13 ERA-CAPS Future-proofing potato: Mechanisms and markers for global-warming tolerant ideotypes

Lead Research Organisation: University of St Andrews
Department Name: Biology

Abstract

Potato is the third most important food crop in the world after rice and wheat. Because of its widely distributed cultivation and high yields, it is considered a critical species in terms of food security in face of a growing world population. However, potato is particularly vulnerable to high temperature during various stages of its life cycle. Elevated temperatures strongly suppress tuberisation, negatively affect storage and shelf life of tubers and reduce fitness of seed potatoes. Breeding new heat-stress tolerant cultivars is an urgent need for sustainable increases in potato production, given the negative impact of the rises in temperature due to global warming.
In this proposal, an integrated approach will be used by combining physiology, genetics, genomics, metabolomics and natural variation studies to analyze the impact of elevated temperatures on (1) sink-source relations of potato plants, (2) potato tuber development, (3) starch accumulation and tuber quality and (4) tuber dormancy. The outcomes of the project will be new knowledge of potato genes and their function that can be used in breeding programmes to increase yields in warmer climates.

Technical Summary

In this proposal, an integrated approach will be used by combining physiology, genetics, genomics, metabolomics and natural variation studies to analyze the impact of elevated temperatures on (1) sink-source relations of potato plants, (2) potato tuber development, (3) starch accumulation and tuber quality and (4) tuber dormancy. To achieve these aims both unbiased and targeted approaches will be employed. The unbiased approaches include the elucidation of phenotypic, biochemical and molecular responses to varying environmental conditions of selected potato genotypes (diploid populations, and a panel of tetraploid varieties and GMPs). Environmental conditions will include elevated and ambient temperatures in combination with different day lengths and light intensities. The plants will be phenotyped with respect to assimilate allocation, tuberisation, tuber yield, quality and dormancy. The genetic approach aims at identifying polymorphisms of candidate genes from diploid populations exhibiting a wide response to elevated temperatures. This will lead to the identification of genes and allelic variants that confer heat tolerance. The targeted approach is based on recent breakthroughs of the partners, which show that two linked regulators (StCDF_V and StSP6A) play a central role in the initiation of tuberisation. Our unpublished work suggests that StSP6A over-expression confers heat tolerance in transgenic potato plants. Therefore, the role of these regulators will be investigated in more detail in order to identify key components of the multiple signal transduction pathway(s). In addition levels of phytohormones known to regulate tuber initiation and dormancy will be manipulated and their impact on heat tolerance will be investigated.

Planned Impact

While the biological questions target the molecular mechanisms underlying the adverse effects of high temperature on potato crop productivity, we expect that the advance in knowledge of these mechanisms to have concrete economic and thus societal impacts. The outcomes of this project will be new knowledge on genes and alleles that can be used in breeding programmes to increase tuber yield under elevated temperatures. The production of EU seed potatoes that will perform well under higher temperatures will enhance exports to warmer climates and future-proof the EU potato industry against expected increases in EU summer temperatures. The plant breeding sector is essential to ensure that the genetic advances we shall make, deliver their full benefits to society. The involvement of several key European breeding companies, prepared to participate actively in this project, will ensure translation of scientific results to commercial production.
It is envisaged that the demand for heat resistant potatoes will intensify with global warming. The value of a heat-tolerant cultivar is difficult to predict, but with many potato-growing regions expected to suffer yield decreases of up to 25% due to heat stress alone, the value of this trait will be significant. In the wider context of a globalised world economy, improved human nutrition anywhere in the world will improve overall global food security and thereby have a benefit in the EU and beyond.
 
