Identification and functional characterization of piwi1 gene in sea cucumber, Apostichopus japonicas

Sun, Zhihui; Wei, Jin-Liang; Cui, Zhouping; Han, Ya-Lun; Zhang, Jian; Jiang, Song; Chang, Yaqing

doi:10.1016/j.cbpb.2020.110536

Cited by 12 publications

(9 citation statements)

References 58 publications

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“…Up to date, few studies have been focused on sex-associated genes in A. japonicus [ 74 ]. To provide insights into the candidate sex-associated gene list, we found a set of genes were associated with sex differentiation or gonadal development.…”

Section: Discussionmentioning

confidence: 99%

Identification of sex determination locus in sea cucumber Apostichopus japonicus using genome-wide association study

Wang

Yang

et al. 2022

BMC Genomics

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Background Sex determination mechanisms are complicated and diverse across taxonomic categories. Sea cucumber Apostichopus japonicus is a benthic echinoderm, which is the closest group of invertebrates to chordate, and important economic and ecologically aquaculture species in China. A. japonicus is dioecious, and no phenotypic differences between males and females can be detected before sexual maturation. Identification of sex determination locus will broaden knowledge about sex-determination mechanism in echinoderms, which allows for the identification of sex-linked markers and increases the efficiency of sea cucumber breeding industry. Results Here, we integrated assembly of a novel chromosome-level genome and resequencing of female and male populations to investigate the sex determination mechanisms of A. japonicus. We built a chromosome-level genome assembly AJH1.0 using Hi-C technology. The assembly AJH1.0 consists of 23 chromosomes ranging from 22.4 to 60.4 Mb. To identify the sex-determination locus of A. japonicus, we conducted genome-wide association study (GWAS) and analyses of distribution characteristics of sex-specific SNPs and fixation index FST. The GWAS analysis showed that multiple sex-associated loci were located on several chromosomes, including chromosome 4 (24.8%), followed by chromosome 9 (10.7%), chromosome 17 (10.4%), and chromosome 18 (14.1%). Furthermore, analyzing the homozygous and heterozygous genotypes of plenty of sex-specific SNPs in females and males confirmed that A. japonicus might have a XX/XY sex determination system. As a physical region of 10 Mb on chromosome 4 included the highest number of sex-specific SNPs and higher FST values, this region was considered as the candidate sex determination region (SDR) in A. japonicus. Conclusions In the present study, we integrated genome-wide association study and analyses of sex-specific variations to investigate sex determination mechanisms. This will bring novel insights into gene regulation during primitive gonadogenesis and differentiation and identification of master sex determination gene in sea cucumber. In the sea cucumber industry, investigation of molecular mechanisms of sex determination will be helpful for artificial fertilization and precise breeding.

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Section: Discussionmentioning

confidence: 99%

Identification of sex determination locus in sea cucumber Apostichopus japonicus using genome-wide association study

Wang

Yang

et al. 2022

BMC Genomics

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“…The middle part of the intestine was used for gene expression analysis. According to previously reports ( Sun et al, 2021 ), the gonads of A. japonicus are classified into four stages, including the early growing stage (Stage 1), growing stage (Stage 2), mature stage (Stage 3) and post-spawning stage (Stage 4). After gonadal histology analysis, gonadal tissues at different development stages were collected to examine the dynamic expression changes in Dmrt1 and SoxE .…”

Section: Methodsmentioning

confidence: 99%

“…The efficiency of primers discovery through standard curve formulation reached 98%. The housekeeping gene NADH was selected as the internal reference gene ( Sun et al, 2021 ). Data were performed from three independent experiments.…”

Section: Methodsmentioning

confidence: 99%

“…Compared with the female sea cucumber, the male might have an advantage in terms of immunocompetence ( Jiang et al, 2017 ; Jiang et al, 2019 ). Recently, several genes related to gonadal development have been identified, such as piwi ( Sun et al, 2021 ), vasa ( Yan M. et al, 2013 ) and foxl2 ( Sun et al, 2022 ), and a hypothetical XX/XY sex determination system has been proposed ( Wei et al, 2021 ). However, the mechanisms underlying sex determination and sex differentiation in the sea cucumber remain a mystery.…”

Section: Introductionmentioning

confidence: 99%

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Identification and functional analysis of Dmrt1 gene and the SoxE gene in the sexual development of sea cucumber, Apostichopus japonicus

Liu

Cong²,

Su³

et al. 2023

Front. Genet.

