Sequence characteristics, SNP screening and association with growth traits of TTLL gene in pearl oyster Pinctada fucata martensii
YIN Shixin, LI Zhiyu, YANG Jingmiao, LIAO Yongshan*, YANG Chuangye, DENG Yuewen, WANG Qingheng
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1.College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China; 2.Pearl Research Institute,Guangdong Ocean University, Zhanjiang 524088, China; 3.Guangdong Technology Research Center for Pearl Aquaculture and Process, Zhanjiang 524088, China
In order to explore the function of tubulin tyrosine ligase-like (TTLL) in pearl oyster Pinctadafucatamartensii, the sequence characteristics, expression patterns and SNP screening of the TTLL gene were analyzed in the pearl oyster. The results showed that the full-length sequence of TTLL cDNA of the pearl oyster cloned by RACE technology was 1 962 bp, including 1 611 bp of open reading frame (ORF) encoding 536 amino acids, 134 bp of 5′UTR and 191 bp of 3′UTR. It is predicted that there was a TTL domain in the N-terminal of TTLL protein sequence in the pearl oyster. Multi-sequence alignments and the construction of phylogenetic tree analysis of its amino acid sequences showed that the TTLL had high homology with TTLL of other species, and was clustered with other bivalves like Pacific oyster Crassostreagigas. qRT-PCR results revealed that TTLL was ubiquitously expressed in mantle, adductor muscle, gonads, hepatopancreas, gills and feet of the pearl oyster, with the maximal expression level in gonads (P<0.05). The TTLL was highly expressed in large-scale populations and hybrid families with better growth traits (P<0.05). There was significantly higher expression level of TTLL gene in eggs and fertilized eggs than that in other developmental stages (P<0.05). SNPs analysis of the TTLL gene revealed that 277 SNP were obtained, in which 109 SNP sites were in accordance with Hardy-Weinberg equilibrium, 6 SNP sites were moderately polymorphic and 103 SNP sites low-level polymorphisms. Haplotype analysis of TTLL gene of the pearl oyster exon region SNPs showed that 4 SNP sites formed 2 haploblocks and 5 haplotypes, in which the haplotype GG was an excellent haplotype, with significantly better growth characteristics than that in the haplotype AA. Analysis of TTLL gene mutation sites revealed that 7 SNP sites were mutated, 4 missense mutations and 3 synonymous mutations, in which locus g.7572350 T>G and locus g.7572368 A>G were significantly correlated with shell width (P<0.05), g.7572405 C>T was significantly correlated with shell length and shell height (P<0.05), without significant correlation with growth traits in other sites (P>0.05). The findings indicated that TTLL gene was involved in the growth and development of the pearl oyster, and its excellent haplotype GG and mutation sites g.7572350 T>G, g.7572368 A>G and g.7572405 C>T were all conducive to the growth of the pearl oyster, which are of great significance to the breeding of the pearl oyster with excellent economic traits.
YIN Shixin, LI Zhiyu, YANG Jingmiao, LIAO Yongshan, YANG Chuangye, DENG Yuewen, WANG Qingheng.
Sequence characteristics, SNP screening and association with growth traits of TTLL gene in pearl oyster Pinctada fucata martensii[J]. Journal of Dalian Fisheries University, 2022, 37(6): 949-959 https://doi.org/10.16535/j.cnki.dlhyxb.2021-321
中图分类号：
S 917.4
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