Analysis of the heavy metal contents' effect on steroidal saponins and the anti-breast cancer activity of Paris polyphylla var. yunnanensis.

Background: P. polyphylla var. yunnanensis, as a near-threatened and ethnic medicine in China, used to be a key ingredient in traditional Chinese medicine in treatment of traumatic injuries, sore throat, snakebites, and convulsions for thousands of years. However, there were no reports on the inverse relationship between the contents of heavy metals and saponins and its anti-breast cancer pharmacological activity in P. polyphylla var. yunnanensis. Methods: The present study aimed to reveal the characteristics of heavy metal contents and saponins and its anti-breast cancer pharmacological activity and their interrelationships in P. polyphylla var. yunnanensis from different production areas. The contents of heavy metal and steroidal saponins in P. polyphylla var. yunnanensis were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) and the high-performance liquid chromatography technique, respectively. The Pearson correlation was used to study the correlation between saponins and heavy metals. 4T1 mouse mammary tumor cells were selected and cultivated for antitumor studies in vitro. Cell Counting Kit-8 (CCK-8) assay, Hoechst staining, and flow cytometry analysis were used for the examination of the proliferation and apoptosis of 4T1 tumor cells. Mouse breast cancer 4T1 cells were subcutaneously injected into BALB/c mice to construct a tumor model to explore the in vivo inhibitory effect on breast cancer. TUNEL assay and immunohistochemistry were used for the examination of the effect of P. polyphylla var. yunnanensis from different origins on cancer cell proliferation and apoptosis induction in 4T1 tumor mice. Results: Heavy metal contents were highly correlated with the content of steroidal saponins. The overall content of 10 metals in the three producing origins was of the order C3 >C2 >C1. The total content of eight steroidal saponins in the extracts of P. polyphylla var. yunnanensis from three different origins was C1 >C2 >C3. The Pearson correlation study showed that in all of the heavy metals, the contents of Cd and Ba were positively correlated with the main steroidal saponins in P. polyphylla var. yunnanensis, while Al, Cr, Cu, Fe, Zn, As, Hg, and Pb showed a negative correlation. In vitro experiments showed that the extracts of P. polyphylla var. yunnanensis from three origins could inhibit the proliferation and induce cell apoptosis of 4T1 cells in a concentration- and time-dependent manner, especially in the C1 origin. In vivo experiments showed that the extract of P. polyphylla var. yunnanensis from the three origins could inhibit the growth of tumors and induce the apoptosis of tumor cells. In the three origins, C1 origin had the lowest total heavy metal level but the highest total steroidal saponin level. Therefore, it showed a better effect in reducing the expression of the human epidermal growth factor receptor 2 (HER2) and Kiel 67 (Ki67) and increasing the expression of p53 in tumor tissues compared to the other origins. In conclusion, in the three origins, C1 origin exhibits antitumor pharmacological effects in vivo and in vitro which are better than those in the other origins. Conclusion: In this study, we found that with the increase of the heavy metal content, the content of steroid saponins and anti-breast cancer activity decreased. The results showed that the high content of the total heavy metals may not be conducive to the accumulation of steroidal saponins in P. polyphylla var. yunnanensis and lead to the low anti-breast cancer activity. The results of this study suggest that the content of heavy metals should be controlled in the artificial cultivation process of P. polyphylla var. yunnanensis.

Effects of plant growth-promoting rhizobacteria on growth indicators and physiological characteristics of Peucedanum praeruptorum Dunn leaves.