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Members of the Doublesex and Mab-3-related transcription factor (Dmrt) gene family handle various vital functions in several biological processes, including sex determination/differentiation and gonad development. Dmrt1 and Sox9 (SoxE in invertebrates) exhibit a very conserved interaction function during testis formation in vertebrates. However, the dynamic expression pattern and functional roles of the Dmrt gene family and SoxE have not yet been identified in any echinoderm species. Herein, five members of the Dmrt gene family (Dmrt1, 2, 3a, 3b and 5) and the ancestor SoxE gene were identified from the genome of Apostichopus japonicus. Expression studies of Dmrt family genes and SoxE in different tissues of adult males and females revealed different expression patterns of each gene. Transcription of Dmrt2, Dmrt3a and Dmrt3b was higher expressed in the tube feet and coelomocytes instead of in gonadal tissues. The expression of Dmrt1 was found to be sustained throughout spermatogenesis. Knocking-down of Dmrt1 by means of RNA interference (RNAi) led to the downregulation of SoxE and upregulation of the ovarian regulator foxl2 in the testes. This indicates that Dmrt1 may be a positive regulator of SoxE and may play a role in the development of the testes in the sea cucumber. The expression level of SoxE was higher in the ovaries than in the testes, and knocking down of SoxE by RNAi reduced SoxE and Dmrt1 expression but conversely increased the expression of foxl2 in the testes. In summary, this study indicates that Dmrt1 and SoxE are indispensable for testicular differentiation, and SoxE might play a functional role during ovary differentiation in the sea cucumber.

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“…Meanwhile, the first-strand cDNAs of each sample were synthesized according to the protocols of RevertAid First Strand cDNA Synthesis Kit (Fermentas). Real-time quantitative PCR (RT-qPCR) was performed on the LightCycler ® 96 Instrument (Roche) according to a previous description ( 41 ), and 18S ribosomal RNA ( 18S rRNA ) was used as the internal control. The relative expression of the target genes was calculated with the 2 −ΔΔCt method.…”

Section: Methodsmentioning

confidence: 99%

Application of SNP in Genetic Sex Identification and Effect of Estradiol on Gene Expression of Sex-Related Genes in Strongylocentrotus intermedius

et al. 2021

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Sea urchin (Strongylocentrotus intermedius) is an economically important mariculture species in Asia, and its gonads are the only edible part. The efficiency of genetic breeding in sea urchins is hampered due to the inability to distinguish gender by appearance. In this study, we first identified a sex-associated single nucleotide polymorphism (SNP) by combining type IIB endonuclease restriction site-associated DNA sequencing (2b-RAD-seq) and genome survey. Importantly, this SNP is located within spata4, a gene specifically expressed in male. Knocking down of spata4 by RNA interference (RNAi) in male individuals led to the downregulation of other conserved testis differentiation-related genes and germ cell marker genes. We also revealed that sex ratio in this validated culture population of S. intermedius is not 1:1. Moreover, after a 58-day feeding experiment with estradiol, the expression levels of several conserved genes that are related to testis differentiation, ovary differentiation, and estrogen metabolism were dynamically changed. Taken together, our results will contribute toward improving breeding efficiency, developing sex-controlled breeding, and providing a solid base for understanding sex determination mechanisms in sea urchins.

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Identification and functional characterization of piwi1 gene in sea cucumber, Apostichopus japonicas

Cited by 12 publications

References 58 publications

Identification of sex determination locus in sea cucumber Apostichopus japonicus using genome-wide association study

Identification of sex determination locus in sea cucumber Apostichopus japonicus using genome-wide association study

Identification and functional analysis of Dmrt1 gene and the SoxE gene in the sexual development of sea cucumber, Apostichopus japonicus

Application of SNP in Genetic Sex Identification and Effect of Estradiol on Gene Expression of Sex-Related Genes in Strongylocentrotus intermedius

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