehensive searching of relevant literature. These sources will be

utilized during the future PPP project, creating MAGIC populations combining a large proportion of

the genetic variation contained in the collection. This gives the opportunity of pyramiding sources

for specific traits or combining several desirable traits in one background. Based on the results of the

ongoing PPP project, we have obtained a detailed and valuable knowledge of the genetic pool

available in the Nordic material right now. In order for us to prepare for the future environmental

challenges it is necessary to introduce new sources thereby increasing the variability of the genetic

pool.

2.2 TASKS

Task 2.1 Techniques for development of homozygous MAGIC population:

Description: Work package 2 deals with the production of homozygous segregating plant lines from

each MAGIC population. These populations will be screened for a broad selection of diseases and

agronomic traits at different locations in the Nordic countries (see WP4 and 5). This requires that

each line is genetically stable, ensuring that it is the exact same genetic pool being screened at the

different locations under various environmental conditions.

Several techniques such as Single Seed Descent (SSD) and Double Haploid (DH) are available for

production of homozygous lines. SSD is a relative uncomplicated method in which conventional

inbreeding involves descending of one seed per plant in each generation. Using this method a

genetically stable (homozygous) population requires 6-7 generations to be reached. Based on the

short timeframe of the PPP project (2015-17) the high number of generations is a critical issue and in

this respect a technique such as Double Haploids would be more suitable. This technique represents

a fast way of creating homozygous lines within one generation, based on androgenizes (anther or

microspore cultures). The DH procedure is a highly specialized technique that requires optimal tissue

culture and growth facilities. Within the PPP group there is a high degree of expertise in the private

breeding companies and the technique is utilized by the companies as an important procedure to

optimize and accelerate the breeding programs.

Time: Month 8-16 (first round of MAGIC populations), month 32-40 (second round of MAGIC

populations).

Deliverables: 250 DH lines from each MAGIC population.

Task 2.2 Multiplication of homozygous MAGIC populations for screening in the field:


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Description: Each DH line will be multiplied in the field in order to produce enough material to

distribute between the partners for screening of selected diseases and agronomic traits. Each single

DH plant will be harvested and the seeds sown in approximately 4-6 one meter rows for

multiplication (SeedMatic format). This will, at a minimum, produce 500 g of seeds for distribution.

Time: Month 16-20 (first round of MAGIC populations), month 40-44 (second round of MAGIC

populations).

Deliverables: Multiplication of seed from each individual DH line. Approximately 0,5 kg of each line.

2.3 PARTNERS

The production of DH lines will be divided between Nordic Seed, Sejet Planteforædling and Boreal

each of these companies has the technique actively running in the laboratory.

Each company will be responsible for the production of 250 DH lines from 1-2 MAGIC populations.

This will be followed by multiplication of each individual line, producing material enough for field

screening in multiple locations. The selection of, Finland and Denmark for multiplication is coupled

to the environmental conditions, thereby selecting locations in which all lines will reach full maturity

before harvest.


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Work package 3: Searching for and screening of new sources

Leader: Outi Manninen; Boreal – Finland

3.1 INTRODUCTION

Screening for new sources of resistance is a continuous process. Pathogen populations are prone to

evolve and break the resistances in varieties. It is not wise to rely on only a few resistance sources in

varieties since this poses a strong selection pressure on the pathogen populations. Durability of

resistance may be enhanced by pyramiding several resistance genes into a variety. Climate change

may also cause quick changes in the pathogen populations and breeding has to be ready to meet

new challenges. We will continue searching new resistance sources and incorporating them to

adapted breeding material. It is important to work on several putative resistance sources since some

resistance genes may affect yield or other agronomic traits negatively and thus are not useful in

breeding programs. The ‘dynamic gene pool of barley’ preserved at the NordGen will also serve as

potential source of donors.

3.2 TASKS (TABLE 3.1)

Task 3.1 Collection of information on resistance sources:

Description: Collecting new publicly available information on resistance sources and making this

information available to the project participants in the form of a content management system

(CMS).

Time: Months 1-3, updates during months 13 and 25.

Deliverables: Information on new putative resistance sources stored at CMS.

Task 3.2 Collection and multiplication of seed:

Description: Ordering and multiplication of seed for potential resistance sources. Distribution of seed

to project participants for screening and crossing.

Time: Months 4-6, and when needed.

Deliverables: Seed lots from potential resistance sources available for project partners.

