1.Chemotactic response of ginseng endophyte to ginseng root exudates.
Xin-Xin ZHANG ; Ai-Hua ZHANG ; Feng-Jie LEI ; Li CAI ; Zhou-Yang XU ; Zhi-Qing LIU ; Lian-Xue ZHANG
China Journal of Chinese Materia Medica 2019;44(24):5358-5362
The ginseng endophytic bacteria F1 is a potential biocontrol agent for ginseng bacterial soft rot. In this paper,the chemotactic response of ginseng endophytic bacteria F1 on 8 kinds of sugar and amino acids was detected by capillary method to explore its biocontrol mechanism. The chemotactic response of F1 strain to 4 kinds of better chemotaxis substances such as glucose,glycine,L-rhamnoseand L-glutamic acid under parameters( concentration,time,temperature and pH) was studied. The results showed that under the same experimental conditions( incubation temperature 25 ℃,incubation time 60 min,chemotaxis concentration 1 mg·L~(-1)),ginseng endophytic bacteria F1 showed different degrees of response to the eight substances tested. The phenomenon of positive chemotaxis of the measured sugars and amino acids was obvious,and the chemotactic response to total ginsenosides was low. The degree of chemotaxis response is positively correlated with the chemotaxis index within a certain range of parameters,but as the temperature,p H,time,concentration and other factors continue to increase,the chemotaxis effect decreases,and F1 optimizes the chemotaxis of the four substances. The parameters are as follows: glucose: 25 ℃,10 mg·L~(-1),45 min,pH 7; glycine: 30 ℃,10 mg·L~(-1),75 min,pH7; L-rhamnose: 30 ℃,1 mg·L~(-1),30 min,pH 6; L-glutamic acid: 25 ℃,0. 1 mg·L~(-1),45 min,pH 8. The chemotactic response is more sensitive to low concentrations of chemotactic substances.
Amino Acids/pharmacology*
;
Bacteria/drug effects*
;
Chemotaxis
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Endophytes/physiology*
;
Ginsenosides/pharmacology*
;
Panax/chemistry*
;
Plant Exudates/pharmacology*
;
Sugars/pharmacology*
2.Identification of endophytic bacteria BZJN1 and research on biological control of root rot of Atractylodes macrocephala.
Jing-Mao YOU ; Kun XIONG ; Sen MU ; Jie GUO ; Xiao-Liang GUO ; Yuan-Yuan DUAN ; Juan LI ; Fan CAO ; Zong-Cheng ZOU ; Hai TAN
China Journal of Chinese Materia Medica 2018;43(3):478-483
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
3.Comparative study on distribution of endophytic fungi in Eucommia barks from different habitats.
Xue-Juan LIANG ; Shui-Han ZHANG ; Ping ZHANG ; Fei PENG ; Jian KE ; Ya-Nan MI
China Journal of Chinese Materia Medica 2014;39(2):204-208
A total of 152 strains of endophytic fungi were isolated from the barks of Eucommia ulmoides in three regions (Lueyang country, Zunyi country, Cili country). Based on morphological characteristics and analysis of ITS sequences, these strains were identified into 8 genera. Thereinto Phomopsis, Diaporthe and Alternaria were common genera to Eucommia barks from different sites. But the dominant genus was different: Alternaria was the dominant genus in the barks from Cili country, and Phomopsis was the dominant genus from Zunyi country, then Diaporthe was the one from Lueyang country. According to the similarity coefficient, the composition of the endophytic fungi was distinctly different between the barks from three sites. The diversity and species richness in Lueyang country and Cili country were found higher than those in Zunyi country. The evenness of endophytic fungi was 0.936 5 in Lueyang county, which was higher than 0.737 1 or 0.641 0 in Cili county or Zunyi county, respectively. After phylogenic analysis and calculating the genetic distances of typical strains belong to Phomopsis and its perfect stage--Diaporthe, there was very high genetic diversity in the two genera from our study. In conclusion, the community structure and diversity of endophytic fungi were significant different in Eucommia barks from the three habitats.
