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Contact: Nicola McCallum

More Information:

• Barley TILLING at SCRI
• Barley Mutagenesis at SCRI

Current status

SCRI have used Ethylene Methyl Sulphonate (EMS) to generate a population of approximately 22,000 lines. EMS introduces methyl groups into the DNA which leads to errors during DNA replication and thus introduces mutations. Typically this means that some C nucleotides within a sequence are replaced with T residues and also G residues with A residues. Within our population this occurs at random sites at a frequency of approximately 1-5 changes in every 100000 nucleotides. SCRI have recently bought three LiCOR sequencing systems which allow the detection of mutants using the Cel 1 digestion method. SCRI are currently funded by BBSRC and the Scottish Government to develop this protocol to provide a service of mutant isolation for the research and industrial community in the UK. We are currently collaborating with many laboratories in the UK and across the EU to identify mutations in candidate genes which may be involved in sprouting (germination), nutrient partitioning, flowering time/vernalisation, pathogen resistance and malting quality.

SCRI has a working screening population of about 5000 barley cv. Optic lines, which will be extended to 10,000 in the next 6 months. This was BBSRC funded in order to establish a 'free' service to the UK community. Currently 22 groups have submitted amplicons for screening, or are working on amplicon design. A submission system has been established via a website. SCRI has 3 LiCOR analysis systems and a Tepnel DNA prep machine. A phenotyping database is available from seed to maturity for 21,000 EMS lines. Characters recorded include seed shape/size, germination, root development, vegetative and inflorescence characters, disease resistance etc.

Costs and time scales

see http://bioinf.scri.ac.uk/barley/

Current work and future directions

The collection of mutants is currently maintained as seed and DNA stocks at SCRI. It is currently used as an in-house resource but is available to collaborators outside of SCRI on a consumables-costs recovery basis (see http://bioinf.scri.ac.uk/barley/) . Given the success of the TILLING screening at JIC, we may consider transferring out LICOR screening to ABI 3730's. A population of 5000 DNAs has been transferred to the genome centre at JIC which pending agreement we would like to extend to 10,000 in the coming months.