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Title: A simplified method for differential staining of aborted and non-aborted pollen grains

item PETERSON, ROSS - University Of Minnesota
item Slovin, Janet
item CHEN, CHANG-BIN - University Of Minnesota

Submitted to: International Journal of Plant Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/16/2010
Publication Date: 7/16/2010
Citation: Peterson, R., Slovin, J.P., Chen, C. 2010. A simplified method for differential staining of aborted and non-aborted pollen grains. International Journal of Plant Biology. 1:13.

Interpretive Summary: The production of pollen by plants is necessary for fruit and seed production. In strawberries, heat results in poor pollen development, leading to fewer fruit being produced per plant and fruit of poor quality. Research to produce strawberry plants that are not affected by heat requires an easy way to tell if pollen is normal or if development of pollen was aborted. A common way to do this is to stain pollen with chemicals that give a different color for dead or live pollen. The best stains available use chemicals that are hazardous or require expensive equipment to see. We have developed a pollen staining technique that is short, safe, and easy to use. This technique will be useful for breeders developing heat tolerant strawberry varieties, and researchers working to determine why heat affects pollen development.

Technical Abstract: The ability to use chemical staining to discriminate aborted from non-aborted pollen grains has well-known practical applications in agriculture. A commonly used technique for assessing pollen vitality, Alexander’s stain, uses chloral hydrate, phenol and mercuric chloride, all of which are highly toxic. We describe here an improved pollen staining technique that avoids the use of a regulated chemical chloral hydrate and two extremely toxic chemicals mercuric chloride and phenol, and requires a much shorter time period for sample preparation and staining. This simplified method is very useful for field studies without high-end equipment such as fluorescence microscopes. Samples can be collected and fixed in the fields, then examined in a laboratory that has simple light microscopes.