As a kind of medicinal plant, Peucedanum praeruptorum Dunn has been over-harvested in the wild population, which leads to its artificial cultivation. The present study aims to analyze the effects of different plant growth-promoting rhizobacteria (PGPR) on the growth and physiological characteristics of P. Praeruptorum leaves. Compared with the CK, the content of malondialdehyde (MDA) was drastically reduced in the leaves of P. Praeruptorum in different treatment groups (P < 0.05), and with S6 showing the most significant reduction in MDA content (content was only about 1/3 that of the CK). The indicators of leaf area, length and width were found to be the highest in group S9, reaching a level that is 3.75, 3.08 and 1.48 times higher than those in group CK, respectively. Group S8 has the largest plant height, which is 1.22 times higher than that in group CK. S2 has the largest stem diameter, which is 1.69 times higher than that in group CK. Group S1 has the largest petiole length, which is 1.74 times higher than that in group CK. Group S6 has the largest chlorophyll content, which is 1.63 times higher than that in group CK. Group S2 has the highest content of soluble sugar and soluble protein, which are 2.02 times and 3.82 times higher than those in group CK. Group S9 exhibits the strongest CAT activity, which is 3.71 times higher than that in group CK. S5 exhibits the strongest SOD activity, which is 2.32 times higher than group CK. Group S1 exhibits the strongest POD activity, which is 5.94 times higher than that in group CK. In conclusion, the inoculation with PGPR is effective in improving the growth of P. Praeruptorum leaves and their physiological indicators, which provides guidance on the application of PGPR to achieve the high quality and yield of P. Praeruptorum.

The rhizospheric bacterial diversity of Fritillaria taipaiensis under single planting pattern over five years.

Rhizospheric microorganisms can profoundly influence the nutritional status of soil and the growth of plant. To reveal the change on the bacterial diversity in the rhizosphere of Fritillaria taipaiensis under long-term single planting, the bacterial community structure in the rhizospheric soils of F. taipaiensis with different cultivation years from 1 to 5 were analyzed. The result showed the Chao1 and the ACE indices of the bacterial community had no significant difference among samples while the Shannon and Simpson indices declined with the cutivation year; the intra group beta diversity of the rhizospheric bacteria increased after a initial decline with the cultivation year; in the sample with 1 year of cultivation, the dominant bacterial genera were mainly the species that can improve the soil nutrient status and promote plant growth while with the increase of cultivation year, the dominant genera in samples then gradually reflected the pathogen accumulation and soil nutrient status deterioration; pH was the most significant factor affected by the bacterial community composition. These results indicated long term continuous cropping changed the bacterial community structure and soil nutritional status in the F. taipaiensis rhizospheric soils, which could badly affect its growth.

Electrochemical Behavior and Direct Quantitative Determination of Paclitaxel.

The electrochemical behavior and direct quantitative determination of paclitaxel, a poorly soluble drug made into microemulsion, were researched by cyclic voltammetry in acetate buffer solutions (pH = 4.0) at a glassy carbon electrode. The results show that the oxidation process is irreversible and controlled by diffusion. Moreover, the effects of anodic peak current (Ipa), anodic peak potential, scan rate, pH, and the electrochemical redox mechanism have been studied. The anodic peak current varied linearly with paclitaxel concentration in the range of 5 × 10-5 mol/L to 5 × 10-4 mol/L, and the detection limit was 9.15 × 10-8 mol/L. The results of RSD (0.90%) and recovery (99.22%-101.69%) were obtained. Additionally, it has been proved that one electron and one proton are involved in the electrochemical redox process. The present research has been successfully used to determine paclitaxel in pure and real samples, which further supported the electrochemical behavior investigation of paclitaxel and direct determination of micro-emulsion.

Characterization and analysis of complete chloroplast genome of Clausena anisum-olens (Blanco) Merrill.

In this study, the chloroplast genome (Cp) of Clausena anisum-olens ((Blanco) Merrill) was sequenced by high-throughput sequencing technology. We found that the length of the chloroplast genome of Clausena anisum-olens was 159,753 bp, and the total GC content was 38.7%, including a large single-copy (LSC) region of 87,623 bp, a small single-copy (SSC) region of 17,986 bp, and 27,072 bp pairs of reverse repeats (IRS). The Cp genome encoded 129 genes, containing 86 protein-coding, 37 tRNA, and 6 rRNA genes. Phylogenetic analysis of the genome sequence showed that Clausena anisum-olens was closely related to Micromelum minutum and Glycosmis mauritiana.

Complete chloroplast genome sequence and phylogenetic analysis of Ilex pernyi Franch.

Ilex pernyi Franch. is a Chinese unique medicinal plant with high value for decoration, human consumption, and as a medicine. In this study, we sequenced the complete chloroplast (cp) genome of I. pernyi Franch. and established its phylogenetic relationship in the Aquifoliaceae family. The total length of the chloroplast genome of I. pernyi Franch. was found to be 157,220 bp, with an overall GC content of 37.7% and a typical quadripartite structure with a pair of inverted repeats (IRs, 29,092 bp), which was separated by a small single-copy (SSC) region of 18,427 bp and a large single-copy (LSC) region of 89,609 bp. The cp genome contained 132 genes: 87 protein-coding, 37 tRNA, and 8 rRNA genes. The phylogenetic analysis indicated that I. pernyi Franch. was closely related to Ilex cornuta.

