description
- WORK PROGRAMME: The main objective of the lab rotation project will be to identify DELLA (RHT-1) interacting proteins that control GA-responsive aleurone responses in the wheat grain. The identity of these components has the potential to provide insights into the molecular mechanisms through which DELLAs regulate GAMYB activity in aleurone cells. To identify RHT-1 interacting proteins, a yeast 2-hybrid screen will be performed using an RHT-1 bait construct to screen an aleurone cDNA prey library (generated by our group). Prey clones encoding potential interactors will be sequenced and bioinformatics analyses performed to investigate function. Confirmation that clones encode DELLA interactors will be performed by retransforming plasmids into yeast and repeating yeast 2-hybrid assays. An additional objective of the rotation project will be the characterisation of a-amylase induction in wheat gamyb mutants. These experiments will involve monitoring the effect of GA on a-amylase induction in all mutant combinations to establish whether the roles of the different homoeologous GAMYB genes in wheat. TRAINING: This project will involve a broad range of techniques and experimental approaches used in plant biology. These will include general molecular biology cloning techniques using bacteria and yeast. The student will also gain considerable experience in using yeast 2-hybrid assays to identify interacting proteins. There will also be a strong bioinformatics element within the project, allowing the student to gain experience using Geneious, QTLnetminer and interrogation of wheat genomic resources. In addition, the student will gain experience in reverse genetics approaches in wheat.