DOI QR코드

DOI QR Code

A Novel Protein Elicitor PeBL2, from Brevibacillus laterosporus A60, Induces Systemic Resistance against Botrytis cinerea in Tobacco Plant

  • Jatoi, Ghulam Hussain (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences) ;
  • Lihua, Guo (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences) ;
  • Xiufen, Yang (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences) ;
  • Gadhi, Muswar Ali (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences) ;
  • Keerio, Azhar Uddin (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences) ;
  • Abdulle, Yusuf Ali (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences) ;
  • Qiu, Dewen (State Key Laboratory of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences)
  • 투고 : 2018.12.08
  • 심사 : 2019.03.28
  • 발행 : 2019.06.01

초록

Here, we reported a novel secreted protein elicitor PeBL2 from Brevibacillus laterosporus A60, which can induce hypersensitive response in tobacco (Nicotiana benthamiana). The ion-exchange chromatography, high-performance liquid chromatography (HPLC) and mass spectrometry were performed for identification of protein elicitor. The 471 bp PeBL2 gene produces a 17.22 kDa protein with 156 amino acids containing an 84-residue signal peptide. Consistent with endogenous protein, the recombinant protein expressed in Escherichia coli induced the typical hypersensitive response (HR) and necrosis in tobacco leaves. Additionally, PeBL2 also triggered early defensive response of generation of reactive oxygen species ($H_2O_2$ and $O_2{^-}$) and systemic resistance against of B. cinerea. Our findings shed new light on a novel strategy for biocontrol using B. laterosporus A60.

키워드

E1PPBG_2019_v35n3_208_f0004.png 이미지

Fig. 4. Phylogenetic tree based on 16S rRNA sequencing, which was generated using the neighbour-joining method. Relationships of. ERM16658.1. sequence (highlighted in yellow) to other Brevibacillus species are shown. Bar, 1 change per nucleotide position.

E1PPBG_2019_v35n3_208_f0006.png 이미지

Fig. 6. ROS burst in tobacco cells after PeBL2 treatment. The brown DAB-stained precipitates represent ROS burst. (A) No ROS generate in tobacco leaves treated with BSA (control). (B) Significant ROS were observed in the recombinant PeBL2-treated areas.

E1PPBG_2019_v35n3_208_f0007.png 이미지

Fig. 7. Induced disease resistance against Botrytis cinerea in tobacco. (A) Representative phenotypes of disease caused by B. cinerea in PeBL2 and BSA-infiltrated tobacco leaves. The sizes of the lesions caused by B. cinerea in PeBL2-treated leaves were smaller than that in BSA at 36 h post-infiltration. (B) Lesion sizes caused by B. cinerea were measured in leaves with PeBL2 or BSA-treated plants. Data presented in (B) were calculated as follows: Inhibition % = [(No (size) of lesions on control leaves − No (size) of lesions on proteintreated leaves)/No (size) of lesions on control leaves] × 100%. (B1) data of treated and untreated plants were plotted showing inhibition percentage rate. (B2) Statistical analysis was performed using Student’s t-test (Circles), Box Plots for BSA (control) and PEBL2 (treated) representing differences among both treatments indicate significant differences between PeBL2 and BSA treatment.

E1PPBG_2019_v35n3_208_f0008.png 이미지

Fig. 1. Purification of the PeBL2 protein. (A) Cation exchange chromatography using a HisTrap SP HP column (2.5 cm × 20 cm). The column was washed with buffer A (25 mM MES-NaOH, pH 6.2) to remove any unbound proteins, and the bound proteins were eluted with a linear gradient of increasing NaCl (0-1 M). Buffer B (25 mM Tris, 200 mM NaCl, 500 mM imidazole, pH 8.0) was used to B. Two fractions (peak a and peak b) were produced. (B) Fractions (peak a and peak b) along with BSA (control) included in the test for HR. Out of these fractions peak a showing HR in tobacco leaves. (C) SDS-PAGE of crude proteins.

