4.6 Article

Rapid separation of developing Arabidopsis seeds from siliques for RNA or metabolite analysis

Journal

PLANT METHODS
Volume 9, Issue -, Pages -

Publisher

BIOMED CENTRAL LTD
DOI: 10.1186/1746-4811-9-9

Keywords

Arabidopsis; Seed; Silique; Gene expression; Dissection; Harvest; Metabolic quench

Funding

  1. U.S. National Science Foundation [DBI-0701919]
  2. Agricultural Research Center at Washington State University

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Background: Protein, starch and oil produced in plant seeds are major renewable sources of food, chemicals and biofuels. Developing Arabidopsis thaliana seeds are commonly utilized as a model for seed crop research. However, due to the very small size of Arabidopsis seeds efficient collection of large amounts of tissue for gene expression or metabolite analysis is very difficult and time consuming. Results/conclusions: Here we describe a method that allows very rapid separation and collection of large amounts of developing Arabidopsis seeds from their encapsulating silique tissue after flash freezing whole siliques in liquid nitrogen. The efficient popping open of the frozen siliques on dry ice and filtering the seeds away from the silique tissue with liquid nitrogen cooled funnels and sieves allows large amounts of developing seeds to be quickly isolated while remaining frozen. This method increases the speed of developing seed collection approximately 10 fold over methods which dissect individual siliques one at a time.

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