To clarify the species, pathogenicity, and distribution of the pathogens causing the root rot of Atractylodes lancea in Hubei province, the tissue separation method was used to isolate the pathogens from root rot samples in the main planting areas of A. lancea in Hubei. Based on the preliminary identification of the Fusarium genus by the internal transcribed spacer(ITS) sequence, three housekeeping genes, EF1/EF2, Btu-F-FO1/Btu-F-RO1, and FF1/FR1, were amplified and sequenced. Subsequently, a phylogenetic tree was constructed based on these TEF gene sequences to classify the pathogens. The pathogenicity of these strains was determined using the root irrigation method. A total of 194 pathogen strains were isolated using the tissue separation method. Molecular identification using the three housekeeping genes identified the pathogens as F. solani, F. oxysporum, F. commune, F. equiseti, F. tricinctum, F. redolens, F. fujikuroi, F. avenaceum, F. acuminatum, and F. incarnatum. Among them, F. solani and F. oxysporum were the dominant strains, widely distributed in multiple regions, with F. solani accounting for approximately 54% of the total isolated strains and F. oxysporum accounting for approximately 34%. Other strains accounted for a relatively small proportion, totaling approximately 12%. The results of pathogenicity determination showed that there were certain differences in pathogenicity among strains. The analysis of the pathogenicity differentiation of the widely distributed F. solani and F. oxysporum strains revealed that these dominant strains in Hubei were mainly highly pathogenic. This study determined the species, pathogenicity, and distribution of the pathogens causing the root rot of A. lancea in Hubei province. The results provide a scientific basis for further understanding the root rot of A. lancea and its epidemic occurrence and scientifically preventing and controlling this disease.
Plant Diseases/microbiology* ; Atractylodes/microbiology* ; Phylogeny ; Plant Roots/microbiology* ; Fusarium/classification* ; China ; Virulence ; Fungal Proteins/genetics*

Microbial detection and control of traditional Chinese medicine(TCM) decoction pieces are crucial for the quality control of TCM preparations. It is also a key area of research in the measurement technology and equipment development for TCM manufacturing. Guided by TCM manufacturing measurement methodologies, this study presented a design of a novel portable microbial detection device, using Atractylodis Macrocephalae Rhizoma decoction pieces as a demonstration. Immunomagnetic separation technology was employed for specific isolation and labeling of target microorganisms. Enzymatic signal amplification was utilized to convert weak biological signals into colorimetric signals, constructing an optical biosensor. A self-developed smartphone APP was further applied to analyze the colorimetric signals and quantify target concentrations. A portable and automated detection system based on Arduino microcontroller was developed to automatically perform target microbial separation/extraction, as well as mimetic enzyme labeling and catalytic reactions. The developed equipment specifically focuses on the rapid and quantitative microbial analysis of TCM active pharmaceutical ingredients, intermediates in TCM manufacturing, and final TCM products. Experimental results demonstrate that the equipment could detect Salmonella in samples within 2 h, with a detection limit as low as 5.1 × 10~3 CFU·mL~(-1). The equipment enables the rapid detection of microorganisms in TCM decoction pieces, providing a potential technical solution for on-site rapid screening of microbial contamination indicators in TCM. It has broad application prospects in measurement technology for TCM manufacturing and offers strong technical support for the modernization, industrialization, and intelligent development of TCM.
Drugs, Chinese Herbal/analysis* ; Atractylodes/microbiology* ; Rhizome/microbiology* ; Biosensing Techniques/methods* ; Medicine, Chinese Traditional ; Colorimetry/instrumentation* ; Quality Control

Soil microorganisms are one of the important biological indictors of soil quality and can reflct the comprehensive ecological environment characteristics of the soil. The research of soil microbial diversity is the key to know the ecological functions and balance with soil. In this paper, high-throughput sequencing on PCR-amplified 16 S rRNA gene V3-V4 fragments was used to determine the bacterial diversity in rhizosphere soil of A. macrocephala under the treatment with BZJN1 or streptoprofen. The results showed that there were no significant differences of the bacteria in A. macrocephala rhizosphere soil of the streptoprofen treatment group and the biocontrol BZJN1 treatment group. All the soil bacteria was classified into 25 categories,67 classes, 108 orders, 167 families and 271 generas, except some unidentified bacteria. Proteobacteria(30.7%-34.8%) was the dominant phylum, of which Alphaproteobacteria(16.8%-18.5%) was the dominant subgroup. Compared with the control group, the relative abundance of multiple phylums bacteria in the rhizosphere soil of A. macrocephala was significantly changed in the streptoprofen treatment group and the biocontrol BZJN1 treatment group. In addition, RDA analysis showed that there was connection with different environmental factors and microbial communities. The abundance of the three genera in the rhizosphere soil of A. macrocephala was significantly positively correlated with Invertase, Urease and AP. PICRUSt function prediction results showed that BZNJ1 could enhance some bacterial functions and promote the plant growth. Biocontrol is a new type of green and safety control pest method. BZNJ1 significantly enhances some bacterial functions on the basis of effectively preventing root rot of A. macrocephala and promoting plant growth, and has no significant effect on the soil bacterial community structure. All the results can provide theoretical support for popularization of BZNJ1.
Atractylodes ; Bacteria ; Rhizosphere ; Soil ; Soil Microbiology