The complete chloroplast genome and phylogenetic analysis of Sida szechuensis matsuda (Malvaceae).

Sida szechuensis Matsuda is an economically and medicinally important plant. Here, we report the first chloroplast (cp) genome of the genus Sida (S. szechuensis). The complete cp genome is 159,878 bp in length with an overall GC content of 36.9% and consists of a large single copy region (LSC, 89,426 bp), a small single copy region (SSC, 114,715 bp), and a pair of inverted repeat regions (IRa and IRb, 25,288 bp). The genome encodes 111 unique genes, including 78 protein-coding genes, 29 tRNA genes, 4 rRNA genes, and 1 pseudogene. Phylogenetic analysis constructed using the maximum likelihood (ML) method showed that Sida was closely related to Malvastrum and Malva.

The complete plastid genome of Cotinus coggygria and phylogenetic analysis of the Anacardiaceae.

Cotinus coggygria Scop. (Anacardiaceae) is an important ornamental tree with beautiful characteristics that is grown in China. In this study, the complete plastid genome of C. coggygria was sequenced and assembled. This genome was 158,843 bp in size and presented a typical tetrad structure, consisting of a large single-copy region (87,121 bp), a pair of inverted repeat regions (26,829 bp), and a small single-copy region (18,064 bp). A total of 134 genes were annotated, including 88 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. We observed a deletion that caused the loss of the rpl32 gene, and a small expansion of IR regions resulted in the trnH gene accessing IR regions; two copies were obtained. Phylogenetic analysis showed that C. coggygria was most closely related to Pistacia, with 100% bootstrap support within Anacardiaceae. In this study, we report the plastid genome of Cotinus species for the first time, which provides insight into the evolution of the plastid genome in Anacardiaceae and promotes the understanding of Cotinus plants.

The complete chloroplast genome sequences and phylogenetic analysis of Crotalaria pallida (Leguminosae).

The complete chloroplast genome of Crotalaria pallida was obtained using the high-throughput sequencing technology in this article. The complete chloroplast genomes of this species were 152,658 bp in length, consisting of a large single-copy region (LSC) of 83,652 bp and a small single-copy region (SSC) of 18,028 bp, which were separated by a pair of inverted repeat (IRs) regions of 25,489 bp. The chloroplast genome contained 111 unique genes, including 77 protein-coding genes, 30 tRNA genes, and four rRNA genes. The phylogenomic relationship analysis suggested that C. pallida was closely related to Lupinus in the family of Leguminosae.

[Screening and identification of potassium-dissolving bacteria from different rhizosphere soil of Paris polyphylla var. yunnanensis].

The study aiming at exploring the potassium-dissolving capacity of rhizosphere potassium-dissolving bacteria from diffe-rent sources and screen the strains with high potassium-dissolving ability, so as to lay a theoretical foundation for cultivation and quality improvement of Paris polyphylla var. yunnanensis sources. The rhizosphere soil of 10 wild and transplanted species from Yunnan, Sichuan and Guizhou provinces was used as the research object. Potassium-dissolving bacteria were isolated and purified, and their potassium-dissolving capacity was determined by flame spectrophotometry, and identified by physiological, biochemical and molecular biological methods. Twenty-six potassium-dissolving bacteria were purified and 13 were obtained from wild and transplanted strains respectively. It was found through the determination of potassium-dissolving capacity that the potassium-dissolving capacity of 26 strains was significantly different, and the mass concentration of K~+ in the fermentation broth were 1.04-2.75 mg·L~(-1), the mcentration of potassium were 0.01-1.82 mg·L~(-1). The strains were identified as Bacillus, Agrobacterium rhizome and Staphylococcus by physiological, biochemical and 16 S rDNA molecular methods, among them Bacillus amylolyticus(4 strains) was the dominant bacterium of Bacillus. The physiology and biochemistry of rhizosphere potassium-dissolving bacteria in P. polyphylla var. yunnanensis rhizosphere were diffe-rent, and the living environment were different, so the potassium-dissolving capacity also changed. Strain Y4-1 with the highest potassium decomposability was Bacillus amylolytic with a potassium increase of 1.82 mg·L~(-1). The potassium-dissolving ability and the distribution of potassium-dissolving bacteria were different in various habitats. The screening of potassium-dissolving bacteria provided a new strain for the preparation of microbial fertilizer. It is expected that B. amyloidococcus Y4-1 can be used as an ideal strain to cultivate mycorrhizal seedlings of P. polyphylla var. yunnanensis.

