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Controlling the location and frequency of meiotic recombination in plants

Principal Supervisor: Dr Eugenio Sanchez Moran - School of Biosciences

Co-supervisor: Chris Franklin

PhD project title: Controlling the location and frequency of meiotic recombination in plants

University of Registration: University of Birmingham

Project outline:

Most of our food arise from plants either directly (crops/vegetables/fruits) or indirectly (animal foods). Plant reproduction plays a key role as it provides the grain and fruit that we eat. In addition, during plant reproduction (meiosis), genetic variation is able to occur. This genetic variation is the base for plant breeding and produce better crops. The challenges of increasing global population (~9billion by 2050) and climate change will constrain the global food security. To overcome these challenges is essential to develop more resilient and higher yield crops. Understanding the processes to control meiotic recombination in plants is essential to provide food security for the 21st century.

Our laboratory has been involved in meiotic recombination research for several years which has allowed us to gain a very good overview of the different molecular pathways and the dynamic interactions of different chromatin components in the control of meiotic recombination (genetic crossovers) in the model plant Arabidopsis thaliana. This knowledge is being recently applied into other plant crops like Brassicas and cereals. We are especially interested in investigating the fundamental molecular mechanisms that are controlling the frequency and distribution of genetic crossovers along chromosomes. We are particularly interested in understanding how the programmed global changes in chromosome organization direct the fate of crossovers and just occur in specific regions. This problem is very important for plant breeders in cereals. Different molecular, biochemical, cytological and microscopy techniques would be applied in this project in order to study different aspects of meiotic recombination and being able to manipulate them to use them as tools in plant breeding techniques.

References:

  1. Sanchez-Moran E, Santos JL, Jones GH and Franklin FCH. 2007. ASY1 mediates AtDMC1-dependent interhomolog recombination during meiosis in Arabidopsis. Genes & Development 21: 2220-2233.
  2. Sanchez-Moran E and Armstrong SJ. 2014. Meiotic chromosome synapsis and recombination in Arabidopsis thaliana: new ways of integrating cytological and molecular approaches. Chromosome Research 22: 179-190

BBSRC Strategic Research Priority: Food Security

Techniques that will be undertaken during the project:

  • Molecular cytogenetics: This is based around a variety of advanced cytological, molecular and microscopy techniques. Confocal microscopy will be used for live imaging (using GFP/RFP recombinant proteins) and quantitative time studies of photobleaching and photoactivation. Super resolution microscopes will be used to analyse the localisation and association of meiotic recombination proteins at molecular level.
  • Molecular Biology: different basic techniques will be used in this project, from PCR amplifications, DNA and protein gel electrophoresis, cloning, production of mutants by T-DNA transformation, RNA interference and proteomic analysis of protein interactions by IP/AP pull downs and spectrophotometry, production of recombinant proteins with fluorescent proteins and different tags for AP analysis. Furthermore genome targeting methods (ZFNs, TALENs, CRISPR/Cas9) will be used to target mutations and to target DSBs formation in sequence specifics sites which could use for genetic recombination.
  • Genetic analysis: Direct and reverse genetic approaches will be used to analyse the role of putative components involved in meiotic recombination. This includes: T-DNA transformation, RNA interference, genome targeting mutagenesis. Complementation and over expression studies will be also performed. Chromatin IP (ChIP) coupled with DNA sequencing will be carried out to correlate meiotic components with the genomic sequence and chromatin context.

Contact: Dr Eugenio Sanchez Moran, University of Birmingham