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Direct analysis of pollen fitness by flow cytometry: implications for pollen response to stress.
- Source :
-
The Plant journal : for cell and molecular biology [Plant J] 2019 Jun; Vol. 98 (5), pp. 942-952. Date of Electronic Publication: 2019 Mar 18. - Publication Year :
- 2019
-
Abstract
- Sexual reproduction in flowering plants depends on the fitness of the male gametophyte during fertilization. Because pollen development is highly sensitive to hot and cold temperature extremes, reliable methods to evaluate pollen viability are important for research into improving reproductive heat stress (HS) tolerance. Here, we describe an approach to rapidly evaluate pollen viability using a reactive oxygen species (ROS) probe dichlorodihydrofluorescein diacetate (i.e. H <subscript>2</subscript> DCFDA-staining) coupled with flow cytometry. In using flow cytometry to analyze mature pollen harvested from Arabidopsis and tomato flowers, we discovered that pollen distributed bimodally into 'low-ROS' and 'high-ROS' subpopulations. Pollen germination assays following fluorescence-activated cell sorting revealed that the high-ROS pollen germinated with a frequency that was 35-fold higher than the low-ROS pollen, supporting a model in which a significant fraction of a flower's pollen remains in a low metabolic or dormant state even after hydration. The ability to use flow cytometry to quantify ROS dynamics within a large pollen population was shown by dose-dependent alterations in DCF-fluorescence in response to oxidative stress or antioxidant treatments. HS treatments (35°C) increased ROS levels, which correlated with a ~60% reduction in pollen germination. These results demonstrate the potential of using flow cytometry-based approaches to investigate metabolic changes during stress responses in pollen.<br /> (© 2019 The Authors The Plant Journal © 2019 John Wiley & Sons Ltd.)
- Subjects :
- Arabidopsis cytology
Arabidopsis metabolism
Arabidopsis physiology
Cell Survival physiology
Flow Cytometry
Flowers cytology
Flowers metabolism
Solanum lycopersicum cytology
Solanum lycopersicum metabolism
Solanum lycopersicum physiology
Oxidative Stress physiology
Pollen cytology
Pollen metabolism
Pollen Tube cytology
Pollen Tube metabolism
Pollen Tube physiology
Reactive Oxygen Species metabolism
Adaptation, Physiological physiology
Flowers physiology
Heat-Shock Response physiology
Pollen physiology
Pollination physiology
Subjects
Details
- Language :
- English
- ISSN :
- 1365-313X
- Volume :
- 98
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- The Plant journal : for cell and molecular biology
- Publication Type :
- Academic Journal
- Accession number :
- 30758085
- Full Text :
- https://doi.org/10.1111/tpj.14286