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Identification, isolation and analysis of stress-induced plant genes in Arabidopsis thaliana by T-DNA tagging and gene fusion

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The main limitation in plant productivity and crop stability both in agriculture and forestry is caused by physical (climatic) and biological stress factors to which the plants are exposed, such as extremes in temperature, drought, pests and diseases. Modern plant biotechnology has provided the means to improve plants by generating genetically engineered plants with new and improved characteristics. Although some economically important traits such as insect-, virus- or herbicide resistance and male sterility have been introduced into some crop plants, only a limited number of useful genes that can be directly applied to crop improvement are readily available. New molecular techniques need to be identified and plant genes cloned, encoding useful but often not so well defined characteristics such as stress tolerance or disease resistance.

Gene tagging, either by insertion of transposable elements or by insertion of foreign DNA (e.g. T-DNA) into the plant genome using the natural gene transfer system of Agrobacterium, provides an effective alternative for gene isolation. Mutations caused by T-DNA insertion are readily identified and the T-DNA along with the flanking chromosomal regions can be recloned. Here the optimisation of T-DNA tagging methods will be done. Tagged genes will therefore be isolated and characterized.

Identification of a particular class of genes by T-DNA tagging can be further facilitated by gene fusions using promoter- or enhancerless reporter genes placed next to the T-DNA border/s which will be expressed only when they come under the control of a particular plant promoter or regulatory elements. Consequently the promoter provides an easy screen for e.g. stress-induced promoters or enhancer elements. New reporter genes and tagging vectors will be developed. Tagging of plant genes by T-DNA mediated gene fusion appears to be widely practicable for molecular analysis of plant genes and it is planned to use this technique for isolation and identification of specific stress-induced genes from Arabidopsis thaliana. The small cruciferae A.thaliana has proved to be an excellent model for many aspects of plant molecular biology. It has a particularly small haploid genome (about 108 bp) containing an estimated 20,000 genes. Hence only a limited number of insertion mutants are required to represent the complete genome. A library of T-DNA added A.thaliana will therefore be generated and the specific tagged lines will be identified.

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University of Helsinki
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00014 Helsinki
Finnland

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Letzte Aktualisierung: 19 September 1996

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Permalink: https://cordis.europa.eu/project/id/INTAS-94-2797/de

European Union, 2024