Sun, Zhuo
(Cultivation Base of State Key Laboratory for Ecological Restoration and Ecosystem Management of JL Province and MOST, College of Chinese Medicinal Materials, Jilin Agricultural University)
,
Yang, Limin
(College of Chinese Medicinal Materials, Jilin Agriculture University)
,
Zhang, Lianxue
(College of Chinese Medicinal Materials, Jilin Agriculture University)
,
Han, Mei
(College of Chinese Medicinal Materials, Jilin Agriculture University)
Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains ...
Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains isolated from ginseng rhizosphere soils were screened using the Kirby-Bauer disk diffusion method and the Oxford cup plate assay. We analyzed the antifungal spectrum of SZ-22 by confronting incubation. To evaluate the efficacy of biocontrol against ginseng black spot and for growth promotion by SZ-22, we performed pot experiments in a plastic greenhouse. Taxonomic position of SZ-22 was identified using morphology, physiological, and biochemical characteristics, 16S ribosomal DNA, and gyrB sequences. Results: SZ-22 (which was identified as Brevundimonas terrae) showed the strongest inhibition rate against A. panax, which showed 83.70% inhibition, and it also provided broad-spectrum antifungal effects. The inhibition efficacies of the SZ-22 bacterial suspension against ginseng black spot reached 82.47% inhibition, which is significantly higher than that of the 25% suspension concentrate azoxystrobin fungicide treatment (p < 0.05). Moreover, the SZ-22 bacterial suspension also caused ginseng plant growth promotion as well as root enhancement. Conclusion: Although the results of the outdoor pot-culture method were influenced by the pathogen inoculum density, the cropping history of the field site, and the weather conditions, B. terrae SZ-22 controlled ginseng black spot and promoted ginseng growth successfully. This study provides resource for the biocontrol of ginseng black spot.
Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains isolated from ginseng rhizosphere soils were screened using the Kirby-Bauer disk diffusion method and the Oxford cup plate assay. We analyzed the antifungal spectrum of SZ-22 by confronting incubation. To evaluate the efficacy of biocontrol against ginseng black spot and for growth promotion by SZ-22, we performed pot experiments in a plastic greenhouse. Taxonomic position of SZ-22 was identified using morphology, physiological, and biochemical characteristics, 16S ribosomal DNA, and gyrB sequences. Results: SZ-22 (which was identified as Brevundimonas terrae) showed the strongest inhibition rate against A. panax, which showed 83.70% inhibition, and it also provided broad-spectrum antifungal effects. The inhibition efficacies of the SZ-22 bacterial suspension against ginseng black spot reached 82.47% inhibition, which is significantly higher than that of the 25% suspension concentrate azoxystrobin fungicide treatment (p < 0.05). Moreover, the SZ-22 bacterial suspension also caused ginseng plant growth promotion as well as root enhancement. Conclusion: Although the results of the outdoor pot-culture method were influenced by the pathogen inoculum density, the cropping history of the field site, and the weather conditions, B. terrae SZ-22 controlled ginseng black spot and promoted ginseng growth successfully. This study provides resource for the biocontrol of ginseng black spot.
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제안 방법
The experiments were conducted under plastic greenhouse conditions (approximately 16 h of sunlight at 14-29°C on average, over a 10-wk period).
A total of three ginseng seedlings were planted into per pot, respectively. Ten replicates of each treatment were performed in a completely randomized block design. All treatment combinations were repeated three times.
To evaluate the influence of the treatment on yield, nine ginseng seedlings were randomly selected after 10 wk; then, the ginseng plant height, whole plant fresh weight, whole plant dry weight, root length, root fresh weight, and root dry mass were measured and recorded.
대상 데이터
Ginseng rhizosphere soil was collected from the Ginseng Standardized Cultivation Base in Fusong county, Jilin Province, China (Y:42°32' N,127°08' E.537 m).
The bacterial PCR amplification universal primers [28] 16 S1F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 16S1R: 5'-TACGGCTACCTTGTTACG-ACTT-3' were used.
데이터처리
The original data were processed using DPS version 9.50 (sinyosoft, China), with Duncan’s multiple range test to analyze the difference.
이론/모형
panax was used as the indicator. The KirbyeBauer disk diffusion method [21] was used for preliminary screening. Colonies with the best fungistatic effect were selected for secondary screening.
성능/효과
The bacterial strain SZ-2 had antifungal activity toward 10 fungi. Strain SZ-2 reduced the radial growth of A. panax, F. graminearum, B. maydis, F. oxysporum, B. sorokiniana, B. cinema, F. bulbigenum, R. cerealis, M. grisea, and D. gregaria from 29.77% to 78.02%, and was not inhibitory toward R. solani, A. brassicae, S. sclerotiorum, and P. capsici. Bacterial strain SZ-22 had antifungal activity toward 12 fungi and showed better broad-spectrum antifungal activity.
1). Ten weeks after treatment, at the highest concentration tested, i.e., 100% (v/v, 108 CFU/mL), the bacterial suspensions of strains SZ-2 and SZ-22 significantly reduced ginseng black spot caused by A. panax, and the average reduction of ginseng black spot were 69.64% and 82.47% compared with the nontreated control, respectively. The bacterial suspension of SZ-22 at a 60% (v/v) concentration had adequate biocontrol activity (p < 0.
In the pot experiments, strains SZ-2 and SZ-22 all showed growth-promoting effects (Table 3). Among these strains, the growth promotion effect of SZ-22 was the most significant, and the ginseng whole plant fresh weight, whole plant dry weight, root fresh weight, and root dry weight increased by 144.03%, 382.94%, 240.28%, and 144.03%, respectively. The strain SZ-22 promoted the increase in ginseng plant height and root length by 83.
At the tested concentrations, i.e., low to 40% (volume/volume) concentrations, the disease severity index of ginseng measured in A or B, was similar to that in D. Under Duncan’s new multiple range test, different lowercase letters in the histogram by significant differences (p < 0.05) were determined.
). Results of the present study showed that SZ-22 bacterial suspension not only has a control effect against ginseng black spot but also has an adequate promoting effect on the growth of ginseng plants, indicating the potential of this bacterium for applications related to the prevention and treatment of ginseng diseases. Through comprehensive morphological, physiological, and biochemical discrimination, as well as using 16S rDNA and gyrB gene sequence alignment, the bacterial strain SZ-22 was observed to be identical to B.
The results have also demonstrated the ability of B. terrae to prevent disease and promote growth, suggesting that B. terrae uses “competition as the main, antagonistic and induction as supplement” as a disease prevention mechanism.
In the present study, using a systematically flat confrontation method, we screened multiple strains of bacteria from the rhizosphere soil of ginseng plants. Our results showed that SZ-22 has a significant inhibitory effect on A. panax, which caused ginseng black spot.
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