Comparing Methods for Detection and Quantification of Plasmodesmal Callose in Nicotiana benthamiana Leaves During Defense Responses
Comparing Methods for Detection and Quantification of Plasmodesmal Callose in Nicotiana benthamiana Leaves During Defense Responses
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DOI：
10.1094/mpmi-09-23-0152-sc
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发表时间：
2024-05-22
影响因子：
3.5
通讯作者：
Burch-Smith，Tessa M.
中科院分区：
文献类型：
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作者：
Sankoh，Amie F.;Adjei，Joseph;Burch-Smith，Tessa M.
胼胝质是一种β-(1,3)-d-葡聚糖聚合物，对通过胞间连丝（PD）调节细胞间运输至关重要。病原体操纵PD定位蛋白，通过移除PD处的胼胝质来实现细胞间运输，或者相反，通过增加PD处的胼胝质积累来限制感染期间的细胞间运输。水杨酸等植物防御激素调节PD定位蛋白，在系统性获得性抗性等免疫防御反应中控制PD和细胞间运输。因此，测量植物PD处的胼胝质沉积已成为评估植物免疫过程中可能的细胞间运输活动的一个流行参数。尽管这个度量很流行，但是对于如何进行这些度量并没有标准。本研究评估了三种常用的苯胺蓝染色法鉴定和定量胞间质胼胝质的方法，以确定其中最有效的方法。结果表明，用苯胺蓝染色法和荧光显微镜法测定固定组织中胼胝质沉积是最可靠的方法。还比较了图像分析的手动或半自动工作流程，发现产生类似的结果，尽管半自动工作流程产生更广泛的数据点分布。版权所有©2024作者。这是一篇在CC BY-NC-ND 4.0国际许可下发布的开放获取文章。
Callose, a β-(1,3)-d-glucan polymer, is essential for regulating intercellular trafficking via plasmodesmata (PD). Pathogens manipulate PD-localized proteins to enable intercellular trafficking by removing callose at PD or, conversely, by increasing callose accumulation at PD to limit intercellular trafficking during infection. Plant defense hormones like salicylic acid regulate PD-localized proteins to control PD and intercellular trafficking during immune defense responses such as systemic acquired resistance. Measuring callose deposition at PD in plants has therefore emerged as a popular parameter for assessing likely intercellular trafficking activity during plant immunity. Despite the popularity of this metric, there is no standard for how these measurements should be made. In this study, three commonly used methods for identifying and quantifying plasmodesmal callose by aniline blue staining were evaluated to determine the most effective in theNicotiana benthamianaleaf model. The results reveal that the most reliable method used aniline blue staining and fluorescence microscopy to measure callose deposition in fixed tissue. Manual or semiautomated workflows for image analysis were also compared and found to produce similar results, although the semiautomated workflow produced a wider distribution of data points.Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.