High-yield isolation of protoplasts from microgram amounts of shoot meristematic tissues and rapid DNA content determination by flow cytometry.
This paper describes a novel approach to rapid cell-cycle analysis of shoot meristematic cells. The method involves fixation and disaggregation of meristems into protoplast suspension and flow-cytometric analysis of these protoplasts stained with fluorescent dyes. We have developed a procedure for a high-yield isolation of protoplasts allowing an accurate flow-cytometric analysis with a few micrograms of meristem tissues. We present here determinations of total DNA content of protoplasts stained with propidium iodide in the dicotyledon Sinapis alba, and the monocotyledon Lolium temulentum.
Houssa
C
C
Département de Botanique, Université de Liège, Belgium.
Bomans
J
J
Greimers
R
R
Jacqmard
A
A
D002453:Cell Cycle; D002458:Cell Fractionation / Q000379:methods; D002467:Cell Nucleus / Q000648:ultrastructure; D004247:DNA / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods; D059828:Plant Cells; D010944:Plants / Q000737:chemistry / Q000648:ultrastructure; D011523:Protoplasts / Q000737:chemistry / Q000648:ultrastructure; D013045:Species Specificity
D004247:DNA