Member

Member

  Biotechnology resources for arable crop transformation (BRACT)

The BRACT research programme aims to provide an efficient transformation capability for the main UK crops, while facilitating further improvements in transformation technology and delivery to the UK research community. It is a collaboration between Rothamsted Research and the John Innes Centre. The work at Rothamsted Research will focus on developing protocols for Agrobacterium-mediated wheat transformation, testing constructs in wheat and on developing ‘clean gene’ transformation methods for wheat.
The key arable crops included in this proposal are wheat, barley and oilseed rape while others are included as secondary crops or addressed in separate proposals from other groups. For the three major crops, the programme will provide all the resources needed to facilitate transformation. These will include constructs requiring the minimum effort to insert genes or promoters of interest, through to efficient transformation methodology that builds in the capacity to produce transgenic plants free of selectable marker genes. To facilitate the production of marker free plants, Agrobacterium tumefaciens-mediated transformation will be used, although biolistic methodology will be available if required.
The principal route to inform users of the resources available will be via a web site. We have acquired the URL ‘www.bract.org’ The new web site will be updated and extended to include new vector sets and details of the transformation capability for the different crops. In developing and delivering a transformation capability for the UK research community, we propose to consult widely with potential users to assess future requirements. Through the involvement of a user group, resources will be tailored to meet demand as far as is practicable. It is envisaged that after three years it will be possible to maintain a transformation resource centre for the benefit of the research community by users building full costs for their transformation needs into grant proposals.

  Exploiting bioactivity of European cereal grains for improved nutrition and health benefits - HEALTHGRAIN

To develop nutritionally enhanced wheat using a combination of improved varieties and innovative processing, with a focus on dietary fibre, resistant starch and phytochemicals. Should have a range of health benefits for consumers including reduced obesity and type 2 diabetes and improved bowel function. May also result in lower cardiovascular disease and reduced incidents of some forms of cancer.

Includes studentship.

  Use of maize and rice Mar sequences to stabilise the expression of transgenes in wheat

The main objective of this project is to develop and improve GM technology in wheat by exploiting matrix attachment regions (MAR) sequences from the cereals maize and rice. The MARs will be used to stabilize transgene expression using a high molecular weight (HMW) glutenin subunit transgene as a marker. A novel approach will be taken in which the Ac/Ds transposable element system is used to generate new insertion sites after the primary transformation process, to ensure that lines with equal insertion numbers can be compared for MAR function and that the transgene loci examined are not biased by subjection to selection pressure.
Transgenic wheat populations will be produced which contain i) a pair of MARs isolated from rice flanking the wheat high molecular weight glutenin subunit promoter and gene (HMW-1Dx5) ii) a pair of MARs isolated from maize flanking the same cassette iii) a pair of control or ‘mock’ MARs flanking the HMW-1Dx5 promoter and gene. These MAR-flanked expression cassettes will in turn be flanked by Dissociator (Ds) transposable elements from the maize Ac/Ds transposon system. When crossed with wheat lines transformed with the maize Activator (Ac) transposable element, the Ds-flanked expression cassettes will ‘jump’ to novel locations which are free from selection pressure. Since Ac/Ds does not replicate sequences prior to transposition, the transgene copy number in each crossed population should remain constant. Levels of expression from the MAR-flanked HMW glutenin cassettes will be determined and compared for MAR-mediated effects on expression.