J Integr Plant Biol. ›› 2018, Vol. 60 ›› Issue (9): 897-906.DOI: 10.1111/jipb.12642

• New Technology • Previous Articles    

Calcium imaging in Arabidopsis pollen cells using G-CaMP5

Min Diao1,2,3, Xiaolu Qu2 and Shanjin Huang2*   

  1. 1Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
    2Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
    3University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2018-01-17 Accepted:2018-02-07 Online:2018-02-09 Published:2018-09-01
  • About author:*Correspondence: Email: Shanjin Huang (sjhuang@tsinghua.edu.cn)

Abstract:

Calcium (Ca2+) signaling has been implicated in pollen germination and pollen tube growth. To date, however, we still know very little about how exactly Ca2+ signaling links to various physiological subcellular processes during pollen germination and pollen tube growth. Given that Ca2+ signaling is tightly related to the cytosolic concentration and dynamics of Ca2+, it is vital to trace the dynamic changes in Ca2+ levels in order to decode Ca2+ signaling. Here, we demonstrate that G‐CaMP5 serves well as an indicator for monitoring cytosolic Ca2+ dynamics in pollen cells. Using this probe, we show that cytosolic Ca2+ changes dramatically during pollen germination, and, as reported previously, Ca2+ forms a tip‐focused gradient in the pollen tube and undergoes oscillation in the tip region during pollen tube growth. In particular, using G‐CaMP5 allowed us to capture the dynamic changes in the cytosolic Ca2+ concentration ([Ca2+]cyt) in pollen tubes in response to various exogenous treatments. Our data suggest that G‐CaMP5 is a suitable probe for monitoring the dynamics of [Ca2+]cyt in pollen cells.

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