Objective
The aim of the investigation is to modify the fatty acid composition, especially the content of hydroxy-unsaturated fatty acids, in rape-seed by introducing alien genes via somatic hybridization.
Somatic hybrids will be produced between rape-seed and Lesquerella fendleri which is a wild species in the family of Brassicaceae known for its high content of lesquerolic acid (14-OH-eicosa-II-enoic acid), a hydroxy-unsaturated fatty acid of large economic importance as a technical oil. Rape-seed and Lesquerella cannot be crossed sexually. Furthermore, since the genes regulating the synthesis of lesquerolic acid are not identified and cloned, transformation techniques cannot be used. Thus the only way to combine the two genomes is via somatic hybridization.
Somatic hybrids will be produced by PEG-induced infusions and the hybrid protoplasts will be selected by flow cytometry and cell sorting using the established techniques developed in the laboratory in Uppsala, Sweden. The produced hybrids will be analysed genetically by calculating chromosome number and determining the presence of hybrid features utilizing RFLP techniques. Furthermore, the progeny obtained via backcrossing to rape-seed and, if possible, via selfing will be analysed with respect to presence of the desired fatty acid. Cytological investigations will be made using in situ hybridization of the hybrid material and the progeny from the hybrids expressing the desired lesquerolic acid to follow and determine whether a stable inheritance and introgression of alien DNA from Lesquerella into the genome of rape-seed takes place.
The project will involve: production of the somatic hybrids and RFLP analysis of the nuclear genomes to determine hybrid features of the plants produced; establishment of the material in greenhouse conditions and analysis of fertility and determination of fatty acid composition in the seeds; cytological investigations including in situ hybridization of species specific DNA from Lesquerella.
Topic(s)
Call for proposal
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75007 Uppsala
Sweden