Task 3.3 Screening:

Description: Screening for new resistance sources. Screening for new resistance sources will first be

done in field conditions and interesting sources will continue to further greenhouse/field tests (Task

4). We will specifically look for new resistance to Ramularia and Fusarium. In addition, based on the

results in the first PPP Barley project we still need to put emphasis on finding resistance to net

blotch and Bipolaris.

Time: Summer 2015, 2016 and 2017.

Deliverables: Preliminary information on the resistance reactions towards Nordic pathogen

populations. Preliminary information of adaptation to the Nordic conditions.


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Task 3.4 Verification:

Description: Verification of resistance sources. We will verify all resistance sources used in current

or new mapping populations with greenhouse and/or field testing in several locations. We have

three types of material: a) parents of the first MAGIC populations, b) potential new resistance

sources (from Task 1 and 2) and c) the parents of the ‘Dynamic Gene Pool’.

a) In the first MAGIC populations under development there are new or unmapped genes for

resistance to leaf rust, scald, Fusarium head blight, Bipolaris spot blotch and net blotch (net

and spot type). These resistance sources were selected based on literature and results of

previous research. The resistance of these sources was tested during 2014 and will be

verified in a second year experiment.

Time: during 2015.

Deliverables: Information on the resistance reactions towards Nordic pathogen populations.

b) New potential resistance sources from the screens will be further tested for resistance in

greenhouse and at several field locations. Best ones will be selected to be parents of the

new mapping crosses (MAGIC, BC or pairwise crosses).

Time: 2016 and 2017.

Deliverables: Information on the resistance reactions towards Nordic pathogen populations.

New sources of resistance to several diseases. New parents for MAGIC or other type of

mapping populations.

c) Dynamic Gene Pool. This collection was produced about 20 years ago with a scheme:

pairwise crosses, diallel, pairwise crosses for 3 generations, diallel. The final material of 276

families provides an interesting population were the genomes of 40 progenitors have been

recombined with six rounds of crosses. Starting material was partly lines and varieties

adapted to Nordic conditions, and partly exotic material. All parents were selected for one

or more resistances to mildew, Bipolaris, rust, net blotch (net or spot type), scald, smut,

stripe or nematodes. The last generation of this material will be multiplied at NordGen

during 2015.

Time: Greenhouse testing during winter 2015/2016, field tests 2016.

Deliverables: Information of the usability of the Dynamic Gene Pool as new Donors for MAGIC

population for mapping resistance genes.

Table 3.1 Participant involvement in tasks in WP3

Boreal Graminor LBHI LUKE Nordic Seed Sejet SLU

Task 3.1 X X X

Task 3.2 X X

Task 3.3 X X X X X

Task 3.4 X X X X X


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Work package 4: Screening of segregation populations for diseases resistance

Leader: Lars Reitan; Graminor - Norway

4.1 INTRODUCTION

The initial project period of three years is too short time for long term pre-breeding activities. The

parties of the project are agreeing to continue the work in a second phase with stronger emphasis

on long term goals. Examples of future perspectives that have been raised are an increased focus on

new and emerging diseases related to climatic change.

To prepare for a next stage of PPP, new sources of resistance was started in the first PPP project

period in ‘WP Preparing for the future’ crosses are initiated and segregating progenies will be

developed .This work will continue by searching in literature, trying new sources and developing

new crosses for testing. All breeding companies participating in this project were involved in the

NordForsk project “Sustainable primary production in a changing climate”, co-ordinated by Rikke

Bagger Jørgensen (RISØ-DTU) and knowledge from this project will likewise be further utilized in the

next stage of PPP, particularly resistance material will be selected and used in crossing for the

development of segregating progenies.

Spot blotch (Bipolaris sorokiniana) is a disease which has increased in the Nordic countries during

the past years, especially in Finland where the pathogen is present in almost all fields. Ramularia leaf

spot caused by Ramularia collo-cygni is another disease on the rise, causing economic losses in

barley in an increasing number of countries. A third disease is Fusarium head blight, also causing

losses due to toxin problems and reduced yields. For these pathogens, more knowledge about the

virulence structure in Northern Europe is needed, but even more important from the breeder’s point

of view are the identification of new sources for resistance and the development of efficient tools

for selection. Access to user-friendly markers would improve the possibilities for successful

pyramiding of different resistance genes to achieve a sustainable resistance. Another example will

be to further elucidate the linkage between Ramularia susceptibility and mlo resistance and

subsequently break this linkage. Our information about the present advanced breeding material

among the Nordic participants paves the way for the next step: to increase the variability and

amount of genes for disease resistance in a longer perspective and under different climatic regime.

