Analysis of genetic diversity in two different shell colors of the giant triton snail (Charonia tritonis) based on mitochondrial COI sequences

The giant triton snail (Charonia tritonis) is widely distributed in tropical coral reefs in the Indo-West Pacific. Its distribution areas in China include the Penghu Islands, Hengchun Peninsula, and Xisha Islands. In this study, we use Multivariate Analysis of Variance (MANOVA) to compare the live weight and shell length between different shell colors of C. tritonis. In addition, we conducted sequence analysis of the mitochondrial cytochrome oxidase (COI) gene to assess the affinity of C. tritonis with two different shell colors based on the sample we obtained. Then, we constructed phylogenetic trees using the maximum likelihood (ML) and Bayesian Inference methods, and constructed haplotype network diagrams. In addition, we performed Tajima’s D and Fu’s neutrality tests. The results show that The Partial mitochondrial COI sequences of 28 C. tritonis were all 603 base pairs in length, and seven haplotypes were detected from the samples, besides, the gene flow was calculated to be 11.78, the genetic differentiation coefficient was 0.02078. Our results indicated that the population size of C. tritonis remained relatively stable. Besides, the genetic and size differentiation between the two different shell colors was small, and individuals of C. tritonis with different shell colors belong to the same genetic clade. In fact, the two morphotypes could not be distinguished by both genetic and morphometric data. The mitochondrial COI gene fragments of the two different shell colors were sequenced and analyzed to accumulate information about the population genetics of C. tritonis and to provide a scientific basis for the conservation of its species resources.The giant triton snail (Charonia tritonis) is widely distributed in tropical coral reefs in the Indo-West Pacific. Its distribution areas in China include the Penghu Islands, Hengchun Peninsula, and Xisha Islands. In this study, we use Multivariate Analysis of Variance (MANOVA) to compare the live weight and shell length between different shell colors of C. tritonis. In addition, we conducted sequence analysis of the mitochondrial cytochrome oxidase (COI) gene to assess the affinity of C. tritonis with two different shell colors based on the sample we obtained. Then, we constructed phylogenetic trees using the maximum likelihood (ML) and Bayesian Inference methods, and constructed haplotype network diagrams. In addition, we performed Tajima’s D and Fu’s neutrality tests. The results show that The Partial mitochondrial COI sequences of 28 C. tritonis were all 603 base pairs in length, and seven haplotypes were detected from the samples, besides, the gene flow was calculated to be 11.78, the genetic differentiation coefficient was 0.02078. Our results indicated that the population size of C. tritonis remained relatively stable. Besides, the genetic and size differentiation between the two different shell colors was small, and individuals of C. tritonis with different shell colors belong to the same genetic clade. In fact, the two morphotypes could not be distinguished by both genetic and morphometric data. The mitochondrial COI gene fragments of the two different shell colors were sequenced and analyzed to accumulate information about the population genetics of C. tritonis and to provide a scientific basis for the conservation of its species resources

Giant Triton Snail Charonia tritonis Macrophage-Expressed Gene 1 Protein Ct-Mpeg1: Molecular Identification, Expression Analysis, and Antimicrobial Activity

Macrophage-expressed gene 1 proteins (Mpeg1/Perforin-2 (PRF2)) are a family of pore-forming proteins (PFPs) which can form pores and destroy the cell membrane of invading pathogens. However, little information is available regarding the function of Mpeg1 in the giant triton snail Charonia tritonis. In this study, a homolog of Mpeg1 (Ct-Mpeg1) was identified in C. tritonis. The predicted protein of Ct-Mpeg1 contains several structural features known in Mpegs, including a membrane attack complex/perforin (MACPF) domain and single transmembrane region. The Ct-Mpeg1 gene was constitutively expressed in almost all tissues examined except in the proboscis, with the highest expression level observed in the mantle. As a typical pore-forming protein, Ct-Mpeg1 has antibacterial activities against Vibrio (including Vibrio alginolyticus and Vibrio parahaemolyticus). In addition, rCt-Mpeg1 challenge to V. alginolyticus represses the expression of most outer membrane protein synthesis-related genes and genes involved in the TCA cycle pathway, which will lead to reduced outer membrane protein synthesis and less energy capacity. This is the first report to characterize the macrophage-expressed gene 1 protein in C. tritonis, and these results suggest that macrophage-expressed gene 1 protein Ct-Mpeg1 is an important immune molecule of C. tritonis that is involved in the bacterial infection resistance of Vibrio, and this study may provide crucial basic data for the understanding of the innate immunity system of C. tritonis

Molecular and Functional Characterization of a Short-Type Peptidoglycan Recognition Protein, Ct-PGRP-S1 in the Giant Triton Snail Charonia tritonis

Peptidoglycan recognition proteins (PGRPs) are a family of pattern recognition receptors (PRRs) involved in host antibacterial responses, and their functions have been characterized in most invertebrate and vertebrate animals. However, little information is available regarding the potential function of PGRPs in the giant triton snail Charonia tritonis. In this study, a short-type PGRP gene (termed Ct-PGRP-S1) was identified in C. tritonis. Ct-PGRP-S1 was predicted to contain several structural features known in PGRPs, including a typical PGRP domain (Amidase_2) and Src homology-3 (SH3) domain. The Ct-PGRP-S1 gene was constitutively expressed in all tissues examined except in proboscis, with the highest expression level observed in the liver. As a typical PRR, Ct-PGRP-S1 has an ability to degrade peptidoglycan (PGN) and was proven to have non-Zn2+-dependent amidase activity and antibacterial activity against Vibrioalginolyticus and Staphylococcus aureus. It is the first report to reveal the peptidoglycan recognition protein in C. tritonis, and these results suggest that peptidoglycan recognition protein Ct-PGRP-S1 is an important effector of C. tritonis that modulates bacterial infection resistance of V. alginolyticus and S. aureus, and this study may provide crucial basic data for the understanding of an innate immunity system of C. tritonis

Molecular and Functional Characterization of a Novel Kunitz-Type Toxin-like Peptide in the Giant Triton Snail Charonia tritonis

It has been reported that the giant triton snail (Charonia tritonis) inserts its large proboscis and then injects venom or acid saliva from its salivary gland into its prey, the crown-of-thorns starfish Acanthaster planci (COTS), paralyzing it. A full-length cDNA sequence of the C. tritonis Ct-kunitzin gene was obtained by RACE PCR based on a transcriptomic database constructed by our laboratory (data not published), which contains an open reading frame (ORF) sequence with a length of 384 bp including a 1–32aa Kunitz domain. The Ct-kunitzin peptide was synthesized by solid-phase polypeptide methods according to its conserved amino acid sequence, with a molecular weight of 3746.0 as well as two disulfide bonds. Renatured Ct-kunitzin was injected into mice ventricles to evaluate its potential function. Compared with the normal control group (physiological saline), the spontaneous locomotor activity of the Ct-kunitzin group decreased significantly. There was a significant effect on Ct-kunitzin on mice grip strength in the grip strength test. In addition, Ct-kunitzin exhibited remarkable biological activity in suppressing pain in the pain thresholds test. There were no significant differences between the Ct-kunitzin group and the normal control group in terms of various hematological indexes and histopathological observations. When tested in COTS, the most significant histological change was the destruction, disorganization, and significant reduction in the amount of COTS tube feet tissues. Altogether, the potential paralyzing effect on mice suggests that Ct-kunitzin is a possible agent for novel drug development