Description A bi-parental diploid potato population (06H1) of 188 genotypes was screened for tuber yield at "normal" and "elevated" temperatures using a model tuberisation system. A QTL explaining 12% of the variation in yield at elevated temperature was identified on Linkage Group 4. A candidate gene encoding a heat shock protein (Hsp70) in the QTL interval was identified. Allelic variants of the Hsp70 gene were identified and a particular allele (A2) was associated with enhanced tuber yield. Time-course experiments indicated that the A2 allele-specific transcript responds more rapidly in both tubers and leaves on exposure to elevated temperature. The A2 allele of the Hsp70 gene was transiently expressed in Nicotiana benthamiana. Exposure of plants expressing the Hsp70 gene demonstrated greatly increased tolerance of elevated temperature assessed by electrolyte leakage assays of leaves, compared with controls. Transgenic potato lines (cv Desiree) have been generated, designed to over-express the Hsp70 gene. These plants are currently being propagated and will be assessed for heat tolerance in the next 2-3 months. In order to increase the resolution of our genetic analysis we are using exome capture analysis. An exome capture array has been developed on the Nimblegen platform, based on ca. 39,000 coding sequences identified in the potato genome. Based on the phenotypic analysis of the 06H1 population three bulks have been assembled based on their differential yield response to elevated temperature. Following exome capture, non-random distribution of sequence read-based polymorphisms between the pools will identify chromosomal regions that co-localise with the heat tolerance and candidate genes will be selected. During the assessment of genotypes with different heat tolerance responses, we observed a very strong acquired thermotolerance response in potato. Exposure to a moderately elevated temperature (25 C) results in protection on subsequent exposure to high temperature (40C). Consequently, we are characterising this response at the molecular level and shall investigate responses in different potato germplasm.
Exploitation Route project is still in progress and further information will be added in future
Sectors Agriculture, Food and Drink,Education,Environment

URL http://www.eracaps.org/joint-calls/era-caps-funded-projects/era-caps-first-call-%E2%80%93-2012/future-proofing-potato-mechanisms
 
Description Potato is the third most important food crop in the world after rice and wheat. Potato is particularly vulnerable to increased temperature, considered to be the most important uncontrollable factor affecting growth and yield. In this project we have found an acquired thermotolerance response in potato whereby treatment at a mildly elevated temperature primes the plant for more severe heat stress. We defined the time course for acquiring thermotolerance and demonstrated that light is essential for the process. In all four commercial tetraploid cultivars that were tested, acquisition of thermotolerance by priming was required for tolerance at elevated temperature. Accessions from several wild type species and diploid genotypes did not require priming for heat tolerance under the test conditions employed, suggesting that useful variation for this trait exists. Using this information we have secured further funding for work in Kenya and Malawi to investigate clones in a population segregating for heat tolerance as well as virus resistance and earliness. The project will also involve training African collaborators in marker assisted selection and modern breeding technologies
First Year Of Impact 2017
Sector Agriculture, Food and Drink,Education
Impact Types Economic

 
Description Global challenge research fund
Amount £89,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2016 
End 02/2018
 
Description Global challenges Research fund
Amount £388,276 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 06/2017 
End 06/2019
 
Title Exome Capture. NimbleGen SeqCap EZ System 
Description NimbleGen technologies allow researchers to selectively target DNA sequences of interest in their genome of interes either through focused exomes or custom designed gene panels, for variant discovery and validation. SeqCap EZ Target Enrichment System is a solution-based capture method that enables enrichment of the whole exome or customer regions of interest in a single test tube. With this approach we could find SNPs associated with the extreme Heat stress phenotypes in the mapping population, and to have better resolution of previously identified QTLs. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Actually, the samples were sent for sequencing (Sequencing Service Provider - Edinburgh Genomics) and we are waiting for the sequencing queue. 
URL https://genomics.ed.ac.uk
 
Title GoldenBraid system_A Digital Toolbox for Plant Synthetic Biology 
Description A number of standard basic DNA pieces, named GBparts, were used as a starting point in the building of Hsc70 multigene structures. Basic GBparts comprised non-coding DNA regions, as the Hsc70 promoter and Patatin promoter and the strong nopaline synthase transcription terminator (Tnos), together with a number of GB-adapted coding sequences required for the assembly of a functional Hsc70. GB-adaptation, also called GB-domestication, consisted of: (i) removal of internal BsaI and BsmBI restriction sites by PCR-directed mutagenesis; (ii) PCR-mediated addition of flanking GB standard bar-codes, which consist of standard labels of 11 nucleotides that serve to facilitate cloning and, ultimately, to specify the relative position of each GBpart in the assembly; and (iii) TA-cloning of the resulting domesticated PCR products into primary and secondary vector 
Type Of Material Biological samples 
Year Produced 2015 
Provided To Others? Yes  
Impact Synthetic biology aims to simplify the process of designing, constructing and modifying complex biological systems. Plants provide an ideal chassis for synthetic biology, are amenable to genetic engineering and have relatively simple requirements for growth 
URL http://www.gbcloning.org
 
