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High yield isolation of mesophyll protoplasts from wheat, barley and rye
Author(s) -
Sarhan Fathey,
Cesar Dismie
Publication year - 1988
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1988.tb05842.x
Subject(s) - protoplast , secale , biology , hordeum vulgare , ascorbic acid , petri dish , botany , incubation , poaceae , horticulture , agronomy , biochemistry , microbiology and biotechnology
Efficient procedures are described for high‐yield isolation of mesophyll protoplasts from spring wheat ( Triticum aestivum L. cv. Glenlea), winter wheat ( Triticum aestivum L. cv. Frederick), barley ( Hordeum vulgare L. cv. Bruce) and rye ( Secale cereale L. cv. Puma). Factors such as plant age, composition of the incubation medium during isolation, purification procedures and culture medium affect protoplast yield, viability and metabolic competence, as measured by light‐dependent CO 2 fixation. Optimal osmolarity of the isolation medium was equivalent to 1.8 times that measured in the leaves of all plant material used. The presence of 2 m M ascorbic acid in the preincubation and isolation medium increased the yield by 50% and conserved viability and metabolic competence. The protoplasts were stable for up to 48 h without loss of either viability or of original activity of CO 2 fixation, which was in the order of 100 μmol CO 2 (mg chl) −1 h −1 . In our MC‐56 liquid medium these protoplasts regenerated cell walls within 72 h and a few divided.