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首次使用牛津纳米孔技术对极度濒危物种厄瓜多尔褐头蜘蛛猴(Ateles fusciceps fusciceps)进行全基因组测序和组装。

First whole-genome sequence and assembly of the Ecuadorian brown-headed spider monkey (Ateles fusciceps fusciceps), a critically endangered species, using Oxford Nanopore Technologies.

机构信息

Laboratorio de Biotecnología Vegetal, Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito (USFQ), Quito 170901, Ecuador.

Instituto Nacional de Biodiversidad (INABIO), Quito 170135, Ecuador.

出版信息

G3 (Bethesda). 2024 Mar 6;14(3). doi: 10.1093/g3journal/jkae014.

Abstract

The Ecuadorian brown-headed spider monkey (Ateles fusciceps fusciceps) is currently considered one of the most endangered primates in the world and is classified as critically endangered [International union for conservation of nature (IUCN)]. It faces multiple threats, the most significant one being habitat loss due to deforestation in western Ecuador. Genomic tools are keys for the management of endangered species, but this requires a reference genome, which until now was unavailable for A. f. fusciceps. The present study reports the first whole-genome sequence and assembly of A. f. fusciceps generated using Oxford Nanopore long reads. DNA was extracted from a subadult male, and libraries were prepared for sequencing following the Ligation Sequencing Kit SQK-LSK112 workflow. Sequencing was performed using a MinION Mk1C sequencer. The sequencing reads were processed to generate a genome assembly. Two different assemblers were used to obtain draft genomes using raw reads, of which the Flye assembly was found to be superior. The final assembly has a total length of 2.63 Gb and contains 3,861 contigs, with an N50 of 7,560,531 bp. The assembly was analyzed for annotation completeness based on primate ortholog prediction using a high-resolution database, and was found to be 84.3% complete, with a low number of duplicated genes indicating a precise assembly. The annotation of the assembly predicted 31,417 protein-coding genes, comparable with other mammal assemblies. A reference genome for this critically endangered species will allow researchers to gain insight into the genetics of its populations and thus aid conservation and management efforts of this vulnerable species.

摘要

厄瓜多尔褐头蜘蛛猴(Ateles fusciceps fusciceps)目前被认为是世界上最濒危的灵长类动物之一,被列为极危物种[国际自然保护联盟(IUCN)]。它面临着多种威胁,最主要的威胁是由于厄瓜多尔西部的森林砍伐导致栖息地丧失。基因组工具是濒危物种管理的关键,但这需要参考基因组,而迄今为止,A. f. fusciceps 的参考基因组尚不可用。本研究报告了使用牛津纳米孔长读长生成的 A. f. fusciceps 的首个全基因组序列和组装。从一只亚成体雄性中提取 DNA,并按照连接测序试剂盒 SQK-LSK112 工作流程制备测序文库。使用 MinION Mk1C 测序仪进行测序。对测序reads 进行处理,生成基因组组装。使用两种不同的组装器使用原始reads 获得草图基因组,其中 Flye 组装被发现更优越。最终组装的总长度为 26.3Gb,包含 3861 个 contigs,N50 为 7560531bp。根据使用高分辨率数据库的灵长类动物直系同源预测,对组装进行了注释完整性分析,结果表明其完成度为 84.3%,重复基因数量较少,表明组装非常精确。该组装的注释预测了 31417 个蛋白质编码基因,与其他哺乳动物组装相当。该极危物种的参考基因组将使研究人员能够深入了解其种群的遗传学,从而有助于保护和管理这一脆弱物种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6127/10917520/e7b75e672082/jkae014f1.jpg

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