The Establishment Of Regeneration System And The Regulation Of Secondary Metabolites Of Pinellia Ternate

In this paper, the modern technology which is tissue culture of Pinellia ternata as test materials was use, regeneration system were built; the effect exogenous additives of Phe, Asp, SNP, ASA on total alkaloids, guanosine and adenosine accumulation in the period of callus and seedling were researched, the activites of nitrogen metabolism key eyzymes and secondary metabolites related eyzymes were measured, and the regulation mechanism was studied. Moreover, the study was the theory base of Phe, Asp, SNP, ASA regulating total alkaloids, guanosine and adenosine production in P. ternate, Also it provided an important method and mean for large area cultivation and industrialized production of P.teranate.The main conclusions are as follows:1. Establishment of regeneration system of P. ternate. The petiole, blade, tuber were taken as explants cultured in vitro to induce callus. The results showed that petiole was the most suitable explants. By adding different hormones to MS medium, the influence of callus induction, proliferation and differentiation of Pinellia Tuber has been observed. The results showed that MS medium containing0.5mg/L2,4-D,1.5mg/L KT and1.0mg/L IAA is the most suitable for callus induction. The MS containing2.0mg/L2,4-D,1.0mg/L KT and1.0mg/L IAA is the optimum medium for callus proliferation under dark condition;0.2mg/L2,4-D and1.5mg/L6-BA is suitable for the budding and rooting of callus, the growth of tissue culture seedlings are good.2. Effects of different exogenous additives on growth and contents of total alkaloid s, guanosine and adenosine in P. ternate callus. Different concentration of Phe, Asp, SNP, ASA were added to culture medium of P.ternate callus respectively. The results showed that precursors of Phe and Asp could inhibit callus growth. However, induc-tion of SNP and ASA promoted the growth of callus in the range of testconcentrati--on. Low concentration of Phe increased the total alkaloid accumulation in callus of P. ternate, while high concentrations of Phe inhibited the accumulation. Different Con--centrations of Asp, SNP, ASA could promote the total alkaloids accumulation, and the best results of additives concentration were50mg/L,150mg/L,1mg/L,1mg/L respectively. Low concentration of Asp and SNP showed positive action on the synthe sis of guanosine in callus, but high concentration of them had adverse effects on the synthesis of guanosine, different concentrations of Asp and SNP could promote the synthesis of guanosine. The concentration of50mg/L,150mg/L,0.1mg/L,1mg/L gave the best effects; The precursors of Phe, Asp were conducine to the synthesis of adenosine in callus of P. ternate, and the concentration of50mg/L,25mg/L gave the best results, but SNP and ASA inhibited the accumulation of adenosine. It could be seen that the effects exogenous additives on the growth and the contents of total alkaloids, guanosine and adenosine in Pinellia callus were related to their types and concentrations3. In the paper, it was revealed that elicited total alkalodis,guanosine and adenosine synthesis was associated with regulated nitrogen-metabolism by SNP, ASA, Phe, Asp alone and two two synergistic effect in callus of P. ternate. on one hand, they could regulate the activities of GOG AT, GDH and GOT. The improving of GOG AT and GDH activities promoted glutamic acid transfering to other amino acids, furthermore, the amino acids synthesize proteins and energy and other nitrogenous compounds which furnished more precursors of secondary metabolism and energies. On the other hand, they induced PAL gene overexpression, increased the activities of PAL, accelerated phenylpropanoid metabolism, promoted the synthesis of alkaoids.1mg/L ASA coope--rate with150mg/L Asp were added to the culture medium, which enhanced the activities of GOGAT, GDH, GOT and PAL, and increased the contents of soluble protein, of which gave the best results.4. Effects of spraying different concentrations of SNP on total alkaloid, guanosine and adenosine contents in regenrated P. ternate. SNP enhanced the activities of PAL, GOT and GOGAT, and increased the contents of total alkaloids, guanosine and adenosine of regenerated P. ternate in different concentration treatments, of which1mmol/L SNP gave the best results and increased alkaloids and guanosine contents by66.1%and188.3%, respectively. SNP promoted the activities of PAL, GOGAT and GOT in regenerated P. ternate, which accelerated the assimulation and conversion efficiency of inorganic nitrogen and furnish more precursor of secondary meta-bolism. Futhermore, the contents of alkaloids and guanosine increased correspondingly.