Description Future-proofing potato: Mechanisms and markers for global-warming tolerant ideotypes 
Organisation Friedrich-Alexander University Erlangen-Nuremberg
Country Germany 
Sector Academic/University 
PI Contribution The molecular basis of genotypic differences in response to elevated temperatures using potato germplasm collections and diploid populations. Using a genetic screen of a potato diploid population a large effect QTL has been identified that explains 12% of tuber yield variation in a model tuberisation system. A candidate gene associated with the QTL encoding a Heat Shock 70 protein has been identified and is being characterised. Alleles of this gene appear to underpin the yield phenotype. Transient expression studies indicate that expression of the Hsp70 gene confers tolerance to heat stress. Stable transgenic potato lines have been developed and will be assessed for effects on temperature stress. Further resolution of QTL for heat tolerance is underway using an exome capture array. A powerful acquired thermotolerance response has been observed in potato and the molecular basis of this response is under test
Collaborator Contribution (FAU): An experimental set-up was established using a "hot/ cold plate" system. This allows in addition to control (22°C) and heat treatment (29°C) for the entire plant to separately apply heat stress to either aerial parts of the plants (leaves) or to the belowground organs (tubers). Working with the cv. Agria, physiological and phenotypic parameters were measured. These data suggest that there are also signals from the tubers which may control heat response. Detailed metabolite and transcript profiles were obtained under the different temperature regimes imposed. These experiments confirmed a negative influence of an altered source/ sink balance on tuber starch content. Interestingly the levels of the signalling metabolite trehalose-6-phosphate (T6P) changed in leaves and tubers. Since T6P was reported to play a role in flowering regulation in Arabidopsis, we searched for candidate genes coding for T6P synthases (TPS by in silico analysis and identified 2 members of the family that show high degree of sequence identity to the active TPS1 from Arabidopsis. Their function and expression pattern is further investigated. Microarray analysis revealed numerous differentially expressed genes. Currently some candidates (FKF1, LOX etc.) are further analysed. Various experimental set-ups such as different day length periods (16h/ 12h or 8h light) were tested which in general confirmed that negative effects on tuber quality (tuber weight, second growth, dormancy) are more pronounced in long day conditions (16h light). Several experiments confirmed a reduced accumulation of starch in tubers when plants were grown under elevated temperatures. The activity of sucrose synthase (SuSy) correlates with starch levels and may serve as a molecular marker. To unravel the molecular mechanisms the heat / cold plate set up is used to identify genes correlating with expression of SuSy or other starch biosynthetic genes. In addition, genomic annotation of genes involved in starch metabolism was improved and this information is being used to identify co-regulated genes. WUR group obtained evidence for the involvement of CYCLING DOF FACTOR 1 (StCDF1) and StCDF3 genes in abiotic stress (heat and drought) sensitivity in potato plants. StCDF1 protein also transcriptionally represses StCO1-3 genes, a set of transcription factors able to indirectly repress the