Ginseng is a traditional medicinal plant harboring diverse communities of bacterial endophytes which play a crucial role for plant growth and development. However, little is known about the communities of bacterial endophytes in mountain cultivated ginseng (MCG) along with their potential activities...
Ginseng is a traditional medicinal plant harboring diverse communities of bacterial endophytes which play a crucial role for plant growth and development. However, little is known about the communities of bacterial endophytes in mountain cultivated ginseng (MCG) along with their potential activities. We investigated the diversity of bacterial endophytes in MCG from 24 different sites in Korea using culturable technique. In total 1,886 culturable bacteria were isolated from different tissues of MCG and identified 252 isolates using 16S rDNA sequence. A mixed composition of endophytic communities was recovered and phylogenetic analysis showed the four clusters of bacterial endophytes: Proteobacteria (82.8%), Actinobacteria (1.9%), Firmicutes (15.0%) and Bacteroidetes (0.3%) phyla. The bacteria belonging to the genus Pseudomonas (58.59%) were predominated. Proteobacteria were the most dominated phylum in all tissues and sites. Actinobacteria and Bacteroidetes were the less dominated phyla. Bacteroidetes were recovered only from the leaf tissue. The genera Pseudomonas, Bacillus, Burkholderia, Cupriavidus, Stenotrophomonas, Sphingomonas, Acinetobacter, Erwinia, Pantoea, Lelliottia and Staphylococcus were highly abundant. Species richness was high in the site of Chungcheong (C) while the low species richness was found in the sites of Gangwon (I), Jeolla (T, U) and Jeju (W). The diversity was obtained the highest in the site C and the lowest diversity was found in the site Chungcheong (J). Species evenness was very high in most of the sites, while was very low in the sites A, F, I, J, O, S and X.
We evaluated 252 bacterial isolates for their potential activities of antimicrobial properties, plant growth promotion, hydrogen cyanide (HCN) production, and ß-glucosidase production. Bacterial isolates showed the positive results for siderophore production (73.4%), phosphate solubilization (46.8%), indole-3-acetic acid (IAA)-like indole derivative production (66.7%), HCN production (12.7%), and ß-glucosidase activity (38.9%), indicating their roles in plant growth promotion and ginsenoside bio-transformation. Proteobacteria were dominant for all potential activities tested. Phosphate solubilization activity was not detected in Actinobacteria and Bacteroidetes, while positive activity for HCN production was found only in Proteobacteria, which is harmful for plant growth.
Twelve bacterial endophytes belonging to ten different species (Bacillus siamensis, Burkholderia stabilis, Pseudomonas koreensis, Bacillus toyonensis, Paenibacillus terrae, Paenibacillus kribbensis, Bacillus amyloliquefaciens, Pseudomonas batumici, Pseudomonas trivialis and Pseudomonas grimontii) were selected based on dual culture assay for their antimicrobial properties against six different ginseng pathogens (Alternaria panax, Botrytis cinerea, Cylindrocarpon destructans, Rhizoctonia solani, Phytophthora cactorum and Pythium sp.). Among them, Burkholderia stabilis EB159 showed overall good (42.3-84.7%) inhibition activity against all the ginseng pathogens. The cell free supernatant (CFS) and ethylacetate (EtOAc) extract of B. stabilis EB159 showed 37.3-100% and 30.6-100% mycelial growth inhibition of ginseng pathogens, respectively. However, the CFS of other isolates or their EtOAc extracts showed very less inhibitory activities against C. destructans. The carbon source fructose along with organic nitrogen source peptone was the best combination for optimization of the bacterial media in terms of growth and antimicrobial compound production of B. stabilis EB159. The metabolite extracted from this optimized media showed strong antimicrobial activities (100%) against all pathogens except C. destructans (16.46%). The minimum inhibitory concentration (MIC) of this extract was 6.25 µg for P. cactorum and Pythium sp.; 50 µg for A. panax and B. cinerea; 100 µg for R. solani, which inhibited the mycelial growth more than 80%. C. destructans was not sensitive to the extract. The extract had strong inhibitory activity against spore germination of B. cinerea and C. destructans. B. stabilis EB159 also had ability to collapse and destroy the hypha of B. cinerea and C. destructans. The growth and virulence related genes (BCG1, PKA1, BOT1 and TRE1) of B. cinerea were also down-regulated by the treatment with CFS of B. stabilis EB159. The defense related genes (PR2, PR3, PR4, PR5, PR6 and PR10) in ginseng were up-regulated either above or below ground parts after 2 weeks of B. stabilis EB159 inoculation. The defense related genes except PR5 in ginseng were up-regulated all time points after inoculation with B. cinerea. In addition, genes involved in growth and virulence (BCG1, BAC, PKA1, CHS3 and TRE1) in B. cinerea were down-regulated in B. stabilis EB159 treated plants compared to control.
Furthermore B. stabilis EB159 also reduced the percent disease area after inoculation with B. cinerea and strongly suppressed B. cinerea leaf spot when used as a biocontrol agent. All these results indicated the ability of B. stabilis EB159 as plant disease resistance inducer and effective biocontrol agent for ginseng against B. cinerea.
