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利用环介导等温扩增技术(LAMP)快速鉴定一种野生雀形目鸟类的性别

Rapid sex determination of a wild passerine species using loop-mediated isothermal amplification (LAMP).

作者信息

Koch Hanna R, Blohm-Sievers Elke, Liedvogel Miriam

机构信息

Max Planck Research Group Behavioral Genomics Max Planck Institute for Evolutionary Biology Plön Germany.

出版信息

Ecol Evol. 2019 Apr 15;9(10):5849-5858. doi: 10.1002/ece3.5168. eCollection 2019 May.

Abstract

Many bird species are sexually monomorphic and cannot be sexed based on phenotypic traits. Rapid sex determination is often a necessary component of avian studies focusing on behavior, ecology, evolution, and conservation. While PCR-based methods are the most common technique for molecularly sexing birds in the laboratory, a simpler, faster, and cheaper method has emerged, which can be used in the laboratory, but importantly also in the field. Herein, we used loop-mediated isothermal amplification (LAMP) for rapid sex determination of blood samples from juvenile European blackcaps, , sampled in the wild. We designed LAMP primers unique to based on the sex chromosome-specific gene, chromo-helicase-DNA-binding protein (CHD), optimized the primers for laboratory and field application, and then used them to test a subset of wild-caught juvenile blackcaps of unknown gender at the time of capture. Sex determination results were fast and accurate. The advantages of this technique are that it allows researchers to identify the sex of individual birds within hours of sampling and eliminates the need for direct access to a laboratory if implemented at a remote field site. This work adds to the increasing list of available LAMP primers for different bird species and is a new addition within the Passeriformes order.

摘要

许多鸟类物种在性别上是单态的,无法根据表型特征来区分性别。快速确定性别通常是鸟类行为、生态、进化和保护研究的必要组成部分。虽然基于聚合酶链反应(PCR)的方法是实验室中对鸟类进行分子性别鉴定最常用的技术,但一种更简单、更快且更便宜的方法已经出现,该方法不仅可在实验室使用,重要的是还可在野外使用。在此,我们使用环介导等温扩增技术(LAMP)对野外采集的欧洲黑头莺幼鸟的血液样本进行快速性别鉴定。我们基于性染色体特异性基因——染色体解旋酶DNA结合蛋白(CHD)设计了针对黑头莺的LAMP引物,对引物进行了实验室和野外应用的优化,然后用它们来检测一部分捕获时性别未知的野生黑头莺幼鸟。性别鉴定结果快速且准确。这项技术的优点是,研究人员在采样后数小时内就能确定个体鸟类的性别,并且如果在偏远的野外地点实施,无需直接进入实验室。这项工作增加了可用于不同鸟类物种的LAMP引物的种类,并且是雀形目鸟类中的一项新成果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/836a/6540700/66029b7d03de/ECE3-9-5849-g001.jpg

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