E1PPBG_2019_v35n3_208_f0009.png 이미지

Fig. 2. Purification of the active fraction. (A) Waters Atlantis T3 C18 reversed-phase column (2.1 mm × 150 mm, 3.5 m, 40℃) was equilibrated with 5% CAN and acetonitrile/2 mM NH4FA/0.1% FA/water. The concentration of ACN in the eluted solvent was raised from 10% (v/v) to 60% (v/v) over 28 min using a linear gradient at a flow rate of 0.2 ml/min. The peaks a, b, c, d, e and f were produced by HPLC. (B) All peaks were checked for HR in tobacco leaves. Out of all peaks peak d displayed an obvious HR.

E1PPBG_2019_v35n3_208_f0010.png 이미지

Fig. 3. Further purification of the active protein with HPLC (A) HPLC was used to further purify fraction D using Zorbax-Eclipse (XDB-C18). (B) After purification, fraction D was checked by Tricine SDS-PAGE to determine the size of the protein, which is shown as about 17 kDa. (C) The HR activation of purified single protein was checked as compared to BSA.

E1PPBG_2019_v35n3_208_f0011.png 이미지

Fig. 5. Expression and purification of protein PeBL2 in E. coli. (A) PCR was used to amplify the full-length DNA sequence encoding PeBL2 from the B. laterosporus A60 strain. The length of the PeBL2 gene is 471 bp, which encodes a protein of 156 amino acids with a theoretical molecular weight of 17 kDa. The PeBL2 gene was cloned and then ligated into pET28a. (B) The recombinant protein PeBL2 was expressed and purified. SDS-PAGE of the purified elicitor protein, PeBL2, displaying a single band by Coomassie Brilliant Blue R-250 staining. (C) The hypersensitive response induced by the recombinant elicitor protein, PeBL2. The response was observed at 24 h post-infiltration. The elicitor treatment (50 μm) and control treatment with BSA are shown.