DNA, Fungal
;
genetics
;
DNA, Intergenic
;
genetics
;
Ecosystem
;
Endophytes
;
classification
;
physiology
;
Eucommiaceae
;
microbiology
;
Fungi
;
classification
;
genetics
;
physiology
;
Phylogeny
;
Plant Bark
;
microbiology
4.Effects of agarwood formation induced by Fusarium sp. A2 on distribution and community composition of endophytic fungi in leaves of Aquilaria sinensis.
Xiao-Xia GAO ; Wei-Ping ZHOU ; Lei WANG ; Wei-Min ZHANG ; Han-Jing YAN
China Journal of Chinese Materia Medica 2014;39(2):197-203
The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.
Cluster Analysis
;
DNA, Fungal
;
genetics
;
Endophytes
;
classification
;
genetics
;
physiology
;
Fusarium
;
classification
;
genetics
;
physiology
;
Phylogeny
;
Plant Leaves
;
metabolism
;
Thymelaeaceae
;
microbiology
;
Wood
;
metabolism
;
microbiology
5.Colonization characteristics of endophytic bacteria NJ13 in Panax ginseng and its biocontrol efficiency against Alternaria leaf spot of ginseng.
Chang-Qing CHEN ; Tong LI ; Xin-Lian LI ; Yun JIANG ; Lei TIAN ; Peng XU
China Journal of Chinese Materia Medica 2014;39(10):1782-1787
To reveal the colonization characteristics in host of endophytic biocontrol bacteria NJ13 isolated from Panax ginseng, this study obtained the marked strain NJ13-R which was double antibiotic resistant to rifampicin and streptomycin through enhancing the method of inducing antibiotic. The colonization characteristics in ginseng and its biocontrol efficiency against Alternaria spot of ginseng in the field were studied. The results showed that the strain could colonize in root, stem and leaf of ginseng and the colonization amount was positive correlated with inoculation concentration. Meanwhile, the strain could infect and then transfer in different tissues of ginseng The colonization amount of strain in roots and leaves of ginseng increased first and then decreased. However, the tendency of colonization amount of strain in stems was ascend at first and then descend slowly, and was more than that in roots and leaves along with time, which had a preference to specific tissue of its host. In field experiment, the endophytic bacteria NJ13 was proved to be effective in controlling Alternaria leaf spot of ginseng. The biocontrol efficiency of fermentation broth at the concentration of 0.76 x 10(8) cfu x mL(-1) reached 75.62%, which was close to the controlling level (73.06%) of 0.67 mg x L(-1) 50% cyprodinil WG.
Alternaria
;
physiology
;
Antibiosis
;
Bacillus
;
growth & development
;
isolation & purification
;
physiology
;
Endophytes
;
growth & development
;
isolation & purification
;
physiology
;
Panax
;
growth & development
;
microbiology
;
Plant Diseases
;
microbiology
6.Effect of different treatment on endophytic bacterial communities in continuous cropping of Chrysanthemum morifoliu.
San-meil PENG ; Bo-lin WANG ; Jian-zhong XU ; Zhi-shan DING ; Xiao-feng YUAN
China Journal of Chinese Materia Medica 2014;39(24):4763-4768
To reveal the effect of rotation cropping and bacterial manure on the growth of Chrysanthemum morifolium and screen the beneficial endophytic, the diversity of endophytic and dominant genera of different treatment groups were analyzed. Four different treatments were continuous cropping, rotation, self-made organic fertilizer and commercially available fertilizer, respectively. Endophytic bacterial diversity and dominant genera in different organs were examined using Terminal Restriction Fragment Length Polymorphism (T-RFLP). The results showed that enzyme Hae III was more appropriate than enzyme Hinfl because the number of TRFs digested by enzyme Hae III was more than that of enzyme Hinfl. In comparison of diversity, the endophytic bacterial communities' diversity index in group of cropping rotation and fertilizer was higher than that of continuous cropping which indicated that the addition of exogenous microorganism in soil could increase the diversity of plant endophyte. 18 dominant species were selected, including 3 kinds of Firmicutes, 4 kinds of Actinomycetes and 11 kinds of Proteobacteria. The results of dominant species comparison showed that the number of dominant species in continuous cropping of Ch. morifolium was significantly less than that of the rotation group. Some dominant bacteria in rotation group and fertilizer group such as Arthrobacter, Streptomyces, Streptomyces, Flavobacterium and Mycobacterium were not found in the continuous cropping of Ch. mortfolium group. Dominant species of fertilizer treatment group was similar with the rotation group, and the continuous cropping group's dominant species was more abundant. It indicates that these bacteria may be able to mitigate hindrance in continuous cropping, especially the Flavobacterium which can decompose the pathogenic fungi is worthy of further attention. Compared with leaves, there are more dominant species in roots and stems. The diversity of edophytic bacterial communities in continuous cropping of Ch. morifolium stays below than that in the rotation of Ch. morifolium, and fertilizer treatment can increase the diversity of continuous cropping so that it could mitigate hindrance in continuous cropping.