4.2 TASKS

Task 4.1 Disease screening of MAGIC populations and Donor lines:

Description: Different diseases have different impact on barley production in different parts of the

Nordic countries, and priority will vary between countries. Some diseases have been investigated for

a long time (e.g. net blotch and scald), while others are ‘new’ and evolving in the field (e.g.

Ramularia, Bipolaris, Fusarium). Our goal is to include diseases with severe impact on crop yields

under current and future climate conditions, and to search for new resistance sources. Based on the

output of the first PPP project and priorities in the initial PPP (‘WP Future’) and the prolongation

year and the creation of MAGIC population the task includes the following diseases:


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1. Scald (Rhynchosporium secalis),

2. Net blotch (Drechlera teres),

3. Ramularia leaf spot (Ramularia collo-cygni),

4. Spot blotch or Bipolaris (Bipolaris sorokiniana),

5. Fusarium head blight (Fusarium spp.)

6. Leaf Rust (Puccinia hordei Otth.)

A brief description of the different disease resistance testing sites and activities (table 4.1):

Scald will be tested for in at least two different environments; in Boreal/LUKE (Fin) and in NOS (DK)

in field conditions with artificial inoculation, each of different local strains. The potential new

resistance sources implemented in the crossings are essential in the testing regime. (In addition,

when possible a natural scald attack on material to be tested for agronomic traits in Iceland, a

registration on scald severity should be done).

Net blotch will be tested in field conditions and artificial inoculated with virulent strains of the two

net blotch fungi Drechlera teres var. teres (D.t.t)/maculate D.t.m). Screening for the disease will be

done at least twice in the season. Greenhouse testing is relevant, although the small- plant

resistance is different from adult plant resistance and should be handled by caution. At least two

sites will be included for testing: at Boreal/LUKE(Fin) (both D.t.t and D.t.m) and at Graminor(N)

(D.t.t).

Ramularia leaf spot will be tested under natural infection in two to three locations; at Graminor

(GN) and in Sejet (DK). Scoring the disease will be done at least two times in the season.

Spot blotch or Bipolaris will be carried out in Boreal/LUKE in both field and greenhouse conditions

with artificial inoculation, and scored according to known practice at the station.

Fusarium head blight will be tested in two locations: in Graminor (N) on artificial Fusarium

graminearum inoculated, mist irrigated field and FHB, yield impact and DON values will be recorded.

At Boreal/LUKE (Fin), a similar setup will be done on F. culmorum.

Leaf Rust will be carried out in two environments (Sejet and Nordic Seed) in Denmark on non-

inoculated fields.


16

Table 4.1: Summing up diseases and testing sites

BOREAL* LUKE GRAMINOR NORDIC SEED SEJET LBHI

1. Scald

(Rhynchosporium secalis),

X

X

X

2.Net blotch

(Drechlera teres),

X

X

3.Ramularia leaf spot

(Ramularia collo-cygni),

X

X

4.Spot blotch or Bipolaris

(Bipolaris sorokiniana),

X

5.Fusarium head blight

(Fusarium spp.)

X

X

6. Leaf Rust

(Puccinia spp.)

X X

*Disease screening is done in a coordinated collaboration with LUKE

Preferably 250 lines will be screened pr. MAGIC population. Screening of the DH lines should be

tested in standard setup for the participant, and even hillplot or ‘headrow’ plots might be relevant

for certain diseases, but it is a big advantage to have bigger plots in order to avoid border effects.

Both field conditions with natural/artificial inoculation, and/or greenhouse small-/adult plants might

be relevant depending on conditions and disease. Even controlled conditions with attached leaves

etc. might be relevant.

In the first year of seed multiplication and pre-screening 500-1000 g of seed should be harvested for

screening in the following season.

Each participant is responsible for the scientific work necessary for the screening of the segregating

material and donors. The work package leader might visit the different experiments and encourage a

good setup.

Donor lines was tested for a set of diseases in previous project, but the seed amount was scarce and

the germplasm hardly had repetitions in the tests. It is of big impact to repeat these tests in time

and space, and adding new potential donors to the list. Also some ordinary cultivars should be

included as controls. Testing of donor lines should be done at least two years in the new project.

The donor lines should be tested in the best possible setup to create as good data as possible, and

perhaps it is necessary to choose bigger plots and more replicates than for the offspring testing in

order to achieve this.

