Rare And Endangered Medicinal Plants Uniflora Tissue Culture Isolated Endophytes

Kingdonia uniflora Balf. f. et W. W. Smith belongs to the Circaeasteraceae and is the only species of Kingdonia. It is one of the most endangered species in China and was ranked as the first grade of National Protected Plants and the key species of Chinese biodiversity. Kingdonia uniflora seeds have largely failed to mature in the field environmental conditions, rely mainly on the top of rhizome elongation reproduction, survive in the harsh environment, poor natural regeneration. Coupled with the destruction of forest vegetation and excavation, result in mutation of the natural environment, its plants declined and shrinking natural distribution While the single mode propagation can not be restored. This experiment was maiuly to probe an effieient way through tissue culture, which offer the theory foundation for enlarging source of Kingdonia uniflora. Meanwhile, endophytic fungi Kingdonia uniflora was isolated and identified in growth of it. As follows:1.The establishment of regeneration system of Kingdonia unifloraIn the experiment, The effects of the different sterilization of explants, different basic medium (MS,1/2MS,1/4MS, B5, N6, White), different hormones, different concentrations and combinations (auxins (NAA,2,4-D, IAA, IBA) and cytokinins (6-BA, KT, ZT, TDZ)), different culture conditions and different explants on the specific role of growth and differentiation of Kingdonia uniflora were debated.(1) The sterilization of explants:Different explants are sterilized at different time by using 0.1% mercuric chloride solution and 2% sodium hypochlorite solution and found that the rhizome with 0.1% mercuric chloride solution,3 min more suitable disinfection, mature laminae with 2% sodium hypochlorite disinfection 1 min more suitable, mature petiole sterilized with 0.1% mercuric chloride solution,1 min or 2 min more suitable, Buds sterilized with 0.1% mercuric chloride solution,4 min more suitable(2) The experiment on induction of Kingdonia uniflora:The different explants are cultured in culture conditions with different basic medium, different hormones, different concentrations and combinations. The results indicated that the 1/4MS media was the most suitable basic medium for the most suitable explants of young petiole of Kingdonia uniflora, and Callus, adventitious buds, adventitious roots are induced. While the induction rates of rhizomes, mature petioles, mature laminae and bud scales were zero. Only 2,4-D can induce callus, but the callus become brown and dead.2. Isolation and identification of endophytic fungi of Kingdonia uniflora (1) Roots surface and inner of Kingdonia uniflora:The surface of Kingdonia uniflora roots were obsered by scanning electron microscopy, and we found that there are two or more regions clustered filaments on the root sruface. Through hand slicing and lactic acid phenolic cotton blue staining,the internal structure of Kingdonia uniflora root was observed and found that hyphae exist in epidermal cells.(2) 5 fungal strains have been isolated. Isolates were identified on the basis of colonial morphology and microscopic characteristics of hpyhae, spores, conceptacle, the results showed that the resulting strains are mainly Ascomycetes and Imperfect fungi, The superior genus is Pencillium...