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Validation of a molecular sex marker in three sturgeons from eastern North America 北美东部三种鲟鱼分子性别标记的验证
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2024-01-27 DOI: 10.1007/s12686-024-01346-6

Abstract

Despite the importance of sex-specific information for sturgeon conservation and management, sex identification has been a major challenge outside of mature adults on spawning grounds. Recent work identified a sex-specific locus (AllWSex2) that appears to be broadly conserved across many Acipenserids, but the assay was not validated for all species within the family. We tested the AllWSex2 marker in three sturgeon taxa (shortnose sturgeon Acipenser brevirostrum, Gulf sturgeon A. oxyrhinchus desotoi, and Atlantic sturgeon A. oxyrhinchus oxyrhinchus) from the Atlantic and Gulf of Mexico Coasts of North America to validate its use for sex identification. Our results indicate AllWSex2 is conserved in all three taxa, presenting a new opportunity to derive sex-specific information from tissue samples, which are routinely collected from these taxa. We found high concordance (range: 97–100%) between genotypic and phenotypic/histological methods, suggesting the assay is broadly effective. However, the small amount of discordance between the methods (< 3%) suggests further refinement may be possible.

摘要 尽管性别特异性信息对于鲟鱼的保护和管理非常重要,但在产卵场成熟成鱼之外的性别鉴定一直是一个重大挑战。最近的研究发现了一个性别特异性位点(AllWSex2),该位点似乎在许多鲟科鱼类中具有广泛的保守性,但该检测方法并未在鲟科的所有物种中得到验证。我们在北美大西洋和墨西哥湾沿岸的三个鲟鱼类群(短吻鲟Acipenser brevirostrum、海湾鲟A. oxyrhinchus desotoi和大西洋鲟A. oxyrhinchus oxyrhinchus)中测试了AllWSex2标记,以验证其在性别鉴定中的应用。我们的研究结果表明,AllWSex2 在所有这三个类群中都是保守的,这为从这些类群常规采集的组织样本中获取性别特异性信息提供了一个新的机会。我们发现基因型和表型/组织学方法之间的一致性很高(范围:97-100%),这表明该检测方法具有广泛的有效性。然而,两种方法之间存在的少量不一致(3%)表明有可能进一步改进。
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引用次数: 0
Evolutionary stamps for adaptation traced in Cervus nippon genome using reduced representation sequencing 利用还原表示测序技术追踪海马基因组中的适应进化印记
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2024-01-27 DOI: 10.1007/s12686-023-01339-x
Nidhi Sukhija, Rangasai Chandra Goli, Mukul Sukhija, Pallavi Rathi, Komal Jaglan, Aishwarya Dash, P. B. Nandhini, K. K. Kanaka

Sika deer is indigenous to East Asia. Currently the species is extinct or are nearing the verge of extinction, in numerous places. Reduced population size is sensitive to climate change and has poor adaptation as a result of inbreeding. In this study genome-wide Single nucleotide polymorphisms (SNPs) were identified using double-digest restriction site associated digestion (ddRAD) approach and annotated further to identify genes related to fitness. A total 85,001 SNPs were retained following the quality control. Composite Likelihood Ratio (CLR) statistics found 13 stronger selective sweeps, identified sweeps overlapped with genes related to adaptation, reproduction, cellular functions. Average heterozygosity and nucleotide diversity were 0.27 and 0.29 respectively. Admixture analysis furnished a single cluster indicating no further sub-divergence of Sika deer. These findings would help fabricate appropriate genetic improvement programs by inclusion in the SNP chips, in order to get deeper insights into deer genetics, evolution, domestication and conservation.

Graphical Abstract

Bioinformatic analysis for unravelling genomic variants and selective sweep regions in Cervus nippon

梅花鹿原产于东亚。目前,该物种在许多地方已经灭绝或濒临灭绝。梅花鹿种群数量的减少对气候变化非常敏感,而且由于近亲繁殖,其适应性很差。在这项研究中,使用双位限制性位点相关消化(ddRAD)方法鉴定了全基因组单核苷酸多态性(SNPs),并进一步注释,以确定与适应性相关的基因。经过质量控制后,共保留了 85 001 个 SNPs。综合似然比(CLR)统计发现了 13 个较强的选择性横扫,所确定的横扫与适应、繁殖和细胞功能相关基因重叠。平均杂合度和核苷酸多样性分别为 0.27 和 0.29。混杂分析得出的单一聚类表明梅花鹿没有进一步的亚分化。这些发现将有助于通过加入 SNP 芯片制定适当的遗传改良计划,从而更深入地了解鹿的遗传学、进化、驯化和保护。 图文摘要生物信息学分析揭示梅花鹿基因组变异和选择性扫描区域
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引用次数: 0
Genome-wide identification of simple sequence repeats and development of polymorphic SSR markers in brown trout (Salmo trutta) 褐鳟(Salmo trutta)简单序列重复序列的全基因组鉴定和多态性 SSR 标记的开发
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-29 DOI: 10.1007/s12686-023-01341-3
Haibing Ma, Tianqing Huang, Wei Gu, Enhui Liu, Gaochao Wang, Bingqian Wang, Jianshe Zhou, Gefeng Xu

