Nature’s Pharmacy under Siege: Investigating Antibiotic Resistance Pattern in Endophytic Bacteria of Medicinal Plants

Antibiotic resistance poses a significant global health threat, necessitating a thorough understanding of its prevalence in various ecological contexts. Medicinal plants, renowned for their therapeutic properties, host endophytic bacteria that produce bioactive compounds. Understanding antibiotic resistance dynamics in these bacteria is vital for human health and antibiotic efficacy preservation. In this study, we investigated antibiotic resistance profiles in endophytic bacteria from five medicinal plants: Thankuni, Neem, Aparajita, Joba, and Snake plant. We isolated and characterized 113 endophytic bacteria, with varying resistance patterns observed against multiple antibiotics. Notably, 53 strains were multidrug-resistant (MDR), with 14 exhibiting extensive drug resistance (XDR). Thankuni-associated bacteria displayed 44% MDR and 11% XDR, while Neem-associated bacteria showed higher resistance (60% MDR, 13% XDR). Aparajita-associated bacteria had lower resistance (22% MDR, 6% XDR), whereas Joba-associated bacteria exhibited substantial resistance (54% MDR, 14% XDR). Snake plant-associated bacteria showed 7% MDR and 4% XDR. Genus-specific distribution revealed Bacillus (47%), Staphylococcus (21%), and Klebsiella (11%) as major contributors to MDR. Our findings highlight diverse drug resistance patterns among plant-associated bacteria and underscore the complexity of antibiotic resistance dynamics in diverse plant environments. Identification of XDR strains emphasizes the severity of the antibiotic resistance problem, warranting further investigation into contributing factors.

Antimicrobial and plant growth-promoting activities of bacterial endophytes isolated from Calotropis procera(Ait.)W.T.Aiton

Bacterial endophytes are beneficial to their hosts as they can fix nitrogen in the soil and make it available to the host.Endophytic bacteria also secrete plant growth-promoting hormones to support their host plants under normal as well as stress conditions.The current study aimed to isolate endophytic bacteria from different parts of Calotropis procera,i.e.,roots,stem and leaves of Calotropis procera(Ait.)W.T.Aiton.Plants were collected from the Lundkhwar,district Mardan.A total of 12 bacterial strains,i.e.,six from roots,three from the stem and three from the leaves were isolated.The strains were screened for their growth-promoting activity in rice plants because rice shows a quick and easy response to the bioactive compounds present in the culture filtrate(CF)of the potent endophytic strains.The rice plants were cultivated in pots containing 30 mL of 0.8%w/v water-agar medium.The pots were placed in a growth chamber,operated at 28±0.3℃ for 14 h(day);and 25±0.3℃ for 10 h(night),at 70%relative-humidity.Among the isolated strains,R1,S1,S3,L1,R5 and R6 showed visible growth promotion in rice plants.The biochemical analysis revealed that the strains were able to produce indole acetic acid(IAA)and flavonoids in higher quantities.Moreover,the strains also produced bioactive compounds that inhibited the growth of Escherichia coli and Aspergillus flavus using the well diffusion method.From the results,it was concluded that these strains can secrete potent compounds that can promote the host plant growth and inhibit the growth of pathogenic microorganisms and,therefore,can be used as bio-fertilizer and bio-control agents.

Plant growth-promoting properties and anti-fungal activity of endophytic bacterial strains isolated from Thymus altaicus and Salvia deserta in arid lands

Endophytes,as crucial components of plant microbial communities,significantly contribute to enhancing the absorption of nutrients such as nitrogen and phosphorus by their hosts,promote plant growth,and degrade pathogenic fungal mycelia.In this study,an experiment was conducted in August 2022 to explore the growth-promoting potential of endophytic bacterial strains isolated from two medical plant species,Thymus altaicus and Salvia deserta,using a series of screening media.Plant samples of Thymus altaicus and Salvia deserta were collected from Zhaosu County and Habahe County in Xinjiang Uygur Autonomous Region,China,in July 2021.Additionally,the inhibitory effects of endophytic bacterial strains on the four pathogenic fungi(Fusarium oxysporum,Fulvia fulva,Alternaria solani,and Valsa mali)were determined through the plate confrontation method.A total of 80 endophytic bacterial strains were isolated from Thymus altaicus,while a total of 60 endophytic bacterial strains were isolated from Salvia deserta.The endophytic bacterial strains from both Thymus altaicus and Salvia deserta exhibited plant growth-promoting properties.Specifically,the strains of Bacillus sp.TR002,Bacillus sp.TR005,Microbacterium sp.TSB5,and Rhodococcus sp.TR013 demonstrated strong cellulase-producing activity,siderophore-producing activity,phosphate solubilization activity,and nitrogen-fixing activity,respectively.Out of 140 endophytic bacterial strains isolated from Thymus altaicus and Salvia deserta,104 strains displayed anti-fungal activity against Fulvia fulva,Alternaria solani,Fusarium oxysporum,and Valsa mali.Furthermore,the strains of Bacillus sp.TR005,Bacillus sp.TS003,and Bacillus sp.TSB7 exhibited robust inhibition rates against all the four pathogenic fungi.In conclusion,the endophytic bacterial strains from Thymus altaicus and Salvia deserta possess both plant growth-promoting and anti-fungal properties,making them promising candidates for future development as growth-promoting agents and biocontrol tools for plant diseases.