참고문헌

  1. Akkopru, A. and Demir, S. 2005. Biological control of fusarium wilt in tomato caused by Fusarium oxysporum f. sp. lycopersici by AMF Glomus intraradices and some rhizobacteria. J. Phytopathol. 153:544-550. https://doi.org/10.1111/j.1439-0434.2005.01018.x
  2. Allen, R. L., Bittner-Eddy, P. D., Grenville-Briggs, L. J., Meitz, J. C., Rehmany, A. P., Rose, L. E. and Beynon, J. L. 2004. Hostparasite coevolutionary conflict between Arabidopsis and downy mildew. Science 306:1957-1960. https://doi.org/10.1126/science.1104022
  3. Anbu, P. 2016. Enhanced production and organic solvent stability of a protease from Brevibacillus laterosporus strain PAP04. Braz. J. Med. Biol. Res. 49:e5178. https://doi.org/10.1590/1414-431X20165178
  4. Asai, S., Ohta, K. and Yoshioka, H. 2008. MAPK signaling regulates nitric oxide and NADPH oxidase-dependent oxidative bursts in Nicotiana benthamiana. Plant Cell 20:1390-1406. https://doi.org/10.1105/tpc.107.055855
  5. Blumwald, E., Aharon, G. S. and Lam, B. C. 1998. Early signal transduction pathways in plant-pathogen interactions. Trends Plant Sci. 3:342-346. https://doi.org/10.1016/S1360-1385(98)01289-8
  6. Boka, K. and Orban, N. 2007. New aspect of $H_2O_2$ signaling. Plant Signal. Behav. 2:498-500. https://doi.org/10.4161/psb.2.6.4582
  7. Cct, R., Vermeulen, J. P., Vels, A., Himmelbach, A., Mascher, M. and Niks, R. E. 2018. Mapping resistance to powdery mildew in barley reveals a large-effect nonhost resistance QTL. Theor. Appl. Genet. 131:1031-1045. https://doi.org/10.1007/s00122-018-3055-0
  8. Chandel, S., Allan, E. J. and Woodward, S. 2010. Biological Control of Fusarium oxysporum f.sp. lycopersici on Tomato by Brevibacillus brevis. J. Phytopathol. 158:470-478. https://doi.org/10.1111/j.1439-0434.2009.01635.x
  9. Chen, M., Zhang, C., Zi, Q., Qiu, D., Liu, W. and Zeng, H. 2014. A novel elicitor identified from Magnaporthe oryzae triggers defense responses in tobacco and rice. Plant Cell Rep. 33:1865-1879. https://doi.org/10.1007/s00299-014-1663-y
  10. Cui, A. L., Hu, X. X., Gao, Y., Jin, J., Yi, H., Wang, X. K., Nie, T. Y., Chen, Y., He, Q. Y., Guo, H. F., Jiang, J. D., You, X. F. and Li, Z. R. 2018. Synthesis and bioactivity investigation of the individual components of cyclic lipopeptide antibiotics. J. Med. Chem. 61:1845-1857. https://doi.org/10.1021/acs.jmedchem.7b01367
  11. de Oliveira, E. J., Rabinovitch, L., Monnerat, R. G., Passos, L. K. and Zahner, V. 2004. Molecular characterization of Brevibacillus laterosporus and its potential use in biological control. Appl. Environ. Microbiol. 70:6657-6664. https://doi.org/10.1128/AEM.70.11.6657-6664.2004
  12. Diez-Navajas, A. M., Wiedemann-Merdinoglu, S., Greif, C. and Merdinoglu, D. 2008. Nonhost versus host resistance to the grapevine downy mildew, Plasmopara viticola, studied at the tissue level. Phytopathology 98:776-780. https://doi.org/10.1094/PHYTO-98-7-0776
  13. Ellis, J. G., Rafiqi, M., Gan, P., Chakrabarti, A. and Dodds, P. N. 2009. Recent progress in discovery and functional analysis of effector proteins of fungal and oomycete plant pathogens. Curr. Opin. Plant Biol. 12:399-405. https://doi.org/10.1016/j.pbi.2009.05.004
  14. Garcia-Brugger, A., Lamotte, O., Vandelle, E., Bourque, S., Lecourieux, D., Poinssot, B., Wendehenne, D. and Pugin, A. 2006. Early signaling events induced by elicitors of plant defenses. Mol. Plant-Microbe Interact. 19:711-724. https://doi.org/10.1094/MPMI-19-0711
  15. Glazebrook, J. 2001. Genes controlling expression of defense responses in Arabidopsis. Curr. Opin. Plant Biol. 4:301-308. https://doi.org/10.1016/S1369-5266(00)00177-1
  16. Haas, D. and Defago, G. 2005. Biological control of soil-borne pathogens by fluorescent pseudomonads. Nat. Rev. Microbiol. 3:307-319. https://doi.org/10.1038/nrmicro1129
