Choi, Jeahyuk
(Department of Biotechnology, Yeungnam University)
,
Park, Euiho
(Department of Biotechnology, Yeungnam University)
,
Lee, Se-Weon
(International Technology Cooperation Center, Rural Development Administration)
,
Hyun, Jae-Wook
(Citrus Research Station, National Institute of Horticultural and Herbal Science, Rural Development Administration)
,
Baek, Kwang-Hyun
(Department of Biotechnology, Yeungnam University)
Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fou...
Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of $100{\mu}g/ml$ suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of $10{\mu}g/ml$. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides.
Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of $100{\mu}g/ml$ suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of $10{\mu}g/ml$. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides.
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문제 정의
In this study we showed that selection of ssAMPs through PS-SPCL can provide us a rich pool of novel antimicrobial peptides with semi target selectivity. And the selected hexapeptides showed good potential to suppress the disease development in citrus leaves when they were co-infiltrated with SR strains.
Effective alternatives to conventional agricultural antibiotics should be devel-oped soon to fight against emerging antibiotics resistant pathogens. This study is designed to develop new antibacterial small synthetic antimicrobial peptides (ssAMPs) against citrus canker pathogen and evaluate the potential as an alternative of streptomycin to control wild type as well as antibiotics resistant citrus canker pathogen.
가설 설정
The arbitrary unit represents average none-grown colonies for each strain in the agar plate. If no colonies were grown within 3 replications, it was counted as 3. If there was a colony with intermediate growth 0.5 was subtracted.
제안 방법
The suppression of citrus canker disease caused by wild type strain XccW1 upon treatment of hexapeptides and streptomycin (ST). Effects of three hexapeptides and ST on the disease progress were evaluated for 17 days by using a disease index: 0 = no symptom, 1 = first symptom, 2 = yellowish halo around the pustule, 3 = prominent pustule, 4 = reddish or brownish pustule appearance of the necrotic tissue, 5 = leaf distortion or leaf drop. The hexapeptides were treated individually (A) or simultaneously with ST (B).
In this study, we performed several PS-SPCLs against a wild type as well as three SR mutants of Xcc. The amino acid information for each position was first integrated into 14 complex hexamers (XC series) that were designed to allow more than one amino acid residue at a position, and then the final 14 individual hexapeptides (BHC series) were synthesized based on information obtained from the assay results of the XC series. The antimicrobial spec-trum of the BHC series was addressed at 100 μg/ml with variety of microbes including a food pathogen, Bacillus cereus (Bc), two plant pathogenic pseudomonads, Pseu-domonas syringe pv.
대상 데이터
One set of the library consisted of 114 tubes (6 positions × 19 amino acids).
The antimicrobial activities of the BHC series on Xcc were tested with two wild types and three SR mutant strains. Three novel hexapeptides (BHC02, BHC06, and BHC11) were selected for further study. They showed antimicrobial activity against various Xcc strains at concentration of 10 μg/ml.
후속연구
It is not clear if BHC7 may have inverse-selectivity against other pseudo-monads. Further study will be needed to understand the antimicrobial selectivity as well as activities in different hexapeptide context.
Co-treatment of streptomycin enhanced the activity of BHC06 and BHC11 but not with KCM21 suggesting sequence-dependent synergic interac-tions with streptomycin. Further study would be need to understand the potential synergic interactions of selected hexapeptides with streptomycin or by themselves. Overall, the selected hexapeptides showed good potential to suppress the disease development in this experimental set-ting.
And the selected hexapeptides showed good potential to suppress the disease development in citrus leaves when they were co-infiltrated with SR strains. However, further studies will be needed to develop alternative agricultural antibiotics based on hexapeptides such as testing the cytotoxicity, the environmental effects on non-pathogenic microbes and the residual periods of hexapeptides in addition to confirm the disease control efficiency in field condition.
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