Molecular cloning and characterization of cDNAs for anionic and neutral peroxidases from suspensioncultured-cells of sweet potato and their differential expression in response to stress

Abstract

Two peroxidase (POD) cDNAs, swpa1 and swpn1, were isolated and characterized from suspension-cultured cells of sweet potato in order to understand the physiological function of POD isozymes. Sequence analysis showed that swpa1 encoded an anionic POD and swpn1 encoded a neutral POD. The swpa1 and swpn1 genes were both highly expressed in suspension-cultured cells in accordance with the high POD activity of these cells. Although both gene transcripts were detected in the stems of intact plants, their transcription levels were much lower than in suspension-cultured cells. During cell growth the pattern of mRNA accumulation of swpa1 differed from that of swpn1, suggesting that expression of these genes is differentially regulated by cell growth stage. In addition, the swpa1 and swpn1 genes responded differently to oxidative stress induced by chilling. The expression of swpa1 was weakly induced by 15° C acclimation and strongly induced by 4° C chilling, whereas the mRNA level of swpn1 was increased by 15° C acclimation and reduced by 4° C chilling. This indicates that the two isozymes encoded by swpa1 and swpn1 might contribute to protection against cold-induced oxidative stress through different signaling pathways. In leaves, both genes were induced by wounding with broadly similar expression patterns. Genomic analysis suggests that the two isozymes are encoded by different loci in the sweet potato genome.