NIAB - National Institute of Agricultural Botany

Community Resource for Wheat Transformation
Information and Application form

The Community Resource for Wheat Transformation provides capacity for 50 genes to be transformed into wheat free of charge. Half of this resource is reserved for researchers working on model crops, to encourage early testing of novel genes in wheat. The remaining half of the resource is reserved for researchers working on UK cereal crops, wheat and barley. More information on previous accepted genes. This project has been funded for 5 years by the BBSRC Bioinformatics & Biological Resources fund.

Who can apply?

Academic researchers from UK who are eligible for BBSRC funding are invited to apply for the 5th and final round of genes to be transformed into wheat. Researchers may apply for several different genes, or for over-expression and silencing by RNAi of the same gene target. Only 1 gene/construct can be included per application, however a researcher may submit more than 1 application per round. Further applications will not be considered by the Advisory Group until outstanding construct(s) from the same researcher, accepted in previous rounds, have been received.

This is a resource for researchers who would like to express a gene in wheat, but do not have the funding to support the wheat transformation. There must be a clear relevance to BBSRC food security targets.

Vectors

We would prefer genes to be supplied in an intermediate Entry construct flanked by aatL sequences, ready for transfer into a NIAB binary vector with an nptII selection cassette via Gateway™ cloning. Some examples of Destination vector T-DNA regions are shown below. Confirmation that the supplied Entry construct is correct will be required at the construct submission stage. Advice on construct design, and bioinformatics support to identify homologous sequences from wheat may be provided.

We use the rice actin promoter for constitutive over-expression (pSc4Act-R1R2-SCV) or knockout via RNAi silencing (pAct-IR-2) of the gene of interest.

Alternatively researchers may need to introduce their own "promoter-gene of interest-terminator" cassette, for example with the gene of interest expressed from the native sequence or an alternative promoter, (pRLF12R1R2).

Other vectors and promoters for traditional cloning approaches or Gateway® cloning may also be available. Please contact us at an early stage to discuss your cloning strategy. Email croptransformation@niab.com

Delivery

Our standard service is to deliver 30 independent transformed plants per construct, either as plantlets at 12 weeks from experiment start, or T1 seed at 6-7 months. Non-transformed plants will be provided as control material. In the majority of cases we will use a US spring hexaploid wheat, Fielder, for this activity.

As part of this project we are currently assessing the transformation efficiency of 50 UK and northern European wheat varieties. These varieties cover bread, biscuit, and feed wheats, and include varieties with specific disease susceptibilities. If germplasm with a specific end-use or disease susceptibility is necessary for your project, please include this information in your application. An adjustment to the number of plants produced with the alternative germplasm may be made.

Post transformation analysis

We will confirm that plantlets are transgenic by PCR and/or QPCR copy number analysis for the presence of the nptII selectable marker gene. Researchers are very strongly encouraged to undertake RNA /protein expression, metabolic or phenotypic analysis on the T0 plants to prioritise lines for further investigation. When plants are grown to seed at NIAB, researchers are welcome to visit us and collect plant samples for such analysis in their own laboratories. The strategy for this analysis must be included as part of the programme of research.

The application & review process

There will be an annual call for applications each year, until the programme is completed. If the number of applications exceeds the capacity available, applications will be reviewed by our external Advisory Group. Applications will be ranked on scientific merit/relevance to BBSRC food security, the quality of the proposal and the strategy for analysis.

The closing date for the 5th and final round of applications is 31st May 2016. We will aim to inform applicants of the Advisory Group's decision by 31st July 2016. Note, researchers sending Entry constructs after 1st January 2017, may receive small rooted wheat plantlets, rather than T1 seed.

The application process for this project is now closed. If you have a requirement for wheat transformation please contact croptransformation@niab.com.