The purpose of this project was to create molecular tools for the study of meiosis in Arabidopsis thaliana and to evaluate their effectiveness. Two types of transgenic plants were created with an intron-spliced hairpin RNA (ihpRNA) construct to target the AHP2 gene for RNA silencing. One had a constitutively expressed promoter; the other’s promoter was inducible with dexamethasone (DEX). Transformations with the constitutively expressed construct gave rise to ahp2RNAi plants with reduced AHP2 transcript levels, abnormal meioses and reduced fertility phenotypes. The creation of plants containing the dexamethasone-inducible construct was confirmed via PCR genotyping, and induction with DEX. However, the induction conditions tested do not appear to silence AHP2 as the transgenics had normal meiotic and reproductive phenotypes. Also a triple-locus, three color, fluorescent protein marker-tagged Arabidopsis line was created that will allow calculation of recombination frequencies for two intervals on chromosome 2 in both wild type and mutant plants.

The purpose of this project was to create molecular tools for the study of meiosis in Arabidopsis thaliana and to evaluate their effectiveness. Two types of transgenic plants were created with an intron-spliced hairpin RNA (ihpRNA) construct to target the AHP2 gene for RNA silencing. One had a constitutively expressed promoter; the other’s promoter was inducible with dexamethasone (DEX). Transformations with the constitutively expressed construct gave rise to ahp2RNAi plants with reduced AHP2 transcript levels, abnormal meioses and reduced fertility phenotypes. The creation of plants containing the dexamethasone-inducible construct was confirmed via PCR genotyping, and induction with DEX. However, the induction conditions tested do not appear to silence AHP2 as the transgenics had normal meiotic and reproductive phenotypes. Also a triple-locus, three color, fluorescent protein marker-tagged Arabidopsis line was created that will allow calculation of recombination frequencies for two intervals on chromosome 2 in both wild type and mutant plants.