Determination of 10 Kinds of Inorganic Elements in Paris polyphylla Smith var. yunnanensis(Franch.) Hand.-Mazz.(and Polygerm Varieties)

[Objectives] To determine the content of 10 kinds of inorganic elements(Pb,Cu,As,Cd,Hg,Zn,Fe,Mn,Sr and Cr) in samples of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( and polygerm varieties) produced in different areas. [Methods] The wet digestion and technique of inductively coupled plasma optical emission spectrometry( ICP-OES) were used. [Results] Under the experimental conditions,elements were not related to each other,and several kinds of elements could be measured at the same time. The content of Pb exceeding the standard only in sample S4 of P. polyphylla Smith var. yunnanensis( and polygerm varieties),Hg was not detected in all samples,and the rest heavy metals did not exceed the limit. [Conclusions] This study was expected to provide a certain reference for the development and utilization of P. polyphylla Smith var. yunnanensis( and polygerm varieties).

Identification of Paris polyphylla Smith var. yunnanensis(Franch.) Hand.-Mazz.(Polygerm Varieties) Produced in Gaoligong Mountain Areas

[Objectives] To make original plant and microscopic identification and study the traits of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( polygerm varieties) produced in Gaoligong Mountain areas,and provide experimental data for their identification and application. [Methods] The original plant powder was used,and paraffin sections and physicochemical properties were identified. [Results]P. polyphylla Smith var. yunnanensis( polygerm varieties) produced in Gaoligong Mountain areas showed obvious microscopic features and physicochemical properties. [Conclusions] This study was expected to provide certain reference for identification of original plant and medicine and formulation of quality standard for P. polyphylla Smith var. yunnanensis( polygerm varieties) produced in Gaoligong Mountain areas,and provided basis for their further research and development.

云南重楼生物量积累与营养分配研究

文章对不同繁殖方式、不同生长年限的云南重楼进行了生物量积累和营养分配研究。结果表明,1～8年生云南重楼根茎切段苗、实生苗生物量逐年递增,未有最高峰值。各营养器官折干率和营养分配高低顺序：根茎〉叶〉茎〉须根。根茎干物质积累及全株总生物量积累分为3个生长阶段：缓慢期、增重快速期、增重加速期;叶、茎干物质积累可分为缓慢期、增重快速期、增重平缓期。

滇重楼最佳授粉期的研究

[目的]优选滇重楼的最佳授粉期。[方法]采用TTC法测定花粉的活力,并采用联苯胺-过氧化氢法测定柱头可授性,根据研究滇重楼花粉活力变化规律及柱头可授性优选滇重楼的最佳授粉期。[结果]滇重楼花粉囊开裂时活力最高为68.2%,前2 d内,花粉活力下降不明显。第4天开始,活力明显下降,但仍然具有一定的活性。柱头的活性在花粉囊开裂后5 d内最强,滇重楼的授粉应选择在这5 d内进行。[结论]该研究优选了滇重楼的最佳授粉期,为滇重楼的进一步开发利用提供了依据。

改良CTAB法提取重楼属3种药用植物干燥根茎DNA

目的:本研究采用改良CTAB法提取重楼属3种植物的DNA,分析了CTAB浓度对其DNA得率以及醋酸钾(KAc)或KAc+20%乙醇对其DNA纯度的影响,为重楼属植物药材的分子标记鉴定研究提供技术和理论依据。方法:采用基于CTAB的6种方法提取重楼属3种植物干燥根茎的DNA,并用紫外分光光度法测定A260和A280的值计算评价DNA得率和纯度。结果:4%CTAB提取3种重楼的DNA得率均高于2%CTAB;添加KAc的方法 2(2%CTAB)、方法 5(4%CTAB)以及添加KAc+20%乙醇的方法 3(2%CTAB)和方法 6(4%CTAB)提取的DNA纯度和质量以及通用引物ITS2F/ITS2R的PCR扩增结果均优于不添加KAc或KAc+20%乙醇的方法 1和方法 4。结论:本研究表明4%CTAB有利于重楼属3种植物干燥根茎DNA得率的提高,在DNA沉淀之前添加KAc或KAc+20%乙醇有利于改善DNA纯度和质量。

