Abstract:Phospholipase C (Phospholipase C, PLC) is a class of enzymes related to membrane proteins, which play an important role in signal transduction pathways. Onchidium reevesii that inhabits the intertidal zone can perceive the local tidal laws. In order to explore the role and molecular mechanism of phospholipase C in O. reevesii in tide sensing, in this experiment, we used the ganglion of O. reevesii as the experimental material. The cDNA sequence of PLC gene was cloned by RACE-PCR technology, and bioinformatics analysis and qRT-PCR experiment were performed. The results showed that the full length of the cDNA of the PLC gene consists of 2 578 base pairs, the open reading frame(ORF) is 1 881 bp, the 5'non-coding (UTR) region is 337 bp, the 3'non-coding region (UTR) is 340 bp, and there is an AATAAA tailing signal at the end, which encodes a total 627 amino acids. The result of amino acid sequence comparison showed that the PLC gene of O. reevesii has the highest match with the phospholipase C-like protein 2 amino acid sequence of Pomacea canaliculata, with a similarity of 88%, followed by the phospholipase C-like protein 2 amino acid sequence of Crassostrea virginica, and it is consistent with the classification of traditional morphology. Using the low frequency sound in the tide in the laboratory to stimulate the O. reevesii, the expression of PLC gene at different frequencies was detected by fluorescence quantitative PCR. The results showed that the expression of PLC was basically stable when the stimulation sound wave frequency was 40 Hz-70 Hz. When the frequency was increased to 160 Hz-310 Hz, it was in a state of low expression. When the sound wave frequency reached 220 Hz and 280 Hz, it was in a state of high expression, indicating that this gene plays an important role in the low-frequency perception of O. reevesii. This experiment lays a theoretical support for further understanding of the tidal sensing function of the O. reevesii.