Ternata Tissue Culture Research

Pinellia ternate(Thunb.)Breit,is an important traditional Chinese herbal medicine,with the function of antitussive,expectorant,analgesic, anti-fertility,anticancer and improve intellect,having good prospect of exploitation.According to statistics,within the 558 kinds of traditional Chinese medicine prescriptions,P.ternate(Thunb.)Breit.has been listed as section 22 at the highest frequency of use of 30 kinds of Chinese medicine,which has a huge market demand.However,under natural conditions,P.ternate(Thunb.)Breit.mainly by bulbil,tubers for vegetative propagation,as breeding coefficient is low, reform of farming systems and the use of chemical herbicides,its wildlife resources have been seriously damaged.In addition,long-term process of vegetative propagation made P.ternate(Thunb.)Breit.easily infected with virus,and its production and medicinal ingredients were markedly decreased.In this paper,P.ternate(Thunb.)Breit.tuber is used as materials.By inducing the aseptic seedling explant in different hormones culture,and discuss the induced effects on P.ternate(Thunb.)Breit.with different hormones.Established the rapid propagation of tissue culture,selected the medium for the osteoporosis callus,thus laid the foundation for P.ternate(Thunb.)Breit.regeneration,and the determination of medicinal ingredients.The main experimental results are as follows:1.P.ternate(Thunb.)Breit.tuber is used as explant,which is cultured in medium MS supplement with different levels and different kinds of hormones.1.0mg/L 2,4-D is optimum for callus induction;0.5mg/L IAA is better for embryoid formation,and suitable for tubercle induction; NAA is mainly to induce callus rooting.MS + 2.0mg/L NAA is optimum rooting medium;MS + 0.5mg/L KT is better for tuber proliferation and multiplication.2.Use leaf blade and petiole of P.ternate(Thunb.)Breit.as explant, cultured on medium MS supplemented with different levels and different kinds of hormones through single-factor-induced and orthogonal experimental designed.The results showed that:Low concentrations of auxin and cytokinin are conducive for tubercle induction.0.2mg/L IAA and 0.5mg/L 6-BA is appropriate;when the concentration up to 1.0mg/L, leaf blade explants induced by IAA can produce the large number of tubercle,However,an excess number of tubercle affected its germination. Both KT and ZT induce white callus,then from white callus differentiated tubercles,which slow down the germination rate of tubercles;when the concentration increased to 2.0mg/L,2,4-D induced large quantity of hair root,NAA have induced dense long root,thereby inhibit the callus inducing to tubercle.Through orthogonal experiments selecte the best medium for plant regeneration:MS + 0.5mg/L IAA + 0.5mg/L KT + 0.5mg/L 6-BA.50 days latter,transplant the regenerated plantlets,two weeks latter,the transplant survival rate is more than 95%.3.By use of two-factor and orthogonal experiment methods, screening the best hormone ratio medium to induced loose callus.In the two factors hormones experiment,3.0mg/L 2,4-D and 1.0mg/L 6-BA induced the loose callus with a high growth speed;In the orthogonal experiment,inoculation the leaves and petiole of Pinellia ternate(Thunb.) Breit.in the medium MS with 1.0mg/L 2,4-D + 1.0mg/L KT + 1.0mg/L 6-BA is better for loose callus induction,which subcultured for 3～4 times, can be transfer to suspension culture.