Preparation and vitality detection of protoplast in Salvia miltiorrhiza Bunge.
- Author:
Nan ZHU
;
Jun LIU
;
Xinyu ZHANG
;
Juan'e DONG
- Publication Type:Journal Article
- MeSH:
Aniline Compounds;
chemistry;
Cell Culture Techniques;
Cellulase;
chemistry;
Fluorescent Dyes;
chemistry;
Protoplasts;
chemistry;
Salvia miltiorrhiza;
growth & development;
Xanthenes;
chemistry
- From:
Chinese Journal of Biotechnology
2014;30(10):1612-1621
- CountryChina
- Language:Chinese
-
Abstract:
We prepared protoplasts from Salvia miltiorrhiza Bunge suspension culture cells. Then, the protoplasts' vitality and functions were tested by fluorescein diacetate staining method and Fluo-3/AM flourescent probe. The optimal condition of protoplast isolation was Cellulase R-10 1.5%, Pectinase Y-23 0.3%, Macerozyme R-10 0.5%, 40 r/min 12 h, 600 r/min 5 min, and the protoplasts yield was 1.1x10(6) cells/g FW, the vitality was more than 95% by using fluorescein diacetate staining method. It has been confirmed that calcium fluorescent probe Fluo-3/AM can be successfully loaded into protoplasts.
