Keyes Samuel D, Gostling Neil J, Cheung Jessica H, Roose Tiina, Sinclair Ian, Marchant Alan
2The Faculty of Engineering and the Environment,The University of Southampton,Southampton,SO17 1BJ,UK.
1The Centre for Biological Sciences,The University of Southampton,Southampton,SO17 1BJ,UK.
Microsc Microanal. 2017 Jun;23(3):538-552. doi: 10.1017/S1431927617000319. Epub 2017 Mar 21.
The use of in vivo X-ray microcomputed tomography (μCT) to study plant root systems has become routine, but is often hampered by poor contrast between roots, soil, soil water, and soil organic matter. In clinical radiology, imaging of poorly contrasting regions is frequently aided by the use of radio-opaque contrast media. In this study, we present evidence for the utility of iodinated contrast media (ICM) in the study of plant root systems using μCT. Different dilutions of an ionic and nonionic ICM (Gastrografin 370 and Niopam 300) were perfused into the aerial vasculature of juvenile pea plants via a leaf flap (Pisum sativum). The root systems were imaged via μCT, and a variety of image-processing approaches used to quantify and compare the magnitude of the contrast enhancement between different regions. Though the treatment did not appear to significantly aid extraction of full root system architectures from the surrounding soil, it did allow the xylem and phloem units of seminal roots and the vascular morphology within rhizobial nodules to be clearly visualized. The nonionic, low-osmolality contrast agent Niopam appeared to be well tolerated by the plant, whereas Gastrografin showed evidence of toxicity. In summary, the use of iodine-based contrast media allows usually poorly contrasting root structures to be visualized nondestructively using X-ray μCT. In particular, the vascular structures of roots and rhizobial nodules can be clearly visualized in situ.
利用活体X射线微型计算机断层扫描(μCT)研究植物根系已成为常规方法,但根系、土壤、土壤水分和土壤有机质之间对比度不佳常常阻碍研究进展。在临床放射学中,对比度欠佳区域的成像常常借助使用不透射线的造影剂来辅助。在本研究中,我们展示了碘化造影剂(ICM)在利用μCT研究植物根系方面的效用证据。通过叶瓣(豌豆)将离子型和非离子型ICM(泛影葡胺370和尼奥普兰300)的不同稀释液灌注到幼年豌豆植株的地上维管系统中。通过μCT对根系进行成像,并采用多种图像处理方法来量化和比较不同区域之间对比度增强的程度。尽管该处理似乎并未显著有助于从周围土壤中提取完整的根系结构,但它确实能使胚根的木质部和韧皮部单元以及根瘤内的维管形态清晰可见。非离子型、低渗造影剂尼奥普兰似乎被植物良好耐受,而泛影葡胺则显示出毒性迹象。总之,使用碘基造影剂能够利用X射线μCT无损地可视化通常对比度欠佳的根系结构。特别是,根系和根瘤的维管结构能够在原位清晰地可视化。