In China, the brown trout is a second-level key aquatic wildlife protection animal in the Tibet Autonomous Region. In order to protect the brown trout germplasm resources in the Tibet Autonomous Region, the Institute of Aquatic Sciences of the Academy of Agricultural and Animal Husbandry Sciences of the Tibet Autonomous Region and the Heilongjiang Institute of the Chinese Academy of Aquatic Sciences collaborated on research on the key technologies of brown trout family construction. A fast in silico simulation method was employed in this study to discover numerous polymorphic SSRs. A total of 1,115,563 SSR loci with two to six nucleotide repeat motifs were mined from 40 chromosomes of the brown trout reference genome by GMATA. The whole genome sequences (30×) of 30 brown trout samples were compared to the reference genome’s chromosome sequence. Then, the HipSTR program was employed to carry out in silico genotyping through comparisons and enumerations of repeat count variants on the motifs of the SSR loci among the samples. The loci with monomorphism and high deletion in the SSR genotype data were removed, and the loci with more than one non-reference allele were retained. The final set of polymorphic SSR loci screened was 106,194. Forty-three SSR markers were randomly selected for characteristic analysis. The range of alleles per locus was 3 to 9 (mean = 5). Observed heterozygosity was in the ranged from 0.167 to 1.000. Expected heterozygosity was in the ranged from 0.155 to 0.823. Twenty-three loci significantly deviated from Hardy-Weinberg equilibrium. These SSR markers will provide valuable information for assessing the genetic diversity of constructed brown trout families and for the genetic improvement of brown trout.

在中国,褐鳟是西藏自治区二级重点保护水生野生动物。为保护西藏自治区褐鳟种质资源,西藏自治区农牧科学院水产研究所与中国水产科学研究院黑龙江研究所合作开展了褐鳟家系构建关键技术研究。该研究采用快速硅模拟方法发现了大量多态性 SSR。利用GMATA从褐鳟鱼参考基因组的40条染色体中挖掘出具有2至6个核苷酸重复基序的共1,115,563个SSR位点。将 30 个褐鳟鱼样本的全基因组序列(30×)与参考基因组的染色体序列进行比较。然后,利用 HipSTR 程序,通过比较和枚举样本中 SSR 位点图案上的重复次数变异,进行硅基因分型。删除 SSR 基因型数据中单态性和高缺失的位点,保留有一个以上非参考等位基因的位点。最终筛选出 106 194 个多态 SSR 位点。随机选择了 43 个 SSR 标记进行特征分析。每个位点的等位基因数范围为 3 至 9(平均值 = 5)。观察到的杂合度在 0.167 至 1.000 之间。预期杂合度在 0.155 至 0.823 之间。23 个位点明显偏离哈代-温伯格平衡。这些 SSR 标记将为评估构建的褐鳟家族的遗传多样性和褐鳟的遗传改良提供有价值的信息。
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引用次数: 0
Isolation and characterization of 55 SNP markers in Triplophysa (Hedinichthys) yarkandensis macroptera 叶尔羌三疣梭子蟹 55 个 SNP 标记的分离与鉴定
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-27 DOI: 10.1007/s12686-023-01345-z

Abstract

Triplophysa (Hedinichthys) yarkandensis macroptera is an endemic species in China, that is mainly distributed in Gansu Province. To date, research on this species has been very limited, focusing mainly on its taxonomy, without any available molecular markers. In the present study, we identified 55 SNP markers in T. (Hedinichthys) yarkandensis macroptera. The minor allele frequency ranged from 0.050 to 0.500, and the observed and expected heterozygosity ranged from 0.167 to 1.000 and 0.155 to 0.508, respectively. The polymorphic information content ranged from 0.141 to 0.375. Among these SNP markers, seventeen loci showed significant departures from Hardy-Weinberg equilibrium after Bonferroni correction. These novel SNP markers will be helpful for future studies on genetic management and population conservation in this species.

