| Plant latin name | Scutellaria baicalensis Georgi |
| Literature code | Scutellaria_baicalensis-Ref-1 |
| Reference | Li H et al, Plant Cell, Tissue and Organ Culture, 62: 169-173 (2000) |
| Summary | Development of an efficient in vitro propagation system for Huang-qin (Scutellaria baicalensis), a traditional
Chinese medicinal plant used in the treatment of a wide range of human ailments, is described. Thidiazuron [TDZ:
N-phenyl-N'-(1,2,3-thidiazol-5-ylurea)] effectively induced regeneration on cultured intact seedlings, etiolated hypocotyl explants and sterile stem segments of Huang-qin. Histological examinations of excised hypocotyl or nodal
explants revealed that adventitious shoots formed through an intermediate callus. Comparison of TDZ-induced
regeneration in the three tissue types indicated that isolation of explants was not essential for optimal regenerative
efficiency. Significantly more regenerants formed along hypocotyls of intact seedlings (20 shoots/explant) than
were observed on excised hypocotyls (9.7 shoots/explant) indicating that endogenous metabolites produced in
adjacent tissues provided resources for the shoot initiation. More than 95% of de novo regenerants formed roots
and then intact plantlets under either sterile culture or greenhous conditions. Regeneration protocols developed
in this study may provide the basis for improvement of this crop through the identification of medicinally active
constituents and eventual development optimized pharmaceutical products. |
| Objectives | Development of an efficient in vitro propagation sysytem for Scutellaria baicalensis |
| Materials | Scutellaria baicalensis seeds obtained from Richter's Herb's Inc. (Goodwood, ON, Canada) |
| Explant | Seeds |
| Initial culture | Seeds were surface sterilized by dipping in 95% ethanol for 30 second, then immersion in 1.5% sodim hypochlorite containing Tween-20 (2drops per 100 ml solution) for 18 min, followed by 3 rises with sterile distilled water. Seeds were cultured in Petri disheds containing 25 ml of culture medium containing Murashige and Skoog (MS) salts, Gamborg B5 (B5) vitamines and 3.0%(w/v) sucrose, hereafter referred to as MSO. The pH was adjusted to 5.75 and 0.3% gellan gum (Gelrite, Schweitzerhall Inc., South Plainfield, NJ, USA) was added before autoclaving at 121℃, 1.4 kg cm-2 for 20 min. The cultures were sealed with Parafilm and incubated in a growth chamber at 24±2 ℃ with 16-h photoperiod provided by cool-white fluoresxent tubes at 30-35 μmol m-2s-1 (Model F40/CW/RS/EW-II Philips Canada, Scarborough, ON, Canada). More than 95% seeds germinated after 7 days of culture. |
| Shoot multiplication | For intact seedling cultures, MSO was supplemented with 0, 2.5, 5.0, 7.5, 10.0, or 20.0 μmol L-1 of thidiazuron (TDZ) and 4 ml L-1 Plant Preservative Mixture (PPM, Plant Cell Technology, Inc., Washinton DC). By day 14, all of the germinated seedlings cultured on TDZ-containing medium showed de novo shoot formation. Cultures on medium with 2.5μmol L-1 TDZ had an average of 19 shoots per seedling, while seeds germinated on medium without TDZ had an average of 2 shoots. There was no significant increase in the number of shoots formed as a result of increasing the concentration of TDZ to levels higher than 2.5μmol L-1. |
| Rooting | De novo shoots were excised and subcultured on either liquid MSO, semi-solid MSO or sterilized peat pellets (Premier Brands Inc. Stratford, ON, Canada) in Magenta boxes after 2 months of culture. More than 95% of excised regenerants formed roots and intact plants. |
| Acclimation | Plantlets were transferred to a misting-bed system under standard greenhouse conditions following incubation on sterile peat pellets for one month. Plants were acclimatized for 2 weeks. |
| Planting | Acclimatized plants were repotted in a greenhouse soil mixture (Promix BX, Plant Products, Brampton, ON, Canada) for growth and maintenance in the greenhouse envirnment. |
| Cultivation conditions | Plants were maintained in the greenhouse environment. |
| Traints of regenerants | All plantlets were eventually grown to maturity under standard greenhouse conditions. |
| Ingredients analyzed | |
| Extraction | |
| Analitical methods | |
| Notes | |