  17. Hazotte, A., Peron, O., Gaudin, P., Abdelouas, A. and Lebeau, T. 2018. Effect of Pseudomonas fluorescens and pyoverdine on the phytoextraction of cesium by red clover in soil pots and hydroponics. Environ. Sci. Pollut. Res. 25:20680-20690. https://doi.org/10.1007/s11356-018-1974-6
  18. Hu, Y., Li, Y., Hou, F., Wan, D., Cheng, Y., Han, Y., Gao, Y., Liu, J., Guo, Y., Xiao, S., Wang, Y. and Wen, Y. Q. 2018. Ectopic expression of Arabidopsis broad-spectrum resistance gene RPW8.2 improves the resistance to powdery mildew in grapevine (Vitis vinifera). Plant Sci. 267:20-31. https://doi.org/10.1016/j.plantsci.2017.11.005
  19. Huang, C. J., Tsay, J. F., Chang, S. Y., Yang, H. P., Wu, W. S. and Chen, C. Y. 2012. Dimethyl disulfide is an induced systemic resistance elicitor produced by Bacillus cereus C1L. Pest Manag. Sci. 68:1306-1310. https://doi.org/10.1002/ps.3301
  20. Huang, X., Tian, B., Niu, Q., Yang, J., Zhang, L. and Zhang, K. 2005. An extracellular protease from Brevibacillus laterosporus G4 without parasporal crystals can serve as a pathogenic factor in infection of nematodes. Res. Microbiol. 156:719-727. https://doi.org/10.1016/j.resmic.2005.02.006
  21. Islam, W., Noman, A., Qasim, M. and Wang, L. 2018. Plant responses to pathogen attack: Small RNAs in focus. Int. J. Mol. Sci. 19:E515. https://doi.org/10.3390/ijms19020515
  22. Jackson, S. N., Wang, H. Y., and Woods, A. S. 2005. Direct profiling of lipid distribution in brain tissue using MALDI-TOFMS. Anal. Chem. 77:4523-4527. https://doi.org/10.1021/ac050276v
  23. Jain, S., Vaishnav, A., Kasotia, A., Kumari, S., Gaur, R. K. and Choudhary, D. K. 2013. Bacteria-induced systemic resistance and growth promotion in Glycine max L. Merrill upon challenge inoculation with Fusarium oxysporum. Proc. Natl. Acad. Sci. India 83:561-567.
  24. Jones, J. D. G. and Dangl, J. L. 2006. The plant immune system. Nature 444:323-329. https://doi.org/10.1038/nature05286
  25. Kloeppe, J. W., Rodriguez-Kabana, R., Zehnder, A. W., Murphy, J. F., Sikora, E. and Fernandez, C. 1999. Plant root-bacterial interactions in biological control of soilborne diseases and potential extension to systemic and foliar diseases. Aust. Plant Pathol. 28:21-26. https://doi.org/10.1071/AP99003
  26. Kukawka, R., Czerwoniec, P., Lewandowski, P., Pospieszny, H. and Smiglak, M. 2018. New ionic liquids based on systemic acquired resistance inducers combined with the phytotoxicity reducing cholinium cation. New J. Chem. 42:11984-11990. https://doi.org/10.1039/C8NJ00778K
  27. Kulye, M., Liu, H., Zhang, Y., Zeng, H., Yang, X. and Qiu, D. 2012. Hrip1, a novel protein elicitor from necrotrophic fungus, Alternaria tenuissima, elicits cell death, expression of defence-related genes and systemic acquired resistance in tobacco. Plant Cell Environ. 35:2104-2120. https://doi.org/10.1111/j.1365-3040.2012.02539.x
  28. Lai, Y. S., Renna, L., Yarema, J., Ruberti, C., He, S. Y. and Brandizzi, F. 2018. Salicylic acid-independent role of NPR1 is required for protection from proteotoxic stress in the plant endoplasmic reticulum. Proc. Natl. Acad. Sci. U.S.A. 115:E5203-E5212. https://doi.org/10.1073/pnas.1802254115
  29. Ma, L., Zhang, H. Y., Zhou, X. K., Yang, C. G., Zheng, S. C., Duo, J. L. and Mo, M. H. 2018. Biological control tobacco bacterial wilt and black shank and root colonization by bio-organic fertilizer containing bacterium Pseudomonas aeruginosa NXHG29. Appl. Soil Ecol. 129:136-144. https://doi.org/10.1016/j.apsoil.2018.05.011
  30. Mao, J., Liu, Q., Yang, X., Long, C., Zhao, M., Zeng, H., Liu, H., Yuan, J. and Qiu, D. 2010. Purification and expression of a protein elicitor from Alternaria tenuissima and elicitormediated defence responses in tobacco. Ann. Appl. Biol. 156:411-420. https://doi.org/10.1111/j.1744-7348.2010.00398.x
  31. Marche, M. G., Mura, M. E. and Ruiu, L. 2016. Brevibacillus laterosporus inside the insect body: Beneficial resident or pathogenic outsider?. J. Invertebr. Pathol. 137:58-61. https://doi.org/10.1016/j.jip.2016.05.002
