Characterization of P4H7 and P4H5 Arabidopsis thaliana T-DNA mutants
2004
Anca, I.-A.
The prolyl 4-hydroxylases is a large family of genes encoding proteins that catalyze the conversion of proline to 4-hydroxyproline using as cofactors: iron, oxygen and ascorbate. The 4 residues are widespread in the plant and animal proteins and their functional significance involves root structure and development and cell wall organization. Preliminary experiments in our laboratory indicated increased levels of expression of P4H7 and P4H5 under 5% O2 in both roots and shoots of Arabidopsis thaliana. The purpose of the present study is the functional characterization of the p4h7d and p4h5a Arabidopsis thaliana T-DNA mutants using Salk T-DNA mutant lines. The presence of the T-DNA insertion was confirmed for both mutants by PCR screening and sequencing. PCR screening of the homozygous p4h7d progeny revealed that the mutation is transmitted in a homozygous manner. Studies of phenotypic differences were concentrated on growth parameters such as root and hypocotyl length. The p4hd plants had shorter roots and longer hvpocotyls in comparison to wild type plants and these differences were statistically significant. The particular phenotype was observed in 7, 10 and 13 day-old plants. The survival assay after anoxia revealed an accentuated bending of the p4h7d mutants compared to the wild type, probably due to the longer hypocotyl. The data obtained in this study indicate the possible involvement of the P4H7 and P4H5 genes in plant growth and further research is required to thoroughly investigate their physiological significance.
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