Deliverables: Phenotypic data from disease screenings. All data will be controlled and handed over

to SLU for central computation and will be used for the association mapping in WP 7 (Therese

Bengtsson). External expertise might also be consulted when necessary, and the budget should

cover this.

Time: Summer 2015, 2016 and 2017


17

Some of the crosses connected to the Magic population development are already done, and others

are already planned in detail and will be carried out during 2014 and beginning of 2015. In autumn

2015 and spring 2016 Double Haploid (DH) offspring from the MAGIC crosses will be produced and

the first pre-screening and multiplication of the offspring will be done in 2016. During autumn 2016

and spring 2017 the material will be genotyped by SNP chip, and a full scale field screening of the DH

offspring will take place in the summer season of 2017. Parallel to this a full set of Donor Screening

will take place in all summers 2015-2017 in all sites.

Donors will screened in 2015, 2016 and 2017

4.3 PARTNERS

The different project partners dealing with testing of segregating material are the following:

BOREAL/LUKE (Fin), GRAMINOR (N), NORDIC SEED (DK), SEJET (DK) LBHI (I)

Apart from these partners also SLU are important participant in doing the calculations on the results

of the testing related to marker association etc. (WP6)


18

Work package 5: Screening of segregating populations for agronomic traits

Leader: Sæmundur Sveinsson; LBHI - Iceland

5.1 INTRODUCTION

The Nordic region spans over diverse climatic conditions caused by a multitude of latitudinal,

altitudinal and climatic (maritime/continental) interactions.

Heat sum over the growth season varies both between locations and years, as well as the length of

the growth season and the precipitation pattern. This yearly variation is expected to increase, and be

less predictable in the future climate (IPCC, 2014). In addition, the number of extreme weather

events is predicted to increase. It is vital to address the upcoming challenges and accelerate the pace

in which we are base broadening our Nordic barley gene pool to sustain both biotic and abiotic

stresses, if we want to maintain a sustainable crop production and resilient bio economy in the

Nordic region.

We need to breed varieties that are robust and have wide tolerance for climatic variations, e.g. a low

genotype by environment interaction for agronomic characters such as height and straw strength. In

order to identify such material, multi-location and year trials are essential. In addition to abiotic

stresses, susceptibility to diseases has effects on certain agronomic traits, e.g. lodging.

Furthermore, it will be essential to fine tune the time of flowering and maturity in order to optimally

utilize the growth season in each location. In previous screenings for heading day in Phase 1 of the

PPP project (17 trials, 2012 – 1014) low environmental effect was seen, with a strong Pearson

correlation between trials (R-values ranging 0.72 – 0.93). For maturity, however, variation was

greater between locations as heat sum requirements for maturity are higher. Length of growth

season and temperature varies greatly in the Nordic region (table 5.1). This calls for locally adapted

cultivars that are adjusted to the local day length and temperature regimes for completing maturity.

Table 5.1: Length of growth season and heat sums of selected locations in the ongoing PPP-project.

Location Year Days from sowing to maturity of

latest line in PPP-set

Heat sum

(Day degrees, >0°C)

Korpa, Iceland 2012 143 1407

2013 Did not reach maturity 12691

Sejet, Denmark 2012 133 1731

2013 111 1756

Staur, Norway 2012 104 1786

2013 112 1964

2014 85 1715 1From sowing to harvesting (142 days)

MAGIC population

In the initial phase of the PPP (2012 – 2014), we identified several markers which were associated

with alleles for earliness and straw length. Some of the identified alleles were rare in the Nordic

barley population. In the case where rare alleles gives only a few lines of contrasting phenotype,

GWAS is an unsuitable method as the statistical power in the model requires at least 10% frequency


19

of each phenotype in the population. Hence, a way to further study the role of rare alleles is to

develop segregating populations based on two or more parental lines, or MAGIC populations . This

will increase the frequency of the rare alleles in the study population which in turn enables GWAS as

a tool for QTL-mapping.

Educational part

In the first phase of the PPP, a PhD student was assigned to LBHI working with the agronomic traits.

The analyses are continuing in the first year of phase two, and are expected to be completed by the

end of 2015. In 2016, a new student will be assigned to LBHI within the framework of the PPP phase

two, to work with trials of the MAGIC earliness population.

5.2 Tasks

Task 5.1 Populations

Description: Screening of the developmental traits heading day, grain filling period, maturity and

height. Screenings should take place in two distinct climatic conditions:

1. Long and cool growth season (Iceland/coastal northern Norway)

2. Early sowing and warm growth season (Denmark/south of Sweden)

Time

A four-way MAGIC has been developed (2012 – 2014).

