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BIONÆR-Bionæringsprogram

Fra laboratorium til feltforsøk: Anvendelse av gen-markører for mer effektiv fremstilling av overvintringssterke jordbærsorter.

Alternative title: From laboratory to field trials: Application of genetic markers for more efficient production of wintering strong strawberry varieties.

Awarded: NOK 2.8 mill.

Project Manager:

Project Number:

244658

Project Period:

2015 - 2020

Funding received from:

Location:

Partner countries:

The project is a collaboration between Graminor AS, NIBIO, UiO, Hedmark University college and the international Indiana Purdue University at Indianapolis, USA. The survival of strawberry plants in areas with low temperatures is highly complex and affected by several physiological responses and a range of biotic (e.g. diseases) and abiotic (e.g. environment -cold, drought) factors. Due to the complexity involved in the regulation and enhancing cold tolerance, low temperature stress in strawberry has been one of our research focus since 2007 in three user driven innovation projects. This project aimed to capitalized on results from the previous projects and proceed to verify the usefulness of the identified biomarkers that are linked to cold tolerance in Graminor?s core breeding material. Our previous work has identified candidate molecules whose accumulation in strawberry cultivars shows strong association with cold tolerance. Two gene products were particularly correlative with cold tolerance; dehydrins, and alcohol dehydrogenase. Nonetheless, we will still need to validate these biomarkers further to gain more confidence in order to employ them in marker-based breeding. To further understand the mechanism of cold tolerance in strawberry, we have investigated the transcript levels of eight genes (i.e. a total of 1296 PCRs) extracted from leaf and crown tissues of three woodland strawberry genotypes that vary in cold tolerance. The transcriptomic results were also compared with proteomics data from the same samples. In this effort, we identified the key cold-responsive transcription factor (CBF) in strawberry and found that CBF transcript expression correlated well with cold tolerance. When we asked whether if protein expression of Xero2 and Cor47 correlated with their respective transcripts, we surprisingly found in the case of COR47, they did not; suggesting caution should be used in routinely applying transcript markers, based solely upon protein data. During the project period we have also utilized RNA sequencing (RNASEQ) to characterize the cold responsive transcriptome in two octoploid strawberry cultivars that differ in relative cold tolerance. In the absence of a reference genome from F x ananassa, various approaches for transcriptome assembly were evaluated prior to performing differential expression analyses to identify cold-responsive transcripts in each cultivar. RNAseq data from 30 samples of two strawberry cultivars that exhibit varying hardiness to cold treatment were assembled by differential de novo transcriptome strategies using the Trinity pipeline. In this direction, we have found marked differences in the extent and timing of changes in genes expression in response to cold treatment in these two cultivars differing in relative cold tolerance. Regulatory mechanisms underlying differential expression of cold tolerance-associated markers are largely expected to be genetic in nature, but may also result from epigenetic responses. In this project we have also preliminary addressed the epigenetic phenomena contribution to winter survival in strawberry. This research still ingoing in a phd study that is supported by Inland University (HiHM). In this project we have also used the woodland model strawberry to identify regions regulating cold tolerance, with an ultimate aim to identify similar regions in the commercial strawberry (F × ananassa) or more ambitiously transfer the important genomic region(s) between species. We have been able to identify a DNA region on linkage group 2 that is associated with cold tolerance in strawberry woodland. This is indeed an important finding for the future application of molecular DNA-based tools to screen for cold tolerance material in our national strawberry breeding. In order to implement the find from this WP in our national breeding program, we still need to validate this DNA region in an independent breeding population. We are hoping to perform this in the near future. In conclusion, the project has been highly successful at scientific and commercial levels. The project has generated enormous amount of novel and applicable results. The impact of this work has also contributed to our existing products and make their analysis more efficient. The outcome of this research project has been a platform for establishing a breeding and research network with national and international institutes, e.g. initiating Nordic collaboration (Public-Private Partnership) in strawberry breeding (project PPP_NORDFRUIT). This project has also created and provided excellent research training opportunities for graduate students at the participating institutions.

The project has been highly successful at scientific and commercial levels. The project has generated enormous amount of novel and applicable results; including: (i) gained immense genetic and molecular knowledge for breeding better cold tolerant strawberries; (ii) investigated and validated candidate molecular biomarkers for strawberry breeding; (iii) identify potential DNA regions regulating cold tolerance in the woodland model strawberry, which can be used to identify similar regions in the commercial strawberry. The impact of this work has also contributed to our existing products and make their analysis more efficient. The outcome of this research project has been a platform for establishing a breeding and research network with national and international institutes, e.g. initiating Nordic Public-Private Partnership in strawberry breeding. This project has also created and provided excellent research training opportunities for graduate students at the participating institutions.

Jordbær er den viktigste veksten innen norsk hagebruk. Et stort problem for dyrkerne er at sortene som dyrkes i dag med ujevne mellomrom blir utsatt for til dels store overvintringsskader. Det påvirker avlingsnivået og dermed også lønnsomheten for dyrkerne. Utvikling av vinterrobuste sorter er derfor et viktig avlsmål for Graminor. Overvintringsevne er dog en svært komplisert egenskap å foredle fram både fordi egenskapen i seg selv trolig er regulert av mange ulike gener og fordi utprøving og utvalg under naturlige frilandsforhold må skje over lang tid og er heftet med stor statistisk unøyaktig. Anvendelse av ny molekylærbiologisk teknologi og ny testmetodikk kan dog gi foredlerne verktøy som øker presisjon og effektiviteten en avlsarbeidet. Graminor har i to tidligere NFR-prosjekter samarbeidet med en forskergruppe bestående av nasjonale og internasjonale eksperter som har lett etter markører/biomarkører for lavtemperaturtoleranse. Gruppen har, ved siden av å publisere flere artikler i internasjonale periodika, også funnet flere biomarkør-kandidater for lavtemperaturtoleranse. Vi søker nå om å få undersøkt om disse biomarkørene kan benyttes i Graminors anvendte jordbærforedlingsprogram. Siden lavtemperaturtoleranse reguleres av mange gener, vil ikke biomarkørene kunne forklare hele variasjonen som observeres i overvintringsevne. Til det trengs flere markører. Nye høykapasitets genotypings-verktøy, skal derfor benyttes på Graminors foredlingslinjer. Sammenholdt med data hvor de samme linjene er testet for lav-temperaturstress/overvintringsevne, vil vi få et helhetlig bilde av hvilke gener/genregioner som påvirker overvintringsevnen og følgelig også ha et verktøy som foredlerne kan benytte i det daglige avlsarbeidet. Dette prosjektet benytter seg av avansert genteknologiske metoder. Vi er dog av den oppfatning at denne delen av prosjektet har lav risiko. Størst risiko knyttes til testingen av foredlingslinjene på en god og reproduserbar måte.

Funding scheme:

BIONÆR-Bionæringsprogram