Investigation on the Antifungal Ingredients of Saccharothrix Yanglingensis Hhs.015, an Antagonistic Endophytic Actinomycete Isolated from Cucumber Plant
Abstract
:1. Introduction
2. Results and Discussion
2.1. Isolation of Antifungal Polyene Macrolides
2.2. Structural Elucidation
2.3. Antifungal Activity of Polyenes against V. mali
2.4. Light Microscopy and Transmission Electron Microscopy Analyses
3. Materials and Methods
3.1. General Procedure
3.2. Microorganisms and Culture Conditions
3.3. Extraction, Isolation, and Structural Elucidation of Antifungal Ingredients
3.4. In Vitro Antifungal Activities of WH01 and WH02 against V. mali
3.5. In Vivo Antifungal Activity of WH01 against V. mali
3.6. Light Microscopy
3.7. Transmission Electron Microscope
3.8. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
16S rRNA | 16S ribosomal RNA |
Fr. | Fragment |
HPLC | High Performance Liquid Chromatography |
UV | Ultraviolet |
HRMS | High Resolution Mass Spectrometer |
NMR | Nuclear magnetic resonance |
DEPT | Distortionless Enhancement by Polarization Transfer |
HSQC | Heteronuclear singular quantum correlation |
COSY | Correlation Spectroscopy |
HMBC | 1H detected heteronuclear multiple bond correlation |
NOESY | Nuclear overhauser effect spectroscopy |
GSA | Gause’s Synthetic Agar medium |
PDA | Potato Dextrose Agar medium |
DMSO | Dimethylsulfoxide |
References
- Cantrell, C.L.; Dayan, F.E.; Duke, S.O. Natural Products As Sources for New Pesticides. J. Nat. Prod. 2012, 75, 1231–1242. [Google Scholar] [CrossRef] [PubMed]
- Laakso, J.A.; Mocek, U.M.; Van Dun, J.; Wouters, W.; Janicot, M. R176502, a New Bafilolide Metabolite with Potent Antiproliferative Activity from a Novel Micromonospora Species. Cheminform 2004, 35, 909. [Google Scholar] [CrossRef]
- Shu-Wei, Y.; Tze-Ming, C.; Joseph, T.; Reena, P.; David, L.; Guodong, C.; Mahesh, P.; Vincent, G.; Birendra, P.; Min, C. A new anthracycline antibiotic micromonomycin from Micromonospora sp. Cheminform 2005, 36, 601–604. [Google Scholar]
- Li, W.; Al, E. Nocathiacins, New Thiazolyl Peptide Antibiotics from Nocardia sp. Part 1. Taxonomy, Fermentation and Biological Activities. Cheminform 2003, 56, 226–231. [Google Scholar] [CrossRef]
- Malet-Cascon, L.; Romero, F.; Espliego-Vazquez, F.; Gravalos, D.; Fernández-Puentes, J.L. IB-00208, a new cytotoxic polycyclic xanthone produced by a marine-derived Actinomadura. II. Isolation, physico-chemical properties and structure determination. J. Antibiot. 2003, 56, 219–225. [Google Scholar]
- Fukami, A.; Mc, K.K.R.; Matsumoto, A.; Takahashi, Y.S.K.; Hayashi, M.; Komiyama, K.; Omura, S.; Nakamura, T. A New Antimicrobial Antibiotic from Actinoplanes capillaceus sp. K95-5561^T. J. Antibiot. 2010, 32, 1212–1214. [Google Scholar]