Endophytic Bacteria Promote the Growth of Suaeda glauca in Saline-Alkali Stress:Regulation of Osmotic Pressure and Antioxidative Defense System

Endophytic bacteria are promising bacterial fertilizers to improve plant growth under adverse environment.For ecological remediation of coastal wetlands,it was necessary to investigate the effect and interaction of endophytes on halophytes under saline-alkali stress.In this study,an endophytic bacterium strain HK1 isolated from halophytes was selected to infect Suaeda glauca under pH(7 and 8)and salinity gradient(150,300 and 450mmolL^(-1)).Strain HK1 was identified as Pantoea ananatis and it had ability to fix nitrogen,dissolve inorganic phosphorus and produce indole-3-aceticacid(IAA).The results showed that strain HK1 could promote the growth of S.glauca seedings when the salinity was less than 300mmolL^(-1),in view of longer shoot length and heavier fresh weight.The infected plants could produce more proline to decrease the permeability of cells,which content increased by 26.2%–61.1%compared to the non-infected group.Moreover,the oxidative stress of infected plants was relieved with the malondialdehyde(MDA)content decreased by 16.8%–32.9%,and the peroxidase(POD)activity and catalase(CAT)activity increased by 100%–500%and 6.2%–71.4%,respectively.Statistical analysis revealed that increasing proline content and enhancing CAT and POD activities were the main pathways to alleviate saline-alkali stress by strain HK1 infection,and the latter might be more important.This study illustrated that endophytic bacteria could promote the growth of halophytes by regulation of osmotic substances and strengthening antioxidant activities.This finding would be helpful for the bioremediation of coastal soil.

Isolation and Identification of Houttuynia cordata Endophytic Bacteria and Study of Antibiotic Activity

[Objectives]To further explore the resources of biocontrol strains and provide new options for the control of pathogenic bacteria.[Methods]Houttuynia cordata endophytic bacteria were isolated and identified,and the antibiotic activity was explored.[Results]There are 8 strains(H1-H8)of endophytic bacteria isolated from the healthy plants of H.cordata,which had an antagonistic effect on the pathogenic bacteria.According to the gene sequence,the 8 strains were identified as Stenotrophomonas,Pseudomonas,and Bacillus,which strongly inhibited Colletotrichum(HC1),Fusarium(HC2),and Rhizoctonia(HC5)of H.cordata.On the PDA culture dish,the inhibition diameter of the 8 strains on Colletotrichum,Fusarium,and Rhizoctonia was 6,8,and 5 mm,respectively.[Conclusions]Pathogenic bacteria and endophytic bacteria inoculated with H.cordata seedlings showed that endophytic bacteria have certain antagonistic activity,and have the potential for development and application.

Diversity Analysis of Endophytes in Wheat Infected by Powdery Mildew

Common wheat (Triticum aestivum L.) is one of the most important food crops. Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most serious diseases on wheat. In this study, the changes of endogenous bacteria in root, stem and leaf tissues of wheat infected and uninfected with powdery mildew were measured based on 16S rDNA. Integration, OTU cluster analysis, taxonomic analysis, diversity index, Shannon-Index curve, Rank-Abundance curve and PCoA analysis were carried out for each sample, and the roots, stems and leaves of different tissue parts were classified and summarized. The results showed that the infection of wheat powdery mildew had a certain effect on endophytic bacteria in stem tissue. There are also differences in the control and treatment of leaf tissue and root tissue. This indicated that endophytic bacteria were distributed differently in different parts of wheat.

