Table_1_An automatic fluorescence phenotyping platform to evaluate dynamic infection process of Tobacco mosaic virus-green fluorescent protein in tobacco leaves.XLSX

Tobacco is one of the important economic crops all over the world. Tobacco mosaic virus (TMV) seriously affects the yield and quality of tobacco leaves. The expression of TMV in tobacco leaves can be analyzed by detecting green fluorescence-related traits after inoculation with the infectious clone of TMV-GFP (Tobacco mosaic virus - green fluorescent protein). However, traditional methods for detecting TMV-GFP are time-consuming and laborious, and mostly require a lot of manual procedures. In this study, we develop a low-cost machine-vision-based phenotyping platform for the automatic evaluation of fluorescence-related traits in tobacco leaf based on digital camera and image processing. A dynamic monitoring experiment lasting 7 days was conducted to evaluate the efficiency of this platform using Nicotiana tabacum L. with a total of 14 samples, including the wild-type strain SR1 and 4 mutant lines generated by RNA interference technology. As a result, we found that green fluorescence area and brightness generally showed an increasing trend over time, and the trends were different among these SR1 and 4 mutant lines samples, where the maximum and minimum of green fluorescence area and brightness were mutant-4 and mutant-1 respectively. In conclusion, the platform can full-automatically extract fluorescence-related traits with the advantage of low-cost and high accuracy, which could be used in detecting dynamic changes of TMV-GFP in tobacco leaves.

烟草是全球重要的经济作物之一。烟草花叶病毒（Tobacco mosaic virus, TMV）会严重影响烟叶的产量与品质。通过接种烟草花叶病毒-绿色荧光蛋白（Tobacco mosaic virus - green fluorescent protein, TMV-GFP）的侵染性克隆，检测烟叶的绿色荧光相关性状，即可分析TMV在烟叶中的表达情况。然而，传统的TMV-GFP检测方法耗时耗力，且大多依赖大量人工操作流程。本研究基于数码相机与图像处理技术，开发了一款基于机器视觉的低成本表型分析平台，用于自动评估烟叶的荧光相关性状。本研究开展了一项为期7天的动态监测实验，以普通烟草（Nicotiana tabacum L.）为试验材料，共设置14份样本，涵盖野生型菌株SR1以及4株经RNA干扰技术创制的突变株系，以此评估该平台的检测效能。结果显示，绿色荧光面积与亮度随时间总体呈上升趋势，且SR1菌株与4个突变株系的变化趋势存在差异：绿色荧光面积与亮度的最大值和最小值分别对应突变株系4与突变株系1。综上，该平台可全自动提取荧光相关性状，具备低成本、高精度的优势，可用于检测烟草叶片中TMV-GFP的动态变化。