[Isolation and identification of phosphatolytic bacteria in Paris polyphylla var. yunnanensis].

The wild resources of Paris polyphylla var. yunnanensis, a secondary endangered medicinal plant, are severely scarce. Introduction and cultivation can alleviate market demand. To screen phosphatolytic bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis and provide data support for the development of high-efficiency microbial fertilizer, in this study, the dilution plate coating method was used to isolate and screen the phosphorus solubilizing bacteria with the ability of mineralizing organic phosphorus from the rhizosphere soil of wild and transplanted varieties of P. polyphylla var. yunnanensis in 10 different locations in Yunnan, Sichuan and Guizhou. After separation and purification, the phosphatolytic capacity was analyzed by qualitative and quantitative analysis. Combined with physiological and biochemical experiments, the strains were identified using 16 S rDNA sequencing analysis. Forty one strains were selected from the rhizosphere soil of P. polyphylla var. yunnanensis from 10 different habitats. Among them, 21 strains were obtained from the rhizosphere soil of the wild variety P. polyphylla var. yunnanensis and 20 strains were obtained from the rhizosphere soil of the transplanted variety. And significance analysis found that 41 organophosphate solubilizing strains had significant differences in their ability to solubilize phosphorus. The amount of phosphate solubilizing was 0.08-67.61 mg·L~(-1), the pH value was between 4.27 and 6.82. The phosphatolytic amount of strain Y3-5 was 67.61 mg·L~(-1), and the phosphorus increase amount was 57.57 mg·L~(-1). All 41 strains were identified as Gram-positive Bacillus. Combining physiological characteristic and phylogenetic trees, Bacillus mobilis Y3-5 was finally selected as the candidate rhizosphere phosphatolytic bacteria of P. polyphylla var. yunnanensis. The distribution of phosphorus solubilizing bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis was different, and there were significant diffe-rences in phosphorus solubility. Organophosphate-dissolving strain Y3-5 is expected to be a candidate strain of P. polyphylla var. yunnanensis microbial fertilizer.

The complete chloroplast genome of Zanthoxylum acanthopodium DC. (Rutaceae) and its phylogenetic analysis.

We sequenced the complete chloroplast genome of Zanthoxylum acanthopodium via high-throughput sequencing technology, and analyzed its structural characteristics and phylogenetic relationships. The chloroplast exhibits a genome length of 158,473 bp, including a pair of inverted repeat regions (IRa and IRb) of 27,369 bp, a small single-copy (SSC) region of 17,629 bp and a large single-copy (LSC) region of 85,570 bp. The annotation analysis identified 112 genes, containing 78 protein-coding genes, 30 tRNA genes and 4 rRNA genes. The phylogenetic analysis showed that Z. acanthopodium was closely related to Z. piperitum and Z. tragodes.

[Correlation analysis of quality,origin and phenotypic characters of Paris polyphylla var. yunnanensis].