Feasibility of enhanced phytoextraction of Zn contaminated soil with Zn mobilizing and plant growth promoting endophytic bacteria

Three bacterial endophytes of Sedum alfredii, VI8L2, II8L4 and VI8R2, were examined for promoting soil Zn bioavailability and Zn accumulation in S. alfredii. Results showed that three strains were re-introduced into S. alfredii rhizosphere soils under Zn stress and resulted in better plant growth, as roots biomass increased from 80% to 525% and shoot biomass from 11% to 47% compared with the uninoculated ones. Strains IVsLz, II8L4 and IVsR2 significantly increased shoot and root Zn concentrations in the ZnCO3 contaminated soil. Inoculation with strain IVsL2 resulted in 44% and 39% higher shoot and root Zn concentrations, while strain IV8R2 significantly decreased shoot Zn concentration in the Zn3（PO4）2 contaminated soils. In the aged contaminated soil, isolates IVsL2, IIsL4 and IVsR2 significantly increased root Zn concentration, but decreased shoot Zn concentration of Sedum alfredii. It suggested that endophytes might be used for enhancing phytoextraction efficiency.

Screening for Siderophore-producing Endophytic Bacteria against Fusarium oxysporum

[Objective] With a strain of endophytic antagonistic bacteria with good disease control efficacy in pot as trial material,the paper was to explore the antagonistic mechanism of siderophore against Fusarium oxysporum.[Method] Whether the strain produced siderophore and its fluorescent property was judged by MSA（Modified Sugar-Aspartic acid） plate.The siderophore activity of strains in liquid MSA medium was determined through the absorbance values at particular wavelength.The inhibition effects of it siderophore against F.oxysporum under different concentrations of Fe3＋ were compared.Then the strain was preliminarily identified based on morphological,physiological and biochemical characteristics [Result] The fluorescent siderophore produced by the endophytic bacteria in MSA medium had a strong inhibition effect against F.oxysporum.With the increasing concentration of Fe3＋,the inhibition effect against the pathogen weakened.The preliminary analysis showed these strains belonged to Bacillus.[Conclusion] Bacillus could compete the absorption of Fe3＋ by secreting siderophore to inhibit the growth of F.oxysporum.

Determination of Growth-promoting and Antagonistic Action of Endophytic Bacteria Strains Itb57 and Itb295 of Tobacco

[ Objective ] The paper was to study the growth-promoting and antagonistic action of endophytic bacteria strains Itb57 and Itb295 of tobacco to explore their functions in biological control. [ Method] The growth-promoting effects of bacterial suspension ~ff endophytic bacteria Itb57 and Itb295 on tobacco seedling un- der different treatment modes were studied using potting method in greenhouse. The antagonistic action of bacterial suspension of endophytic bacteria Itb57 and Itb295 on Phytophthora nicotianae, Alternaria alternata and Botrytis cinerea were measured by duel culture method. [ Result] Bacterial suspensions of enduphytic bacteria Itb57 and Itb295 had certain growth-promoting effects on tobacco seedling, which could significantly increase the fresh weight and dry weight in aerial part; the growth-promoting effect of soaking ＋ spraying and irrigating treatment was the best. Itb57 strain had good antagonistic action against P. nicotianae. A. alterna- ta and B. cinerea, while Itb295 strain only had good antifungal effect against P. nicotianae. [ Conclusion] The results provided basis for the study and application of tobacco endophytic bacteria strains Itb57 and Itb295 in biocontrol of tobacco diseases.

Impacts of Irrigation with Reclaimed Water on Endophytic Bacteria in Chinese Cabbage in Urumqi City

[Objective] This study aimed to investigate the feasibility of agricultural irri- gation with reclaimed water in Urumqi City. [Method] Chinese cabbages were used as experimental materials and irrigated with control water, 50% reclaimed water and 100% reclaimed water, and then the number of endophytic bacteria in Chinese cab- bages was measured. [Result] Using 50% reclaimed water, the fresh weight of Chi- nese cabbage was improved by 68.94%; however, with the deepening internalization of exogenous microorganism internalization, the total number of endophytic bacteria and coliform flock in Chinese cabbages treated with 50% reclaimed water was sig- nificantly higher than the other two treatment groups （P〈0.05）. [Conclusion] Re- claimed water enhances both the weight of Chinese cabbage and the number of endophytic bacteria, which is unsuitable for irrigation of edible crops such as Chi- nese cabbage. Strengthening disinfection during the process of sewerage treatment could solve this problem.

