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贝曼漏斗法快速分离野生线虫。

Rapid Isolation of Wild Nematodes by Baermann Funnel.

机构信息

Department of Biology and Center for Genomics & Systems Biology, New York University.

Department of Biology and Center for Genomics & Systems Biology, New York University;

出版信息

J Vis Exp. 2022 Jan 31(179). doi: 10.3791/63287.

Abstract

Beyond being robust experimental model organisms, Caenorhabditis elegans and its relatives are also real animals that live in nature. Studies of wild nematodes in their natural environments are valuable for understanding many aspects of biology, including the selective regimes in which distinctive genomic and phenotypic characters evolve, the genetic basis for complex trait variation, and the natural genetic diversity fundamental to all animal populations. This manuscript describes a simple and efficient method for extracting nematodes from their natural substrates, including rotting fruits, flowers, fungi, leaf litter, and soil. The Baermann funnel method, a classical nematology technique, selectively isolates active nematodes from their substrates. Because it recovers nearly all active worms from the sample, the Baermann funnel technique allows for the recovery of rare and slow-growing genotypes that co-occur with abundant and fast-growing genotypes, which might be missed in extraction methods that involve multiple generations of reproduction. The technique is also well suited to addressing metagenetic, population-genetic, and ecological questions. It captures the entire population in a sample simultaneously, allowing an unbiased view of the natural distribution of ages, sexes, and genotypes. The protocol allows for deployment at scale in the field, rapidly converting substrates into worm plates, and the authors have validated it through fieldwork on multiple continents.

摘要

除了作为强大的实验模式生物外,秀丽隐杆线虫及其相关物种也是生活在自然界中的真正动物。研究自然环境中的野生线虫对于理解生物学的许多方面很有价值,包括独特的基因组和表型特征进化的选择机制、复杂性状变异的遗传基础,以及所有动物种群所必需的自然遗传多样性。本文描述了一种从其自然基质中提取线虫的简单而有效的方法,这些基质包括腐烂的水果、花朵、真菌、落叶和土壤。贝尔曼漏斗法是一种经典的线虫学技术,可从基质中选择性地分离出活跃的线虫。由于它从样本中几乎回收了所有活跃的线虫,因此贝尔曼漏斗技术允许回收与丰富和快速生长的基因型共同存在的稀有和生长缓慢的基因型,这些基因型可能会在涉及多代繁殖的提取方法中被遗漏。该技术也非常适合解决宏遗传学、种群遗传学和生态学问题。它同时捕获样本中的整个种群,使人们能够公正地观察年龄、性别和基因型的自然分布。该方案允许在野外大规模部署,快速将基质转化为线虫板,作者已经通过在多个大陆的实地工作验证了该方案的有效性。

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