ISSN:
1432-2048
Keywords:
Amyloplast purification
;
Endosperm
;
Triticum (amyloplast purification)
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract A rapid method for the isolation and purification of amyloplasts from the endosperm of developing grains of Triticum aestivum L. has been developed. Cell-free 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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00198226
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