Microbial Fertilizer: A Sustainable Strategy for Medicinal Plants Production

Medicinal plants have aroused considerable interest as an alternative to chemical drugs due to the beneficial effects of their active secondary metabolites.However,the extensive use of chemical fertilizers and pesticides in pursuit of yield has caused serious pollution to the environment,which is not conducive to sustainable devel-opment in thefield of medicinal plants.Microbial fertilizers are a type of“green fertilizer”containing specific microorganisms that can improve the soil microbial structure,enhance plant resistance to biological and abiotic stresses,and increase the yield of medicinal plants.The root exudates of medicinal plants attract different micro-organisms to the rhizosphere,which then migrate further to the plant tissues.These microbes can increase the levels of soil nutrients,and improve the physical and chemical properties of soil through nitrogenfixation,and phosphorus and potassium solubilization.In addition,soil microbes can promote the synthesis of hormones that increase plant growth and the accumulation of active compounds,eventually improving the quality of med-icinal plants.In 2022,the total value of the global microbial fertilizer market was$4.6 billion and is estimated to reach$10.36 billion by 2030.In this review,we have summarized the types of microbial fertilizers,the coloniza-tion and migration of microorganisms to plant tissues,and the beneficial effects of microbial fertilizers.In addi-tion,the prospects of developing microbial fertilizers and their application for medicinal plants have also been discussed.It aims to provide a reference for the rational application of microbial fertilizers in thefield of med-icinal plants and the green and sustainable development of medicinal plant resources.

Genetic Diversity of Antagonistic Endophytic Bacteria Isolated from Sophora alopecuroide

[ Objective ] The paper was to study and analyze the genetic diversity of 48 endophytic bacteria isolated from Sophora alopecuroide with strong antagonis- tic effect against Fusarium oxysporum and Verticillium dahliae, and carry out sequence measurement and phylogenetic analysis on 10 representative strains. [ Meth- od] Using total DNA ERIC-PCR and 16S rRNA sequencing analysis method, a total of 48 endophytic bacteria were carried out genetic diversity and phylogenetic analysis. [Result] ERIC-PCR fingerprint map showed that 48 endophytic bacteria were divided into 6 ERIC groups and 2 strains with independent groups at Wat- son distance of 0.31. Representative strains were selected from each group to determine 16S rRNA gene for phylogenetic analysis, and the results showed that these strains belonged to Bacillus atrophaeus, Bacillus subtilis, Bacillus cereus, Bacillus megaterium, Bacillus pumilus and Serratia marcescens, respectively. [ Conclu- sion ] The genetic diversity of 48 strains of bacteria was obvious. Determination of 48 strains of bacteria on diversity and phylogenetic status could lay foundation for study on mechanism of F. oxysporum and V. dahliae, which could also provide new strain sources for biological control of diseases in cotton production.

Determination of Endophytic Bacteria Resistance against Rifampin and UV-mutagenesis

The study aimed to isolate and screen endophytic bacteria from the plant in order to understand their colonization dynamics in plants. [Method] 5 kinds of endophytic bacteda including H1, DP1, CJL1, DJL12 and YC1 were selected as the original strains, and conducted UV mutagenesis studies on H1 and DJL12 with weak resistance, while the resistance of them against rifampicin was confirmed. [Result] Through preliminary screening, five kinds of endophytic bacteria could grew well in the plate without rifampin, among them, H1 and DLJ12 were unable to survive under 10 μg/ml concentration of rifampicin, and the other three strains could survive under 50 μg/ml concentration of rifampicin. After UV mutagenesis, DLJ12 strain with dfampicin resistance concentration of 80 μg/ml was obtained. [ Conclusion] The study could provide theoretical and practical basis for development and utilization of endophytic bacteria.

