Abstract
Large numbers of vacuoles (106-107) have been isolated from Sorghum bicolor protoplasts and analyzed for the cyanogenic glucoside dhurrin. Leaves from light-grown seedlings were incubated for 4 hours in 1.5% cellulysin and 0.5% macerase to yield mesophyll protoplasts which then were recovered by centrifugation, quantitated by a hemocytometer, and assayed for cyanogenic glucosides. Mature vacuoles, released from the protoplasts by osmotic shock, were purified on a discontinuous Ficoll gradient and monitored for intactness by their ability to maintain a slightly acid interior while suspended in an alkaline buffer as indicated by neutral red stain. Cyanide analysis of the protoplasts and the vacuoles obtained there from yielded equivalent values of 11 μmoles of cyanogenic glucoside dhurrin per 107 protoplasts or 107 vacuoles. This work supports an earlier study from this laboratory which demonstrated that the vacuole is the site of accumulation of the cyanogenic glucoside in Sorghum.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beier H., Bruening G. The use of an abrasive in the isolation of cowpea leaf protoplasts which support the multiplication of cowpea mosaic virus. Virology. 1975 Mar;64(1):272–276. doi: 10.1016/0042-6822(75)90099-9. [DOI] [PubMed] [Google Scholar]
- Butcher H. C., Wagner G. J., Siegelman H. W. Localization of Acid hydrolases in protoplasts: examination of the proposed lysosomal function of the mature vacuole. Plant Physiol. 1977 Jun;59(6):1098–1103. doi: 10.1104/pp.59.6.1098. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Indge K. J. The isolation and properties of the yeast cell vacuole. J Gen Microbiol. 1968 May;51(3):441–446. doi: 10.1099/00221287-51-3-441. [DOI] [PubMed] [Google Scholar]
- Leigh R. A., Branton D. Isolation of Vacuoles from Root Storage Tissue of Beta vulgaris L. Plant Physiol. 1976 Nov;58(5):656–662. doi: 10.1104/pp.58.5.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakamura K. D. The isolation of vacuoles from Candida utilis. Prep Biochem. 1973;3(6):553–561. doi: 10.1080/00327487308061537. [DOI] [PubMed] [Google Scholar]
- Saunders J. A., Conn E. E. Subcellular localization of the cyanogenic glucoside of sorghum by autoradiography. Plant Physiol. 1977 Apr;59(4):647–652. doi: 10.1104/pp.59.4.647. [DOI] [PMC free article] [PubMed] [Google Scholar]