Large-scale isolation of intact vacuoles and isolation of chloroplasts from protoplasts of mature plant tissues

Journal Article · Fri Dec 26 00:00:00 EST 1975 · Science; (United States)

DOI:https://doi.org/10.1126/science.190.4221.1298· OSTI ID:7199543

Wagner, G J; Siegelman, H W

Intact plant vacuoles were prepared in large numbers (10/sup 6/) from protoplasts of mature leaves, flower petals, stems, pedicels, filaments, styles, and young fruits. Treatment of protoplasts with 0.2 molar K/sub 2/HPO/sub 4/-HCl, pH 8, with slow stirring resulted in gentle osmotic rupture of the protoplasts and release of intact vacuoles. Particulate components of the protoplast, less the vacuole, were largely shed as an aggregate, which was removed by filtration. Vacuoles were recovered from the filtrate by low-speed centrifugation. The general procedure was also used to isolate chloroplasts with a high degree of integrity and excellent photochemical activity. (auth)

• Research Organization: Brookhaven National Lab., Upton, NY
• OSTI ID: 7199543
• Journal Information: Science; (United States), Vol. 190:4221
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CHLOROPLASTS
PLANT CELLS
SEPARATION PROCESSES
PLANTS
TISSUES
BIOMASS
BODY
CELL CONSTITUENTS
ENERGY SOURCES
RENEWABLE ENERGY SOURCES
550800* - Morphology