Endophytic bacteria associated with endangered plant Ferula sinkiangensis K. M. Shen in an arid land： diversity and plant growth-promoting traits

Ferula spp. are traditional medicinal plants found in arid land. Large-scale excavation for extracting bioactive compounds from the plants in arid regions of Xinjiang over the last few years has, however, significandy decreased their distributions. Due to the urgent need for preservation of these plant resources, along with the need of searching for alternative source of the useful metabolites, it is important to screen the endophytic microbial resources associated with the plant Ferula sinkiangensis K. M. Shen. In the study, a total of 125 endophytic bacteria belonging to 3 phyla, 13 orders, 23 families, and 29 genera were isolated based on 16S rRNA gene sequencing data. Among the different isolates, three strains isolated from roots were potential novel species of the genera Porphyrobacter, Paracoccus and draycofatopsis. In this study, 79.4% and 57.1% of the total isolates were capable of producing indole-3-acetic acid （IAA） and siderophore, respectively. And, 40.6% of the strains inhibit the growth of fungal pathogen Afternaria alternata, 17.2% and 20.2% strains were positive for antagonism against Vertidllium dahlia 991 and V. dahlia 7, respectively. These results demonstrated that E sinkiangensis is a rich reservoir of endophytic bacterial resources with potential for production of biologically important functions such as plant growth-promoting factors.

Antimicrobial Activity of Ulva reticulata and Its Endophytes

Seaweeds are known to exhibit various antimicrobial properties, since it harbours an enormous range of indigenous bioactive compounds. The emergence of drug resistant strains has directed to the identification of prospective metabolites from seaweed and its endophytes, thereby exploiting the properties in resisting bacterial diseases. The current study was aimed to assess the antimicrobial activity of extracts obtained from Ulva reticulate, for which metabolites of Ulva reticulata and its endophytes were extracted and assessed against human pathogens like Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and Bacillus subtilis. It was observed that the hexane extract of isolate VITDSJ2 was effective against all the tested pathogens but a significant inhibition was observed for Staphylococcus aureus and Escherichia coli. Further, Gas chromatography coupled with Mass spectroscopy （GC-MS） revealed the existence of phenol, 3, 5-bis （1, 1-dimethylethyl） in the crude hexane extract which is well-known to possess antibacterial activity. The effective isolate VITDSJ2 was identified to be the closest neighbour of Pseudomonas smtzeri by phenotypic and genotypic methods. The crude extracts of the seaweed Ulva reticulata was also screened for antibacterial activity and the hexane extract was effective in showing inhibition against all the tested pathogens. The compound in the crude extract of Ulva reticulata was identified as hentriacontane using GC-MS. The extracts obtained from dichloromethane did not show significant activity in comparison with the hexane extracts. Hence the metabolites of Ulva reticulata and the bacterial secondary metabolites of the endophytes could be used in the treatment of bacterial infections.

The diversity and potential function of endophytic bacteria isolated from Kobreasia capillifolia at alpine grasslands on the Tibetan Plateau, China

A total of 50 endophytic bacterial isolates were obtained from Kobreasia capillifolia at alpine grasslands in the Eastern Qilian Mountains on the Tibetan Plateau in China. Based on the sequencing and phylogenetic analysis of 16 S r DNA genes, all isolates phylogenetically related closely to Bacillus, Acinetobacter, Stenotrophomonas, Brevundimonas, Arthrobacter, Curtobacterium, Paenibacillus, Plantibacter, Promicromonospora, Serratia, and Microbacterium, among which Bacillus was the predominant genus（47.8% of the total number of endophytic isolates）. These isolates possessed different biological functions. In 50 endophytic bacteria, 42 isolates produced indole acetic acid（IAA） on King medium. There were seven isolates showing potency of mineral phosphate solubilization in Pikovaskaia＇s（PKO） liquid medium. Seven isolates exhibited antagonistic effect against Fusarium avenaceum, Colletotrichum coccodes and Phoma foveata. This was the first report on diversity and plant growth promotion of endophytic bacteria from K. capillifolia on alpine grassland in the Eastern Qilian Mountains, Chain. It is essential for revealing the relationship among plant, microorganism, and the special environment because the potential function of endophytic bacteria made a contribution to the interactions of plants and endophytic bacteria.

Endophytic bacteria associated with halophyte Seidlitzia rosmarinus Ehrenb.ex Boiss.from saline soil of Uzbekistan and their plant beneficial traits