potato tuberigen StSP6A. Transgenic plants with reduced expression levels of StCDF1 showed high level of tolerance to drought and higher tolerance to heat stress. Interestingly, plants over-expressing StCDF3, another gene of the same family repressed by StCDF1 also presented a higher tolerance to drought and clearly improved growth and yield during heat treatments. Plants with different levels of StCDF1 presented a strong stomatal conductance phenotype, being lower in plants with low StCDF1 expression. Furthermore, in early potato genotypes, with more stable StCDF1, stomatal conductance is also greatly reduced. Transgenic plants with low expression of StCDF1 also have a decreased basal transpiration rate and they are extremely resistant to drought stress. On the other hand, a decrease in the transpiration rate has a negative effect on yield and plant production under heat stress. They explain the antagonism between treatments as a consequence of the miss-regulation in the StCDF1 gene expression, and concomitant incapacity of the plant to properly coordinate the induction of tuberisation with the needs of photosynthetic capacity under circumstances of environmental challenges. If transpiration is interrupted by stomatal closure due to StCDF1/StCDF3 miss-regulation, the major plant cooling mechanism is lost. They have established Chromatin Immune precipitation (ChIP) for potato. As a first approach we set up the experiment with the StCDF1 target genes StCO1-3. We show that StCDF1 protein binds close to ORF in StCO genes. Consistent with the ChIP result the StCDF1 binding site (AAAG) is preferentially present close to, or in 5´UTR ORF chromatin regions. They now also generated a SUC2 regulated GFP-tagged StCDF1 protein in potato. These plants will be used to carry out ChIP-seq analyse of the regulatory activity of StCDF1 throughout the genome. CSIC: Transgenic SP6A OE lines (35S promoter) in cv. Spunta and Sylvana backgrounds have been generated and characterized. Compared to wild-type plants an improved tuber yield was observed at elevated temperature together with reduced secondary growth. OE- lines accumulated more tuber starch and lower soluble sugars. Tubers from SP6A OE-lines sprouted later than controls during storage at 4°C. Microarray experiments with the transgenic lines have been conducted in collaboration with FAU and data analysis is ongoing. An initial heat stress experiment was performed including potato plants with increased levels of gibberellic acids (OE of AtGA20-ox1) and reduced amounts of strigolactones (CCD8-RNAi lines).These lines were obtained from JHI/StA. Compared to their respective wildtypes AtGA20-ox-OE lines did not exhibit significant differences in tuber number/ weight and dormancy period. CCD8-RNAi lines formed already under control conditions less tubers. Interestingly, these plants tended to form aerial tubers under the conditions used which will be further analysed.
Impact During this first 18-month period collaboration was established with all the groups and companies in the HotSol project. In this regard agreements were made to share information and project progress of all teams via DropBox. Protocols and experiments performance are also shared. We agreed to have two project meetings per year. Thus far, we had four meetings (see engagement activities) with attendance of all partners, associate partners and representatives of each company. In addition, monthly online meetings were conducted between all groups and companies.
Start Year 2016
 
