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Isolation of functional chloroplasts from the sacoglossan mollusc Elysia viridis Montague
Author(s) -
WILLIAMS MICHAEL L.,
COBB A. H.
Publication year - 1989
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/j.1469-8137.1989.tb04701.x
Subject(s) - biology , chloroplast , botany , differential centrifugation , biochemistry , gene
SUMMARY A novel technique has been developed in which functional chloroplasts have been isolated and purified from the symbiotic sacoglossan mollusc Elysia viridis , Montague. This entailed the progressive filtration and gel‐filtration of a hyaluronidase‐incubated homogenate. Isopycnic centrifugation of the resultant crude chloroplast suspension reduced the cytoplasmic/mitochondrial and microbody contamination by approx. 70 and 90% respectively. Purified chloroplast suspensions were capable of linear rates of 14 CO 2 fixation for at least 25 min at 50 μmol m 2 s −1 (PPFD), even though approx. 54% of assimilate was being released into the bathing medium. Values for CO 2 fixation and assimilate release were similar to those obtained for the intact mollusc.