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ISOLATION AND PURIFICATION OF GLANDULAR SECRETORY CELLS FROM ARTEMISIA TRIDENTATA (SSP. VASEYANA) BY PERCOLL DENSITY GRADIENT CENTRIFUGATION
Author(s) -
Slone J. Henry,
Kelsey Rick G.
Publication year - 1985
Publication title -
american journal of botany
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.218
H-Index - 151
eISSN - 1537-2197
pISSN - 0002-9122
DOI - 10.1002/j.1537-2197.1985.tb08402.x
Subject(s) - percoll , biology , differential centrifugation , centrifugation , trichome , density gradient , artemisia , chromatography , botany , concentration gradient , microbiology and biotechnology , biochemistry , chemistry , physics , quantum mechanics
Glandular trichome heads (the secretory cells), obtained by mechanical homogenization of young floral buds and subtending leaves of Artemisia , were isolated and partially purified in discontinuous and continuous Percoll density gradients. With discontinuous gradients, the mixed‐cell suspension was fractionated on four layers of Percoll with increasing densities: 0, 1.048, 1.068, and 1.084 g/ml. Gland heads banded primarily at the 0/1.048 interface, mesophyll cells at the 1.068/1.084 interface, and the hairs and hair fragments pelleted at the bottom of the tube. Twenty to thirty percent of the cells in the 0/1.048 band were intact gland heads, which represented about half of those recovered from the gradient. Hairs were the major contaminant. Over 90% of the gland heads excluded Evan's blue dye and were apparently viable. Similar results were obtained from preliminary experiments using continuous density gradients. The whole procedure for either method requires 3–6 hr, depending on the amount of starting material.