  32. Nurnberger, T. 1999. Signal perception in plant pathogen defense. Cell. Mol. Life Sci. 55:167-182. https://doi.org/10.1007/s000180050283
  33. Ongena, M., Jourdan, E., Adam, A., Paquot, M., Brans, A., Joris, B., Arpigny, J. L. and Thonart, P. 2007. Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants. Environ. Microbiol. 9:1084-1090. https://doi.org/10.1111/j.1462-2920.2006.01202.x
  34. Pieterse, C. M. J. and van Loon, L. C. 1999. Salicylic acid-independent plant defence pathways. Trends Plant Sci. 4:52-58. https://doi.org/10.1016/S1360-1385(98)01364-8
  35. Pieterse, C, M. J., Leon-Reyes, A., Van der Ent, S. and Van Wees, S. C. M. 2009. Networking by small-molecule hormones in plant immunity. Nat. Chem. Biol. 5:308-316. https://doi.org/10.1038/nchembio.164
  36. Qiu, D., Mao, J., Yang, X. and Zeng, H. 2009. Expression of an elicitor-encoding gene from Magnaporthe grisea enhances resistance against blast disease in transgenic rice. Plant Cell Rep. 28:925-933. https://doi.org/10.1007/s00299-009-0698-y
  37. Rahantaniaina, M. S., Li, S., Chatel-Innocenti, G., Tuzet, A., Mhamdi, A., Vanacker, H. and Noctor, G. 2017. Glutathione oxidation in response to intracellular $H_2O_2$: key but overlapping roles for dehydroascorbate reductases. Plant Signal. Behav. 12:e1356531. https://doi.org/10.1080/15592324.2017.1356531
  38. Riera, N., Wang, H., Li, Y., Li, J., Pelz-Stelinski, K. and Wang, N. 2018. Induced systemic resistance against citrus canker disease by rhizospheric bacteria. Phytopathology 108:1038-1045. https://doi.org/10.1094/PHYTO-07-17-0244-R
  39. Ryu, C. M., Pare, F. P. W. and Kloepper, J. W. 2005. Invisible signals from the underground: Bacterial volatiles elicit plant growth promotion and induce systemic resistance. Plant Pathol. J. 21:7-12. https://doi.org/10.5423/PPJ.2005.21.1.007
  40. Saikia, R., Gogoi, D. K., Mazumder, S., Yadav, A., Sarma, R. K., Bora, T. C. and Gogoi, B. K. 2011. Brevibacillus laterosporus strain BPM3, a potential biocontrol agent isolated from a natural hot water spring of Assam, India. Microbiol. Res. 166:216-225. https://doi.org/10.1016/j.micres.2010.03.002
  41. Saitou, N. and Nei, M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4:406-425.
  42. Shilina, J. V., Gushcha, M. I., Molozhava, O. S., Litvinov, S. V., and Dmitriev, A. P. 2018. Erratum to: "Induction of Arabidopsis thaliana resistance to pathogenic bacteria by lipopolysaccharide and salicylic acid". Cytol. Genet. 52:322. https://doi.org/10.3103/S0095452718040114
  43. Smolinska, U. and Kowalska, B. 2018. Biological control of the soil-borne fungal pathogen Sclerotinia sclerotiorum - a review. J. Plant Pathol. 100:1-12. https://doi.org/10.1007/s42161-018-0023-0
  44. Song, Z., Liu, K., Lu, C., Yu, J., Ju, R. and Liu, X. 2011. Isolation and characterization of a potential biocontrol Brevibacillus laterosporus. Afr. J. Microbiol. Res. 5:2675-2681. https://doi.org/10.5897/AJMR11.335
  45. Somasundaram, V., Basudhar, D., Bharadwaj, G., No, J. H., Ridnour, L. A., Cheng, R., Fujita, M., Thomas, D. D., Anderson, S. K., and McVicar, D. W. and Wink, D. A. 2018. Molecular mechanisms of nitric oxide in cancer progression, signal transduction and metabolism. Antioxid. Redox Signal. 30:1124-1143.
  46. Srivastava, N., Gonugunta, V. K., Puli, M. R. and Raghavendra, A. S. 2009. Nitric oxide production occurs downstream of reactive oxygen species in guard cells during stomatal closure induced by chitosan in abaxial epidermis of Pisum sativum. Planta 229:757-765. https://doi.org/10.1007/s00425-008-0855-5
  47. Thordal-Christensen, H., Zhang, Z., Wei, Y. and Collinge, D. B. 1997. Subcellular localization of $H_2O_2$ in plants. $H_2O_2$ accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J. 11:1187-1194. https://doi.org/10.1046/j.1365-313X.1997.11061187.x
