NIAB - National Institute of Agricultural Botany

Community Resource for Wheat and Rice Transformation
Information and Application form

The Community Resource for Wheat and Rice Transformation provides capacity for 100 genes to be transformed into wheat and rice free of charge. We envisage that 75 genes will be transformed into wheat and 25 genes into rice. In the event that wheat or rice transformation is not fully subscribed, additional slots will be made available in the other species. We will encourage researchers working with genes from model crops to test their hypotheses in wheat and rice as appropriate to the gene studied. This will enable new genes to be evaluated more quickly in crops essential to food security worldwide, and allow researchers to amass crucial data which can be used as a basis to seek follow-on funding.

This project also provides funded capacity to characterise 50 regulatory elements in wheat and rice. Promoter and terminator sequences will be included for their expression in a wide range of tissue types, biotic and abiotic stress induced elements will also be included. Researchers can either nominate promoters to be included in this project, or provide constructs which are compatible with our vectors. Please contact us for further details on this aspect of the project.

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 transformation of their nominated genes. 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 or rice, but do not have the funding to support the 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 appropriate attL sequences, ready for transfer into a NIAB binary vector with a selection cassette via Gateway™ cloning. Some examples of typical destination vector T-DNA regions for wheat transformation are shown below. We have similar constructs for rice transformation. Confirmation by sequencing that the supplied Entry-type construct is correct will be required at the construct submission stage. Advice on construct design can 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. Goldengate constructs can also be accepted if the gene cassettes are flanked by attL sites. 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 T0 wheat plants per construct, as plantlets at 12 weeks from experiment start to the researcher to grow in their own facilities. Non-transformed plants will be provided as control material. In the majority of cases we will use a US soft spring hexaploid wheat, Fielder, for this activity, but if spring or winter wheat 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.

Rice transformation will use Nipponbare, Kitaake or Donjin germplasm, with 15-20 T0 plants per construct at about 8 weeks from experiment start, provided to the researcher to grow in their own facilities.

Post transformation analysis

All plantlets will be analysed by qPCR at the small rooted plantlet stage, to verify that they are transgenic and to provide information on the number of T-DNA copies introduced. Researchers will receive small rooted T0 plants and are very strongly encouraged to undertake RNA /protein expression, metabolic or phenotypic analysis on these T0 plants to prioritise lines for further investigation. The strategy for this analysis must be included in the application, together with confirmation that the researcher has the appropriate facilities to grow the plants to maturity.

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 1st round of applications is 24th June 2019.

Application Form

Please fill in all fields. We may contact you for further information before a final decision is made by the Advisory Group.

Applicants details. If this is a joint/collaborative application please include the names of all applicants, with affiliations, plus full contact details for the primary applicant whose name should be underlined.

Name and
affiliation
Address
Telephone
Email


Project title. Maximum of 20 words.

Brief details of project for inclusion on CRWT website if the application is successful. Maximum of 80 words.

Details of proposed gene and programme of research. Maximum of 800 words.
The strategy for characterising the T0 plants and subsequent generations of transgenic wheat or rice must be included. If a wheat germplasm other than Fielder is more suitable for your project, please include the justification for the end-use (nabim) group, disease susceptibility or other attribute requested.

How will this research address BBSRC food security targets? Maximum of 400 words

Please describe the resources, including both staff and growth facilities, which you have in place to evaluate the wheat or rice plants transformed with your candidate gene, and length of time the resources are available. Please also include your justification for applying for this resource rather than applying directly for grant funding. Maximum of 200 words.

Is the gene from a cereal species? Yes No
Please include the species from which your gene has originated:

Is an over-expression, RNAi or genome editing construct required? OEX RNAi genome editing

If over-expression construct, is the intermediate construct available? Yes No
If not, please indicate when it will be available for transfer to NIAB:

If RNAi/genome editing construct, do you have wheat homeologous sequences identified? Yes No
If RNAi construct, do you have wheat sequences cloned? Yes No

If not, please include strategy and timescale for identifying and cloning wheat sequence Maximum of 200 words

Are you submitting other applications in this round:

  • For over-expression and silencing by RNAi of the same gene? Yes No
  • For a different gene? Yes No