AUTHOR=Zhang Zhidong , Sun Xuefeng , Chen Aihua , Yang Jiaxin , Chen Suhua , Zhang Yu , Cao Yi , Peng Yuheng , Zhu Yanqing , Wu Yangping 

TITLE=Molecular bioinformatic and functional analysis of Enolase4 gene during siphonal autotomy and regeneration of razor clam Solen grandis

JOURNAL=Frontiers in Marine Science

VOLUME=10

YEAR=2023

URL=https://www.frontiersin.org/journals/marine-science/articles/10.3389/fmars.2023.1195253

DOI=10.3389/fmars.2023.1195253

ISSN=2296-7745

ABSTRACT=<p>For the razor clam <italic>Solen grandis</italic>, siphonal autotomy and regeneration is a necessary means of facing external stress, which is accompanied by a large amount of energy consumption. To explore the function of the <italic>ENO4</italic> gene in the siphonal autotomy and regeneration of <italic>S. grandis</italic>, the full-length cDNA of the <italic>ENO4</italic> in <italic>S. grandis</italic> was cloned and characterized. The full-length cDNA of <italic>SgENO4</italic> was 2,773 base pairs (bp), with an 1,851 bp coding sequence (CDS) encoding 616 amino acids. ​Bioinformatic analysis revealed a slightly lower codon usage bias and a relatively stable nucleotide composition in the <italic>SgENO4</italic> sequence. Three conserved motifs were found from the SgENO4 sequence. These conserved motifs vary in length from 14 to 27 amino acids. Moreover, in the siphonal pre-autotomy of razor clams, expression level of the <italic>SgENO4</italic> gene was significantly higher in the hepatopancreas and siphonal base than other tissues (<italic>P</italic> &lt; 0.05). However, the expression of the <italic>SgENO4</italic> gene in the siphonal base decreased significantly in the siphonal post-autotomy 7 hours and gradually recovered as the regeneration process continued. ​Linear correlation analysis showed that there was a significant linear correlation between specific weight of siphon and relative expression levels of <italic>SgENO4</italic> (R<sup>2 </sup>= 0.826). RNA interference (RNAi) result showed that the specific weight (the proportion of siphonal weight to body weight) is significantly lower than other groups on the seventh day (<italic>P</italic> &lt; 0.05), suggesting that interfering with the expression of the <italic>SgENO4 gene</italic> could inhibit the siphonal regeneration of <italic>S. grandis</italic>. These results further demonstrate that <italic>SgENO4</italic> is a conserved sequence and that it plays a crucial role in the siphonal autotomy and regeneration of <italic>S. grandis</italic>, advancing further understanding in exploring molecular mechanisms in the siphonal autotomy and regeneration.</p>