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A rapid method for the isolation of purified amyloplasts from wheat endosperm

✍ Scribed by Ian J. Tetlow; Kerry J. Blissett; Michael J. Emes

Publisher
Springer-Verlag
Year
1993
Tongue
English
Weight
423 KB
Volume
189
Category
Article
ISSN
0032-0935

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✦ Synopsis

A rapid method for the isolation and purification of amyloplasts from the endosperm of developing grains of Triticum aestivum L. has been developed. Cellfree amyloplasts were mechanically isolated from plasmolysed tissue, and then purified by low-speed centrifugation through a single layer of Nycodenz sedimenting onto a cushion of agar. Recovery of amyloplasts was greater than 20% with less than 1% contamination by cytosol, 0.2% by mitochondria, 0.5% by endomembrane system and no contamination by microbodies. This method yields preparations which are routinely 55-65% intact up to 2 h after extraction. Amyloplast integrity was shown to depend upon the external sorbitol concentration, and amyloplastic enzymes in intact preparations were protected from digestion by trypsin.

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