In this study， an endophytic bacteria strain BZJN1 was isolated from Atractylodes macrocephala， and identified as Bacillus subtilis by physiological and biochemical tests and molecular identification. Strain BZJN1 could inhibit the growth of mycelia of Ceratobasidium sp. significantly， and the inhibition rate was more than 70%. The mycelium growth deformity with bulge as spherical and partially exhaustible in apex or central with microscopic observation. The inhibitory rates under 3% and 6% concentrations of the cell free fermentation were 22.7% and 38.7% expectively. The field test proved that the control efficacy of treatment of 1×10⁸ cfu·mL⁻¹ is 75.27% and 72.37% after 10 and 20 days. All the treatments of strain BZJN1 was able to promote the growth of A. macrocephala， the treatment of 1×10⁸ cfu·mL⁻¹ could able to increase the yield to 14.1%.
Atractylodes ; microbiology ; Bacillus subtilis ; physiology ; Basidiomycota ; pathogenicity ; Biological Control Agents ; Endophytes ; classification ; isolation & purification ; Plant Diseases ; microbiology ; prevention & control

In order to transform main active ingredient of volatile oil, endophytic fungi were screened from the root of Atractylodes lancea. Transformation method was used in vitro. The changes of volatile oil were traced by gas chromatography. One endophytic fungus (strain ALG-13) which could uitilize volatile oil selectively was screened. Single factor experiment were conducted for exploring the effects of various factors that including kinds of carbon source, speed, liquid volume, pH and concentration of plant tissue on degradation by this strain. Subsequently, the main affecting factors carbon source, speed, pH and liquid volume were optimized using orthogonal array design. Results showed that endophytic fungus ALG-13 selectively used the volatile oil, change the relative percentage of the main components of volatile oil, Atractylon and Atractydin were increased, While, beta-eudesmol and Atractylol decreased. After selectively degradation by fungus, volatile oil components percentage were closer to the geo-herbs. Strain ALG-13 was identified as Bionectria ochroleuca according to its morphological characteristics and systematic analysis of ITS sequence. The optimal conditions were as follows: sucrose used as carbon source, rotating speed was 200 r x min(-1), initial pH for medium was 4.5, 50 mL liquid was added in 250 mL flask. The endophytic fungus ALG-13 could degrade the volatile oil selectively, which was benefit for forming geoherbs A. lancea volatile oil composition.
Atractylodes ; chemistry ; microbiology ; Fungi ; classification ; genetics ; isolation & purification ; metabolism ; Oils, Volatile ; chemistry ; metabolism ; Phylogeny ; RNA, Ribosomal, 18S ; genetics

OBJECTIVETo investigate the effects of endophytic fungal elicitors on the growth and atractylodin accumulation of cell suspension cultures of Atractylodes lancea.

METHODThe endophytic fungal elicitors were added to the medium with different concentrations and culture period. Their effects on biomass, atractylodin content and relevant enzyme activities in suspension cultured cells were studied.

RESULTThe cell growth was not affected by elicitors at low concentration and obviously inhibited at high concentration. Inhibition rate reached 46.7% by 100 mg L(-1) elicitor. In addition, six strains from A. lancea, among which Rhizoctonia SP1 activity was higher, had distinctly promoted the accumulation of atractylodin. Atractylodin biosynthesis was notably promoted by 20-60 mg L(-1) Rhizoctonia SP1 elicitor. When 40 mg L(-1) Rhizoctonia SP1 elicitor was added in the medium at the 12 day, the maximum content of atractylodin was 28.06 microg L(-1) at the 21 day with 48.3% higher than that of the control and PPO, POD and CAT activities remarkably increased.

CONCLUSIONAdding the endophytic elicitors to the medium is able to be effective approaches to enhance atractylodin yield in the suspension culture cell of A. lancea.

