Lee, Byung Dae
(Department of Genetic Engineering, Sungkyunkwan University)
,
Dutta, Swarnalee
(Division of Agricultural Microbiology, National Academy of Agricultural Sciences, Rural Development Administration)
,
Ryu, Hojin
(Department of Biology, Chungbuk National University)
,
Yoo, Sung-Je
(Division of Agricultural Microbiology, National Academy of Agricultural Sciences, Rural Development Administration)
,
Suh, Dong-Sang
(Department of Genetic Engineering, Sungkyunkwan University)
,
Park, Kyungseok
(Division of Agricultural Microbiology, National Academy of Agricultural Sciences, Rural Development Administration)
Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention...
Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.
Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.
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문제 정의
On application in the field, various factors such as acclimatization to the environment and competition with the native strains lead to the inability of potential strains to survive in the rhizosphere. Therefore, the search continues for potential bacterial candidates preferably of native origin to combat diseases and improve plant growth. Bacillus is the most abundant genus in the rhizosphere and is preferred for bioformulations due to advantageous traits such as a multilayered cell wall, endospore formation, the production of extracellular enzymes, antibiotic peptides, and peptide signal molecules, which contribute toward their survival under varied environmental conditions [19, 20], The present work evaluated native rhizosphere bacilli from P.
제안 방법
For field studies, ginseng was sown in 0.64-m2 area seed beds with a seedling gap of3 cm x 3 cm in a completely randomized block design. A total of 700 replicates were maintained for this study.
First, bacterial treatment was given at 1 week after transfer to pots and another treatment was carried out after an additional week of growth. The treated plants were grown for 2 additional weeks and then leaf samples were subjected to square plate assay against P. cactorum to analyze the priming effect.
대상 데이터
64-m2 area seed beds with a seedling gap of3 cm x 3 cm in a completely randomized block design. A total of 700 replicates were maintained for this study. Ten liters of HK34 suspension (1 x 106 cfu/mL) was applied to each bed after 2 months of plant growth as soil drench.
Specific primers used for PgPR5 were 5'-AAC CGA CTG CAA CIT CGA CT-3' and 5'-GGC ACA TTA AAC CCATCC AC-3, [26]. For the gene PgCATl, the primers used were 5'-CAA GGA TGG GAA AGC ACA CT-3, and 5, -TGG TTA CAT CGA GTG GGT CA-3Z. The housekeeping gene that encoded P-actin (AY907207) was used as a control and was amplified with the primers 5'-CGT GAT CTT ACA GAT AGC TTG ATG A-3' and 5'-AGA GAA GCT AAG ATT GAT CCT CC-3'.
이론/모형
After 1 month of growth, roots were collected and the adherent soil was removed by shaking. Then the roots (1 g) were excised and ground from root tip to estimate the HK34 population using the serial dilution method. Plants treated with sterile distilled water (SDW) were taken as control.
성능/효과
A total of 78 different bacilli were isolated from the roots of wild ginseng plants (Fig. 1) with Bacillus thuringiensis (20.51%) being the bacteria with the highest percentage, followed by Bacillus cereus (14.10%) and Bacillus weihenstephanensis (11.54%) along with species of Paenibacillus, Viridibacillus, Lysinibacillus, and Brevibacterium. All the strains were studied for their efficacy in disease control in ginseng plants against P.
Analysis of defense-related genes for ISR against P. cactorum resulted in a significant upregulation of PgPR5, PgPRIO, and PgCATin leaves of plants treated with HK34 compared with untreated controls and only pathogen-treated plants (Fig. 9). Relative expression of PgPRW was highest (approximately 5-fold increase) in HK34-treated plants compared with the control followed by PgPR5 (approximately 4-fold increase) and PgCAT (approximately 2-fold increase).
In conclusion, it can be stated that native B. amyloliquefaciens isolate HK34 from ginseng is a potential candidate for biocontrol of P. cactorum because it upregulated the defense-related marker genes PgPR5, PgPRIO, and PgCAT in treated ginseng plants, and thus elicits ISR. The efficacy was consistent under greenhouse and fieldgrowing plants, indicating the potency to survive and act under varied conditions.
6 and 7) compared with the control. The consistent results of field studies as in greenhouse samples signify the ability of HK34 as a potential isolate against P. cactorum.
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