- Petkovic, H.; Lill, R.E.; Sheridan, R.M.; Wilkinson, B.; McCormick, E.L.; McArthur, H.A.I.; Staunton, J.; Leadlay, P.F.; Kendrew, S.G. A novel erythromycin, 6-desmethyl erythromycin D, made by substituting an acyltransferase domain of the erythromycin polyketide synthase. J. Antibiot. 2003, 56, 543–551. [Google Scholar] [CrossRef]
- Wang, X.; Zang, R.; Yin, Z.; Kang, Z.; Huang, L. Delimiting cryptic pathogen species causing apple Valsa canker with multilocus data. Ecol. Evol. 2014, 4, 1369–1380. [Google Scholar] [CrossRef]
- Wang, L.; Gao, Z.P.; Huang, L.L.; Wei, J.L.; Zang, R.; Kang, Z.S. Screening fungicide for pathogen inhibition and disease control of apple tree Valsa canker. Acta Phytopathol. Sin. 2009, 39, 549–554. [Google Scholar]
- Miles, L.A.; Lopera, C.A.; González, S.; De García, M.C.C.; Franco, A.E.; Restrepo, S.; García, M.C.C. Exploring the biocontrol potential of fungal endophytes from an Andean Colombian Paramo ecosystem. BioControl 2012, 57, 697–710. [Google Scholar] [CrossRef]
- Labeda, D.P.; Testa, R.T.; Lechevalier, M.P.; Lechevalier, H.A. Saccharothrix: A New Genus of the Actinomycetales Related to Nocardiopsis. Int. J. Syst. Bacteriol. 1984, 34, 426–431. [Google Scholar] [CrossRef]
- Lamari, L.; Zitouni, A.; Boudjella, H.; Badji, B.; Sabaou, N.; Lebrihi, A.; Lefebvre, G.; Seguin, E.; Tillequin, F. New dithiolopyrrolone antibiotics from Saccharothrix sp SA 233 - I. Taxonomy, fermentation, isolation and biological activities. J. Antibiot. 2002, 55, 696–701. [Google Scholar] [CrossRef] [PubMed]
- Rabiaa, M.; Noureddine, B.; Yannick, C.; Florence, M.; Nasserdine, S.; Ahmed, L. New dithiolopyrrolone antibiotics induced by adding sorbic acid to the culture medium of Saccharothrix algeriensis NRRL B-24137. Fems Microbiol. Lett. 2011, 318, 41–46. [Google Scholar]
- Abdelghani, Z.; Hadjira, B.; Florence, M.; Nasserdine, S.; Ahmed, L. Mutactimycin PR, a new anthracycline antibiotic from Saccharothrix sp. SA 103. I. Taxonomy, fermentation, isolation and biological activities. J. Antibiot. 2004, 57, 367–372. [Google Scholar]
- Murakami, R.; Shinozaki, J.; Kajiura, T.; Kozone, I.; Takagi, M.; Shin-Ya, K.; Seto, H.; Hayakawa, Y. Ammocidins B, C and D, new cytotoxic 20-membered macrolides from Saccharothrix sp. AJ9571. J. Antibiot. 2009, 62, 123–127. [Google Scholar] [CrossRef]
- Gao, Z.P.; Ke, X.W.; Wei, J.L.; Chen, Y.C.; Kang, Z.S.; Huang, L. Biocontrol efficacy of apple tree valsa canker by endophytic actinomycetes. Acta Phytophylacica Sin. 2009, 36, 410–416. [Google Scholar]
- Yan, X.; Huang, L.-L.; Tu, X.; Gao, X.-N.; Kang, Z.-S. Saccharothrix yanglingensis sp nov., an antagonistic endophytic actinomycete isolated from cucumber plant. Antonie Van Leeuwenhoek 2012, 101, 141–146. [Google Scholar] [CrossRef]
- Fan, D.; Li, Y.; Zhao, L.; Li, Z.; Huang, L.; Yan, X. Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR. PLOS ONE 2016, 11, e0162174. [Google Scholar] [CrossRef]