Plants harbor diverse communities of bacterial endophytes which play a crucial role for plant growth and development. Ginseng is traditional medicinal plants but little is known about the communities of bacterial endophytes associated with it. We investigated the diversity of bacterial endophytes associated with mountain cultivated ginseng (MCG) from 24 different sites in Korea using culturable technique. In total 1,886 culturable bacteria were isolated from different tissues of MCG and identified 252 isolates using 16S rDNA sequence. The bacteria belonging to the genus Pseudomonas (58.59% of total bacteria) were predominated. A mixed composition of endophytic communities was recovered and phylogenetic analysis showed the 4 clusters of bacterial endophytes: Proteobacteria (82.8%), Firmicutes (15.0%), Bacteroidetes (
Ginseng is a traditional medicinal plant harboring diverse communities of bacterial endophytes which play a crucial role for plant growth and development. However, little is known about the communities of bacterial endophytes in mountain cultivated ginseng (MCG) along with their potential activities. We investigated the diversity of bacterial endophytes in MCG from 24 different sites in Korea using culturable technique. In total 1,886 culturable bacteria were isolated from different tissues of MCG and identified 252 isolates using 16S rDNA sequence. A mixed composition of endophytic communities was recovered and phylogenetic analysis showed the four clusters of bacterial endophytes: Proteobacteria (82.8%), Actinobacteria (1.9%), Firmicutes (15.0%) and Bacteroidetes (0.3%) phyla. The bacteria belonging to the genus Pseudomonas (58.59%) were predominated. Proteobacteria were the most dominated phylum in all tissues and sites. Actinobacteria and Bacteroidetes were the less dominated phyla. Bacteroidetes were recovered only from the leaf tissue. The genera Pseudomonas, Bacillus, Burkholderia, Cupriavidus, Stenotrophomonas, Sphingomonas, Acinetobacter, Erwinia, Pantoea, Lelliottia and Staphylococcus were highly abundant. Species richness was high in the site of Chungcheong (C) while the low species richness was found in the sites of Gangwon (I), Jeolla (T, U) and Jeju (W). The diversity was obtained the highest in the site C and the lowest diversity was found in the site Chungcheong (J). Species evenness was very high in most of the sites, while was very low in the sites A, F, I, J, O, S and X.
We evaluated 252 bacterial isolates for their potential activities of antimicrobial properties, plant growth promotion, hydrogen cyanide (HCN) production, and ß-glucosidase production. Bacterial isolates showed the positive results for siderophore production (73.4%), phosphate solubilization (46.8%), indole-3-acetic acid (IAA)-like indole derivative production (66.7%), HCN production (12.7%), and ß-glucosidase activity (38.9%), indicating their roles in plant growth promotion and ginsenoside bio-transformation. Proteobacteria were dominant for all potential activities tested. Phosphate solubilization activity was not detected in Actinobacteria and Bacteroidetes, while positive activity for HCN production was found only in Proteobacteria, which is harmful for plant growth.
Twelve bacterial endophytes belonging to ten different species (Bacillus siamensis, Burkholderia stabilis, Pseudomonas koreensis, Bacillus toyonensis, Paenibacillus terrae, Paenibacillus kribbensis, Bacillus amyloliquefaciens, Pseudomonas batumici, Pseudomonas trivialis and Pseudomonas grimontii) were selected based on dual culture assay for their antimicrobial properties against six different ginseng pathogens (Alternaria panax, Botrytis cinerea, Cylindrocarpon destructans, Rhizoctonia solani, Phytophthora cactorum and Pythium sp.). Among them, Burkholderia stabilis EB159 showed overall good (42.3-84.7%) inhibition activity against all the ginseng pathogens. The cell free supernatant (CFS) and ethylacetate (EtOAc) extract of B. stabilis EB159 showed 37.3-100% and 30.6-100% mycelial growth inhibition of ginseng pathogens, respectively. However, the CFS of other isolates or their EtOAc extracts showed very less inhibitory activities against C. destructans. The carbon source fructose along with organic nitrogen source peptone was the best combination for optimization of the bacterial media in terms of growth and antimicrobial compound production of B. stabilis EB159. The metabolite extracted from this optimized media showed strong antimicrobial activities (100%) against all pathogens except C. destructans (16.46%). The minimum inhibitory concentration (MIC) of this extract was 6.25 µg for P. cactorum and Pythium sp.; 50 µg for A. panax and B. cinerea; 100 µg for R. solani, which inhibited the mycelial growth more than 80%. C. destructans was not sensitive to the extract. The extract had strong inhibitory activity against spore germination of B. cinerea and C. destructans. B. stabilis EB159 also had ability to collapse and destroy the hypha of B. cinerea and C. destructans. The growth and virulence related genes (BCG1, PKA1, BOT1 and TRE1) of B. cinerea were also down-regulated by the treatment with CFS of B. stabilis EB159. The defense related genes (PR2, PR3, PR4, PR5, PR6 and PR10) in ginseng were up-regulated either above or below ground parts after 2 weeks of B. stabilis EB159 inoculation. The defense related genes except PR5 in ginseng were up-regulated all time points after inoculation with B. cinerea. In addition, genes involved in growth and virulence (BCG1, BAC, PKA1, CHS3 and TRE1) in B. cinerea were down-regulated in B. stabilis EB159 treated plants compared to control.
Furthermore B. stabilis EB159 also reduced the percent disease area after inoculation with B. cinerea and strongly suppressed B. cinerea leaf spot when used as a biocontrol agent. All these results indicated the ability of B. stabilis EB159 as plant disease resistance inducer and effective biocontrol agent for ginseng against B. cinerea.
Plants harbor diverse communities of bacterial endophytes which play a crucial role for plant growth and development. Ginseng is traditional medicinal plants but little is known about the communities of bacterial endophytes associated with it. We investigated the diversity of bacterial endophytes associated with mountain cultivated ginseng (MCG) from 24 different sites in Korea using culturable technique. In total 1,886 culturable bacteria were isolated from different tissues of MCG and identified 252 isolates using 16S rDNA sequence. The bacteria belonging to the genus Pseudomonas (58.59% of total bacteria) were predominated. A mixed composition of endophytic communities was recovered and phylogenetic analysis showed the 4 clusters of bacterial endophytes: Proteobacteria (82.8%), Firmicutes (15.0%), Bacteroidetes (
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