  48. Tian, B., Li, N., Lian, L., Liu, J., Yang, J. and Zhang, K. Q. 2006. Cloning, expression and deletion of the cuticle-degrading protease BLG4 from nematophagous bacterium Brevibacillus laterosporus G4. Arch. Microbiol. 186:297-305. https://doi.org/10.1007/s00203-006-0145-1
  49. Torres, M. A. 2010. ROS in biotic interactions. Physiol. Plant. 138:414-429. https://doi.org/10.1111/j.1399-3054.2009.01326.x
  50. Ueki, A., Kaku, N. and Ueki, K. 2018. Role of anaerobic bacteria in biological soil disinfestation for elimination of soil-borne plant pathogens in agriculture. Appl. Microbiol. Biotechnol. 102:6309-6318. https://doi.org/10.1007/s00253-018-9119-x
  51. van Loon, L. C., Bakker, P. A. and Pieterse, C. M. 1998. Systemic resistance induced by rhizosphere bacteria. Annu. Rev. Phytopathol. 36:453-483. https://doi.org/10.1146/annurev.phyto.36.1.453
  52. Wang, H., Yang, X., Guo, L., Zeng, H. and Qiu, D. 2015. PeBL1, a novel protein elicitor from Brevibacillus laterosporus strain A60, activates defense responses and systemic resistance in Nicotiana benthamiana. Appl. Environ. Microbiol. 81:2706-2716. https://doi.org/10.1128/AEM.03586-14
  53. Wang, N., Liu, M., Guo, L., Yang, X. and Qiu, D. 2016. A novel protein elicitor (PeBA1) from Bacillus amyloliquefaciens NC6 induces systemic resistance in Tobacco. Int. J. Biol. Sci. 12:757-767. https://doi.org/10.7150/ijbs.14333
  54. Wei, C., Zhu, L., Wen, J., Yi, B., Ma, C., Tu, J., Shen, J. and Fu, T. 2018. Morphological, transcriptomics and biochemical characterization of new dwarf mutant of Brassica napus. Plant Sci. 270:97-113. https://doi.org/10.1016/j.plantsci.2018.01.021
  55. Wei, Z. M., Laby, R. J., Zumoff, C. H., Bauer, D. W., He, S. Y., Collmer, A. and Beer, S. V. 1992. Harpin, elicitor of the hypersensitive response produced by the plant pathogen Erwinia amylovora. Science 257:85-88. https://doi.org/10.1126/science.1621099
  56. Wu, G., Liu, Y., Xu, Y., Zhang, G., Shen, Q. R. and Zhang, R. 2018. Exploring elicitors of the beneficial rhizobacterium Bacillus amyloliquefaciens SQR9 to induce plant systemic resistance and their interactions with plant signaling pathways. Mol. Plant-Microbe Interact. 31:560-567. https://doi.org/10.1094/MPMI-11-17-0273-R
  57. Zhang, H., Yu, P., Zhao, J., Jiang, H., Wang, H., Zhu, Y., Botella, M. A., Samaj, J., Li, C. and Lin, J. 2018b. Expression of tomato prosystemin gene in Arabidopsis reveals systemic translocation of its mRNA and confers necrotrophic fungal resistance. New Phytol. 217:799-812. https://doi.org/10.1111/nph.14858
  58. Zhang, W., Yang, X., Qiu, D., Guo, L., Zeng, H., Mao, J. and Gao, Q. 2011. PeaT1-induced systemic acquired resistance in tobacco follows salicylic acid-dependent pathway. Mol. Biol. Rep. 38:2549-2556. https://doi.org/10.1007/s11033-010-0393-7
  59. Zhang, Y., Yan, X., Guo, H., Zhao, F. and Huang, L. 2018a. A novel protein elicitor BAR11 from Saccharothrix yanglingensis Hhs.015 improves plant resistance to pathogens and interacts with catalases as targets. Front. Microbiol. 9:700. https://doi.org/10.3389/fmicb.2018.00700
  60. Zhang, Y. H., Yang, X. F., Liu, Q., Qiu, D. W., Zhang, Y. L., Zeng, H. M., Yuan, J. J. and Mao, J. J. 2010. Purification of novel protein elicitor from Botrytis cinerea that induces disease resistance and drought tolerance in plants. Microbiol. Res. 165:142-151. https://doi.org/10.1016/j.micres.2009.03.004
  61. Zhao, J., Davis, L. C. and Verpoorte, R. 2005. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol. Adv. 23:283-333. https://doi.org/10.1016/j.biotechadv.2005.01.003
  62. Zhao, J., Guo, L., Zeng, H., Yang, X., Yuan, J., Shi, H., Xiong, Y., Chen, M., Han, L. and Qiu, D. 2012. Purification and characterization of a novel antimicrobial peptide from Brevibacillus laterosporus strain A60. Peptides 33:206-211. https://doi.org/10.1016/j.peptides.2012.01.001