Description Future-proofing potato: Mechanisms and markers for global-warming tolerant ideotypes 
Organisation Spanish National Centre for Biotechnology
Country Spain 
Sector Public 
PI Contribution The molecular basis of genotypic differences in response to elevated temperatures using potato germplasm collections and diploid populations. Using a genetic screen of a potato diploid population a large effect QTL has been identified that explains 12% of tuber yield variation in a model tuberisation system. A candidate gene associated with the QTL encoding a Heat Shock 70 protein has been identified and is being characterised. Alleles of this gene appear to underpin the yield phenotype. Transient expression studies indicate that expression of the Hsp70 gene confers tolerance to heat stress. Stable transgenic potato lines have been developed and will be assessed for effects on temperature stress. Further resolution of QTL for heat tolerance is underway using an exome capture array. A powerful acquired thermotolerance response has been observed in potato and the molecular basis of this response is under test
Collaborator Contribution (FAU): An experimental set-up was established using a "hot/ cold plate" system. This allows in addition to control (22°C) and heat treatment (29°C) for the entire plant to separately apply heat stress to either aerial parts of the plants (leaves) or to the belowground organs (tubers). Working with the cv. Agria, physiological and phenotypic parameters were measured. These data suggest that there are also signals from the tubers which may control heat response. Detailed metabolite and transcript profiles were obtained under the different temperature regimes imposed. These experiments confirmed a negative influence of an altered source/ sink balance on tuber starch content. Interestingly the levels of the signalling metabolite trehalose-6-phosphate (T6P) changed in leaves and tubers. Since T6P was reported to play a role in flowering regulation in Arabidopsis, we searched for candidate genes coding for T6P synthases (TPS by in silico analysis and identified 2 members of the family that show high degree of sequence identity to the active TPS1 from Arabidopsis. Their function and expression pattern is further investigated. Microarray analysis revealed numerous differentially expressed genes. Currently some candidates (FKF1, LOX etc.) are further analysed. Various experimental set-ups such as different day length periods (16h/ 12h or 8h light) were tested which in general confirmed that negative effects on tuber quality (tuber weight, second growth, dormancy) are more pronounced in long day conditions (16h light). Several experiments confirmed a reduced accumulation of starch in tubers when plants were grown under elevated temperatures. The activity of sucrose synthase (SuSy) correlates with starch levels and may serve as a molecular marker. To unravel the molecular mechanisms the heat / cold plate set up is used to identify genes correlating with expression of SuSy or other starch biosynthetic genes. In addition, genomic annotation of genes involved in starch metabolism was improved and this information is being used to identify co-regulated genes. WUR group obtained evidence for the involvement of CYCLING DOF FACTOR 1 (StCDF1) and StCDF3 genes in abiotic stress (heat and drought) sensitivity in potato plants. StCDF1 protein also transcriptionally represses StCO1-3 genes, a set of transcription factors able to indirectly repress the potato tuberigen StSP6A. Transgenic plants with reduced expression levels of StCDF1 showed high level of tolerance to drought and higher tolerance to heat stress. Interestingly, plants over-expressing StCDF3, another gene of the same family repressed by StCDF1 also presented a higher tolerance to drought and clearly improved growth and yield during heat treatments. Plants with different levels of StCDF1 presented a strong stomatal conductance phenotype, being lower in plants with low StCDF1 expression. Furthermore, in early potato genotypes, with more stable StCDF1, stomatal conductance is also greatly reduced. Transgenic plants with low expression of StCDF1 also have a decreased basal transpiration rate and they are extremely resistant to drought stress. On the other hand, a decrease in the transpiration rate has a negative effect on yield and plant production under heat stress. They explain the antagonism between treatments as a consequence of the miss-regulation in the StCDF1 gene expression, and concomitant incapacity of the plant to properly coordinate the induction of tuberisation with the needs of photosynthetic capacity under circumstances of environmental challenges. If transpiration is interrupted by stomatal closure due to StCDF1/StCDF3 miss-regulation, the major plant cooling mechanism is lost. They have established Chromatin Immune precipitation (ChIP) for potato. As a first approach we set up the experiment with the StCDF1 target genes StCO1-3. We show that StCDF1 protein binds close to ORF in StCO genes. Consistent with the ChIP result the StCDF1 binding site (AAAG) is preferentially present close to, or in 5´UTR ORF chromatin regions. They now also generated a SUC2 regulated GFP-tagged StCDF1 protein in potato. These plants will be used to carry out ChIP-seq analyse of the regulatory activity of StCDF1 throughout the genome. CSIC: Transgenic SP6A OE lines (35S promoter) in cv. Spunta and Sylvana backgrounds have been generated and characterized. Compared to wild-type plants an improved tuber yield was observed at elevated temperature together with reduced secondary growth. OE- lines accumulated more tuber starch and lower soluble sugars. Tubers from SP6A OE-lines sprouted later than controls during storage at 4°C. Microarray experiments with the transgenic lines have been conducted in collaboration with FAU and data analysis is ongoing. An initial heat stress experiment was performed including potato plants with increased levels of gibberellic acids (OE of AtGA20-ox1) and reduced amounts of strigolactones (CCD8-RNAi lines).These lines were obtained from JHI/StA. Compared to their respective wildtypes AtGA20-ox-OE lines did not exhibit significant differences in tuber number/ weight and dormancy period. CCD8-RNAi lines formed already under control conditions less tubers. Interestingly, these plants tended to form aerial tubers under the conditions used which will be further analysed.
Impact During this first 18-month period collaboration was established with all the groups and companies in the HotSol project. In this regard agreements were made to share information and project progress of all teams via DropBox. Protocols and experiments performance are also shared. We agreed to have two project meetings per year. Thus far, we had four meetings (see engagement activities) with attendance of all partners, associate partners and representatives of each company. In addition, monthly online meetings were conducted between all groups and companies.
Start Year 2016
 
