High-Throughput Assays of Critical Thermal Limits in Insects
High-Throughput Assays of Critical Thermal Limits in Insects
复制标题
DOI：
10.3791/61186
复制
发表时间：
2020-06-01
影响因子：
1.2
通讯作者：
Teets, Nicholas M.
中科院分区：
文献类型：
--
作者：
Awde, David N.;Fowler, Tatum E.;Teets, Nicholas M.
植物和动物的热上限和下限是其性能、生存和地理分布的重要预测因素，对于预测对气候变化的反应至关重要。这项工作描述了两种用于测量昆虫热极限的高通量协议：一种用于评估临界热最小值（CTmin），另一种用于评估响应静态热应激源的热击倒时间（KDT）。在 CTmin 测定中，个体被放置在丙烯酸夹套柱中，经受逐渐降低的温度梯度，并使用红外传感器在它们从栖息处掉落时进行计数。在热 KDT 测定中，个体被放置在 96 孔板中，放置在设置为有压力的高温培养箱中，并录制视频以确定它们不再保持直立和移动的时间。这些协议比常用技术具有优势。两种检测成本都很低，并且可以相对较快地完成（大约 2 小时）。 CTmin 测定减少了实验者误差，并且可以同时测量大量个体。热 KDT 协议会生成每次检测的视频记录，从而消除实验者偏差以及持续实时监测个体的需要。
Upper and lower thermal limits of plants and animals are important predictors of their performance, survival, and geographic distributions, and are essential for predicting responses to climate change. This work describes two high-throughput protocols for measuring insect thermal limits: one for assessing critical thermal minima (CTmin), and the other for assessing heat knock down time (KDT) in response to a static heat stressor. In the CTmin assay, individuals are placed in an acrylic-jacketed column, subjected to a decreasing temperature ramp, and counted as they fall from their perches using an infrared sensor. In the heat KDT assay, individuals are contained in a 96 well plate, placed in an incubator set to a stressful, hot temperature, and video recorded to determine the time at which they can no longer remain upright and move. These protocols offer advantages over commonly used techniques. Both assays are low cost and can be completed relatively quickly (similar to 2 h). The CTmin assay reduces experimenter error and can measure a large number of individuals at once. The heat KDT protocol generates a video record of each assay and thus removes experimenter bias and the need to continuously monitor individuals in real time.