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基于组织分辨率数据的植物茎尖分生组织高通量三维表型分析

High-Throughput 3D Phenotyping of Plant Shoot Apical Meristems From Tissue-Resolution Data.

作者信息

Åhl Henrik, Zhang Yi, Jönsson Henrik

机构信息

Sainsbury Laboratory, University of Cambridge, Cambridge, United Kingdom.

Department of Applied Mathematics and Theoretical Physics, University of Cambridge, Cambridge, United Kingdom.

出版信息

Front Plant Sci. 2022 Apr 18;13:827147. doi: 10.3389/fpls.2022.827147. eCollection 2022.

Abstract

Confocal imaging is a well-established method for investigating plant phenotypes on the tissue and organ level. However, many differences are difficult to assess by visual inspection and researchers rely extensively on manual quantification techniques and qualitative assessment. Here we present a method for quantitatively phenotyping large samples of plant tissue morphologies using triangulated isosurfaces. We successfully demonstrate the applicability of the approach using confocal imaging of aerial organs in . Automatic identification of flower primordia using the surface curvature as an indication of outgrowth allows for high-throughput quantification of divergence angles and further analysis of individual flowers. We demonstrate the throughput of our method by quantifying geometric features of 1065 flower primordia from 172 plants, comparing auxin transport mutants to wild type. Additionally, we find that a paraboloid provides a simple geometric parameterisation of the shoot inflorescence domain with few parameters. We utilise parameterisation methods to provide a computational comparison of the shoot apex defined by a fluorescent reporter of the central zone marker gene with the apex defined by the paraboloid. Finally, we analyse the impact of mutations which alter mechanical properties on inflorescence dome curvature and compare the results with auxin transport mutants. Our results suggest that region-specific expression domains of genes regulating cell wall biosynthesis and local auxin transport can be important in maintaining the wildtype tissue shape. Altogether, our results indicate a general approach to parameterise and quantify plant development in 3D, which is applicable also in cases where data resolution is limited, and cell segmentation not possible. This enables researchers to address fundamental questions of plant development by quantitative phenotyping with high throughput, consistency and reproducibility.

摘要

共聚焦成像技术是一种成熟的用于在组织和器官水平研究植物表型的方法。然而,许多差异通过肉眼检查很难评估,研究人员广泛依赖手动量化技术和定性评估。在此,我们提出一种使用三角化等值面定量分析植物组织形态大样本的方法。我们通过对拟南芥地上器官进行共聚焦成像成功证明了该方法的适用性。利用表面曲率作为生长的指标自动识别花原基,可实现发散角的高通量量化以及对单朵花的进一步分析。我们通过量化172株植物的1065个花原基的几何特征,将生长素运输突变体与野生型进行比较,展示了我们方法的通量。此外,我们发现抛物面能用较少参数对茎尖花序域进行简单的几何参数化。我们利用参数化方法对由中央区标记基因的荧光报告基因定义的茎尖与由抛物面定义的茎尖进行计算比较。最后,我们分析了改变机械性能的突变对花序穹顶曲率的影响,并将结果与生长素运输突变体进行比较。我们的结果表明,调节细胞壁生物合成和局部生长素运输的基因的区域特异性表达域在维持野生型组织形状方面可能很重要。总之,我们的结果表明了一种在三维空间中对植物发育进行参数化和量化的通用方法,该方法在数据分辨率有限且无法进行细胞分割的情况下也适用。这使研究人员能够通过高通量、一致性和可重复性的定量表型分析来解决植物发育的基本问题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8e43/9062647/84a43fba55bf/fpls-13-827147-g001.jpg

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