马氏珠母贝Kelch-like基因的序列及其SNP位点分析

doi:10.16535/j.cnki.dlhyxb.2021-146

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大连海洋大学学报 ›› 2022, Vol. 37 ›› Issue (3) : 386-393. DOI: 10.16535/j.cnki.dlhyxb.2021-146

马氏珠母贝Kelch-like基因的序列及其SNP位点分析

杨创业，张丞澍，黄静，郝瑞娟，师尚丽，黄荣莲，周芮，邱金玉，颜翼龙，邓岳文

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Sequence and SNP analysis of Kelch-like gene from pearl oyster Pinctada fucata martensii

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摘要

为了探究马氏珠母贝Pinctada fucata martensii Kelch样基因(Kelch-like)的功能，对Kelch-like基因序列及其SNP位点进行了分析。结果表明：马氏珠母贝Kelch-like序列全长为2 127 bp，其中，5′UTR为58 bp，3′UTR为44 bp，其开放式阅读框长度为2 025 bp，可编码674个氨基酸；预测其Kelch-like氨基酸序列含有1个BTB结构域、1个BACK结构域和2个Kelch结构域；马氏珠母贝Kelch-like与美洲牡蛎Kelch-like氨基酸序列的一致性最高(35%)，其次为虾夷扇贝(32.05%)；马氏珠母贝Kelch-like氨基酸序列的BTB、BACK保守性较高，且Kelch具有保守的GG位点；Kelch-like基因在马氏珠母贝边缘膜组织的相对表达量显著高于其他组织(P<0.05)，且贝壳损伤后6、12、120 h时该组织中的基因表达量显著上调(P<0.05)；马氏珠母贝Kelch-like基因共有56个SNP位点，其中，31个SNP位点为中度多态，其余位点为低度多态；10个SNP位点形成了1个单倍块，为4种单倍型，其中单倍型AATCTGTCGG为优异单倍型，其壳质量性状显著大于单倍型AGCCTACCGA和单倍型AGCCTACTTA(P<0.05)。研究表明，马氏珠母贝Kelch-like可能参与贝壳棱柱层的形成，其单倍型AATCTGTCGG具有作为以壳质量为目标的育种标记的潜力，该结果对于选育具有优良经济性状的马氏珠母贝具有重要意义。

Abstract

In order to explore the function of Kelch-like gene in pearl oyster Pinctada fucata martensii,the full-length sequence and SNPs of Kelch-like were analyzed in the present study.It was found that the full-length sequence of Kelch-like was 2 127 bp containing a 5′UTR of 58 bp and a 3′UTR of 44 bp.The open reading frame had length of 2 025 bp encoding 674 amino acids.The Kelch-like sequence contained a BTB domain,a BACK domain and two Kelch domains.The pearl oyster Kelch-like had high homology with Kelch-like of other species in amino acid sequence.Multi-sequence alignment indicated that the BTB and BACK domains were highly conservative,and Kelch domain held a conservative GG site.Kelch-like gene showed significantly higher expression in the marginal zone of mantle (P<0.05),especially high expression (up-regulated) level in the 6,12 h and 120 h in the repair process of the pearl oyster with damaged mantles (P<0.05).SNPs analysis of Kelch-like exon region revealed that 31 SNP sites in 56 SNP obtained were moderately polymorphic and the rest were low-level polymorphisms.Among the 56 SNPs,10 SNP loci formed a haplotype set containing,4 haplotypes,of which the haplotype AATCTGTCGG was an excellent haplotype,with significantly greater shell weight than the haplotype AGCCTACCGA and haplotype AGCCTACTTA (P<0.05).These findings showed that Kelch-like may participated in the shell prismatic layer and haplotype AATCTGTCGG hold the potential as the molecular maker of shell weight in the breeding process,which are of great significance to the breeding of pearl oyster with excellent economic traits.

导出引用

杨创业, 张丞澍, 黄静, 郝瑞娟, 师尚丽, 黄荣莲, 周芮, 邱金玉, 颜翼龙, 邓岳文. 马氏珠母贝Kelch-like基因的序列及其SNP位点分析[J]. 大连海洋大学学报, 2022, 37(3): 386-393 https://doi.org/10.16535/j.cnki.dlhyxb.2021-146

YANG Chuangye, ZHANG Chengshu, HUANG Jing, HAO Ruijuan, SHI Shangli, HUANG Ronglian, ZHOU Rui, QIU Jinyu, YAN Yilong, DENG Yuewen. Sequence and SNP analysis of Kelch-like gene from pearl oyster Pinctada fucata martensii[J]. Journal of Dalian Fisheries University, 2022, 37(3): 386-393 https://doi.org/10.16535/j.cnki.dlhyxb.2021-146