Description Future-proofing potato: Mechanisms and markers for global-warming tolerant ideotypes 
Organisation University of Wageningen
Country Netherlands 
Sector Academic/University 
PI Contribution The molecular basis of genotypic differences in response to elevated temperatures using potato germplasm collections and diploid populations. Using a genetic screen of a potato diploid population a large effect QTL has been identified that explains 12% of tuber yield variation in a model tuberisation system. A candidate gene associated with the QTL encoding a Heat Shock 70 protein has been identified and is being characterised. Alleles of this gene appear to underpin the yield phenotype. Transient expression studies indicate that expression of the Hsp70 gene confers tolerance to heat stress. Stable transgenic potato lines have been developed and will be assessed for effects on temperature stress. Further resolution of QTL for heat tolerance is underway using an exome capture array. A powerful acquired thermotolerance response has been observed in potato and the molecular basis of this response is under test
Collaborator Contribution (FAU): An experimental set-up was established using a "hot/ cold plate" system. This allows in addition to control (22°C) and heat treatment (29°C) for the entire plant to separately apply heat stress to either aerial parts of the plants (leaves) or to the belowground organs (tubers). Working with the cv. Agria, physiological and phenotypic parameters were measured. These data suggest that there are also signals from the tubers which may control heat response. Detailed metabolite and transcript profiles were obtained under the different temperature regimes imposed. These experiments confirmed a negative influence of an altered source/ sink balance on tuber starch content. Interestingly the levels of the signalling metabolite trehalose-6-phosphate (T6P) changed in leaves and tubers. Since T6P was reported to play a role in flowering regulation in Arabidopsis, we searched for candidate genes coding for T6P synthases (TPS by in silico analysis and identified 2 members of the family that show high degree of sequence identity to the active TPS1 from Arabidopsis. Their function and expression pattern is further investigated. Microarray analysis revealed numerous differentially expressed genes. Currently some candidates (FKF1, LOX etc.) are further analysed. Various experimental set-ups such as different day length periods (16h/ 12h or 8h light) were tested which in general confirmed that negative effects on tuber quality (tuber weight, second growth, dormancy) are more pronounced in long day conditions (16h light). Several experiments confirmed a reduced accumulation of starch in tubers when plants were grown under elevated temperatures. The activity of sucrose synthase (SuSy) correlates with starch levels and may serve as a molecular marker. To unravel the molecular mechanisms the heat / cold plate set up is used to identify genes correlating with expression of SuSy or other starch biosynthetic genes. In addition, genomic annotation of genes involved in starch metabolism was improved and this information is being used to identify co-regulated genes. WUR group obtained evidence for the involvement of CYCLING DOF FACTOR 1 (StCDF1) and StCDF3 genes in abiotic stress (heat and drought) sensitivity in potato plants. StCDF1 protein also transcriptionally represses StCO1-3 genes, a set of transcription factors able to indirectly repress the potato tuberigen StSP6A. Transgenic plants with reduced expression levels of StCDF1 showed high level of tolerance to drought and higher tolerance to heat stress. Interestingly, plants over-expressing StCDF3, another gene of the same family repressed by StCDF1 also presented a higher tolerance to drought and clearly improved growth and yield during heat treatments. Plants with different levels of StCDF1 presented a strong stomatal conductance phenotype, being lower in plants with low StCDF1 expression. Furthermore, in early potato genotypes, with more stable StCDF1, stomatal conductance is also greatly reduced. Transgenic plants with low expression of StCDF1 also have a decreased basal transpiration rate and they are extremely resistant to drought stress. On the other hand, a decrease in the transpiration rate has a negative effect on yield and plant production under heat stress. They explain the antagonism between treatments as a consequence of the miss-regulation in the StCDF1 gene expression, and concomitant incapacity of the plant to properly coordinate the induction of tuberisation with the needs of photosynthetic capacity under circumstances of environmental challenges. If transpiration is interrupted by stomatal closure due to StCDF1/StCDF3 miss-regulation, the major plant cooling mechanism is lost. They have established Chromatin Immune precipitation (ChIP) for potato. As a first approach we set up the experiment with the StCDF1 target genes StCO1-3. We show that StCDF1 protein binds close to ORF in StCO genes. Consistent with the ChIP result the StCDF1 binding site (AAAG) is preferentially present close to, or in 5´UTR ORF chromatin regions. They now also generated a SUC2 regulated GFP-tagged StCDF1 protein in potato. These plants will be used to carry out ChIP-seq analyse of the regulatory activity of StCDF1 throughout the genome. CSIC: Transgenic SP6A OE lines (35S promoter) in cv. Spunta and Sylvana backgrounds have been generated and characterized. Compared to wild-type plants an improved tuber yield was observed at elevated temperature together with reduced secondary growth. OE- lines accumulated more tuber starch and lower soluble sugars. Tubers from SP6A OE-lines sprouted later than controls during storage at 4°C. Microarray experiments with the transgenic lines have been conducted in collaboration with FAU and data analysis is ongoing. An initial heat stress experiment was performed including potato plants with increased levels of gibberellic acids (OE of AtGA20-ox1) and reduced amounts of strigolactones (CCD8-RNAi lines).These lines were obtained from JHI/StA. Compared to their respective wildtypes AtGA20-ox-OE lines did not exhibit significant differences in tuber number/ weight and dormancy period. CCD8-RNAi lines formed already under control conditions less tubers. Interestingly, these plants tended to form aerial tubers under the conditions used which will be further analysed.
Impact During this first 18-month period collaboration was established with all the groups and companies in the HotSol project. In this regard agreements were made to share information and project progress of all teams via DropBox. Protocols and experiments performance are also shared. We agreed to have two project meetings per year. Thus far, we had four meetings (see engagement activities) with attendance of all partners, associate partners and representatives of each company. In addition, monthly online meetings were conducted between all groups and companies.
Start Year 2016
 
