Abstract

Cereal seeds are agriculture's most important renewable resource for food and industrial raw material. Conventional methods for the improvement of quality and yield have produced spectacular advances in the past. Future developments, however, will involve tapping the vast potential of the modern technology of plant modification. The chances of successful modification increase if the activity of a transgene can be precisely directed to the targeted cells. This requires the isolation and characterisation of many more cell type-specific promoters than are currently available. SeedDesign aims to isolate at least two functional promoter sequences for each of the cereal seed compartments, thus allowing the efficient modification of seed composition. This will be of enormous potential benefit for both European farmers and consumers by improving the quality and diversity of products. In addition, this technology will allow the EU seed industry to keep abreast of global competition.

Objectives

The principal objective of SeedDesign is to provide the European seed industry with a set of regulatory elements that can be used to genetically modify cereal seeds in a directed and efficient way. The project has been broken down into four workpackages (WP). In WP1 and WP2 the consortium will characterise promoter sequences specific for different compartments and developmental stages of the maize kernel. In WP3, promoters shown to be functional in maize will be tested in wheat. In WP4, the possibility of modifying maize seed composition using SeedDesign promoters will be tested in two practical situations.

Description of Work

WP1 and WP2 aim to complete the current list of compartment and developmental stage specific promoters in maize:

WP1 Isolation of genes specifically expressed in various seed compartments: Different methods, from cDNA display to high-density array hybridisation will be used. The expression pattern of the candidate genes will be analysed in a collaborative effort in which each partner will be in charge of one specific aspect of the characterisation process. Each partner will clone the promoter sequence of the tissue-specific genes that s/he initially isolated.

WP2 Cloning of the promoter sequences of compartment-specific genes that partners of SeedDesign have already isolated in maize: Promoters isolated in WP1 and WP2 will be used to prepare promoter-GUS constructs for maize transformation (Partner 5). Partners 1 to 4 will characterise the transgenic lines by determining transgene copy number, expression pattern of the reporter gene and, whenever possible, transmission stability.

WP3 Analyses the performance of SeedDesign promoters in the wheat system: Partner 6 (subcontractor of P1) will introduce Promoter-GUS constructs shown to be compartment-specific in the maize system into wheat. The partner that prepared the promoter-GUS construct will be responsible for the characterisation of the transgenic lines, as in WP1 and WP2.

WP4 Testing SeedDesign promoters: Promoters isolated in WP2 and in prior work of SeedDesign members will be fused to relevant genes and introduced into maize by partner 5. Plants will be analysed for transgene expression and backcrossed to elite cultivars to analyse its influence in agronomic performance.

Deliverables

• At least 10 maize promoters that direct the expression of a transgene to a specific compartment and developmental stage of the seed.
• Evaluation of the maize promoters in wheat, including maize promoters previously characterised by members of SeedDesign.
• Maize transgenic lines overexpressing three different genes in 5 different seed compartments.
• Generic know-how and intellectual property.

Contacts

Coordinator

• Gregorio HUEROS SOTO / Universidad de Alcala Depto. Biologia Celular y Genética / SPAIN

Participant

• Pascual PEREZ / Biogemma FRANCE
• Max-Planck-Institut für Züchtungsforschung GERMANY
• Peter ROGOWSKY / Reproduction et Développement des Plantes FRANCE
• Wolfgang WERR / Universitaet zu Köln Institut für Entwicklungsbiologie / GERMANY