Isolation of pollutant (pine needle ash)-responding genes from the seaweed Porphyra yezoensis tissue

Abstract

Genetic responses of the aquaculturable seaweed Porphyra yezoensis to pine needle ash have been compared using differential display technique. The tissue viability was assessed to evaluate the stress level with triphenyltetrazolium chloride. Total RNA, from tissues challenged in seawater containing ash, was reverse transcribed and amplified by PCR with arbitrary primers. The genetic fragments responded by the stress were selectively isolated from agarose gel and sequenced with DNA auto sequencer. According to sequence analysis by the NCBI BLASTX program, ash-suppressed genes (1065 bp, 347 bp) were putative glutaredoxin-like protein and hypothetical protein. Pine needle ashinducible genes (939 bp, 556 bp) were recognized as hypothetical protein and phosphoenolpyruvate-protein phosphotransferase, respectively.