In 2015, crosses with four additional sources of earliness, semi-dwarfing and lodging resistance will

be initiated. The eight-way MAGIC will require three rounds of crosses which will be performed in

2015. Development of DH lines will take place in spring 2016, with the population ready for field

trials in 2017.

Deliverables

A four-way MAGIC earliness population is completed, to SSD5 (F6 generation). This population will

be screened in row sowings in 2015, and in plot screenings 2016.

An eight-way MAGIC earliness will be developed to further diversify the sources of earliness. This

population will be screened in field trials in 2017.

Task 5.2 Education

Deliverables: Education of one PhD student (2012 – 2015) completed. Education of a new student

(2016 - ) initiated.

5.3 Partners

Development of MAGIC populations: LBHI. Field trials taking place at LBHI and Sejet.


20

Work package 6: Data management and association mapping

Leader: Therese Bengtsson (SLU-Sweden)

6.1 INTRODUCTION

One of the main goals of the second PPP project is to identify markers linked to new, different and

effective disease resistance genes and agronomical traits such as maturity, earliness etc. that the

breeders can incorporate in their breeding programs. The use of MAGIC populations will allow the

use of both linkage and association analysis without the restrictions encountered with highly

structured populations. DNA will be extracted from the five MAGIC populations and sent to

TraitGenetics (Gatersleben, Germany) for SNP genotyping with the 9K Illumina Chip containing

approximately 6950 functional SNPs. Genome-Wide-Association-Analysis (GWAS) will then be

performed, linking the phenotyping data for diseases resistance and agronomic traits obtained for

the MAGIC population in WP3, WP4 and WP5 to the data obtained from high-throughput SNP

genotyping. The R package GAPIT (Genome Association and Prediction Integrated Tool) will be used,

that performs GWAS and genome prediction based on the state-of-the-art methods for statistical

genetics. One of the objectives for WP6 is to provide the partners with the phenotyping and

genotyping results as well as the outcome of the GWAS via the existing content management system

(CMS) from a server at SLU. Markers found in the GWAS will be transformed to easy-to-use PCR-

based markers such as Kompetitive Allele Specific PCR (KASP) and the sequences and protocols will

be made available to the partners via the CMS. The information about the linked markers will also be

used in WP7 for marker assisted backcrossing (MAB) and for genomic selection (GS).

6.2 TASKS

Task 6.1 Information on CMS:

Description: Making all information and results available for the partners via the internal CMS.

Time: Whole period

Deliverables: Maintenance and updating of the CMS.

Task 6.2 Genotyping:

Description: Collecting the material from the five MAGIC populations produced within the project

and isolate DNA for genotyping of high throughput Single Nucleotide Polymorphism.

Time: Autumn 2016 and autumn 2017.

Deliverables: SNP information of the five MAGIC populations available in CMS.

Task 6.3 GWAS:

Description: Association between the field data from WP3, WP4 and WP5 and the marker data with

Genome Wide Association Analysis (GWAS).

Time: Autumn 2016 and autumn 2017


21

Deliverables: Making the information obtained from the association analysis available to all partners

in the project via the CMS.

Task 6.4 User friendly DNA markers:

Description: Developing easy-to-use PCR-based markers (KASP markers) utilizing new genes

identified by the association analysis when this is necessary and possible.

Time: spring 2016 and spring 2017

Deliverables: Making the sequences and protocols available to all partners in the project via the

CMS.

Task 6.5 Publication:

Description: Making the results from the project available to the public through publication(s) in a

peer-reviewed paper upon agreement with all project partners and in accordance with the co-

operation agreement.

Time: 2017

Deliverables: Manuscript accepted for publication in a peer-reviewed paper.

6.3 PARTNERS

The different project partners contributing with field data for the association analysis are the

following:

Finland: BOREAL and LUKE; Norway: GRAMINOR; Denmark: NORDIC SEED and SEJET; Iceland: LBHI.


22

WP 7: Transfer of resistance genes in breeding program Leader: Ahmed Jahoor; Nordic Seed – Denmark 7.1 INTRODUCTION

Barley is attacked by several pathogens. In addition, due to climate changes, some diseases such as

Fusarium Head Blight, Ramularia and Bipolaris are becoming more prominent in Nordic countries. To

combat with diseases in barley, fungicides are often used. The fungicides can create environmental

concern and they are also problematic for human health. Therefore, breeding for diseases resistance

is the most environmental as well as human friendly methods to combat this problem in barley.

