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Title Identification of molecular markers for the detection of bio-regulators that enhance plant productivity and quality
   
Acronym Bioregulators
   
Duration 1 August 2007 - 1 August 2010
   
Project leader Pascal von Koskull-Döring, Bayer Crop Science AG [Company], Germany
   

Other project participants

John Graham Waples, BIOTEK Agriculture [Company], France
Laurent Guerreiro, ARVALIS - Institute du végétal [Company] , France
Stefan Schillberg, Frauenhofer Institute for Molecular Biology and Applied Ecology, Germany
Paul Christou, Lleida University, Spain
José Luis Guardiola, Technical University of Valencia, Spain
  
Funding National Institute for Agricultural Research (INRA), France 
  Research Centre Juelich – Project Management Juelich (FZJ-PTO) on behalf of the Federal Ministry of Education and Research (BMBF), Germany
  Ministry Science and Innovation (MICINN), Spain
   
Total granted budget  € 2,179,822
   
Abstract  

The objective of this project is to exploit the potential of biologically active chemicals (bioregulators) that positively influence plant growth and productivity by identifying transcripts, proteins and metabolites that are up- or down-regulated when the chemicals are applied under stress and non-stress conditions. So far stress models were successfully established for Arabidopsis (cold, salt and drought) and Rice (salt and drought). (1) Stress assays in the presence of bioregulators were initiated and several promising results obtained already. (2) To identify the responsible gene products we started with transcriptome, proteome and metabolome profiling of these plants. (3) The identified marker genes will be used to establish a cell-based high-throughput assay. (4) The corresponding promoters driving the marker genes will be fused to a fluorescent protein and plant expression cassettes will be introduced into Arabidopsis suspension cells. (5) The transformation procedure with pDsRed and its detection could be established already. (6) This cell-based fluorescence assay will be used to screen for novel or superior compounds that induce the same pathway responsible for enhanced crop yield and quality. (7) Furthermore, an important aspect of the project is the performance of bioregulators on yield and quality of crops in the field under abiotic stress conditions. Addressing this point, salt stress models for tomato and lettuce were established and the establishment of drought stress trials in plastic tunnels as well as wheat and corn field trials is well advanced.

 
Progress

Stress assays for different plant species have been established. The growth of Arabidopsis plants both on soil and in hydroponic cultures under stress-free conditions has been established and monitored. Stress assays for cold (on soil), salt and drought stress (hydroponic) are established as well. Experiments with the application of BioRegulators on cold stress plants have been started.

We have also been able to set up very reliable drought and salt stress assays for rice plants and in vitro cultures. At least two BioRegulators showed discernable effects in preliminary experiments.

Salt stress assays in two lettuce and three tomato cultivars have been established as well. Different growth parameters (e.g. number of leaves, biomass) and photosynthetic parameters (net CO2 fixation, stomatal conductance, chlorophyll fluorescence) are being evaluated.

The conditions for sample preparation for transcriptome analyses of Arabidopsis and rice plants have been set up. We are using the Arabidopsis ATH1 and the Rice Genome Array from Affymetrix to analyse the changes in gene expression of stressed/non-stressed plants with/without application of BioRegulators.

2D electrophoresis using fluorescent protein tags (DIGE, differential in gel electrophoresis) will be used for the identification of protein biomarkers which are indicative for BioRegulator effects in stressed plants. A first comparative study of the leaf proteome with regards to the effect of cold stress over a period of 7 days has been performed using the DIGE system. After 144 hrs of cold stress a total of 112 protein spots were found to be differentially regulated. Of these, 64 spots increased and 48 spots decreased in abundance. In total ~ 1300 spots were detected on the 2D map.

To establish a cell based screening assay for new Bioregulators an Arabidopsis cell suspension cultures has been initiated. The method for stable transformation of cells via co-cultivation with Agrobacterium tumefaciens has been used to produce cells expressing the fluorescent marker protein DsRed and HSA. Transformed cells are currently characterized with regards to their growth and protein production.

In 2008 several field trials with wheat, corn, tomato and lettuce under abiotic stress conditions are being performed to test the effect of Bioregulators in crops in the field.