In order to provide guidance for the protection and utilization of resources,quality control and breeding of improved varieties,we compared the main phenotypic characters and quality of wild and transplanted Paris polyphylla var. yunnanensis collected from different producing areas. Seven phenotypic characters of 33 samples of P. polyphylla var. yunnanensis collected from Yunnan,Guizhou and Sichuan were determined by conventional methods,and the principal component analysis and cluster analysis were used to analyze the diversity of the samples. The parissaponin( polyphyllin Ⅰ,Ⅱ,Ⅵ,Ⅶ) content of the samples were detected by HPLC,and analyzed by cluster analysis. Correlation analysis of the phenotypic characters and the parissaponin content was performed. There were significant differences in seven phenotypic characters between wild and transplanted samples of P. polyphylla var. yunnanensis from different habitats,with high phenotypic diversity and abundant genetic variation. The results of principal component analysis showed that leaf shape index was the main factor of morphological variation of P. polyphylla var. yunnanensis. Cluster analysis showed that the phenotypic characters of wild and transplanted P. polyphylla var. yunnanensis could not be completely separated. The content of saponins in wild and transplanted samples from different habitats was quite different. Saponins content of 93. 94% samples met the criterion of Chinese Pharmacopoeia 2015 edition,and the overall quality was relatively steady. The results of independent sample t-test showed that there was no significant difference of all the active ingredient between wild and transplanted samples,and it couldn't be used to distinguish between wild and transplanted samples. It is the same as the results of cluster analysis. The results of correlation analysis showed that the phenotypic traits of P. polyphylla var. yunnanensis were correlated with its medicine quality,and the total content of saponins was positively correlated with leaf length and leaf shape index( r = 0. 389,0. 441; P<0. 05). Yunnan,Guizhou and Sichuan are suitable for the growth of P. polyphylla var. yunnanensis. And the transplaned P. polyphylla var. yunnanensis can be used as the same as the wild ones completely. The results provide reference for the protection and selective breeding of P. polyphylla var. yunnanensis.

Damage on functional state of intestinal barrier by microgravity stress in nematode Caenorhabditis elegans.

Due to short life cycle, nematode Caenorhabditis elegans is a suitable animal model for assessing the effect of long-term simulated microgravity treatment on organisms. We here investigated the effect of simulated microgravity treatment for 24-h on development and functional state of intestinal barrier in nematodes. Simulated microgravity treatment not only caused a broadened intestinal lumen, but also enhanced intestinal permeability. Intestinal overexpression of SOD-2, a mitochondrial Mn-SOD protein, prevented the damage on functional state of intestinal barrier by simulated microgravity and induced a resistance to toxicity of simulated microgravity, suggesting the crucial role of oxidative stress in inducing the damage on functional state of intestinal barrier in simulated microgravity treated nematodes. For the molecular basis of damage on functional state of intestinal barrier, we observed significant decrease in expressions of some genes (acs-22, erm-1, and hmp-2) required for maintenance of functional state of intestinal barrier in simulated microgravity treated nematodes. Our results highlight the potential of long-term simulated microgravity treatment in inducing intestinal damage in animals.

[Correlation between rhizospheric microorganisms distribution and alkaloid content of Fritillaria taipaiensis].

In order to reveal the relationship between the amount of soil microorganisms and the quality of Fritillaria taipaiensis, both cultivated and wild F. taipaiensis were collected from Chongqing, Wuxi at different stages of their growth as objects of the research. The mycorrhizal infection rate and colonization intensity, peimisine and total alkaloid content in bulbs, the amount of microorganisms and biomass carbon content in rhizospheric soil were all determined using common methods. The results showed that the typical arbuscular-vesicle roots were formed after the AM fungi infected the F. taipaiensis roots which were collected from different origins. The mycorrhizal infection rates were ranged from 78.74% to 98.68% and the colonization intensities were ranged from 13.29% to 37.06%. The rhizospheric microorganisms of F. taipaiensis showed abundant resources. The distribution rule of them in the rhizospheric soil was as follows: the amount of bacteria>the amount of actinomycetes>the amount of fungi. The rhizospheric bacteria, decomposition inorganic phosphorus bacteria, decomposition organic phosphorus bacteria, actinomycetes amount and the total number of microbes increased first and then decreased with the increase of years, while decomposition potassium bacteria showed decreasing trend and fungi showed gradual increasing trend. The soil microbial flora content in the soil changed from "bacterial type" with a high fertility to "fungal type" with a low fertility. The mass fraction of peimisine and total alkaloid content increased first and then decreased with the increase of over the years, the same trend of culturable rhizosphere soil bacteria and actinomycetes indicated that the growth years affected the quality of soil and medicinal materials on different levels. Therefore, the diversity of microbial communities in rhizosphere soil reduced with the increase of years leading to the continuous cropping obstacles and the destruction of medicinal quality of F. taipaiensis.