Endophytic bacteria of halophytic plants play essential roles in salt stress tolerance.Therefore,an understanding of the true nature of plant-microbe interactions under extreme conditions is essential.The current study aimed to identify cultivable endophytic bacteria associated with the roots and shoots of Seidlitzia rosmarinus Ehrenb.ex Boiss.grown in the salt-affected soil in Uzbekistan and to evaluate their plant beneficial traits related to plant growth stimulation and stress tolerance.Bacteria were isolated from the roots and the shoots of S.rosmarinus using culture-dependent techniques and identified by the 16S rRNA gene.RFLP(Restriction Fragment Length Polymorphism)analysis was conducted to eliminate similar isolates.Results showed that the isolates from the roots of S.rosmarinus belonged to the genera Rothia,Kocuria,Pseudomonas,Staphylococcus,Paenibacillus and Brevibacterium.The bacterial isolates from the shoots of S.rosmarinus belonged to the genera Staphylococcus,Rothia,Stenotrophomonas,Brevibacterium,Halomonas,Planococcus,Planomicrobium and Pseudomonas,which differed from those of the roots.Notably,Staphylococcus,Rothia and Brevibacterium were detected in both roots and shoots,indicating possible migration of some species from roots to shoots.The root-associated bacteria showed higher levels of IAA(indole-3-acetic acid)synthesis compared with those isolated from the shoots,as well as the higher production of ACC(1-aminocyclopropane-1-carboxylate)deaminase.Our findings suggest that halophytic plants are valuable sources for the selection of microbes with a potential to improve plant fitness under saline soils.

Community Structure of Endophytic Bacteria in Chestnut, Castanea mollissima

[ Objective] The paper was to detect the species of endophytic bacteria in Chestnut （Castanea mollissima）. [ Method ] The 16S rDNA-V4 region of endophytic bacteria in chestnut （sample name BL） was sequenced by Illumina MiSeq high-throughput sequencing technology. The number of sequences and op- erational taxonomic units （OTUs） for each sample was sorted and calculated using Qiime and Mothur software. The abundance and α-diversity of species were analyzed. [ Result] The number of effective sequences and OTUs were 21 167/191. The rarefaction curves showed that adequate sampling was achieved, and the number of OTUs was close to saturation. The endophytic bacteria of chestnut belonged to 10 genera, including Koribacter （3.48%） , Solibacter （2.59%） , Bra- d^hizobium （ 3. 57% ）, Rhodoplanes （ 7. 41% ）, Methylobacterium （ 4. 20% ）, Agrobacterium （ 19. 73% ）, Kaistobacter （ 2. 05% ）, Sphingomonas （18.66%）, Acidovorax （35.98%）, and Methylibium （2.32%）. The dominant species were Acidovorax, Agrobacterium and Sphingomonas. [ Conclusion] Illumina high-throughput sequencing technology provided more accurate and scientific data resources for the study of endophytic bacteria in chestnut.

Isolation of Antagonistic Bacteria against Fusarium graminearum and Its Antagonistic Activity

[ Objective] To develop biological control resources of endophytic bacteria in wheat, the bacteria with antagonistic effect against Fusarium graminearum were screened and their antagonistic activity was studied. [ Method] The endophytic bacteria of wheat with antagonistic effect were screened using plate confrontation method; the antagonistic bacteria were identified through physiological and biochemical characteristics and 16S rDNA sequence analysis; the antagonistic spectrum of antagonistic bacteria was studied using plate confrontation method. [ Result] The antagonistic bacterium XY-3 was isolated from wheat leaves, and was identified as Bacillus amyloliquefaciens according to the physiological and biochemical characteristics and 16S rDNA sequence analysis. [ Conclusion] XY-3 had antagonistic effect against four kinds of common pathogenic fungi, presenting broad spectrum antagonistic activity against plant pathogenic fungi.

Analyses on Community Composition and Diversity of Endophytic Bacteria in Different Tissues of Anoectochilus roxburghii

[Objective] This study aimed to investigate the diversity of community composition and diversity of endophytic bacteria in Anoectochilus roxburghii.[Method] By using Illumina MiSeq high-throughput sequencing technology, the 16S rDNA-V4 region of endophytic bacteria in ^4. roxburghii was sequenced. Qiime and Mothur software packages were used to sort and count the number of sequences and operational taxonomic units （OTUs） and analyze the species abundance and distribution, Alpha diversity and differences of species abundance among samples. [Result] The number of effective sequences/OTUs for each sample was 110485/247 （stem） and 98524/259 （leaf） , respectively. The rarefaction curve showed that adequate sampling was performed. The number of OTUs had almost reached a saturation point. Endophytic bacteria in A. roxburghii mainly belonged to the genera Acidovorax, Pseudomonas, Sphingomonas and Halomonas. [Conclusions] Illumi-na MiSeq high-throughput sequencing technology provides more accurate and scientific data resources for the study of endophytic bacteria in A. roxburghii.