Description Abiotic stress in potato 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Raised awareness, led to several discussion points,

several follow up discussions with press and media interest in crop genomics
Year(s) Of Engagement Activity 2015
URL http://www.hutton.ac.uk/events/potatoes-practice-2015
 
Description ERA-CAPS HotSol meeting_Dundee 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact Almudena Trapero Mozos presented the progress from StA/JHI. Further work has been directed at elucidating the gene underlying a Chr 4 QTL that explains ca. 12% of the variance in yield in a tuber nodal cutting assay. A Hsc70 gene is located in the región to which the QTL maps. Genotypes were selected from the population that exhibited extreme sensitivity or tolerance to heat stress. The transcript level of the the HSc70 gene was determined in leaves and tubers of these lines under low and high temperatures and the tolerant lines consistently expressed HSc70 at much higher levels. Constructs for HSc70 o/e have been generated using the GoldenBraid cloning system and transgenic lines are being re-generated. HSc70 expression levels in accessions from the Commonwealth Potato Collection have been measured and the aim is to look for correlations with heat sensitivity using the nodal cutting assay. A potato exome capture array has been designed on the Roche Nimblegen platform and the aim is to apply this to the 06H1 population to improve resolution of QTLs

Experiments in fields and new action points were designed
Year(s) Of Engagement Activity 2015
 
Description ERA-CAPS HotSol meeting_Nurnberg 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact We presented the recent results from the QTL mapping of heat tolerance traits in the segregating diploid potato population 06H1. Several QTLs were found including one on chromosome 4 that explains around 12% of the variation. A candidate gene identified as HSc70 was found mapping below the QTL on chromosome 4. We described the allelic composition of this locus and identified specific allele combinations that appear to be correlated with the heat tolerance effect. Also, We were able to show from transcriptional analysis, that HSc70 gene are induced under high temperature in potato further strengthening the possible role in the heat tolerance phenotype

Different actions points were designed for the next project meeting
Year(s) Of Engagement Activity 2014
 
Description Genomics, Gene Discovery and Genome Editing in Crops 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact This UK-Peru joint workshop greatly enhanced the capacity of Peruvian scientists to implement modern genomics and biotechnological methods in their crop breeding programs for rapid and targeted improvement of their respective crops. Also, it opened the possibility of collaboration between UK plant research centres and key Peruvian agricultural institutes.
Year(s) Of Engagement Activity 2016
URL http://www.hutton.ac.uk/events/genomics-gene-discovery-and-genome-editing-crops
 