摘要 叶尔羌鲑(Triplophysa (Hedinichthys) yarkandensis macroptera)是中国特有物种,主要分布于甘肃省。迄今为止,对该物种的研究非常有限,主要集中在其分类学方面,没有任何可用的分子标记。在本研究中,我们鉴定了55个SNP标记。小等位基因频率范围为 0.050 至 0.500,观察杂合度和预期杂合度范围分别为 0.167 至 1.000 和 0.155 至 0.508。多态性信息含量为 0.141 至 0.375。经 Bonferroni 校正后,这些 SNP 标记中有 17 个位点明显偏离 Hardy-Weinberg 平衡。这些新的SNP标记将有助于该物种未来的遗传管理和种群保护研究。
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引用次数: 0
Development of a multiplex real time-PCR assay to estimate the distribution of the genus Silurus in Lake Biwa using environmental DNA 利用环境 DNA 开发一种多重 real-time-PCR 检测方法,以估算琵琶湖中蚕蛾属的分布情况
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-19 DOI: 10.1007/s12686-023-01340-4
Tetsuro Inamoto, Yuu Kishimoto, Hisashi Okuyama, Ryoichi Tabata, Koichiro Nakano, Jun-ichi Takahashi

Three species of catfish, Silurus biwaensis, Silurus lithophilus, and Silurus asotus, are identified in Lake Biwa. Notably, S. biwaensis and S. lithophilus are endemic species in Lake Biwa; however, their detailed distribution and ecology almost remain unknown. In this study, we developed catfish species-specific primer probe sets based on their whole genome. Using these, we analyzed the water samples collected from three tanks where S. biwaensis, S. lithophilus, and S. asotus were experimentally reared, respectively, and established a new detection system after confirming each primer probe set to be effective. Finally, we conducted environmental DNA analysis for catfishes in Lake Biwa using field samples. Overall, our results showed that environmental DNA analysis is a useful monitoring method for confirming the presence of endemic catfishes in Lake Biwa.

在琵琶湖中发现了三种鲶鱼:琵琶鲶(Silurus biwaensis)、石斑鲶(Silurus lithophilus)和鲇鱼(Silurus asotus)。值得注意的是,S. biwaensis 和 S. lithophilus 是琵琶湖的特有物种,但它们的详细分布和生态几乎仍然未知。在本研究中,我们根据鲶鱼的全基因组开发了鲶鱼物种特异性引物探针组。利用这些探针组,我们分析了分别从实验饲养 S. biwaensis、S. lithophilus 和 S. asotus 的三个水箱中采集的水样,并在确认每个引物探针组有效后建立了新的检测系统。最后,我们利用野外样本对琵琶湖中的鲶鱼进行了环境 DNA 分析。总之,我们的结果表明,环境 DNA 分析是确认琵琶湖特有鲶鱼是否存在的有效监测方法。
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引用次数: 0
Development and validation of 98 SNP markers in Japanese anchovy, Engraulis japonicus 日本鳀鱼 98 个 SNP 标记的开发与验证
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-19 DOI: 10.1007/s12686-023-01342-2
Chenghui Song, Yang Liu, Congcong Wang

The Japanese anchovy, Engraulis japonicus, is an important economic fish that is distributed in the northwest Pacific Ocean. The effective assessment and management of the Engraulis japonicus fishery requires reliable information regarding its population’s genetic structure. The recent development of double digest restriction-site associated DNA sequencing (ddRAD-Seq) methods may contribute to the discovery of SNPs and the assessment of genetic structure. In our study, 98 single nucleotide polymorphism (SNP) markers were developed using ddRAD-Seq. The observed heterozygosity (Ho) and expected heterozygosity (He) ranged from 0.3333 to 0.8000 and 0.2778 to 0.5000, respectively. The polymorphism information content (PIC) ranged from 0.239198 to 0.375. All loci have been substantiated to follow the Hardy–Weinberg equilibrium. These novel polymorphic SNP markers will play an important role in the genetic research of E.japonicus, which will be beneficial to the development and utilization of E.japonicus resources. This study aims to provide technical support for the research of genetic diversity of E. japonicus populations through the development of SNP markers.

