U.S. National Arboretum - Floral and Nursery Plants Research Unit, U.S. Department of Agriculture/Agricultural Research Service, Beltsville, MD 20705.
Plant Dis. 2021 Jan;105(1):164-168. doi: 10.1094/PDIS-09-19-2053-RE. Epub 2020 Nov 16.
and are the causal agents of boxwood blight, a devastating disease of boxwood that has caused significant economic impact on the nursery and landscape industries in the U.S. and in Europe. The two species are genetically distinct and are found in different geographic areas but are difficult to distinguish based on morphology and pathogenicity. Fast, accurate, and inexpensive methods to detect and differentiate these species is critical in stopping the spread of the disease. We designed primer pairs based on available sequences of four conserved regions-calmodulin, histone H3, internal transcribed spacer, and β-tubulin-and tested their ability to differentiate the two species. Here we report three primer pairs derived from sequence differences in the histone H3 region that can be used to specifically detect , , or both species. Specificity of these primers was tested against nine isolates of , three isolates of , 13 other species, and five isolates from related genera using conventional and real-time PCR. These are the first primers available that can be used with either a multiplexed conventional PCR or SYBR-based real-time PCR to specifically detect and differentiate the two fungal species.
和 是黄杨枯萎病的病原体，这是一种严重的黄杨病害，已对美国和欧洲的苗圃和景观产业造成重大经济影响。这两个物种在遗传上是不同的，分布在不同的地理区域，但基于形态和致病性很难区分。快速、准确和廉价的方法来检测和区分这两个物种对于阻止疾病的传播至关重要。我们设计了基于四个保守区域（钙调蛋白、组蛋白 H3、内部转录间隔区和β-微管蛋白）的可用序列的引物对，并测试了它们区分这两个 物种的能力。在这里，我们报告了三个源自组蛋白 H3 区域序列差异的引物对，可用于特异性检测 、 或这两个物种。使用常规 PCR 和实时 PCR，针对 9 个 菌株、3 个 菌株、13 个其他 种和 5 个来自相关属的菌株，对这些引物的特异性进行了测试。这些是第一个可用于多重常规 PCR 或基于 SYBR 的实时 PCR 的引物，可特异性检测和区分这两种真菌物种。
