IN VITRO METHODS FOR THE INDUCTION AND MAINTENANCE OF PLANT CELL LINES AS SINGLE SUSPENSION CELLS WITH INTACT CELL WALLS, AND TRANSFORMATION THEREOF

摘要

The subject invention provides simple and consistent methods to break sus pension cell aggregates to single cells with intact primary cell walls. The subject invention relates in part to cell separation of suspension cell aggr egates cultured in medium containing pectin- degrading enzymes or tubulin de -polymerizing compounds including colchicine. The subject invention also rel ates to novel uses of compounds for such purposes. Another aspect of the sub ject invention relates to transformation of the subject, isolated cells. Suc h processes simplify and integrate single-cell-based transformation and sele ction processes into transgenic and transplastomic event-generation work pro cesses. The subject invention also removes technical constraints and produce s marker- free and uniformly expressing transgenic lines in a high throughpu t fashion to support various needs of animal health, biopharma, and trait an d crop protection platforms.