The pronounced morphological variability exhibited by Speyeria butterflies can hinder the identification of these taxa, and thus presents a challenge to their management and conservation. While several studies have documented declines in American populations of Speyeria due to habitat change, the response of Canadian populations of Speyeria to widespread anthropogenic pressures remains poorly understood due to their difficult identification. Here, we assessed the utility of morphological characters recommended in regional field guides for the identification of Speyeria butterflies in Canada, focusing on three morphologically variable subspecies that inhabit boreal forests subject to widespread oil sands extraction: S. aphrodite manitoba, S. atlantis hollandi, and S. hesperis beani. We scored 159 specimens for 11 morphological characters, and complimented this data by sequencing the barcode region of the COI gene for 15 of these specimens. Our results indicate a high level of intraspecific variability in several characters, and the COI gene revealed that initial morphological identifications were incorrect. A further assessment of character reliability identified sets of morphological characters that, in combination with specimen sex and species natural history, improved subspecies identification. Notably, we found that a relatively novel field marker—in vivo eye color—consistently distinguished S. aphrodite manitoba from S. hesperis beani and S. atlantis hollandi. Our results emphasize the importance of using an integrative approach for the accurate identification of morphologically variable species, particularly in situations where molecular methods are not readily available, such as citizen science programs.

中文翻译：

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“肉眼”鉴定：综合性状评估为大贝母蝴蝶（蛱蝶科：Speyeria）的区域现场鉴定提供信息

Speyeria 蝴蝶表现出的显着形态变异会阻碍这些分类群的识别，从而对其管理和保护提出挑战。虽然有几项研究记录了由于栖息地变化导致美国斯佩氏菌种群下降，但由于难以识别，加拿大斯佩尔菌属种群对广泛的人为压力的反应仍然知之甚少。在这里，我们评估了区域实地指南中推荐的形态特征的效用，用于识别加拿大的 Speyeria 蝴蝶，重点关注三个形态可变的亚种，它们栖息在受广泛油砂开采的北方森林中：S. aphrodite manitoba、S. atlantis hollandi、和 S. hesperis beani。我们对 159 个标本的 11 个形态特征进行了评分，并通过对其中 15 个样本的 COI 基因的条形码区域进行测序来补充这些数据。我们的结果表明在几个性状中存在高度的种内变异性，并且 COI 基因表明最初的形态学鉴定是不正确的。对特征可靠性的进一步评估确定了多组形态特征，结合标本性别和物种自然历史，改进了亚种识别。值得注意的是，我们发现一种相对新颖的场标记——体内眼睛颜色——始终如一地将 S. aphrodite manitoba 与 S. hesperis beani 和 S. atlantis hollandi 区分开来。我们的结果强调了使用综合方法准确识别形态可变物种的重要性，特别是在分子方法不易获得的情况下，