Effects Of Amino Acids,PGRs And E₀ Gene On Accumulations Of Secondary Metabolisms In Asarum Heterotropoides Fr.Schmidt Var.Mandshuricum （Maxim.） Kitag.

A. heterotropoide, a kind of Traditional Chinese Medicine, is widely used in clinic because of its variety pharmacological activities. However, the usage, doses and position of A. heterotropoides is limited a lot, because it contains Aristolochic Acid I（AAI） which can cause kidney injury. Thereby, it becomes a hot topic about how to increase the active ingredients and decrease toxic ingredients. Exogenous amino acids, plant growth regulator（PGRs） and E₀ gene have been proved to promote plant growth. In addition, they could regulate plant secondary metabolism. Nevertheless, it lacks of the researches about the biosynthesis of AAI which is the toxic component and Asarinin（ASA） which is the ingredient component. Thus, this study was done. The results were as follows:1. Effect of exogenous amino acids on accumulations of secondary metabolites in A. heterotropoides: The experiment processing results of effects of exogenous amino acids on living tissue demonstrated that the 2 aromatic amino acid L-tyrosine（L-Tyr） and L-phenylalanine（L-Phe） could increase both AAI and ASA concentrations（P<0.05）; L-histidine（L-His） heterocyclic amino acid could decrease AAI concentration and increase ASA concentration at the same time（P<0.05）; L-proline（L-Pro） sub-heterocyclic amino acids and L-glycine（L-Gly） aliphatic amino acid could decrease AAI concentration without affecting ASA. The results of field experiment of exogenous amino acids application showed that proper concentrations and sprayed times of L-Gly, L-Pro or L-His could decrease AAI accumulation in A. heterotropoides leaves（P<0.05）. After 5 days of the treatment of 50 mM L-Pro for 3 times, AAI concentration in A. heterotropoides leaves was reduced 23.83% which was the maximum of reduced value; ASA accumulation was unchanged. After 5 days of the treatment of 100 mM L-His for 2 times, AAI concentration was reduced 12.74% and ASA concentration was increased by 17.98% in A. heterotropoides leaves.2. Effect of exogenous PGRs on accumulations of secondary metabolites in A. heterotropoides: The experiment processing results of effects of exogenous amino acids on living tissue demonstrated that the high concentration（0.2 mg·L-1） of TDZ could increase ASA accumulation（P<0.05） without affecting AAI accumulation（P>0.05）. IBA could increase the accumulation of AAI and ASA, among which the increase of ASA was higher（P<0.05）. The results of field experiment of exogenous PGRs application showed that after 5 days of the 3 times treatments of 10 mg·L-1 IBA could increase AAI by 17.00% and ASA by 71.91% in A. heterotropoides leaves.3. Effect of exogenous E₀ gene on accumulations of secondary metabolites in A. heterotropoides: GM Asarum was obtained successfully through pollen-tube pathway. E₀ gene made the ASA concentration of GM Asarum seeds remarkably higher than the non-GMO Asarum seeds（P<0.05） without affecting AAI concentration（P>0.05）. E₀ gene stimulated the roots growth of A. heterotropoides.（P<0.05）.