Actinobacteria
;
physiology
;
Agriculture
;
Biodiversity
;
Chrysanthemum
;
growth & development
;
microbiology
;
Deoxyribonucleases, Type II Site-Specific
;
Endophytes
;
Fertilizers
;
Gram-Positive Bacteria
;
physiology
;
Phylogeny
;
Plant Leaves
;
Plant Roots
;
microbiology
;
Polymorphism, Restriction Fragment Length
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Proteobacteria
;
physiology
;
RNA, Ribosomal, 16S
;
chemistry
;
genetics
;
Soil
;
Soil Microbiology
7.Fermentation of Bacillus subtilis ge25 strain and preliminary study on its antagonistic substances.
Chen-Yun HU ; Yong LI ; Min LIU ; Wan-Long DING ; Min-Jian QIN
China Journal of Chinese Materia Medica 2014;39(14):2624-2628
Panax ginseng is one of the most important traditional Chinese herbal medicine, soil borne diseases influenced the yield and quality severely. In our previous work, endophytic Bacillus subtilis ge25 strain was isolated from ginseng root, and which showed significant antagonistic activity against several most destructive ginseng phytopathogens. In the present work, crude protein and lipopeptid extracts were prepared from LB and Landy supernate by salting out, acid precipitation methods respectively. The antagonistic activity of crude extracts and stability to temperature and protease digestion were examined by ginseng phytopathogen Alternaria panax. Results showed that, the antagonistic activity of crude protein extracts from LB culture was complete and partially lost when treated by high temperature and proteinase K. However, crude lipopeptid from Landy culture showed significant stabile antagonistic activity to them. Acid-hydrolyzation and TLC-bioautography analysis showed, that the crude lipopeptide contained at least one cyclic lipopeptide. In consideration of the stability and perfect antagonistic activity of ge25, further researches will promote the biocontrol of ginseng diseases in the field.
Alternaria
;
drug effects
;
physiology
;
Bacillus subtilis
;
metabolism
;
physiology
;
Bacterial Proteins
;
isolation & purification
;
metabolism
;
pharmacology
;
Endopeptidase K
;
metabolism
;
Endophytes
;
metabolism
;
physiology
;
Fermentation
;
Lipopeptides
;
isolation & purification
;
pharmacology
;
Panax
;
microbiology
;
Plant Roots
;
microbiology
;
Temperature
8.Screening and taxonomic identification of endophytic fungi with antitumor and antioxidant activities from Artemisia lactiflora.
Yi-Xin QIAN ; Ji-Chuan KANG ; Bang-Xing LEI ; Lu WANG ; Ying HUANG
China Journal of Chinese Materia Medica 2014;39(3):438-441
Artemisia lactiflora is an important medicinal plant in China. The antitumor and antioxidant activities of the extracts of 54 endophytic fungi from the plant were screened via MTT assay and DPPH scavenging radical assay, respectively. The bioactive strains were identified based on similarity of 5.8S gene and internal transcribed spacer (ITS) sequences. The results showed that extracts from ten (18.5%) isolates exhibited antitumor activity, and which from two (3.7%) isolates exhibited antioxidant activity. The Alternaria sp. GYBH47 strain was simultaneously having antagonistic activity against HL-60 leukemia, MCF-7 breast and COLO205 colon cell lines, and Phomopsis sp. GYBH42 strain having cytotoxic and antioxidant activities. The results indicated that endophytic fungi from Artemisia lactiflora are potential resources to find valuable bioactive components.
