Nguyen, Hoa Thi
(Department of Agricultural Chemistry, Institute of Environmentally Friendly Agriculture, College of Agriculture and Life Sciences, Chonnam National University)
,
Yu, Nan Hee
(Department of Agricultural Chemistry, Institute of Environmentally Friendly Agriculture, College of Agriculture and Life Sciences, Chonnam National University)
,
Park, Ae Ran
(Department of Agricultural Chemistry, Institute of Environmentally Friendly Agriculture, College of Agriculture and Life Sciences, Chonnam National University)
,
Park, Hae Woong
(R&D Division, World Institute of Kimchi)
,
Kim, In Seon
(Department of Agricultural Chemistry, Institute of Environmentally Friendly Agriculture, College of Agriculture and Life Sciences, Chonnam National University)
,
Kim, Jin-Cheol
(Department of Agricultural Chemistry, Institute of Environmentally Friendly Agriculture, College of Agriculture and Life Sciences, Chonnam National University)
This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) wi...
This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibition concentration (MIC) value of $250{\mu}g/ml$. Among the three solvent layers obtained from the methanol extract of P. nil seeds, only the butanol layer displayed the activity with an MIC value of $125{\mu}g/ml$ against Xap. An antibacterial fraction was obtained from P. nil seeds by repeated column chromatography and identified as pharbitin, a crude resin glycoside, by instrumental analysis. The antibacterial activity of pharbitin was tested in vitro against 14 phytopathogenic bacteria, and it was found to inhibit Ralstonia solanacearum and four Xanthomonas species. The minimum inhibitory concentration values against the five bacteria were $125-500{\mu}g/ml$ for the n-butanol layer and $31.25-125{\mu}g/ml$ for pharbitin. In a detached peach leaf assay, it effectively suppressed the development of bacterial leaf spot, with a control value of 87.5% at $500{\mu}g/ml$. In addition, pharbitin strongly reduced the development of bacterial wilt on tomato seedlings by 97.4% at $250{\mu}g/ml$, 7 days after inoculation. These findings suggest that the crude extract of P. nil seeds can be used as an alternative biopesticide for the control of plant diseases caused by R. solanacearum and Xanthomonas spp. This is the first report on the antibacterial activity of pharbitin against phytopathogenic bacteria.
This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibition concentration (MIC) value of $250{\mu}g/ml$. Among the three solvent layers obtained from the methanol extract of P. nil seeds, only the butanol layer displayed the activity with an MIC value of $125{\mu}g/ml$ against Xap. An antibacterial fraction was obtained from P. nil seeds by repeated column chromatography and identified as pharbitin, a crude resin glycoside, by instrumental analysis. The antibacterial activity of pharbitin was tested in vitro against 14 phytopathogenic bacteria, and it was found to inhibit Ralstonia solanacearum and four Xanthomonas species. The minimum inhibitory concentration values against the five bacteria were $125-500{\mu}g/ml$ for the n-butanol layer and $31.25-125{\mu}g/ml$ for pharbitin. In a detached peach leaf assay, it effectively suppressed the development of bacterial leaf spot, with a control value of 87.5% at $500{\mu}g/ml$. In addition, pharbitin strongly reduced the development of bacterial wilt on tomato seedlings by 97.4% at $250{\mu}g/ml$, 7 days after inoculation. These findings suggest that the crude extract of P. nil seeds can be used as an alternative biopesticide for the control of plant diseases caused by R. solanacearum and Xanthomonas spp. This is the first report on the antibacterial activity of pharbitin against phytopathogenic bacteria.
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문제 정의
The methanol extract of Pharbitis nil seeds showed the strongest activity against Rs and Xap. Therefore, this study aimed to isolate and characterize the antibacterial metabolites contained in this extract, and to examine their in vitro antibacterial activity against various plant pathogenic bacteria. Additionally, their disease control efficacy was evaluated against bacterial leaf spots on peach leaves in a detached leaf bioassay, and bacterial wilt on tomato seedlings.
This study focused on the identification of an antibacterial principle in the MeOH extract of P. nil seeds, and the evaluation of its in vitro and in vivo activities against phytopathogenic bacteria. The hevein-like proteins from P.
제안 방법
The minimum inhibitory concentration (MIC) was defined as the lowest concentration that completely inhibited bacterial growth. The assay was repeated three times in triplicate for each sample, against the individual bacterial species at all test concentrations.
