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从圈养中国林麝(Moschus berezovskii)转录组中获取和应用 SSR 标记

SSR Marker Acquisition and Application from Transcriptome of Captive Chinese Forest Musk Deer (Moschus berezovskii).

机构信息

Shaanxi Key Laboratory for Animal Conservation, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, 710069, Xi'an, China.

Northwest Institute of Endangered Zoological Species, Shaanxi Institute of Zoology, 710032, Xi'an, China.

出版信息

Biochem Genet. 2024 Aug;62(4):3215-3230. doi: 10.1007/s10528-023-10595-3. Epub 2023 Dec 14.

Abstract

Forest musk deer (Moschus berezovskii) is one of the most endangered medicinally important wild animals in the world. Forest musk deer farming is the main way of production of musk. However, the single provenance and lack of genetic information lead to reduced genetic diversity of forest musk deer. Therefore, more SSR markers need to be developed to identify forest musk deer germplasm. In this study, bone marrow derived mesenchymal cells were used to construct cDNA library for transcriptome sequencing. The datasets were de novo assembled and annotated. 9 polymorphic simple sequence repeat (SSR) markers were finally identified and used to detect population genetic diversity. 6.07 Gb clean data were generated using Illumina sequencing technology, and de novo assembled into 138,591 transcripts and 81,553 unigenes. 5,777 simple sequence repeats (SSRs) were identified, in which there were 578 repeating motif types, with mono-nucleotide and tri-nucleotides comprising 55.88% and 25.60%, respectively. 100 primer pairs were designed to validate amplification and polymorphism using DNA from fecal samples. 9 polymorphic SSRs were developed and used to detect population genetic diversity of 122 forest musk deer in 2 farms. The average number of alleles per locus varied from 4 to 15 (average = 8.3). The observed heterozygosity (H) per locus ranged from 0.102 to 0.941, while the expected heterozygosity (H) per locus was from 0.111 to 0.651. All loci deviated significantly from the Hardy-Weinberg equilibrium (p < 0.001). The polymorphism information content (PIC) of these loci varied from 0.108 to 0.619. 9 polymorphic SSR markers were developed in this research. These sites can be used for breeding planning and conservation of germplasm resources.

摘要

林麝(Moschus berezovskii)是世界上最濒危的药用野生动物之一。林麝养殖是麝香的主要生产方式。然而,单一来源和缺乏遗传信息导致林麝遗传多样性降低。因此,需要开发更多的 SSR 标记来鉴定林麝种质资源。本研究采用骨髓间充质细胞构建转录组测序 cDNA 文库。对数据集进行从头组装和注释。最终鉴定出 9 个多态性简单序列重复(SSR)标记,并用于检测种群遗传多样性。使用 Illumina 测序技术生成 6.07 Gb 清洁数据,从头组装成 138591 个转录本和 81553 个基因。鉴定出 5777 个简单序列重复(SSR),其中有 578 个重复基序类型,单核苷酸和三核苷酸分别占 55.88%和 25.60%。设计 100 对引物,使用粪便样本 DNA 验证扩增和多态性。开发并使用 9 个多态性 SSR 标记检测 2 个养殖场 122 只林麝的种群遗传多样性。每个位点的等位基因数从 4 到 15 不等(平均值为 8.3)。每个位点的观测杂合度(H)范围从 0.102 到 0.941,而预期杂合度(H)范围从 0.111 到 0.651。所有位点均显著偏离哈迪-温伯格平衡(p<0.001)。这些位点的多态信息含量(PIC)从 0.108 到 0.619 不等。本研究开发了 9 个多态性 SSR 标记。这些位点可用于林麝的育种规划和种质资源保护。

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