A robust and rapid pollen viability test using impedance flow cytometry for high throughput screening of heat tolerant wheat (Triticum aestivum) germplasm

Anowarul I. Bokshi1, Daniel K.Y. Tan1, Richard M. Trethowan1

1The University of Sydney, Plant Breeding Institute, Sydney Institute of Agriculture, School of Life and Environmental Sciences, NSW 2006, anowarul.bokshi@sydney.edu.au

Abstract:

Pollen viability is an important physiological character for the screening of heat tolerant wheat germplasm. A robust and rapid analysis is essential to determine pollen thermotolerance. Our pre-breeding program assessed the Ampha Z32 impedance flow cytometer from Amphasys as a tool for rapid determination of pollen viability. The Ampha Z32 detects viable pollen based on the electric properties of cells which react to an applied alternating current, simultaneously generating information on cell size, membrane integrity and cytoplasmic conductivity. This study tested the hypothesis that pollen viability assessment using the conventional method by staining and counting under a microscope is comparable with the Ampha Z32 for rapid analysis of wheat pollen viability under high temperature stress. Pollen viability of three wheat genotypes was examined after heat stress during meiosis including an untreated control, using both the Ampha Z32 and conventional Lugol’s solution (KI)-stain-count. The cytometer provided comparable data and positive correlations with the conventional staining method.. The cytometer-generated data can enable much larger populations to be screened at lower cost, thus enhancing selection of wheat pollen for heat tolerance.

Host

The Australian Society of Agronomy is the professional body for agronomists in Australia. It has approximately 500 active members drawn from government, universities, research organisations and the private sector.

Photo Credits

David Marland Photography david_marland@hotmail.com Graham Centre for Agricultural Innovation, Charles Sturt University

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