Identification and sorting of plant heterokaryons
AssigneeBoard of Regents of the University of Nebraska
InventorDavid W. Galbraith
Life of this patent
Estimating the term…
Abstract
To label heterokaryons formed by fusion of different somatic tissue cells, somatic tissue cells from one plant are immersed in a first solution including polysaccharidase and fluorescein isothiocyanate and somatic tissue cells from a different plant are immersed in a second solution including polysaccharidase and rhodamine isothiocyanate for a period of from 12 to 18 hours, after which protoplasts emerge, with those from one plant fluorescing green-yellow and those from the other fluorescing orange-red. The protoplasts are then fused in a polyethylene glycol solution. The heterokaryons are identified and sorted by a commercial cell sorter which relies upon the fluorescence from both markers.
Claims17
A method of parasexual hybridization of plants comprising the steps of:
- removing the cell wall from and marking a first group of cells that are able to regenerate and are taken directly from intact plant organs to release marked protoplasts;
- said step of removing the cell wall from cells and marking a first group of cells comprising the step of removing the cell wall from the cells and marking the protoplasts in steps which occur at least in part concurrently;
- removing the cell wall from and marking a second group of cells that are able to regenerate to release marked protoplasts;
- the step of removing the cell wall from and marking a second group of cells comprising the step of removing the cell wall from the cells and marking the protoplasts in steps that are performed at least in part concurrently;
- fusing the marked protoplasts from said first group of cells with the marked protoplasts from said second group of cells;
- and
- identifying heterokaryons from said markings.
Dependencies are inferred from the claim text.