Description Poster Session in Plant and Animal Genome 2017 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Poster included in Antibiotic stress session of Plant and Animal Genome XXV in San Diego, California
Year(s) Of Engagement Activity 2017
URL http://www.intlpag.org/
 
Description Project Meeting_Dundee_31.03.2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Almudena Trapero Mozos presented the progress from StA/JHI. Further work has been directed at elucidating the gene underlying a Chr 4 QTL that explains ca. 12% of the variance in yield in a tuber nodal cutting assay. A Hsc70 gene is located in the región to which the QTL maps. Genotypes were selected from the population that exhibited extreme sensitivity or tolerance to heat stress. The transcript level of the the HSc70 gene was determined in leaves and tubers of these lines under low and high temperatures and the tolerant lines consistently expressed HSc70 at much higher levels. Constructs for HSc70 o/e have been generated using the GoldenBraid cloning system and transgenic lines are being re-generated. HSc70 expression levels in accessions from the Commonwealth Potato Collection have been measured and the aim is to look for correlations with heat sensitivity using the nodal cutting assay. A potato exome capture array has been designed on the Roche Nimblegen platform and the aim is to apply this to the 06H1 population to improve resolution of QTLs.
Year(s) Of Engagement Activity 2015
 
Description Project Meeting_Madrid_13.10.2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Study participants or study members
Results and Impact Almudena Trapero Mozos presented the progress from StA/JHI. A Hsc70 gene was identified previoulsy from a QTL related to heat response. Genotypes were selected from the population that exhibited extreme sensitivity or tolerance to heat stress. Further work confirmed that Hsc70 is highly expressed under heat stress. Constructs for HSc70 o/e have been generated using the GoldenBraid cloning system and transgenic lines are being re-generated. She used transient expression of the gene in Nicotiana benthamiana as a tool to confirm its role in conferring increased heat tolerance. During her work she observed an higher tolerance of plants towards increased temperatures when the plants were allowed to acclimate to 25°C before shifting them to 40°C treatment. The temperature shift experiment was perfomed with different cultivated and wild species showing different responses which will be further investigated.
In addition she reported about a first Exon capture (using Nimblegene design) experiment perfomed in JHI with the 06H1 population segrating for different yield under heat stress

New experiments
Year(s) Of Engagement Activity 2015
 
Description Project Meeting_Nurnberg_23.09.2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Almudena (Post doc, JHI, UK) presented the recent results from the QTL mapping of heat tolerance traits in the segregating diploid potato population 06H1. Several QTLs were found including one on chromosome 4 that explains around 12% of the variation. A candidate gene identified as HSc70 was found mapping below the QTL on chromosome 4. Almundena described the allelic composition of this locus and identified specific allele combinations that appear to be correlated with the heat tolerance effect. Also, she was able to show from transcriptional analysis, that HSc70 gene are induced under high temperature in potato further strengthening the possible role in the heat tolerance phenotype
Year(s) Of Engagement Activity 2014
 
Description Visit to Xisen Potato Company Ltd 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Visit to Xisen potato company where discussions were held with Dr Xie (R&D director), Dr Hu (Managing director) and Mr Liang (owner). L Torrance gave a talk to company technical staff and visited sites in Shendong province, Inner Mongolia and Beijing.
Year(s) Of Engagement Activity 2017
 
Description Workshop on Potato's technology Nairobi Kenya 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The workshop was held to raise awareness of new innovations in potato technologies (results of research, traits of different potato cvs, pest and disease problems) among Kenyan farmers, plant health officials, seed producers and NGO.
Year(s) Of Engagement Activity 2018
 
Description presentation at European potato conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at European Association of Potato Research triennial meeting APPROACHES TO UNDERSTANDING HEAT TOLERANCE IN POTATO 20th EAPR Triennial Conference (EAPR 2017) Paris, July 2017
Year(s) Of Engagement Activity 2017