Department of Botany & Plant Pathology, Oregon State University, Corvallis, Oregon, USA.
Center for Genome Research and Biocomputing, Oregon State University, Corvallis, Oregon, USA.
Plant J. 2021 Apr;106(2):566-579. doi: 10.1111/tpj.15166. Epub 2021 Mar 19.
High-throughput phenotyping systems are powerful, dramatically changing our ability to document, measure, and detect biological phenomena. Here, we describe a cost-effective combination of a custom-built imaging platform and deep-learning-based computer vision pipeline. A minimal version of the maize (Zea mays) ear scanner was built with low-cost and readily available parts. The scanner rotates a maize ear while a digital camera captures a video of the surface of the ear, which is then digitally flattened into a two-dimensional projection. Segregating GFP and anthocyanin kernel phenotypes are clearly distinguishable in ear projections and can be manually annotated and analyzed using image analysis software. Increased throughput was attained by designing and implementing an automated kernel counting system using transfer learning and a deep learning object detection model. The computer vision model was able to rapidly assess over 390 000 kernels, identifying male-specific transmission defects across a wide range of GFP-marked mutant alleles. This includes a previously undescribed defect putatively associated with mutation of Zm00001d002824, a gene predicted to encode a vacuolar processing enzyme. Thus, by using this system, the quantification of transmission data and other ear and kernel phenotypes can be accelerated and scaled to generate large datasets for robust analyses.
高通量表型分析系统功能强大,极大地改变了我们记录、测量和检测生物现象的能力。在这里,我们描述了一种具有成本效益的组合,即定制的成像平台和基于深度学习的计算机视觉管道。使用低成本且易于获得的部件构建了一个最小化的玉米(Zea mays)穗扫描仪。扫描仪旋转玉米穗,同时数码相机捕获穗表面的视频,然后将其数字化地展平成二维投影。在穗投影中可以清楚地区分 GFP 和花青素种仁表型,并且可以使用图像分析软件进行手动注释和分析。通过使用迁移学习和深度学习目标检测模型设计和实现自动化种仁计数系统,提高了通量。计算机视觉模型能够快速评估超过 390000 个种仁,识别出广泛 GFP 标记突变等位基因中的雄性特异性传递缺陷。这包括一个以前未描述的缺陷,推测与Zm00001d002824 基因突变有关,该基因预测编码液泡加工酶。因此,通过使用该系统,可以加速和扩展传输数据和其他穗和种仁表型的量化,以生成用于稳健分析的大型数据集。