Author Correction: Toxicity evaluation of Wanzhou watershed of Yangtze Three Gorges Reservoir in the flood season in Caenorhabditis elegans.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Author Correction: Biosafety assessment of water samples from Wanzhou watershed of Yangtze Three Gorges Reservoir in the quiet season in Caenorhabditis elegans.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Biosafety assessment of water samples from Wanzhou watershed of Yangtze Three Gorges Reservior in the quiet season in Caenorhabditis elegans.

We here employed a model animal of Caenorhabditis elegans to perform toxicity assessment of original surface water samples collected from Three Gorges Reservoir (TGR) in the quiet season in Wanzhou, Chongqing. Using some sublethal endpoints, including lifespan, body length, locomotion behavior, brood size, and intestinal reactive oxygen species (ROS) induction, we found that the examined five original surface water samples could not cause toxicity on wild-type nematodes. Nevertheless, the surface water sample collected from backwater area induced the significant increase in expressions of genes (sod-2 and sod-3) encoding Mn-SODs in wild-type nematodes. Among the examined five original surface water samples, exposure to the original surface water sample collected from backwater area could further cause the toxicity in decreasing locomotion behavior and in inducing intestinal ROS production in sod-3 mutant nematodes. Moreover, the solid phase of surface water sample collected from backwater area might mainly contribute to the observed toxicity in sod-3 mutant nematodes. Our results are helpful for understanding the potential effects of surface water in the TGR region in the quiet season on environmental organisms.

Toxicity evaluation of Wanzhou watershed of Yangtze Three Gorges Reservior in the flood season in Caenorhabditis elegans.

Three Gorges Reservoir (TGR) in the upper stream of Yangtze River in China is a reservoir with the largest and the longest yearly water-level drop. Considering the fact that most of safety assessments of water samples collected from TGR region were based on chemical analysis, we here employed Caenorhabditis elegans to perform in vivo safety assessment of original surface water samples collected from TGR region in the flood season in Wanzhou, Chongqing. Among the examined five original surface water samples, only exposure to original surface water sample collected from backwater area could induce the significant intestinal ROS production, enhance the intestinal permeability, and decrease the locomotion behavior. Additionally, exposure to original surface water sample collected from backwater area altered the expressions of sod-2, sod-5, clk-1, and mev-1. Moreover, mutation of sod-2 or sod-5 was susceptible to the potential toxicity of original surface water sample collected from backwater area on nematodes. Together, our results imply that exposure to surface water sample from the backwater area may at least cause the adverse effects on intestinal function and locomotion behavior in nematodes.

[Effects of Different AM Fungi on Quantity and Enzyme Activity of Rhizosphere Soil Microorganism of Paris polyphylla var. yunnanensis].

Objective: To investigate the rhizosphere soil microorganism and enzyme activity after Paris polyphylla var. yunnanensis inoculated Arbuscular Mycorrhizal( AM) fungi,which provide the technological condition for artificial cultivation of Paris polyphylla var. yunnanensis. Methods: By the method of combining the greenhouse pot inoculation trials with laboratory analysis, The effects of inoculation with 28 AM fungi on rhizosphere soil microorganism and soil enzyme activities of Paris polyphylla var. yunnanensis under sterilized soil conditions were observed. Results: There were certain mutualistic selection between Paris polyphylla var. yunnanensis and AM fungi. After induction treatment with AM fungi, different AM fungi had various effects on the quantity, microbial biomass carbon and soil enzyme activities of rhizosphere soil microorganism of Paris polyphylla var. yunnanensis. With application of AM fungi,the amount of culturable fungi from rhizosphere soil of Paris polyphylla var. yunnanensis were decreased,while the culturable bacteria and actinomycetes showed a rising trend,and increased the diversity index,the content of biomass carbon and protease,urease,phosphatase,catalase and invertase activities of rhizosphere soil microorganism. Conclusion: Applying AM fungi when artificial cultivate Paris polyphylla var. yunnanensis,by which can impel rhizosphere microorganism of Paris polyphylla var. yunnanensis to transform from low-fertile"fungus types"to high-fertile"bacterial forms",and raise fungi diversity and enzymatic activity of rhizosphere soil microorganism,it is helpful to maintain the stability and harmoniousness of microecosystem on rhizosphere of Paris polyphylla var. yunnanensis,which certify the efficiency and the possibility of spreading AM inoculum during artificial cultivation in the field.