데이터처리
Lowercase letters indicate that values are not significantly different from other values with the same letter according to Duncan’s HSD test for multiple comparisons (p ≤ 0.05) with the SAS software (ver. 9.3; SAS Institute, USA).
The data obtained in this study were evaluated by ANOVA, and statistical analysis of the significance of mean differences was performed using Duncan’s HSD test for multiple comparisons (p ≤ 0·05) with the SAS software (ver. 9.3; SAS Institute, USA).
이론/모형
In vitro antibacterial activity was determined in 96-well microtiter plates using the broth microdilution method [6]. The EtOAc, BuOH, and aqueous layers were dissolved in acetone, MeOH, and distilled water, respectively, at a concentration of 500 mg/ml.
후속연구
nil seed extract to be a promising candidate for the development of a new biopesticide for the control of plant diseases caused by Rs and Xanthomonas spp. However, further studies to develop an optimum extraction process and formulation, as well as to investigate the toxicity and determine the effective dose in field crops, are necessary before such a product can be commercialized.
참고문헌 (42)
Kawaguchi A, Inoue K, Inoue Y. 2014. Biological control of bacterial spot on peach by nonpathogenic Xanthomonas campestris strains AZ98101 and AZ98106. J. Gen. Plant Pathol. 80: 158-163.
Stefani E. 2010. Economic significance and control of bacterial spot/canker of stone fruits caused by Xanthomonas arboricola pv. pruni. J. Plant Pathol. 92: S1.99-S1.103.
Wallis FM, Truter SJ. 1978. Histopathology of tomato plants infected with Pseudomonas solanacearum with emphasis on ultrastructure. Physiol. Plant Pathol. 13: 307-310.
Vu TT, Kim J-C, Choi YH, Choi GJ, Jang KS, Choi TH, et al. 2013. Effect of gallotannins derived from Sedum takesimense on tomato bacterial wilt. Plant Dis. 97: 1593-1598.
Le Dang Q, Shin TS, Park MS, Choi YH, Choi GJ, Jang KS, et al. 2014. Antimicrobial activities of novel mannosyl lipids isolated from the biocontrol fungus Simplicillium lamellicola BCP against phytopathogenic bacteria. J. Agric. Food Chem. 62: 3363-3370.
Nguyen HT, Yu NH, Jeon SJ, Lee HW, Bae CH, Yeo JH, et al. 2016. Antibacterial activities of penicillic acid isolated from Aspergillus persii against various plant pathogenic bacteria. Lett. Appl. Microbiol. 62: 488-493.
Kawasaki T, Okabe H, Nakatsuka I. 1971. Studies on resin glycosides. I. Reinvestigation of the components of pharbitin, a resin glycoside of the seeds of Pharbitis nil Choisy. Chem. Pharm. Bull. 19: 2394-2403.
Lee OS, Lee B, Park N, Koo JC, Kim YH, Prasad DT, et al. 2003. Pn-AMPs, the hevein-like proteins from Pharbitis nil confers disease resistance against phytopathogenic fungi in tomato, Lycopersicum esculentum. Phytochemistry 62: 1073-1079.
Bensky D, Gamble A. 1993. Chinese Herbasl Medicine, Revised Ed. Materia Media, Eastland Press, Seattle.
Eich E. 2008. Solanaceae and Convolvulaceae: Secondary Metabolites, Springer Verlag, Berlin Heidelberg.
Okabe H, Kawasaki T. 1970. Structures of pharbitic acids C and D. Tetrahedron Lett. 36: 3123-3126.
Lee TH, Choi JJ, Kim DH, Choi S, Lee KR, Son M, et al. 2008. Gastroprokinetic effects of DA-9701, a new prokinetic agent formulated with Pharbitis Semen and Corydalis Tuber. Phytomedicine 15: 836-843.
Koo JC, Lee SY, Chun HJ, Cheong YH, Choi JS, Kawabata S-I, et al. 1998. Two hevein homologs isolated from the seeds of Pharbitis nil L. exhibit potent antifungal activity. Biochim. Biophys. Acta 1382: 80-90.
Koo JC, Chun HJ, Park HC, Kim MC, Koo YD, Koo SC, et al. 2002. Over-expression of a seed specific hevein-like antimicrobial peptide from Pharbitis nil enhances resistance to a fungal pathogen in transgenic tobacco plants. Plant Mol. Biol. 50: 441-452.
