The Optimization Of Inducing Conditions And Studies On The Contents Of Alkaloids Of Pinellia Ternata Tuber

Pinellia ternata (Thunberg) Tenore ex Breitenbach, which belongs toAraceae, is a perennial herbaceous plant. The medicinal part of P. ternata is its tuber.The fertility of cultivated P. ternata was low, and the quality of P. ternata oftendegenerated due to virus infected, resulting P. ternata provenance was often unable tomeet the demand. At present, due to the lack of P. ternata tuber molecular mechanismof secondary metabolites, the development of its tubers grown artificial regulationtechnology was very slowly, resulting in the development of P. ternata production hasbeen severely constrained. Therefore, it was necessary to carry out related research ofP. ternate.Through this study, we obtained the following results:(1) After the experiment of P. ternata tissue culture and the plant regenerationsystem, we obtained a result that: we could acquire explants fast with the medium ofMS+6-BA1.0mg L-1+IAA0.5mg L-1. With this explant culture medium, the P.ternata tubers could soon form at both ends of the P. ternata petiole, which wasmeaningful for a quick acquisition of the P. ternata tubers. After the formation of theP. ternata tubers, tubers amplified and soon began to differentiate into leaves. Then,we cut the new P. ternata leaves. When the plant regrowthed, it could grow faster andacquire more leaves, and its roots could grow rapidly. This method allowsed the P.ternata tissue culture seedlings to grow leaves and roots in a short time. Researchingconducted by the end of this experiment needed small tubers that formed induced bythe P. ternata petioles. The P. ternata tissue culture seedling that cultured at this timecould provide enough P. ternata petioles. Therefore, the optimization study of P.ternata regeneration system was particularly important.(2) Through the experiment of P. ternata tuber induction, it was concluded that:in order to obtain undifferentiated P. ternata tubers, the experiment used inverted P.ternata petioles. With the accumulation of the time, P. ternata tubers gradually formed.P. ternata tubers was the best when it was cultured for25days. When it was culturedfor30days, P. ternata tubers began to differentiate and grew white roots. At this time,P. ternata tubers were not suitable as suspension culture materials. Therefore, P.ternata tubers that cultured for25days were the best experimental materials. In MSmedium, inverted P. ternata petioles could obtain P. ternata tubers and maintain greenand stout petioles. The P. ternata petioles could not obtain P. ternata tubers when theywere cultured without inversion, and the P. ternata petioles would gradually wither.(3) Through the experiment of the establishment of P. ternata tubers suspensionculture system under the salicylic acid stimulation, it was concluded that: the freshweight and dry weight of the suspension P. ternata tubers of the control group was increasing as the time increased, which indicated that the biomass of the suspension P.ternata petioles were accumulating. The fresh weight and dry weight of thesuspension P. ternata tubers of the treatment group were increasing slowly in0~10days. Through this, it could speculate that P. ternata tubers could not adapt well to thechange of the environment by adding exogenous salicylic acid, causing the growthinhibition of the suspension cultured P. ternata tubers and leading to a slow growth ofthe fresh weight and dry weight of the P. ternata tubers. It could conclude that: theadded exogenous salicylic acid had some certain influence to the growth of the P.ternata tubers.(4) Through the experiment of the P. ternata tubers single alkaloid contentdetermination of guanosine and inosine under the salicylic acid stimulation by highperformance liquid chromatography (HPLC) method, it was concluded that: in theselection of the mobile phase, methanol: ultra pure water=5:95was the best choice.In the selection of the detection wavelength,254nm wavelength was the best choice.We measured the content of the guanosine and inosine under the stimulation ofsalicylic acid using the Agilent1200HPLC, it was concluded that: P. ternata tubers,which were cultivated for10days, in the salicylic acid concentration of50μmol L-1,their guanosine content reached a maximum of1.353mg g-1. P. ternata tubers, whichwere cultivated for30days, in the salicylic acid concentration of200μmol L-1, theirinosine content reached a maximum of0.149mg g-1.