【作者】 刘园；

【导师】 凌征柱； 杨美纯；

【作者基本信息】 广西大学 ， 植物学， 2007， 硕士

【摘要】 通过对珐菲亚进行组织培养技术的研究，建立珐菲亚无性系，以期解决珐菲亚引种栽培种源短缺问题。试验材料用顶芽、腋芽、幼叶、茎段为外植体，进行了芽的诱导及继代增殖培养、愈伤组织的诱导及分化培养、根的分化和组培苗假植等方面的试验，试验结果如下：1、顶芽和腋芽的诱导培养用MS、N₆、B₅、White四种基本培养基附加不同浓度的6-BA和NAA对顶芽和腋芽进行诱导，结果表明，较适宜的基本培养基为MS，激素配比以6-BA1.5mg／L+NAA0.2mg／L为好，芽的增殖系数较高，为3.22。2、芽的继代增殖培养以MS为基本培养基，添加不同浓度的6-BA、KT、ZT和NAA对珐菲亚丛生芽进行继代增殖培养，综合各个因素得出较为优化的继代培养基是MS+6-BA1.2mg／L+NAA0.2mg／L，芽的增殖系数达3.92，芽生长健壮。3、愈伤组织的诱导培养幼叶及茎段均能诱导愈伤组织，但以茎段诱导效果比叶片好，较适宜的培养基配方为：MS+2，4-D1mg／L+20％CW。4、根的诱导培养在1／2MS培养基中加入一定浓度的NAA、IBA、PP₃₃₃，均能诱导小苗生根。用PP₃₃₃促使植株矮化、根系粗壮，有利于提高幼苗假植存活率。较为适宜的生根培养基是1／2MS+NAA1.0mg／L和1／2MS+PP₃₃₃0.2mg／L。5、组培苗假植假植用河沙、蛭石、土壤3种不同基质，结果表明，假植河沙基质的组培苗的成活率较高；生根阶段用PP₃₃₃壮苗促根过的苗，在河沙中假植的组培苗成活率在90％以上。6、化学成分分析珐菲亚组培苗种植一年后，对其收获的根部药材与原产地药材的主要化学成分、总皂苷、氨基酸及微量元素进行比较分析，可以确认组培苗栽植后根部主要成分已接近原产地，所含微量元素和氨基酸的种类一致。总皂苷含量为6.54％，比原产地药材的总皂苷含量的7.32％略低；组培苗的氨基酸总含量为2.6％，比原产地药材的氨基酸总含量的2.2％略高一些。更多还原

【Abstract】 The research on Pfaffia paniculata quick-propagation in vitro culture was done to solve the shortage of Pfaffia paniculata seed source for introduction and cultivation. The stem apex, tender leaf, stem of Pfaffia paniculata were used as explants. The experiments are including shoot induction and multiplication, callus induction and differentiation, roots differentiation, rooting plantlets transplantation and etc. The results are shown as below:1 The culture of stem apex inductionThe stem apex of Pfaffia paniculata were used as explants and cultivated in the basic medium MS, N₆, B₅, White supplied with different concentration of hormone. The result shows that the cluster buds were obtained after forty-day induction culture in the medium of MS+BA1.5mg/L+NAA0.5mg/L, and the rate of shoot multiplication was 3.22.2 The culture of shoot multiplicationThe cluster buds were cut into a single bud and cultivated in the sub-enrichment culture medium MS adding different concentration of 6-BA, KT, ZT and NAA. The best medium is MS supplemented with concentration of 6-BA 1.2mg/L and NAA0.2 mg/L. The rate of shoot multiplication was 3.92 and the buds grow healthy and strong.3 The culture of callus inductionCallus can be inducted by tender leaf and stem, but the callus inducted by tender leaf is better than that of stem. MS+2, 4-Dlmg/L+20% CW medium is suitable for callus induction.4 The culture of rooting1/2MS medium supplemented with proper concentration of NAA, IBA or PP₃₃₃ caninduce rooting. When rooting induced by PP₃₃₃, plantlets are short and strong, the roots are burly, and the numbers of roots are less. PP₃₃₃ can prove the survival rate of transplantation. 1/2MS+NAA1.0mg/L and 1/2MS+PP₃₃₃0.2mg/L medium are suitable for inducing roots.5 Plantlets transplantationThe plantlet was took out, washed the medium of plantlet’s roots and cultivated in sand, vermiculite or soil, the result is that the survival rate cultivated in sand is higher than any other medium. The survival rate of rooting plantlets treated by PP₃₃₃ could be 90%.6 The analysis of chemistry compositionAfter plantlets grown for one year, the comparative analysis was done by major chemistry composition, total mogroside content, amino acids and trace elements. The saponin constituents of culture with twelve month Pfaffia paniculata and provenance were same. Total mogroside content is 6.54%, slightly less than original Pfaffia paniculata 7.32%; the both plantlets in vitro culture and original Pfaffia paniculata had the same kinds of amino acids and trace elements, and the content is 2.6%, original Pfaffia paniculata is 2.2%, slightly more than original.更多还原