Identifying the proximal cue(s) for pupal color variation in the bordered patch butterfly, Chlosyne lacinia (Geyer 1837; Lepidoptera: Nymphalidae)

Color is a multifaceted trait with many functions such as predator defense, thermoregulation, and immune response. We investigated pupal color variation in Chlosyne lacinia pupae, focusing on identifying the cue for increased melanization. Through laboratory experiments, we demonstrated pupae reared on black backgrounds exhibited significantly higher melanization compared to those on white backgrounds. Additionally, black pupae experienced longer developmental periods, suggesting a trade-off between defense and developmental time. Our findings support crypsis as a likely evolutionary driver for increased melanization in response to substrate color. We discuss potential implications for predator avoidance, immune response, and developmental costs associated with melanization. This study provides insights into the adaptive significance of pupal melanization in response to environmental cues, shedding light on the complex interplay between life history traits in butterflies., Identification of Black Pigment
Using the protocol for cuticular pigment extraction in Debecker et al. (2015), we used pupal exuviae from colony individuals to verify the black pigment as melanin. The pigment was extracted by adding 1 M NaOH/10% DMSO, of which the volume was adjusted to the mass of each exuvia (extraction volume in μL = 200 × exuvial mass in mg) after the exuviae had been homogenized twice for seven seconds. The samples were incubated at 80°C for two hours in a water bath and then centrifuged at 12,000 g for 10 minutes. The absorbance of the samples was read at 380 nm and compared to standard absorbance curves for melanin.
Cue Identification
Colony larvae of the third/fourth instar were selected for use in multiple experiments to identify the cue for pupal melanization. First, we conducted a shading experiment. We placed larvae in 32oz Styrofoam cups with white or black mesh over the opening held in place by the lid with a hole cut in the middle. A moist paper towel was..., , # Identifying the proximal cue(s) for pupal color variation in the bordered patch butterfly, Chlosyne lacinia (Geyer 1837; Lepidoptera: Nymphalidae)

## Author Information

A. Principal Investigator Contact Information
Name: Jamie Phelps
Institution: Louisiana State University
Address: Baton Rouge, LA USA
Email: [[e-mail]](mailto:[e-mail])

B. Associate or Co-investigator Contact Information
Name: Luis Santiago-Rosario
Institution: University of Minnesota
Address: St. Paul, MN USA
Email: [[e-mail]](mailto:[e-mail])

C. Associate or Co-investigator Contact Information
Name: Morgan Kelly
Institution: Louisiana State University
Address: Baton Rouge, LA USA
Email: [[e-mail]](mailto:[e-mail])

Date of data collection: 2022-2023

Geographic location of data collection: South Texas, USA &
Baton Rouge, LA USA

## SHARING/ACCESS INFORMATION

Recommended citation for this dataset: Phelps, JM, Santiago-Rosario, LY, and Kelly MW. (2024). Ident...

体色是一类兼具多重功能的复杂性状，可参与天敌防御、体温调节与免疫应答等生理生态过程。本研究以边框斑蛱蝶（Chlosyne lacinia）的蛹为研究对象，旨在探究诱导蛹黑化作用（melanization）程度升高的触发因素。通过室内实验，我们发现相较于在白色背景下饲养的蛹，黑色背景中饲养的蛹其黑化程度显著更高。此外，黑化程度更高的黑色蛹发育周期更长，这暗示防御与发育时长之间存在权衡（trade-off）关系。本研究结果支持“隐蔽色假说”，即基底颜色可作为进化驱动力，推动蛹黑化程度提升。我们还讨论了黑化作用在天敌规避、免疫应答以及相关发育成本中的潜在意义。本研究为蝴蝶蛹期黑化响应环境信号的适应性意义提供了新见解，有助于揭示蝶类生活史性状间的复杂相互作用。

# 黑色色素鉴定
参照Debecker等人(2015)的表皮色素提取方案，我们利用饲养种群个体的蛹蜕来验证黑色色素为黑色素（melanin）。具体操作如下：将蛹蜕两次匀浆7秒后，加入1 M NaOH/10% DMSO混合液，提取体积（单位：μL）按照公式「提取体积(μL) = 200 × 蛹蜕质量(mg)」进行调整。随后将样品置于80℃水浴锅中孵育2小时，再以12,000 g离心10分钟。最后在380 nm波长下读取样品吸光度，并与黑色素标准吸光度曲线进行比对。

# 触发因素鉴定
本研究选取三/四龄期的饲养种群幼虫开展多项实验，以鉴定诱导蛹黑化的触发因素。首先开展遮光实验：将幼虫置于32盎司泡沫塑料杯中，杯口覆盖白色或黑色网纱，并用开有中心孔的杯盖固定。同时放置湿纸巾……

# 边框斑蛱蝶（Chlosyne lacinia，Geyer 1837；鳞翅目：蛱蝶科）蛹体色变异的近端触发因素鉴定

## 作者信息
A. 项目负责人联系方式
姓名：杰米·菲尔普斯（Jamie Phelps）
所属机构：路易斯安那州立大学
地址：美国路易斯安那州巴吞鲁日
邮箱：[e-mail]

B. 副研究员/合作研究者联系方式
姓名：路易斯·圣地亚哥-罗萨里奥（Luis Santiago-Rosario）
所属机构：明尼苏达大学
地址：美国明尼苏达州圣保罗
邮箱：[e-mail]

C. 副研究员/合作研究者联系方式
姓名：摩根·凯利（Morgan Kelly）
所属机构：路易斯安那州立大学
地址：美国路易斯安那州巴吞鲁日
邮箱：[e-mail]

数据采集时间：2022-2023年
数据采集地理区域：美国南德克萨斯州及路易斯安那州巴吞鲁日

## 共享与获取信息
本数据集推荐引用格式：Phelps, JM, Santiago-Rosario, LY, 及 Kelly MW. (2024). 识别...