Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (5): 596-604.DOI: 10.11983/CBB20016

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

An Improved Protocol for Whole Mount Clearing of Plant Root Tip

Long Ma, Guilin Li, Shipeng Li, Su Jiang*()   

  1. Qilu Normal University, Jinan 250200, China
  • Received:2020-02-05 Accepted:2020-06-05 Online:2020-09-01 Published:2020-09-03
  • Contact: Su Jiang

Abstract: Whole mount clearing is a routine method in morphological study, which allows observation of plant internal structure without section. Using high refractive index materials as medium, clearing techniques reduce light scattering, acquire enhancive light quantity and increase depth of field and vertical planes in a particular focal plane, to facilitate the samples transparency for observation. Nevertheless, clearing materials may disturb the osmosis and pH of sample medium, which is adverse to cells morphology. So far, the effective clearing techniques have been widely used in several studies with ovule and leaf. However, the current protocol is not reliable enough for root tip clearing, because the thin cell wall is vulnerable under the treatment of clearing solutions, resulting in abnormal root tips and cells plasmolysis. To achieve a stable and optimized clearing method for root tip, we established a standard protocol via evaluation of root tip morphology, plasmolysis and cells clarity in Arabidopsis thaliana. With these improved clearing methods, we developed an optimized clearing observation system (including clearing time, pH and composition) for root tip, which could provide a reliable technique for vulnerable tissues clearing.

Key words: morphogenesis, whole mount clearing, root tip, Arabidopsis thaliana