Therefore, many major race specific resistance gens controlled by very few genes for barley

important diseases e.g. powdery mildew, yellow brown rust, scald and net blotch have been

introduced in the recently released varieties. However, these major race specific resistance genes

are frequently overcome by newly developed pathogen races through mutations or recombination.

Therefore, it is extremely important to search for new sources of resistance. Since major race

specific resistance does not always provide durable resistance, we aim to identify new race specific

major gens as well race non-specific minor genes for diseases resistance in MAGIC populations. The

race non-specific resistance genes are mostly controlled by several genes with minor effect. While

the major race specific resistance genes provides complete resistance against a specific pathogen

race, race non-specific resistance controlled by minor genes provides wider spectrum of resistance

and slows down the occurrence of mutation in the pathogen populations. Due to these facts, it is

important to develop strategies for sustainable use and management of resistance genes in breeding

programs.

The sources of resistance for different diseases that have or are being used in the development of

MAGIC populations will be available for all project partners. The MAGIC population will provide the

DNA markers linked to diseases resistance as well as to agronomic traits. In addition, these

populations will possess combined resistance to different diseases in a single line. For each plant

breeding company involved in the project, the original sources of resistance, DNA markers linked to

resistance genes as well as lines with combined resistance genes will be available. Here, each

company will incorporate these sources of resistance in their breeding program. Each company will

select crossing parent on their own choice in order to incorporate these sources of resistance in their

breeding program with the aim to develop resistance varieties. Theses crossing parents will not be

disclosed to other partners. However, the applicability of the DNA markers and the effectiveness of

resistance resources should be shared.

7.2 TASKS

Task 7.1 Marker assisted back crossing

Time: month 18-36

Deliverables: Providing linked DNA markers


23

Task 7.2 Pyramiding new sources of resistance

Time: month 18-36

Deliverable: Lines possessing combined resistance to several diseases originating from MAGIC

populations

7.3 PARTNERS

Finland: BOREAL; Norway: GRAMINOR; Denmark: NORDIC SEED and SEJET; Iceland: LBHI.

GANTT diagram "Combining Knowledge from Field and from Laboratory for Pre-breeding in Barley II”

Year / task 2015 Spring

2015 Summer

2015 Autumn

2016 Spring

2016 Summer

2016 Autumn

2017 Spring

2017 Summer

2017 Autumn

2018 Spring

2018 Summer

2018 Autumn

2019 Spring

2019 Summer

2019 Autumn

WP1 Task 1.1 crosses

WP1 Task 1.2 crosses

WP2 Task 2.1 DH production

WP2 Task 2.2 Multiplication

WP3 Task 3.1 Information on donors

WP3 Task 3.2 Collection of donors

WP3 Task 3.3 Screening

WP3 Task 3.4 Verification

WP4 Task 4.1 Screening

WP5 Task 5.1 Populations

WP5 Task 5.2 Education

WP6 Task 6.1 Information on CMS

WP6 Task 6.2 Genotyping

WP6 Task 6.3 GWAS

WP6 Task 6.4 DNA markers

WP6 Task 6.5 Publication

WP7 Task 7.1 Marker assisted backcrossing

WP7 Task 7.2 Pyramiding of resistance

Curriculum Vitae

Ahmed JahoorHome address

Solsortevej 21, 4000 Roskilde, Denmark

Tel: (+45) 29 13 47 57

Work address

Nordic seed A/S – Lolland.

Højbygårdvej 14, 4960 Holeby

Tel: (+45) 79 21 23 24

e-mail: [email protected] http://www.nordicseed.com

EducationMSc, 1982, in Agronomy, University Prague, Czech RepublicPhD, 1987, in Plant breeding, TU München, Lehrstuhl für Pflanzenbau und Pflanzenzüchtung, Germany

Appointments2012- Adjunct professor for genetics and plant breeding at the Swedish Agricultural University2008- Breeding manager at Nordic Seed.2005-2008 Senior Research Scientist, The Royal Veterinary and Agricultural University, Department of agricultural Sciences

now, University Copenhagen, Faculty Science1997-2005 Senior Research Scientist at Risø National Laboratory, Plant Research Department.1995-1997 Wissenschaftlicher Oberassistent (C-2)1990-1995 Wissenschaftl. Assistent (C-1) at Lehrstuhl für Pflanzenbau und Pflanzenzüchtung , TU München1987-1990 Wissenschaftlicher Angestellter (BAT IIa) at Lehrstuhl für Pflanzenbau und Pflanzenzüchtung der TU München

