Sequence characteristics of V-ATPase-d gene in Pinctada fucata martensii and its relationship with low temperature tolerance
V-ATPase plays an important role in biological response to various environmental stress,in order to explore the role of V-ATPase in the low temperature adaptation of Pinctada fucata martensii,the V-ATPase-d gene of P.fucata.martensii was identified,and the changes in the expression of Pm-V-ATPase-d under low temperature stress were analyzed.Screening and comparative analysis of the gene in the low temperature resistant breeding line(R)F3 and Beibu Gulf wild population(W)SNP loci in the exon region.Sequence analysis showed that Pm-V-ATPase-d has a total length of 1 473 bp and open reading frame(ORF)of 1 140 bp,encoding 379 amino acids,with a typical ATP domain,Pfam vATP-synt_AC39.The results of motif analysis and tertiary structure prediction showed that Pm-V-ATPase-d was highly conserved,and it clustered with Crassostrea gigas in the phylogenetic tree.Whole tissue fluorescence quantitative results showed that Pm-V-ATPase-d was expressed in all the tested tis-sues,and the expression level was higher in hepatopancreas,gonads and gill tissues(P＜0.05).Under low temperat-ure stress,the expression level of Pm-V-ATPase-d gene increased first and then decreased with the extension of time,and the expression level of the low temperature group was significantly higher than the control group(P＜0.05),indicating that Pm-V-ATPase-d may be involved in the response of P.fucata martensii to temper-ature stress.The SNP analysis of the exon region of Pm-V-ATPase-d resulted in a total of 35 SNPs,which 34 were synonymous mutations,only one site was a non-synonymous mutation,and 26 SNPs were significantly different between genotypes and alleles in W and R populations(P＜0.05),the results of haplotype linkage disequilibrium analysis showed that the Pm-V-ATPase-d gene SNPs could form 6 haplotype blocks and 14 haplotypes,the haplo-types GAAT,CGC,TC,TG and AG were significantly correlated with low temperature resistance traits of P.fucata martensii(P＜0.05).These research data indicate that Pm-V-ATPase-d may be a candidate gene involved in regulating the adaptation to low temperature of P.fucata martensii.This study can provide a research basis for the adaptation mechanism of P.fucata martensii to low temperature.The SNPs and haplotypes associated with low temperature resistance traits can be used in molecular assisted breeding.
Pinctada fucata martensiiV-ATPase-d genesingle-nucleotide polymorphism(SNP)low temperature
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