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Development of a routine system for Agrobacterium-mediated transformation of barley - CE0159(1)

Description
There is an urgent need for a routine, high frequency, barley transformation system to facilitate both fundamental research and crop improvement. The barley transformation systems which are currently available, tend to be based on the biolistic method which has many shortfalls including dependence on high-cost instrumentation, low transformation frequency, and complex re-arrangement of multiple copies of transgenes in the transgenic plants which are produced. Agrobacterium-mediated DNA delivery promises to be an attractive alternative, which, when optimised, will permit facile, routine, low cost, high frequency transformation of barley. In this project, we plan to establish such a transformation system. We will concentrate upon the development of highly regenerable cultures from immature embryos and microspores as target materials for Agrobacterium and upon the optimisation of co-cultivation procedures, with special emphasis upon the assessment of the effect of wounding and vir gene induction on the transformation frequency. In close collaboration with scientists at the John Innes Centre, IACR-Rothamsted and IGER, we will define an efficient binary vector system and optimise the procedures for growth of highly potent Agrobacterium strains. The new transformation system will facilitate advances in fundamental molecular genetics and barley crop improvement and will also have relevance to corresponding research in other cereals and grasses.
Objective
01: To establish regenerable barley cultures as targets for inoculation by Agrobacterium including:
01-1: the establishment of immature embryo cultures
01-2: the establishment of microspore and anther cultures
01-03: the establishment of cell suspension cultures from 01-01 and 01-02.

02: To establish an effective inoculation procedure by:
02-01: determining the optimum growth conditions and inoculation densities of various Agrobacterium strains
02-02: using established vectors and investigating the effects of virulence induction by virG mutants (provided by D Lonsdale) and vir gene inducers on transformation frequency
02-03: assessing the effect of wounding of target materials on transformation frequency

03: To select and screen transgenic plants

04: To identify transgenic plants and undertake a genetic analysis of their progeny.
Time-Scale and Cost
From: 1998

To: 2001

Cost: £242,563
Contractor / Funded Organisations
University - De Montfort
Keywords
Arable Farming              
Biotechnology              
Cereal Production              
Crop Improvement              
Farming              
Genetically modified food and crops              
GM Food              
Fields of Study
Arable Crops