Sequencing Herbarium Specimens of a Common Detrimental Plant Disease (Powdery Mildew)
Sequencing Herbarium Specimens of a Common Detrimental Plant Disease (Powdery Mildew)
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DOI：
10.1094/phyto-04-20-0139-per
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发表时间：
2020-07-01
期刊：
影响因子：
3.2
通讯作者：
Tobin, Patrick C.
中科院分区：
文献类型：
--
作者：
Bradshaw, Michael;Tobin, Patrick C.
白粉病（白粉菌科）是一种发生在多种重要经济作物上的有害植物病害。白粉病由超过873种真菌病原体组成，影响超过10，000种植物物种。利用核核糖体RNA基因簇的内转录间隔区（ITS）和大亚基（LSU）区域完成白粉病的遗传鉴定。白粉病的ITS和LSU区域可用于生态学、流行病学、系统发育和分类学研究。然而，对这些区域进行测序并非没有挑战。例如，白粉病序列经常被植物和/或真菌DNA污染。此外，往往有一个有限的数量和旧标本的DNA可以随着时间的推移片段。测序白粉病的成功往往取决于用于运行聚合酶链反应（PCR）的引物。引物需要足够宽，以匹配大多数白粉病DNA，但又足够特异，以不与其他生物体对齐。对白粉病的分类学和系统发育进行了综述，重点是ITS + LSU基因组区域的测序。此外，我们引入了一种新的巢式引物方案来对白粉病植物标本进行测序，其中包括六种新的白粉病特异性引物。提出的新测序方案允许对长达130年的标本进行一致的测序。植物标本测序对于解决包括白粉病在内的多种植物病原系统中的许多生态学、流行病学、系统发育和分类学问题非常有用。
Powdery mildew (Erysiphaceae) is a detrimental plant disease that occurs on a variety of economically important crops. Powdery mildew consists of over 873 species of fungal pathogens that affect over 10,000 plant species. Genetic identification of powdery mildew is accomplished using the internal transcribed spacer (ITS) and large subunit (LSU) regions of the nuclear ribosomal RNA gene cluster. The ITS and LSU regions of powdery mildews can be useful in ecological, epidemiological, phylogenetic, and taxonomic investigations. However, sequencing these regions is not without its challenges. For example, powdery mildew sequences are often contaminated with plant and/or fungal DNA. Also, there tends to be a limited amount and older specimens' DNA can fragment over time. The success of sequencing powdery mildew often depends on the primers used for running polymerase chain reaction (PCR). The primers need to be broad enough that they match the majority of powdery mildew DNA yet specific enough that they do not align with other organisms. A review of the taxonomy and phylogeny of the powdery mildews is presented with an emphasis on sequencing the ITS + LSU genomic regions. Additionally, we introduce a new nested primer protocol for sequencing powdery mildew herbarium samples that includes six new powdery mildew-specific primers. The new sequencing protocol presented allows specimens up to 130 years old to be sequenced consistently. Sequencing herbarium specimens can be extremely useful for addressing many ecological, epidemiological, phylogenetic, and taxonomic problems in multiple plant pathogenic systems including the powdery mildews.