Antineoplastic Agents
;
chemistry
;
pharmacology
;
Artemisia
;
microbiology
;
Biphenyl Compounds
;
metabolism
;
Cell Line, Tumor
;
Endophytes
;
chemistry
;
classification
;
physiology
;
Free Radical Scavengers
;
chemistry
;
pharmacology
;
Fungi
;
classification
;
physiology
;
Humans
;
Picrates
;
metabolism
9.Screening and identification of endophytic fungi with growth promoting effect on Dendrobium officinale.
Xiao-qiang HOU ; Shun-xing GUO
China Journal of Chinese Materia Medica 2014;39(17):3232-3237
The endophytic fungi with plant growth promoting effects were screened by co-culture of each endophytic fungus and seedlings of Dendrobium officinale. Anatomical features of the inoculated roots were studied by paraffin sectioning. Morphological characteristics and rDNA ITS1-5. 8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. The results showed that 8 strains inoculated to D. officinale seedlings greatly enhanced plant height, stem diameter, new roots number and biomass. According to the anatomical features of the inoculated roots, each fungus could infect the velamina of seedlings. The hyphae or pelotons were existed in the exodermis passage cells and cortex cells. The effective fungi could not infect the endodermis and vascular bundle sheath, but which was exception for other fungi with harmful to seedlings. Combined with classic morphologic classification, 2 effective strains were identified which were subjected to Pestalotiopsis and Eurotium. Six species of fungi without conidiophore belonged to Pyrenochaeta, Coprinellus, Pholiota, Alternaria, Helotiales, which were identified by sequencing the PCR-amplified rDNA ITS1-5. 8S-ITS2 regions. The co-culture technology of effective endophytic fungi and plant can apply to cultivate the seedlings of D. officinale. It is feasible to shorten growth cycle of D. officinale and increase the resource of Chinese herbs.
Biomass
;
DNA, Fungal
;
genetics
;
DNA, Ribosomal
;
genetics
;
DNA, Ribosomal Spacer
;
genetics
;
Dendrobium
;
cytology
;
growth & development
;
microbiology
;
Endophytes
;
classification
;
genetics
;
physiology
;
Fungi
;
classification
;
genetics
;
physiology
;
Host-Pathogen Interactions
;
Hyphae
;
physiology
;
Plant Roots
;
cytology
;
growth & development
;
microbiology
;
Plants, Medicinal
;
growth & development
;
microbiology
;
RNA, Ribosomal, 5.8S
;
genetics
;
Seedlings
;
cytology
;
growth & development
;
microbiology
;
Species Specificity
10.Microscopic observation on mycorrhiza of rare herb Dysosma versipellis.
Xiao-Ming TAN ; Li-Ying YU ; Ya-Qin ZHOU ; Xiao-Lei ZHOU ; Ying WEI
China Journal of Chinese Materia Medica 2013;38(23):4044-4046
Endophytic fungi played an important role in the growth of its host plant. To investigate the mycorrhizal characteristics and the distribution of fungi in the root, an endangered wild plant-Dysosma versipellis was collected and observed by electron microscope. The results showed that the host was closely associated with endophytic fungi. The fungi were mainly distributed in the epidermis and cortex. The aseptate and septate fungi with swollen hyphae were observed in some cell of the cortex. The result provides a reference for the study of mycorrhizal structure of Dysosma genus and the interaction between the fungi and its host.
Berberidaceae
;
microbiology
;
ultrastructure
;
Endangered Species
;
Endophytes
;
physiology
;
ultrastructure
;
Fungi
;
physiology
;
ultrastructure
;
Microscopy, Electron
;
Plant Roots
;
microbiology
;
ultrastructure

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