日本鳀鱼(Engraulis japonicus)是一种重要的经济鱼类,分布于西北太平洋。对日本鳀鱼渔业的有效评估和管理需要有关其种群遗传结构的可靠信息。最近发展起来的双消化限制位点相关 DNA 测序(ddRAD-Seq)方法可能有助于发现 SNPs 和评估遗传结构。在我们的研究中,利用 ddRAD-Seq 开发了 98 个单核苷酸多态性(SNP)标记。观察到的杂合度(Ho)和预期杂合度(He)分别为 0.3333 至 0.8000 和 0.2778 至 0.5000。多态性信息含量(PIC)为 0.239198 至 0.375。所有位点均符合哈代-温伯格平衡。这些新的多态性SNP标记将在日本鳗鲡遗传研究中发挥重要作用,有利于日本鳗鲡资源的开发和利用。本研究旨在通过SNP标记的开发,为日本鳗鲡种群遗传多样性研究提供技术支持。
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引用次数: 0
An amplicon genotyping panel suitable for species identification and population genetics in sauger (Sander canadensis) and walleye (Sander vitreus) 适用于梭鱼(Sander canadensis)和马眼鱼(Sander vitreus)物种鉴定和种群遗传学的扩增子基因分型面板
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-19 DOI: 10.1007/s12686-023-01329-z

Abstract

Sauger (Sander canadensis) and walleye (Sander vitreus) are closely related North American fish species that are often managed by fishery agencies throughout their ranges. However, genotyping resources for sauger are presently limited to a small set of microsatellite loci. We evaluated whether primers in an existing walleye genotyping-in-thousands panel could amplify single nucleotide polymorphism loci (SNPs) in sauger. We identified 71 primer pairs that amplify 118 SNPs in both species. Allele frequency differences were large enough to confidently distinguish the species and identify hybrids. Additionally, we identified 41 loci with observed heterozygosity > 0.1 in sauger; these markers may be useful for simple population genetic analyses and parentage analysis when few contributors are present and for differentiating highly structured populations.

摘要 长尾鳕(Sander canadensis)和马黑鱼(Sander vitreus)是密切相关的北美鱼类物种,渔业机构经常对它们的整个分布区进行管理。然而,目前针对长尾鳕的基因分型资源仅限于一小部分微卫星位点。我们评估了现有的马口鱼基因分型千人小组中的引物是否能扩增长尾鳕的单核苷酸多态性位点(SNPs)。我们确定了 71 对引物,可在两种鱼类中扩增 118 个 SNP。等位基因频率差异很大,足以区分物种和鉴定杂交种。此外,我们还在长尾鲈中发现了 41 个杂合度大于等于 0.1 的位点;这些标记可能有助于进行简单的种群遗传分析和亲子鉴定分析(当贡献者较少时),也可能有助于区分高度结构化的种群。
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引用次数: 0
Genetic diversity assessment for reintroduced Przewalski’s horse (Equus ferus) based on newly developed SSR markers 基于新开发的 SSR 标记对重新引入的普氏马(Equus ferus)进行遗传多样性评估
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-18 DOI: 10.1007/s12686-023-01330-6

Abstract

Przewalski’s horse (Equus ferus) is a national protected species in China. It first became extinct in the wild in 1976, and was reintroduced to the Kalamaili National Park (KNP) in Xinjiang, China in 2001. This study aimed to analyze the genetic diversity and structure of the reintroduced population of Przewalski’s horse in KNP. Here, we developed and designed microsatellite molecular markers using 48 feces samples and combined this with published genomic data from the species. A preliminary analysis of genetic diversity within the Przewalski’s horse population in the KNP was conducted using 16 pairs of simple sequence repeat (SSR) markers with high levels of polymorphism and high stability. A total of 199 alleles were amplified using 16 SSR primer pairs, averaging 12.438 at each locus. In the sample of 48 genomes, the average effective number of alleles detected was 2.695, the average expected heterozygosity was 0.577 and the inbreeding coefficient was 0.232. Twelve loci showed significant deviation from Hardy–Weinberg equilibrium following Bonferroni sequential corrections. Our research finds that the genetic diversity of the Przewalski’s horse population in the KNP remains high. This is the first research using newly developed SSR markers to describe the genetic diversity and structure of the reintroduced Przewalski’s horse population in KNP, China. As such, it can inform future reintroductions of the species and conservation measures.

摘要 普氏原马(Equus ferus)是中国的国家一级保护动物。普氏野马于 1976 年首次在野外灭绝,2001 年被重新引入中国新疆卡拉麦里国家公园(KNP)。本研究旨在分析新疆卡拉麦里国家公园普氏马重引种群的遗传多样性和结构。在此,我们利用 48 份粪便样本开发和设计了微卫星分子标记,并将其与已发表的该物种基因组数据相结合。我们使用 16 对具有高多态性和高稳定性的简单序列重复(SSR)标记,对九寨沟普氏马种群的遗传多样性进行了初步分析。使用 16 对 SSR 引物共扩增出 199 个等位基因,平均每个位点有 12.438 个等位基因。在 48 个基因组样本中,检测到的等位基因平均有效数目为 2.695,平均预期杂合度为 0.577,近交系数为 0.232。经 Bonferroni 序列校正后,有 12 个基因位点明显偏离 Hardy-Weinberg 平衡。我们的研究发现,KNP 地区普氏马种群的遗传多样性仍然很高。这是首次使用新开发的 SSR 标记来描述中国昆明核电厂普氏马重引入种群的遗传多样性和结构的研究。因此,该研究可为今后该物种的再引入和保护措施提供参考。
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引用次数: 0
Characterization and cross-species amplification of 33 SNP markers for Liobagrus angullicauda (Teleostei, Siluriformes:Amblycipitidae) Liobagrus angullicauda (Teleostei, Siluriformes:Amblycipitidae) 33 个 SNP 标记的特征和跨物种扩增
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-08 DOI: 10.1007/s12686-023-01328-0
Xiang-Yang Jia, Wei Li, Jun Yan, Ling-Li Xia

