| Paris polyphylla var. chinensis is a varietas of Paris polyphylla Smith in Rees of Trilliaceae Paris. Paris polyphylla is a traditional Chinese medicine, which roots can be used as a medicine, and has the effect of clearing heat antitoxicant, relieves swelling and pain, and Cool the liver calm the frightened etc. Medicinal Paris polyphylla var yunnansis and Paris polyphylla var. chinensis are included in Chinese pharmacopoeia.Due to the Paris polyphylla’s poor growth, long regeneration cycle, and the increasing quantity demanded, the imbalance between supply and demand is on the rise. Tissue culture has many advantages of less sampling, high propagation coefficient, anniversary production and so on, and will have great practical significance while it is successful on Paris polyphylla.Polyphylla rhizome has been taken a domesticated research in laboratory conditions. And the beneficial exploration was preceded on many questions of Paris polyphylla var. chinensis isolated culture, whose main research results are as follows:1. Six kinds of soil was used for Polyphylla rhizome’s domesticated research in laboratory conditions which were sand, rotten leaf soil, loam and sandy:rotten leaf soil=1:1, sand:loam=1:1, rotten leaf soil:loam=1:1. And the result shows that:the best kind soil is rotten leaf soil:loam=1:1, which mortality, incidence of root and seedling emergence are9.44%,45.00%and77.8%; the worst kind of soil is sand, which mortality, incidence of root and seedling emergence are38.34%、16.67%aand38.89%。2. After drawing materials the polyphylla rhizomes were inoculated respectively after a week, two weeks, a month, three months, six months, and the results showed that: different place time has a great influence on the setting up of polyphylla rhizome aseptic cultivation system, and the influence is that, with the growth of placing time, the contamination rate and browning rate continues to increase, while the survival rate is declining; Pollution rate of placing one month increased by36.50%compared with a week; Pollution rate of placing three months is close to80%, and the polyphylla rhizomes are no longer suitable for tissue culture; The pollution rate and browning rate continue to increase with extension of incubation time, at the same time survival rate corresponding drops, and the survival rate has dropped to12.70%a month later, less than3.2%three months later, while the polyphylla rhizomes were no longer suitable for tissue culture.3. In order to research the influence of different pretreatment to the establish of polyphylla rhizome aseptic cultivation system, we designed five broad spectrum antibacterial agent (carbendazim, chlorothalonil, allicin antivirus alum, thiophanate, b) wiht a total of ten concentrations and four kinds of antibiotics (amikacin, gentamicin, streptomycin, cephalosporins he organism), with a total of eight concentration. The results showed that:the best effect appeared at broad spectrum antibacterial agent with0.5%carbendazim bacteriostatic, whose fungus contamination rate, bacterial contamination rate and total pollution rate respectively decreased19.04%,7.93%,26.98%compared with CK1; the worst effect appeared at0.1%antivirus vanadium. The best effect appeared at100mg/L cephalosporins in the eight antibiotics concentration whose fungus contamination rate, bacterial contamination rate and total pollution rate respectively decreased39.68%,11.11%,50.79%, while the effect of the other three groups of antibiotics are not ideal.4. In order to research the influence of polyphylla rhizome callus induction, we used MS as basic medium with different concentration of6-BA, NAA, IB A, a total of12kinds of hormone proportion. Results show that:the best callus induction on polyphylla rhizome appeared at the hormone proportion of6-BA1.0mg/L+1.0mg/L NAA, whose browning rate, startup rate and callus induction rate were39.68%,60.31%and39.68%; Followed by6-BA2.0mg/L+IBA1.0mg/L, whose browning rate, startup rate and callus induction rate were52.38%,65.08%and52.38%respectively; The worst result happened at the hormone proportion of6-BA4.0mg/L+NAA0.5mg/L.5. The effect has a very big difference using different parts of the Paris polyphylla as explant callus induction, which is that, as explants crown and roots could induce the callus, while the ground area (leaf, sepal, stem segment, flower stalk) of Paris polyphyllat could not. The start-up rate and callus induction rate of terminal bud were the highest, respectively were34.86%,15.00%.6. In order to research the influence of polyphylla rhizome callus induction, we designed the different ways of cutting. The result shows that:the pollution rate of terminal bud was falling and the startup rate and the callus induction rate difference was not significant with the sliced position downing; Bisection could significantly improve the startup rate and the callus induction rate, but also increase the pollution rate by a certain degree.7. In order to research the influence of polyphylla rhizome seedling induction, we used MS as basic medium with different concentration of6-BA, NAA, a total of6kinds of hormone proportion. Results showed that:the hormone proportion of6-BA1.0mg/L+0.2mg/L NAA has a best performance on polyphylla rhizome seedling induction, which mortality, and seedling induction rate were19.05%,80.93%, and the seedlin was gdark green with about3.5cm high, robust growth; the hormone proportion of6-BA2.0mg/L+0.4mg/L NAA is the worst, which mortality, and seedling induction rate were42.86%,57.14%, and the seedlin was yellow-green with about2cm high, thin growth. |
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