多叶重楼与云南重楼中4种重楼皂苷的积累差异

目的考察多叶重楼Paris polyphylla Smith var.polyphylla与云南重楼Paris polyphylla Smith var.yunnanensis(Franch.)Hand.-Mazz.中重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ的积累差异。方法 HPLC法测定多叶重楼根、叶、茎中4种重楼皂苷含有量,建立20批样品的指纹图谱,聚类分析对2种植物不同部位中各重楼皂苷的积累差异进行评价。结果不同居群多叶重楼中各重楼皂苷含有量及其总量均有显著差异(P<0.05,P<0.01)。根中未检测到重楼皂苷Ⅵ,重楼皂苷Ⅰ、Ⅱ平均含有量最高;叶中重楼皂苷Ⅶ平均含有量和总含有量最高;茎中仅检测到重楼皂苷Ⅶ,总含有量最低。多叶重楼和云南重楼根的相似度较高,叶和茎较低。根据不同部位重楼皂苷总含有量,样品可分为3类。结论可考虑将多叶重楼作为云南重楼的替代品。

Arbuscular mycorrhiza and plant growth promoting endophytes facilitates accumulation of saponin under moderate drought stress

Objective: Paris polyphylla var. yunnanensis, one of the important medicinal plant resources in Yunnan,China, usually takes 6–8 years to be harvested. Therefore, it is urgent to find a method that can not only shorten its growth years, but also improve its quality. In this study, we examined the effects of a combination treatment of arbuscular mycorrhizal fungi(AMF) and plant growth-promoting endophytes(PGPE)and drought stress on the accumulation of saponins in it.Methods: P. polyphylla var. yunnanensis was infected with a mixture of AMF and PGPE under drought stress. The content of saponins, as well as morphological, physiological, and biochemical indicators, were all measured. The UGTs gene related to saponin synthesis was obtained from transcriptome data by homologous comparison, which were used for RT-PCR and phylogenetic analysis.Results: Regardless of water, AMF treatment could infect the roots of P. polyphylla var. yunnanensis, however double inoculation with AMF and PGPE(AMF + PGPE) would reduce the infection rate of AMF. Plant height, aboveground and underground fresh weight did not differ significantly between the single inoculation AMF and the double inoculation treatment under different water conditions, but the inoculation treatment significantly increased the plant height of P. polyphylla var. yunnanensis compared to the noninoculation treatment. Single inoculation with AMF considerably increased the net photosynthetic rate,stomatal conductance, and transpiration rate of P. polyphylla var. yunnanensis leaves under various water conditions, but double inoculation with AMF + PGPE greatly increased the intercellular CO2concentration and chlorophyll fluorescence parameter(Fv/Fm). Under diverse water treatments, single inoculation AMF had the highest proline content, whereas double inoculation AMF + PGPE may greatly improve the amount of abscisic acid(ABA) and indoleacetic acid(IAA) compared to normal water under moderate drought. Double inoculation AMF + PGPE treatment improved the proportion of N, P, and K in the rhizome of P. polyphylla var. yunnanensis under various water conditions. Under moderate drought stress,AMF + PGPE significantly enhanced the contents of P. polyphylla var. yunnanensis saponins I, II, VII, and total saponins as compared to normal water circumstances. Farnesyl diphosphate synthase(FPPS),Geranyl pyrophosphate synthase(GPPS), Cycloartenol synthase(CAS), and Squalene epoxidase(SE1) were the genes that were significantly up-regulated at the same time. The amount of saponins was favorably linked with the expression of CAS, GPPS, and SE1. Saponin VI content and glycosyl transferase(UGT)010922 gene expression were found to be substantially associated, as was saponin II content and UGT010935 gene expression.Conclusion: Under moderate drought, AMF + PGPE was more conducive to the increase of hormone content, nutrient absorption, and total saponin content in P. polyphylla var. yunnanensis, and AMF + PGPE could up regulate the expression of key genes and UGTs genes in one or more steroidal saponin synthesis pathways to varying degrees, thereby stimulating the synthesis and accumulation of steroidal saponins in the rhizome of P. polyphylla var. yunnanensis. The combination of AMF and PGPE inoculation, as well as adequate soil drought, reduced the buildup of saponins in P. polyphylla var. yunnanensis and increased its quality.