Abstract
The study of plant anatomy, which can be traced back to the seventeenth century, advanced hand in hand with light microscopy technology and relies on traditional histologic techniques, which are based on serial two‐dimensional (2D) sections. However, these valuable techniques lack spatial arrangement of the tissue and hence provide only partial information. A new technique of whole‐mount three‐dimensional (3D) imaging termed high‐resolution episcopic microscopy (HREM) can overcome this obstacle and generate a 3D model of the specimen at a near‐histological resolution. Here, we describe the application of HREM technique in plants by analyzing two plant developmental processes in woody plants: oil secretory cavity development in citrus fruit and adventitious root formation in persimmon rootstock cuttings. HREM 3D models of citrus fruit peel showed that oil cavities were initiated schizogenously during the early stages of fruitlet development. Citrus secretory cavity formation, shape, volume, and distribution were analyzed, and new insights are presented. HREM 3D model comparison of persimmon rootstock clones, which differ in their rooting ability, revealed that difficult‐to‐root clones failed to develop adventitious roots due to their inability to initiate root primordia.
Highlights
Traditional histological techniques using serial two‐dimensional (2D) sections which are stained and visualized under light micros‐ copy were already applied in the seventeenth century to study plant morphology and anatomy (Bolam, Jones, & Paton, 1973)
Statement We describe the application of a technique for whole‐ mount three‐dimensional imaging termed high‐resolution episcopic microscopy (HREM) in plants
Analysis of secre‐ tory cavity development and adventitious root formation demonstrated the potential use of the HREM application for studying developmental processes in plants
Summary
Traditional histological techniques using serial two‐dimensional (2D) sections which are stained and visualized under light micros‐ copy were already applied in the seventeenth century to study plant morphology and anatomy (Bolam, Jones, & Paton, 1973). We describe the appli‐ cation of the HREM technique in studying two plant developmental processes: oil cavity development in citrus fruit peel and adventi‐ tious root formation in persimmon cuttings. Analysis of secre‐ tory cavity development and adventitious root formation demonstrated the potential use of the HREM application for studying developmental processes in plants. We utilized the HREM 3D imaging to study Citrus limon secretory cavity anatomy during different stages of fruit development. To study the relationship between stem anatomy and rooting ability, easy‐ and difficult‐to‐root persimmon (Dyospyros virginiana) rootstock clones (Izhaki et al, 2018) were analyzed by HREM and detailed 3D histology images of the cuttings were generated. Comparison of the 3D models revealed that difficult‐ to‐root clones failed to develop adventitious roots due to their inability to initiate root primordia
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