Using Arabidopsis TILLING alleles of COMATOSE to understand germination potential

Abstract

The CTS locus in Arabidopsis thaliana was identified to promote the transition from embryo development to germination. Mutation of this locus results in reduction of germination potential. This is caused by the disruption of the (3-oxidation pathway which is essential in breaking down fatty acids to provide energy for embryo expansion and radicle protrusion for the germination process. Mutant of CTS show much ability to elongate roots on medium with O.SxMS plus 0.5% sucrose plus varying concentrations of 2,4-DB as compared to their relative wild types. The purpose of this study was to try to define an allelic series of cts to help structure function studies and to find out more about the specific role of CTS. This was done using TILLING alleles of CTS. These have point mutations caused by ethy methanesulphate on the exons of the 'CTS locus.. The TILLING mutants were determined with the aid of PCR analysis. To achieve the objectives phenotypic analysis experiments were conducted. Root elongation phenotypic analysis results after 8 days on medium with O.SxMS plus 0.5% sucrose plus 0.05ug/ml 2,4-DB after 8 days showed that the TILLING mutants had a higher mean root elongation length than the wild type, Columbia big mama. The TILLING mutants C22 and C60. exhibited significant root elongation (students, t- test, P< 0.05). The other known mutant of CTS, cts-1 had a much higher significant mean root elongation length when compared to its wild type Ler (students, t- test, P< 0.01). The results obtained indicated that the TILLING mutants have a slightly defective poxidation pathway when compared to the other known mutants of CTS