Selected refereed articles 2004-2012Complete List of Publications: Peer reviewed publications. Total published papers: 67 (23.04.2014, web ofscience) Sum of the time cited: 2.281. Average citation per item: 34,04. H-index: 25

Nielsen NH, Backes G, Stougaard J, Andersen SU, Jahoor A (2014) Genetic Diversity andPopulation Structure Analysis of European Hexaploid Bread Wheat (Triticum aestivum L.) Varieties.PLoS ONE 9(4): e94000. doi:10.1371/journal.pone.0094000

Jihad Orabi, Ahmed Jahoor and Gunter Backes. 2014. Changes in allelic frequency over time inEuropean bread wheat (Triticum aestivum L.) varieties revealed using DArT and SSR markers.Euphytica. DOI: 10.1007/s10681-014-1080-x.

Zeratsion Abera Desta, Jihad Orabi, Ahmed Jahoor, Gunter Backes. 2014. Genetic diversity andstructure found in samples of Eritrean bread wheat. Plant Genetic Resources. 12,01: 151-155DOI:10.1017/S1479262113000324

Seeholzer S, Tsuchimatsu T, Jordan T, Bieri S, Pajonk S, Yang WX, Jahoor A, Shimizu KK, KellerB, Schulze-Lefert P (2010). Diversity at the Mla powdery mildew resistance locus from cultivatedbarley reveals sites of positive selection. Molecular Plant-Microbe Interaction, 23:497-509

Orabi J, Jahoor A, Backes B. 2009. Genetic diversity and population structure of wild and cultivatedbarley from West Asian and north Africa. Plant Breeding, 128:332-336

Lababidi S, Mejlhede N, Rasmussen SK, Backes G, Al-Said M, Baum M, Jahoor A (2009).Identification of barley mutants in the cultivar “Lux” at the Dhn loci through TILLING. PlantBreeding, 128:332-336

Backes G, Orabi J, Wolday A, YahyaouiA, Jahoor A 2009. High genetic diversity revealed in barleycollected from small-scale farmers fields in Eritrea. Genetic Resour Crop Evol, 56:85-97

Dayteg C, Rasmussen M, Tuvensson T, Merker A, Jahoor A 2008. Development of an ISSR-derivedPCR marker linked to nematode-resistance 8Ha2) in spring barley. Plant Breeding, 127:24-27

Vitamvas P, Saalbach G, Prasil IT, Capkova V, Opatrna J, Jahoor A (2007). WCS120 protein familyand proteins soluble upon boiling in cold-acclimated winther wheat. Journal of plant Physiology,164:1197-1207

Gahoonia TS, Ali R, Malhotra RS, Jahoor A, Rahman MM (3007). Variation in root morphologicaland physiological traits and nutrition uptake of chickpea genotypes. Journal of Plant Nutrition,30:829-841

Dayteg C, Tuvensson T, Merker A, Jahoor A, Kolodinska-Brantestam A (2007). Automation ofDNA marker analysis for molecular breeding in crops: practical experience of a plant breedingcompany. Plant Breeding, 126:410-415

Participant:

Grand total

Budget Item PPP own contrib. PPP own contrib. PPP own contrib. PPP own contrib. % contrib.

Personal Cost 110.000kr. 70.000kr. 70.000kr. 250.000kr. -kr. 0% 250.000kr.

Consumable 40.000kr. 40.000kr. 80.000kr. -kr. 0% 80.000kr.

Travel 10.000kr. 10.000kr. 10.000kr. 30.000kr. -kr. 0% 30.000kr.

Overhead 30.000kr. -kr. 30.000kr. -kr. 30.000kr. -kr. 90.000kr. -kr. 0% 90.000kr.

sum 150.000kr. -kr. 150.000kr. -kr. 150.000kr. -kr. 450.000kr. -kr. 0% 450.000kr.

Participant:

Grand total


Personal Cost 75.000kr. 110.000kr. 75.000kr. 110.000kr. 75.000kr. 110.000kr. 225.000kr. 330.000kr. 59% 555.000kr.

Consumable 5.000kr. 5.000kr. 5.000kr. 15.000kr. -kr. 0% 15.000kr.