Ko SG, Koh SH, Jun CY, Nam CG, Bae HS, Shin MK. 2004. Induction of apoptosis by Saussurea lappa and Pharbitis nil on AGS gastric cancer cells. Biol. Pharm. Bull. 27: 1604-1610.
Hao B, Liu GL, Hu XG, Wang GX. 2012. Bioassay-guided isolation and identification of active compounds from Semen pharbitidis against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus). Vet. Parasitol. 187: 452-458.
Lee HJ, Jo EJ, Kim NH, Chae Y, Lee S-W. 2011. Disease responses of tomato pure lines against Ralstonia solanacearum strains from Korea and susceptibility at high temperature. Res. Plant Dis. 17: 326-333.
Koh YJ, Kim GH, Jung JS, Lee YS, Hur JS. 2010. Outbreak of bacterial canker on Hort16A (Actinidia chinensis Planchon) caused by Pseudomonas syringae pv. actinidiae in Korea. N. Z. J. Crop Hort. Sci. 38: 275-282.
Ono M, Takigawa A, Mineno T, Yoshimitu H, Nohara T, Ikeda T, et al. 2010. Acylated glycosides of hydroxyl fatty acid methyl esters generated from the crude resin glycoside (pharbitin) of seeds of Pharbitis nil by treatment with indium(III) chloride in methanol. J. Nat. Prod. 73: 1846-1852.
Ono M, Noda N, Kawasaki T, Miyahara K. 1990. Resin glycosides. VII. Reinvestigation of the component organic and glycosidic acids of pharbitin, the crude ether-insoluble resin glycoside ("convolvulin") of Pharbitidis Semen (seeds of Pharbitis nil). Chem. Pharm. Bull. 38: 1892-1897.
Socquet-Juglard D, Patocchi A, Pothier JF, Christen D, Duffy B. 2012. Evaluation of Xanthomonas arboricola pv. pruni inoculation techniques to screen for bacterial spot resistance in peach and apricot. J. Plant Pathol. 94: S1.91-S1.96.
Winstead NN, Kelman A. 1952. Inoculation techniques for evaluating resistance to Pseudomonas solanacearum. Phytopathology 42: 628-634.
Bieber LW, da Silva Filho AA, Correa Lima RMO, de Andrade Chiappeta A, do Nascimento SC, de Souza IA, et al. 1986. Anticancer and antimicrobial glycosides from Ipomoea bahiensis. Phytochemistry 25: 1077-1081.
Reynolds WF, Yu M, Enriquez RG, Gonzalez H, Leon I, Magos G, et al. 1995. Isolation and characterization of cytotoxic and antibacterial tetrasaccharide glycosides from Ipomoea stans. J. Nat. Prod. 58: 1730-1734.
Okabe H, Koshito N, Tanaka K, Kawasaki T. 1971. Studies on resin glycosides. II. Unhomogeneity of "pharbitic acid" and isolation and partial structures of pharbitic acids C and D, the major constituents of "pharbitic acid". Chem. Pharm. Bull. 19: 2394-2403.
Pontes N de C, Kronka AZ, Moraes MFH, Nascimento AS, Fujinawa MF. 2011. Incorporation of neem leaves into soil to control bacterial wilt of tomato. J. Plant Pathol. 93: 741-744.
Yuan G-Q, Li Q-Q, Qin J, Ye Y-F, Lin W. 2012. Isolation of methyl gallate from Toxicodendron sylvestre and its effect on tomato bacterial wilt. Plant Dis. 96: 1143-1147.
Pradhanang PM, Momol MT, Olson SM, Jones JB. 2003. Effects of plant essential oils on Ralstonia solanacearum population density and bacterial wilt incidence in tomato. Plant Dis. 87: 423-427.
Ji P, Momol MT, Olson SM, Pradhanang PM, Jones JB. 2005. Evaluation of thymol as biofumigant for control of bacterial wilt of tomato under field conditions. Plant Dis. 89: 497-500.
Lee YH, Choi CW, Kim SH, Yun JG, Chang SW, Kim YS, et al. 2012. Chemical pesticides and plant essential oils for disease control of tomato bacterial wilt. Plant Pathol. J. 28: 32-39.
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