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Ca2+ participates in programmed cell death by modulating ROS during pollen cryopreservation.

Ruifen Ren1,2, Hao Zhou1, Lingling Zhang1

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Cryopreservation increases pollen calcium (Ca2+) levels, impacting reactive oxygen species (ROS) and programmed cell death (PCD), leading to reduced viability. Understanding Ca2+ regulation is key to improving pollen cryopreservation success.

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Area of Science:

  • Plant reproductive biology
  • Cellular and molecular biology
  • Cryobiology

Background:

  • Programmed cell death (PCD) contributes to reduced pollen viability after cryopreservation.
  • The specific role of calcium ions (Ca2+) in pollen PCD during cryopreservation remains largely unexplored.

Purpose of the Study:

  • To investigate the effects of Ca2+ fluctuations on PCD and reactive oxygen species (ROS) in cryopreserved Paeonia lactiflora 'Fen Yu Nu' pollen.
  • To elucidate the mechanism by which Ca2+ influences PCD during pollen cryopreservation.

Main Methods:

  • Cryopreservation of Paeonia lactiflora 'Fen Yu Nu' pollen.
  • Measurement of Ca2+ content, PCD indices (e.g., mitochondrial membrane potential, caspase activity, apoptosis rate), and ROS levels.
  • Application of Ca2+ carrier (A23187) and Ca2+-chelating agent (EGTA) to modulate Ca2+ levels.
  • Analysis of pro-PCD and anti-PCD gene expression.

Main Results:

  • Cryopreservation led to increased Ca2+ content and decreased pollen viability.
  • Ca2+ modulation significantly affected PCD indices, with A23187 accelerating PCD and EGTA showing opposite effects.
  • Ca2+ levels influenced ROS production and clearance systems, with A23187 increasing ROS and EGTA decreasing it.
  • Gene expression analysis revealed altered regulation of PCD-related genes.

Conclusions:

  • Increased Ca2+ content after cryopreservation is linked to enhanced ROS production and subsequent PCD in pollen.
  • Ca2+ plays a critical role in regulating ROS homeostasis and PCD pathways during pollen cryopreservation.
  • Targeting Ca2+ signaling pathways presents a potential strategy to improve pollen cryopreservation outcomes.