Project 3 - Identification and analysis of genes controlling early embryo and endosperm development

1. Planned Outcome / Aims

• Identification and characterisation of genes controlling the early stages of development of the embryo, endosperm and other grain tissues

2. Contracted Output(s)

• Mapped positions of candidate genes relative to genetic loci controlling grain quality characteristics in wheat and barley
  
  
• Transgenic lines over-expressing or under-expression candidate genes
  
  
• Phenotypic evaluation of transgenic lines
  
  
• Patented genes of defined function and identification of tissue and organ specific promoter sequences.

3. Research Methodology

1. Standard gene analysis including:
   
   
   • isolation of full length clones (most cDNA and AFLP clones will be partial)
     
     
   • in situ localisation to confirm the cells expressing the genes
     
     
   • For selected genes expression in heterologous systems (usually E. coli), antibody preparation and in situ localisation of protein.
   
   
   
2. Genetic analysis through mapping to existing wheat and barley mapping populations.  Five fully mapped barley and three wheat doubled haploid populations are currently available.  Linkage to loci previously identified as being associated with components of grain quality will be sought.  The barley and wheat populations have been extensively analysed for a wide range of characteristics, for example over 200 traits (largely related to grain quality) have been scored to the wheat population Halberd x Cranbrook.
   
   
3. Transgenesis.  The applicants have extensive experience in the transformation of both wheat and barley.  Target gene will be over-expressed or expression will be suppressed in transgenic lines and assayed for altered phenotype.  In some cases, transformation of rice, tobacco or Arabidopsis will also be used since this is easier and faster than wheat and barley transformation.
   
   

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