[Preparation and vitality detection of protoplast in Salvia miltiorrhiza Bunge].
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.
Nan
N
Zhu
Jun
J
Liu
Xinyu
X
Zhang
Juan'e
J
Dong
D000814:Aniline Compounds / Q000737:chemistry; D018929:Cell Culture Techniques; D002480:Cellulase / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D011523:Protoplasts / Q000737:chemistry*; D027561:Salvia miltiorrhiza / Q000254:growth & development*; D014966:Xanthenes / Q000737:chemistry
D000814:Aniline Compounds; D005456:Fluorescent Dyes; D014966:Xanthenes; C059715:Fluo-3; D002480:Cellulase