- Li, Z.; Gao, X.; Fan, D.; Xia, Y.; Huang, L. Saccharothrix yanglingensis Strain Hhs.015 Is a Promising Biocontrol Agent on Apple Valsa Canker. Plant Dis. 2015, 100, 510–514. [Google Scholar] [CrossRef]
- Lu, Y.; Wang, N.; He, J.; Li, Y.; Gao, X.; Huang, L.; Yan, X. Expression and characterization of a novel chitinase with antifungal activity from a rare actinomycete, Saccharothrix yanglingensis Hhs.015. Protein Expr. Purif. 2017, 143, 45. [Google Scholar] [CrossRef]
- Zhang, Y.; Yan, X.; Guo, H.; Zhao, F.; Huang, L. A Novel Protein Elicitor BAR11 From Saccharothrix yanglingensis Hhs.015 Improves Plant Resistance to Pathogens and Interacts With Catalases as Targets. Front. Microbiol. 2018, 9, 700. [Google Scholar] [CrossRef] [PubMed]
- Wang, X.; Huang, L.; Kang, Z.; Buchenauer, H.; Gao, X. Optimization of the Fermentation Process of Actinomycete Strain Hhs.015T. J. Biomed. Biotechnol. 2010, 2010, 1–10. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yan, X.; Huang, Y.-C.; Lan, X.; Gao, X.-N.; Zhou, T.-N.; Kang, Z.-S.; Huang, L.-L. Properties and preliminary identification of antimicrobial active substances of endophytic actinomycete strain Hhs.015. J. Northwest A&F Univ.- Nat. Sci. Ed. (In Chinese) 2014, 42, 175–183. [Google Scholar]
- Noguchi, H.; Harrison, P.H.; Arai, K.; Nakashima, T.T.; Trimble, L.A.; Vederas, J.C. Biosynthesis and full NMR assignment of fungichromin, a polyene antibiotic from Streptomyces cellulose. J. Am. Chem. Soc. 1988, 110, 2938–2945. [Google Scholar] [CrossRef]
- Li, Y.; Aioub, A.A.; Lv, B.; Hu, Z.; Wu, W. Antifungal activity of pregnane glycosides isolated from Periploca sepium root barks against various phytopathogenic fungi. Ind. Crop. Prod. 2019, 132, 150–155. [Google Scholar] [CrossRef]
- Wei, J.L.; Huang, L.L.; Gao, Z.P.; Ke, X.W.; Kang, Z. Laboratory evaluation methods of apple Valsa canker disease caused by Valsa ceratosperma sensu Kobayashi. Acta Phytopathol. Sin. 2010, 40, 14–20. [Google Scholar]
- Liu, C.; Fan, D.; Li, Y.; Chen, Y.; Huang, L.; Yan, X. Transcriptome analysis of Valsa mali reveals its response mechanism to the biocontrol actinomycete Saccharothrix yanglingensis Hhs.015. BMC Microbiol. 2018, 18, 90. [Google Scholar] [CrossRef]
- Kang, Z.; Buchenauer, H. Cytology and ultrastructure of the infection of wheat spikes by Fusarium culmorum. Mycol. Res. 2000, 104, 1083–1093. [Google Scholar] [CrossRef]
Sample Availability: Samples of the compounds WH01 and WH02 are available from the authors. |
Compounds | EC50 (95% Confidence Interval) mg/L | Toxicity Regression Equation (Y = a + bx) | R2 |
---|---|---|---|
WH01 | 5.24 (4.36~6.21) | Y = −1.52 + 2.12x | 0.99 |
WH02 | 4.92(4.07~5.87) | Y = −1.42 + 2.05x | 0.99 |
1,2-Benzisothiazolin-3-One | 8.85(7.09~11.33) | Y = −2.26 + 2.42x | 0.98 |