Liobagrus angullicauda is an endemic species to streams along the southeast coast in China. Due to over fishing and habitat destruction, the natural population of L. angullicauda has decreased dramatically in recent years. Insufficient molecular markers have limited the effective conservation and management of this species. In the present study, we reported the isolation and characterization of 33 SNP markers in L. angullicauda. The minor allele frequency ranged from 0.083 to 0.507, and the observed and expected heterozygosities ranged from 0.177 to 0.624 and 0.290 to 0.712, respectively. Polymorphic information content ranged from 0.118 to 0.408. Among these SNPs, only four loci showed significant departures from the Hardy–Weinberg equilibrium (P < 0.05). Cross-amplification was tested in related species Liobagrus nigricauda, Liobagrus marginalus and Liobagrus marginatoides. The novel polymorphic SNPs should be helpful for the future study on genetic management and population conservation for this species.

黄颡鱼(Liobagrus angullicauda)是中国东南沿海溪流的特有物种。由于过度捕捞和栖息地的破坏,近年来Liobagrus angullicauda的自然种群数量急剧下降。分子标记的不足限制了对该物种的有效保护和管理。在本研究中,我们分离并鉴定了 33 个 L. angullicauda 的 SNP 标记。小等位基因频率范围为 0.083 至 0.507,观察到的和预期的杂合率范围分别为 0.177 至 0.624 和 0.290 至 0.712。多态信息含量在 0.118 至 0.408 之间。在这些 SNPs 中,只有四个位点明显偏离 Hardy-Weinberg 平衡(P < 0.05)。交叉扩增在相关物种 Liobagrus nigricauda、Liobagrus marginalus 和 Liobagrus marginatoides 中进行了测试。这些新的多态 SNPs 将有助于今后对该物种进行遗传管理和种群保护研究。
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引用次数: 0
Development of 60 SNP markers in Pelodiscus sinensis based on transcriptome sequencing 基于转录组测序的60个SNP标记的开发
IF 1.1 4区 环境科学与生态学 Q4 BIODIVERSITY CONSERVATION Pub Date : 2023-12-02 DOI: 10.1007/s12686-023-01335-1
Mingsong Xiao, Bing Gong, Huimin Xu, Dandan Ju

The Chinese softshell turtle (Pelodiscus sinensis) is a kind of subaquatic reptile with high nutritional value, medicinal value and economic value. However, the number of wild turtles has dropped dramatically due to excessive fishing and habitat destruction. Artificial breeding of the turtle is facing the dilemma of germplasm degradation and lack of seedlings, which seriously hinders the sustainable development of the turtle industry. In the present study, 60 novel single nucleotide polymorphism (SNP) markers were developed based on high-throughput sequencing. Genetic diversity analysis revealed that the values of observed heterozygosity (Ho) and expected heterozygosity (He) varied from 0.0312 to 0.7500 and from 0.0312 to 0.4841, respectively, and the minor allele frequency ranged from 0.3333 to 0.5000. Only four loci showed significant deviation from Hardy–Weinberg equilibrium. The present study will provide a valuable tool for population study and resource conservation.

中华软壳龟(Pelodiscus sinensis)是一种具有较高营养价值、药用价值和经济价值的水下爬行动物。然而,由于过度捕捞和栖息地破坏,野生海龟的数量急剧下降。龟类人工养殖面临着种质退化和种苗不足的困境,严重阻碍了龟类产业的可持续发展。本研究基于高通量测序,开发了60个新的单核苷酸多态性(SNP)标记。遗传多样性分析表明,观察杂合度(Ho)和期望杂合度(He)分别为0.0312 ~ 0.7500和0.0312 ~ 0.4841,次要等位基因频率为0.3333 ~ 0.5000。只有4个位点明显偏离Hardy-Weinberg平衡。本研究将为种群研究和资源保护提供有价值的工具。
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引用次数: 0
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