Total RNAs were isolated from cultured roots of Scopolia parviflora, poly(A)+ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.
Optimal culture conditions for efficient in vitro propagation and polysaccharide production of Orostachys japonicus were established. The highest growth yield was achieved in 1/2 MS medium, while the lowest growth yield was obtained in 4 MS medium. The patterns of polysaccharide formation were a little similar in all cases, but on MB5 medium, the po]ysaccharide contents of plant were higher than others. Among the modified nitrate levels, effective growth level were obtained in 1/4 N and 1/2 N. High contents of polysaccharide were obtained in 4 N. Different concentration of potassium and calcium did not improve the growth and polysaccharide production. The micropropagated shoots were successfully acclimatized artificial soils.