Oryzalin-induced Chromosome Doubling In Scutellaria Barbata D.Don. In Vitro

Scutellaria barbata D.Don for lamium radix scutellariae genus of medicinal plants, a perennial herb with all the grass medicine was studied medicinal value for anticancer, antipyretic, anti-inflammatory, antibacterial, antiviral effect and antioxidant. This study used tissue culturing seedlings of polyploidy induced by oryzalin; Then, identified of the polyploidy in the morphology, cytology and molecular biology. Finally the polyploids were determined in medicinal ingredients of flavonoids content and indexes of physiological resistant.This would cultivate the new medicinal value high germplasm.First, Scutellaria barbata with stem segments auxiliary buds and shoot tips as explants, used of oryzalin which was less harmful to the environment as inducer, the effects of different concentrations and time ducation was studied using the method of drifting treatment. The results showed that using stem segments with axillaries as explants, the appropriate combination was 10μmol/L oryzalin induced adventitious buds for 12 hour; but the appropriate combination was 10μmol/L oryzalin induced 24 h using shoot tips. The use of stem segments with axillary buds as explants induced better than shoot tips.Second, plant morphology, stomata, guard cells, the number of guard cell chloroplasts, stomatal density, chromosome number, nuclear DNA content were compared between the variation plants and control. The results showed that: compared with the control plants, variation plants were slower growth, compact plant type, leaves larger and darker green, stem shorter and thicker, smaller leaf index, stomatal length, stomatal width, guard cell length, width of guard cells, the number of pores within the chloroplast of variant plants were significantly increased, but stomatal density was significantly reduced. The variants identified by the observation of external morphology and stomatal, used root tip squash method for identification of chromosome number. The resules showed that the treated variation plants which existed a tetraploid chromosome number 2n=4x=52; and control was a normal diploid 2n=2x=26. Identification of flow cytometry also got the same conclusion.Finally, the doubling plantlets were transplanted to determine the medicinal ingredient of total flavonoids and resistance to commonly used physiological indicators of superoxide dismutase (Superoxide dismutase, SOD), catalase (Catalase, CAT), malondialdehyde (MDA) and soluble sugar content. The results showed that the three polyploid lines of total flavonoids were significantly higher than the control of diploid plants. Lines one, lines three, lines four increased the total flavonoid content 34.16%,37.13%,40%. Three polyploid lines of SOD, CAT activity were significantly different than the control diploid plants. SOD activity of lines three were 44.7% higher than diploid and CAT activity 16.8% higher than the diploid. Three polyploid lines of MDA were significantly lower than the diploid plants, which lines three of MDA were 47.6% lower than the diploid lines. Three polyploid lines of soluble sugar decreased than the control, which lines three of soluble sugar were 29.5% lower than the diploid lines but lines one were no significant difference whith the diploid lines.