Assessing hatching rates and the timing of hatching from plankton resting stages—an accurate and cost effective high throughput approach

Assessing hatching rates and the timing of hatching from plankton resting stages—an accurate and cost effective high throughput approach
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评估浮游生物静止阶段的孵化率和孵化时间——一种准确且具有成本效益的高通量方法

DOI:
10.1002/lom3.10125
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发表时间:
2016
期刊:
Limnology and Oceanography: Methods
影响因子:
--
通讯作者:
De Meester
De Meester
中科院分区:
--
文献类型:
--
作者:
Czypionka;Reeves;Vanhamel;De Meester

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滞育生物的孵化率和滞育终止时间是与滞育生物生态学和进化密切相关的性状。从休眠阶段的休眠生物孵化的分析是一个活跃的研究领域,这已经在纬度上的物候调整的背景下进行了广泛的研究。对临时池塘种群的研究揭示了由各种孵化高峰组成的时间孵化曲线,这被认为是一种风险分散策略。对突出的科学问题的研究,如风险分散策略的演变,遗传变异在生态进化背景下孵化的重要性,以及孵化表型的基因组基础,都需要非常大的样本量。因此,我们开发了一个高通量的方法,从甲壳类浮游动物的孵化率和孵化剖面的休息阶段的评估。我们的方法包括在休息阶段的摄影监测孵化试验和使用免费软件的生物图像分析的自动化检测孵化。我们比较了我们的新方法与传统的视觉评估孵化获得的数据的性能。我们的研究结果表明,我们的方法是高度准确的(>95%),并提高了10倍的效率相比,非自动化的视觉评估。因此,我们新的高通量方法允许将时间孵化概况和孵化成功率的分析扩大到样本量,这将使未来能够进行大规模基因组研究或大规模表型分析并监测这些高度相关的性状。
Hatching rates and the timing of diapause termination are traits of high relevance in ecology and evolution of diapausing organisms. The analysis of hatching from dormant stages of planktonic organisms is an active area of research, which has been extensively studied in the context of phenology adjustment over latitude. Research on populations from temporary ponds has revealed temporal hatching profiles consisting of various hatching peaks, which is considered a risk‐spreading strategy. Studies on outstanding scientific questions such as the evolution of risk‐spreading strategies, the importance of genetic variation in hatching in an eco‐evolutionary context, and the genomic underpinning of hatching phenotypes all require very large sample sizes. We therefore developed a high throughput approach for the assessment of hatching rates and temporal hatching profiles of resting stages from crustacean zooplankton. Our method consists in the photographic monitoring of resting stages hatching assays and the detection of hatchlings using free software for the automation of biological image analysis. We compared the performance of our new method with data obtained by traditional visual assessment of hatching. Our results indicate that our approach is highly accurate (>95%) and raises efficiency by a factor of 10 compared to non‐automated visual assessment. Our new high throughput approach therefore allows to upscale the analysis of temporal hatching profiles and hatching success to sample sizes that will enable large‐scale genomic studies or massive phenotyping and monitoring of these highly relevant traits in the future.
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