Overhead 27.500kr. 27.500kr. 27.500kr. 27.500kr. 27.500kr. 27.500kr. 82.500kr. 82.500kr. 50% 165.000kr.

sum 137.500kr. 137.500kr. 137.500kr. 137.500kr. 137.500kr. 137.500kr. 412.500kr. 412.500kr. 50% 825.000kr.

Participant:

Grand total


Personal Cost 175.500kr. 380.000kr. 175.500kr. 390.000kr. 175.500kr. 380.000kr. 526.500kr. 1.150.000kr. 69% 1.676.500kr.

Consumable 40.000kr. 30.000kr. 40.000kr. 30.000kr. 40.000kr. 30.000kr. 120.000kr. 90.000kr. 43% 210.000kr.

Travel 10.000kr. 15.000kr. 10.000kr. 15.000kr. 10.000kr. 15.000kr. 30.000kr. 45.000kr. 60% 75.000kr.


sum 281.875kr. 531.250kr. 281.875kr. 543.750kr. 281.875kr. 531.250kr. 845.625kr. 1.606.250kr. 66% 2.451.875kr.

Participant:

Grand total


Personal Cost 242.400kr. 477.600kr. 489.600kr. 1.209.600kr. -kr. 0% 1.209.600kr.

Consumable 68.000kr. 500.000kr. 68.000kr. 636.000kr. -kr. 0% 636.000kr.


Overhead 151.200kr. 297.600kr. 304.800kr. 753.600kr. -kr. 0% 753.600kr.

sum 497.600kr. -kr. 1.311.200kr. -kr. 898.400kr. -kr. 2.707.200kr. -kr. 0% 2.707.200kr.

SLU

2015 2016 2017 Sum

Sejet Plant Breeding

2015 2016 2017 Sum

LBHI (Breeding)

2015 2016 2017 Sum

LBHI (University)

2015 2016 2017 Sum

Participant: MTT Agrifood Research Finland (LUKE Natural Resources Institute Finland from 1.1.2015)

Grand total


Personal Cost 113.519kr. -kr. 113.519kr. -kr. 113.519kr. -kr. 340.558kr. -kr. 0% 340.558kr.

Consumable 14.888kr. -kr. 14.888kr. -kr. 14.888kr. -kr. 44.663kr. -kr. 0% 44.663kr.

Travel 14.888kr. -kr. 14.888kr. -kr. 14.888kr. -kr. 44.663kr. -kr. 0% 44.663kr.

Overhead -kr. 151.492kr. -kr. 151.492kr. -kr. 151.492kr. -kr. 454.475kr. 100% 454.475kr.

sum 143.295kr. 151.492kr. 143.295kr. 151.492kr. 143.295kr. 151.492kr. 429.885kr. 454.475kr. 51% 884.360kr.

Participant:

Grand total






sum 468.750kr. 887.500kr. 468.750kr. 900.000kr. 468.750kr. 887.500kr. 1.406.250kr. 2.675.000kr. 66% 4.081.250kr.

Participant:

Grand total


Personal Cost 143.741kr. 290.000kr. 143.741kr. 295.000kr. 143.741kr. 290.000kr. 431.223kr. 875.000kr. 67% 1.306.223kr.

Consumable 1.500kr. 1.500kr. -kr. 1.500kr. 1.500kr. 50% 3.000kr.




Participant:

Grand total







Participant:

Grand total


Personal Cost 1.200.160kr. 1.790.000kr. 1.395.360kr. 1.825.000kr. 1.407.360kr. 1.790.000kr. 4.002.881kr. 5.405.000kr. 57% 9.407.881kr.

Consumable 274.388kr. 81.500kr. 744.888kr. 80.000kr. 312.888kr. 80.000kr. 1.332.163kr. 241.500kr. 15% 1.573.663kr.


Overhead 450.635kr. 638.057kr. 596.660kr. 646.432kr. 603.860kr. 637.682kr. 1.651.156kr. 1.922.170kr. 54% 3.573.326kr.

sum 2.138.071kr. 2.584.317kr. 2.949.796kr. 2.626.192kr. 2.536.996kr. 2.582.442kr. 7.624.864kr. 7.792.950kr. 51% 15.417.814kr.

All

2015 2016 2017

Boreal Plant Breeding Ltd.

2015 2016 2017

Sum

Graminor

2015 2016 2017 Sum

Sum

Nordic Seed

2015 2016 2017 Sum

2015 2016 2017 Sum

project proposal for public private partnership in …...biotic and abiotic stresses in the future....

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