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Accelerated Development of multiple-stress tolerAnt PoTato

Periodic Reporting for period 3 - ADAPT (Accelerated Development of multiple-stress tolerAnt PoTato)

Période du rapport: 2023-07-01 au 2024-10-31

Potato is one of the most important food crops worldwide. Despite its high yield potential and high water and nutrient use efficiency, potato is sensitive to environmental stresses such as heat, drought and flooding. Stress acclimation requires metabolic reprogramming triggered by signalling pathways, which are mostly still not understood in potato. ADAPT developed novel approaches to understand the dynamics of complex signalling and response mechanisms combining high throughput phenotyping and novel tools for live-imaging of cellular signalling responses. This guided in-depth analysis of key developmental stages for tuber formation under stress conditions. Key findings of ADAPT were: 1. Detailed understanding potato stress acclimation to heat, drought, and waterlogging. 2. Collection of data how different potato genotypes handle combined abiotic stress occurring under realistic conditions. 3. Establishment of advanced phenotyping with high-throughput in glass houses and field trials using drones and environmental sensors. 4. Monitoriing of critical moments during potato growth and tuber formation to identify specific signatures and molecular responses to be explored in future breeding. 5. Development of powerful data analysis pipelines for processing and analysis of huge data sets from field trials and high-throughput studies. 6. Improved understanding of stress reactions as a basis for fine-phenotyping and marker development for future potato breeding. Knowledge gained in ADAPT will directly be used by relevant stakeholders and end-users feeding into breeding programmes and guiding technology development for improved crop management strategies.
To explore the genetic variety present in current breeding lines, we performed two stress-related field trials, each consisting of 54 potato varieties, selected based on previous projects for differential sensitivity to heat and drought stress. To obtain insights into molecular events during stress acclimation, glasshouse studies under controlled stress conditions were performed in the PlantScreenTM phenotyping platform at the PSI Research Center in Brno (CZ). This allowed us to define growth conditions, stress application, experimental design, and phenotyping protocols. We assessed the morpho-physiological traits by image-based sensors and harvested samples for downstream analyses of hormone, metabolite, marker gene and proteomics analyses by different partners. Understanding the dynamics of the stress response and the molecular pathways involved will be critical to screen selected potato cultivars out of the larger panel used in the field trials. Additionally, 30 different varieties were screened for their response to heat- and drought stress revealing genetic variance in the stress-response of different varieties and suitable parameters to rank them.

Most important for stability of potato yield under stress conditions is the amount and the quality of potato tubers generated. Thus, the overall aim was to unravel the complex relationship between the tuberisation signalling pathways and responses to abiotic stresses at the molecular level. A detailed transcriptomic analysis of transgenic potato lines revealed how SP6A expression is related to plant hormone signalling pathways. Transgenic reporter lines have been developed and demonstrated in detail the control of SP6A as major tuberization signal under environmental stress conditions.
Insights into early signalling (sensory) events in potato that regulate the molecular and physiological acclimation to stress were gained using two types of transgenic plants were generated: (i) sensor lines to measure secondary messengers (Ca2+ and ROS); and (ii) gene reporter lines to measure ROS, drought-stress responses, and stress-induced plant hormones as important regulators determining plant resistance to drought and water logging.

A key issue arising from large-scale and high-throughput studies is the analysis of large data sets. Therefore, we focused in the first period on setting up the methodological basis for the analysis and have developed the StressKnowledgeMap App, which allows both visualisation of current knowledge in network format, reproducible insertion of novel knowledge and easy linkage to mechanistic modelling. We compiled several data analysis protocols that are now available for use by ADAPT and the broader scientific community.
Field trials were done in Austria, The Netherlands, Serbia and Spain using newest technologies monitored plant growth and development with sensors for 3D plant modelling (LIDAR), photosynthetic performance (hyperspectral camera), transpiration (IR sensor). This led to the identification of varieties with a wide spectrum of responses and sensitivity to abiotic stress for optimal yield at different sites. It allowwed to correlate drone data and obtained yield, indicating promising possibilities to develop methods for yield prediction from a set of 44 commercial varieties and 10 diploid clones.

Tailor-made tools and high-throughput techniques identified the morphological, physiological and molecular changes that determine potato resilience to abiotic stresses (flooding, heat and drought). An advanced phenotyping protocol was developed for multiple scale integration of single- and combined stresses responses, from observable growth phenotypes to specific molecular responses identifying specific stress signatures and molecular insights for future selection of traits.

Studiedies of potato tuber formation and growth, which is typically shut down under stress conditions were performed. Molecular key regulators of tuberisation are now understood at unprecedented level providing novel tools to predict impact on yield. These were the first systematic approaches to study how tuberisation signals are affected by the environment.

To study molecular signalling events during stress acclimation, we developed sensor and gene reporter lines to monitor dynamic changes in stress signalling in real time and under HTP conditions. These tools enable the detection of rapid and early plant adaptive responses in potato, which can be applied as very early markers for selection of stress resilience.

Data Integration and modelling were performed to identify novel molecular master regulators and physiological signatures associated to the performance of the potato in the field environment. Data analysis pipelines enabled systematic and fast integration of complex data sets from multiple experiments and multiple molecular levels. They form the basis for future yield prediction as well as toolboxes to be released to the public for data analysis and modelling, applicable to similar studies in other crops. The open access StressKnowledgeMap tool provides a comprehensive framework for further molecular analyses and mechanistic understanding of complex stress responses.

A survey of European potato farmers revealed that they feel the impact of climate change in their daily work and urgently need this research and better adapted varieties. Software tools for implementation of novel phenotyping technologies involving High-Throughput-Phenotyping (HTP) and analysis of drone data will be adjusted to the needs of the breeders and other end-users with the aim to develop procedures for yield prediction and gained knowledge will feed into optimizing support/decision making tools, available to grower’s through local support agencies.
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