Atractylodes ; growth & development ; microbiology ; physiology ; Cell Culture Techniques ; Cells, Cultured ; Fungi ; physiology ; Furans ; metabolism ; Symbiosis

OBJECTIVETo find the development rules of microbial community in rhizoma sphere of the cultivated Atractylodes lancea.

METHODTotal bacteria, fungi and actinomyces were counted by CFU x g(-1) though dilution plate method. And genomic DNA of microbes were extracted and amplified by primers of E. coli's 27f and 1492r to get the 16S rDNA, then the restriction endonuclease Hinf was used to digest the 16S rDNA.

RESULTTotal bacteria, fungi and actinomyces in 2-year old soil were lower than in 1-year old soil, they decreased 46. 14%, 49. 25%, 31.88% respectively and made the ratio of themselves changed. At the same time, all the 8 soil samples got fine 16S rDNA bands, which were about 1500 bp. And the main bands of most of the samples were found at 1000 bp, but the weak bands of each were different although most bands in the same year samples were more similar than in different year ones.

CONCLUSIONIt is indicated that the change of soil microbial community may has some relation to the continous cropping barrier of A. lancea.

Actinomyces ; genetics ; isolation & purification ; Atractylodes ; growth & development ; Bacteria ; genetics ; isolation & purification ; Biodiversity ; Colony Count, Microbial ; DNA, Ribosomal ; genetics ; Fungi ; genetics ; isolation & purification ; Plants, Medicinal ; growth & development ; RNA, Ribosomal, 16S ; genetics ; Rhizome ; growth & development ; Soil Microbiology

OBJECTIVETo study the effects of Arbuscular Mycorrhizae on cultivated Atractylodes lancea.

METHODPot experiment of A. lancea, with (code as AM) or without (code as CK) Glomus mosseae (GM) was conducted 5 times respectively, then the biomass, essential oil, and soil nutrition, soil organism, soil microbial were detected after A. lancea were harvested.

RESULT(1) Mycorrhizal dependency of A. lancea was 245%, and height of individuals, numbers of leaves, leaf area, biomass of A. lancea were all higher in AM than in CK (P < 0.05). (2) GC-MS analysis with cluster analysis and principal components analysis showed that there were no differences in essential oil of A. lancea between AM and CK. (T3) Total N, available N, available P and available K in AM soil were all lower than in CK soil. (4) GC-MS analysis showed organic matters changed differenly in AM soil and CK soil, components 5,6 in AM soil were higher than that in CK soil, but component 9, 10, 11 were lower in AM soil than that in CK soil. (5) Biolog detect showed AWCD of AM soil microbe were higher than that of CK soil throughout the incubation, and AWCD of the former was 0.66, and the later was 0.46 after 192 h incubation. and t-test showed, Shannon seven indices and McIntosh'seven indices were same both at 72 h and 192 h, and diversity indices of Shannon and McIntosh were also same at 72 h, but AM soil microbe were higher than CK soil microbe at 168 h (P < 0.05).

CONCLUSIONAM could promote nutrition uptake, improve the function diversity and activity of microbe in rhizosphere of A. lancea, influence the composition of the organic matter, that lead the growth of A. lancea, but not to the quality.

Atractylodes ; growth & development ; metabolism ; microbiology ; Mycorrhizae ; physiology ; Nitrogen ; analysis ; Oils, Volatile ; metabolism ; Phosphorus ; analysis ; Plants, Medicinal ; growth & development ; metabolism ; microbiology ; Potassium ; analysis ; Soil ; analysis ; Soil Microbiology

Disinfection means killing or removing pathogenic microorganisms in media to realize a harmless process. A disinfectant, which is also referred to as a disinfection medicine in relevant regulations, is the medicine used to kill microorganisms for the purpose of disinfection. The disinfectants prepared from plants (including traditional Chinese herbal medicines) and the extracts thereof are called herbal disinfectants. China has a long history of using herbal disinfectants. As early as in 533 A.D., the use of Cornel to sterilize well water was recorded in Necessary Techniques for Qi People by Jia Enxie of the Beiwei Dynasty. During the Dragon Boat Festival, people often use fumigants made of traditional Chinese herbal medicines like Chinese Atractylodes, Argy Wormwood Leaf and Red Arsenic Sulfide to smoke their houses, so as to ward off plagues and drive away evils. In fact this is now a kind of disinfection practice.
Arsenicals ; pharmacology ; Atractylodes ; chemistry ; China ; Disinfectants ; pharmacology ; Fumigation ; Medicine, Chinese Traditional ; Mucous Membrane ; microbiology ; Plant Extracts ; chemistry ; pharmacology ; Plant Leaves ; chemistry ; Skin Diseases, Infectious ; microbiology ; prevention & control ; Sulfides ; pharmacology