Chemicals | mg/L | Protective Effect (%) |
---|---|---|
WH01 | 250 | 44.44% |
500 | 74.07% | |
1000 | 81.48% | |
2000 | 96.30% |
WH01,(Fungichromin) | WH02 | |||
---|---|---|---|---|
Position | δ C | δ H | δ C | δ H |
1 | 170.6 | 172.6 | ||
2 | 58.2 | 2.48(m) | 51.9 | 2.25(ddd, J = 11.0, 8.0, 3.5 Hz) |
3 | 69.1 | 3.68(m) | 70.3 | 3.62(m) |
4 | 39.6 | 1.37(m), 1.56(m) | 39.2 | 1.37(m), 1.56(m) |
5 | 69.8 | 3.89(m) | 69.2 | 3.87(m) |
6 | 43.4 | 1.38(m) | 43.0 | 1.31(m), 1.49(m) |
7 | 69.6 | 3.77(m) | 69.0 | 3.68(m) |
8 | 43.5 | 1.31(m), 1.49(m) | 43.3 | 1.31(m), 1.49(m) |
9 | 68.8 | 3.87(m) | 68.7 | 3.87(m) |
10 | 42.4 | 1.32(m), 1.47(m) | 42.1 | 1.32(m), 1.47(m) |
11 | 70.1 | 3.88(m) | 70.1 | 3.88(m) |
12 | 38.2 | 1.37(m), 1.56(m) | 38.1 | 1.37(m), 1.56(m) |
13 | 67.9 | 3.14(m) | 67.9 | 3.14(m) |
14 | 76.1 | 3.49(d, J = 9.0 Hz ) | 76.0 | 3.49(d, J = 9.0 Hz) |
15 | 77.6 | 3.69(d, J =9.0 Hz) | 77.4 | 3.69(dd, J=9.0, 2.5 Hz) |
16 | 138.3 | 138.2 | ||
17 | 126.7 | 5.94(d, J = 11.0 Hz) | 126.5 | 5.94(m) |
18 | 127.4 | 6.45(dd, J = 15.0, 11.0 Hz) | 127.4 | 6.45(dd, J = 15.0, 11.0 Hz) |
19 | 130.8 | 6.26(m) | 131.0 | 6.27(m) |
20 | 128.6 | 6.26(m) | 128.6 | 6.26(m) |
21 | 132.6 | 6.35(m) | 132.2 | 6.36(m) |
22 | 131.8 | 6.26(m) | 131.9 | 6.28(m) |
23 | 132.7 | 6.35(m) | 132.4 | 6.32(m) |
24 | 132.8 | 6.35(m) | 132.4 | 6.32(m) |
25 | 134.6 | 6.02(dd, J = 15.0, 4.0 Hz) | 134.4 | 5.95(m) |
26 | 70.7 | 3.96(m) | 70.9 | 3.96(dd, J = 9.0, 5.0 Hz) |
27 | 72.7 | 4.63(m) | 72.4 | 4.63(m) |
28 | 17.3 | 1.19(d, J = 6.5) | 16.9 | 1.22(d, J = 6.0Hz) |
29 | 11.2 | 1.69(s) | 11.2 | 1.69(s) |
1′ | 70.6 | 3.97(m) | 28.5 | 1.44(m),1.65(m) |
2′ | 33.7 | 1.27(m), 1.36(m) | 25.9 | 1.14(m),1.21(m) |
3′ | 24.1 | 1.25(m), 1.43(m) | 27.9 | 1.23(m) |
4′ | 30.8 | 1.23(m) | 30.4 | 1.22(m) |
5′ | 21.6 | 1.25(m) | 21.3 | 1.25(m) |
6′ | 13.5 | 0.86(t, J = 7.5Hz) | 13.3 | 0.85(t, J = 7.5Hz) |
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Wang, H.; Tian, R.; Tian, Q.; Yan, X.; Huang, L.; Ji, Z. Investigation on the Antifungal Ingredients of Saccharothrix Yanglingensis Hhs.015, an Antagonistic Endophytic Actinomycete Isolated from Cucumber Plant. Molecules 2019, 24, 3686. https://doi.org/10.3390/molecules24203686
Wang H, Tian R, Tian Q, Yan X, Huang L, Ji Z. Investigation on the Antifungal Ingredients of Saccharothrix Yanglingensis Hhs.015, an Antagonistic Endophytic Actinomycete Isolated from Cucumber Plant. Molecules. 2019; 24(20):3686. https://doi.org/10.3390/molecules24203686
Chicago/Turabian StyleWang, Hua, Runze Tian, Qizhen Tian, Xia Yan, Lili Huang, and Zhiqin Ji. 2019. "Investigation on the Antifungal Ingredients of Saccharothrix Yanglingensis Hhs.015, an Antagonistic Endophytic Actinomycete Isolated from Cucumber Plant" Molecules 24, no. 20: 3686. https://doi.org/10.3390/molecules24203686