Chinese Bulletin of Botany ›› 2017, Vol. 52 ›› Issue (5): 608-614.DOI: 10.11983/CBB16196

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Fluorescence Imaging of the Extracellular Zinc Distribution in Plants by Using a Highly Specific Fluorescent Probe

Hongwei Yao1, Yang Liu2, Yulai Cheng2, Haiyang Yu3, Zhiliang Liu3, Ju Yang1   

  1. 1College of Life Sciences, Inner Mongolia University, Hohhot 010021, China
    2Research Institute of Inner Mongolia Hesheng Ecological Science and Technology, Hohhot 010010, China
    3Chemical Institute of Chemical Industry, Inner Mongolia University, Hohhot 010021, China
  • Received:2016-10-10 Accepted:2017-03-07 Online:2017-09-01 Published:2017-07-10
  • Contact: Ju Yang

Abstract: We chose Arabidopsis thaliana and Setaria italic as material to study the character of HL1 that can specifically combine with zinc, and the distribution of extracellular Zn2+ by using fluorescence spectrophotometry, isothermal titration calorimetry (ITC200) and inverted fluorescence microscopy. Fluorescence intensity for HL1 was greatly enhanced with the addition of Zn2+ but not other ions. The dissociation constant (KD=7.02×10-4 mol·L-1) exhibited product stability with the combined reaction of HL1 and Zn2+. In A. thaliana, the distribution of extracellular free Zn2+ was mainly located in leaf intercellular space and surface of the trichome where fluorescence intensity was corresponding to the concentration of Zn2+. The distribution of Zn2+ were located in intercellular space and fibrovascular tissue in the leaf of S. italic. The root of elongation zone was existing the blue fluorescence corresponded to the presence and concentration of Zn2+. The root elongation zone relates to Zn2+ transportation, and the leaf intercellular space and trichome surface are related to Zn2+ storage. In conclusion, Investigation of extracellular free Zn2+ by using HL1 is efficient.

Key words: zinc, blue fluorescence, leaf trichome, vascular tissue, intercellular space