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快速干燥对植物基因组大小估计的影响。

The effects of rapid desiccation on estimates of plant genome size.

机构信息

Department of Integrative Biology, University of Guelph, Guelph, ON N1G2W1, Canada.

出版信息

Chromosome Res. 2011 Aug;19(6):825-42. doi: 10.1007/s10577-011-9232-5.

Abstract

Flow cytometry has become the dominant method for estimating nuclear DNA content in plants, either for ploidy determination or quantification of absolute genome size. Current best practices for flow cytometry involve the analysis of fresh tissue, however, this imposes significant limitations on the geographic scope and taxonomic diversity of plants that can be included in large-scale genome size studies. Dried tissue has been used increasingly in recent years, but largely in the context of ploidy analysis. Here we test rapid tissue drying with silica gel as a method for use in genome size studies, potentially enabling broader geographic sampling of plants when fresh tissue collection is not feasible. Our results indicate that rapid drying introduces comparatively minor error (<10%), which is similar to the error introduced by other common methodological variations such as instrument. Additionally, the relative effect of drying on genome size and data quality varied between species and buffers. Tissue desiccation provides a promising approach for expanding our knowledge of plant genome size diversity.

摘要

流式细胞术已成为估计植物核 DNA 含量的主要方法,无论是用于倍性测定还是绝对基因组大小的定量。目前流式细胞术的最佳实践涉及新鲜组织的分析,然而,这对可以包含在大规模基因组大小研究中的植物的地理范围和分类多样性施加了重大限制。近年来,干燥组织的使用越来越多,但主要是在倍性分析的背景下。在这里,我们测试了硅胶的快速组织干燥作为基因组大小研究的一种方法,当新鲜组织收集不可行时,可能能够更广泛地对植物进行地理采样。我们的结果表明,快速干燥引入的错误相对较小(<10%),与仪器等其他常见方法变化引入的错误相似。此外,干燥对基因组大小和数据质量的相对影响因物种和缓冲液而异。组织干燥为扩大我们对植物基因组大小多样性的认识提供了一个很有前途的方法。

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