Acta Scientific Microbiology (ASMI) (ISSN: 2581-3226)

Research Article Volume 3 Issue 6

Cocculus hirsutus Trypsin Inhibitor Confers Resistance to Ralstonia solanacearum: In silico Analysis

Manushree V1,2, Udit NM1,3, Manjunath BJ1, Devaraj VR 2 and Theertha Prasad D1*

1Department of Plant Biotechnology, University of Agricultural Sciences, GKVK, Bangalore, India
2Department of Biochemistry, Bangalore University, Central College Campus, Bangalore, India
3Department of Biochemistry and Crop physiology, MS Swaminathan School of Agriculture, Centurion University of Technology and management, Paralakhemundi, Odisha, India.

*Corresponding Author: D Theertha Prasad, Professor, Department of Plant Biotechnology, University of Agricultural Sciences, GKVK, Bangalore, India.

Received: April 08, 2020; Published: May 13, 2020

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Abstract

  Genetic engineering has proven to be an efficient strategy for the development of Disease resistant crops. Gram negative bacteria Ralstonia solanacearum, the causative agent of wilt has been causing devastating economic losses in tomato crop yield worldwide. In this study Cocculus hirsutus trypsin inhibitor has demonstrated for its bactericidal activity. Transgenic plants expressing ChTI were developed conferring resistance to Gram negative bacteria, particularly R. solanacearum. In vitro and In vivo studies revealed the bactericidal activity of ChTI with mortality rate upto 42% and 48% in E. coli and R. solanacearum at 500 TIU/mg respectively. Transgenic plants expressing ChTI also developed resistance to the wilt. Whereas non transgenic plants developed symptoms of the disease. Further Strong interaction patterns of ChTI-Lipid and ChTI-membrane proteins revealed the possible mechanism of action of these PIs. These results demonstrate the effectiveness of ChTI as bactericidal agent and is able to enhance resistance in transgenic plants against bacterial pathogens.

Keywords: R. Solanacearum; Cell Membrane Binding Protein; Cocculus hirsutus Trypsin Inhibitor; Evans Blue Assay; Spot Dilution Bacterial Assay; In Silico Protein-Lipid Interactions

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References

  1. Ingmer H and Brondsted L. “Proteases in bacterial pathogenesis”. Research in Microbiology 160 (2009): 704-710.
  2. Yu GL., et al. “Arabidopsis mutations at the RPS2 locus result in loss of resistance to Pseudomonas syringae strains expressing the avirulence gene avrRpt2”. Molecular Plant-Microbe Interactions 6 (1993): 434- 443.
  3. Guttman DS and Greenberg JT. “Functional analysis of the type III effectors AvrRpt2 and AvrRpm1 of Pseudomonas syringae with the use of a single-copy genomic integration system”. Molecular Plant-Microbe Interactions 14 (2001): 145-155.
  4. Bardoel BW., et al. “Pseudomonas evades immune recognition of flagellin in both mammals and plants”. PLOS Pathogens 7 (2011): e1002206.
  5. Feng T., et al. “Characterization of an extensin-modifying metalloprotease: N-terminal processing and substrate cleavage pattern of Pectobacterium carotovorum Prt1”. Applied Microbiology and Biotechnology 98 (2014): 10077-10089.
  6. Ustun S., et al. “The Xanthomonas campestris type III effector XopJ targets the host cell proteasome to suppress salicylic-acid mediated plant defence”. PLOS Pathogens 9 (2013): e1003427.
  7. Ustun S., et al. “HopZ4 from Pseudomonas syringae, a member of the HopZ type III effector family from the YopJ superfamily, inhibits the proteasome in plants”. Molecular Plant-Microbe Interactions 27 (2014): 611-623.
  8. Gimenez-Ibanez S., et al. “The bacterial effector HopX1 targets JAZ transcriptional repressors to activate jasmonate signaling and promote infection in Arabidopsis”. PLOS Biology 12 (2014): e1001792.
  9. Nawrot R., et al. “Plant antimicrobial peptides”. Folia Microbiologica 59 (2014): 181-196.
  10. Kumar P., et al. “Antimicrobial Peptides: Diversity, Mechanism of Action and Strategies to Improve the Activity and Biocompatibility In Vivo”. Biomolecules 19 (2018): 8.
  11. Satheesh LP and Murugan K. “Antimicrobial activity of protease inhibitors from leaves of Coccinia grandis (L.) Voigt”. Indian Journal of Experimental Biology 49 (2011): 366-374.
  12. Rakashanda S., et al. “Antibacterial activity of a trypsin-chymotrypsin-elastase inhibitor isolated from Lavatera cashmeriana seeds”. The Journal of Animal and Plant Sciences 22 (2012): 983-986.
  13. Shamsi T and Fatima S. “Protease Inhibitors as Antibiotics”. Journal of Pharmaceutical Sciences 3 (2016): 131-137.
  14. Zasheva D and Lyudmila S. “Antitumour potential of plant protease inhibitors”. Journal 'Genetics and Plant Physiology 7 (2017): 147-159.
  15. Souza LdC., et al. “Effects of an Anticarcinogenic Bowman-Birk Protease Inhibitor on Purified 20S Proteasome and MCF-7 Breast Cancer Cells”. PLoS One 9 (2014): e86600.
  16. Srikanth S and Chen Z. “Plant protease inhibitors in therapeutics-focus on cancer therapy”. Frontiers in Pharmacology 7 (2016): 470.
  17. Fear G., et al. “Protease inhibitors and their peptidomimetic derivatives as potential drugs”. Pharmacology and Therapeutics 113 (2007): 354-368.
  18. Bijina Sreeja Chellappan., et al. “Protease inhibitor from Moringa oleifera with potential for use as therapeutic drug and as seafood preservative”. Saudi Journal of Biological Sciences 18 (2011): 273-281.
  19. Bhattacherjee C., et al. “Purification of a trypsin inhibitor from Cocculus hirsutus and identification of its biological activity”. Journal of Crop Science and Biotechnology 12 (2010): 253-260.
  20. Manushree V., et al. “Expression of Cocculus hirsutus trypsin inhibitor promotes endogenous defensive response against Helicoverpa armigera and enhanced levels of antioxidants”. African Journal of Plant Science 14 (2020): 65-82.
  21. March SC., et al. “A simplified method for cyanogen bromide activation of agarose for affinity chromatography”. Analytical Biochemistry 60 (1974): 149-152.
  22. Yan Y., et al. “HDOCK: a web server for protein-protein and protein-DNA/RNA docking based on a hybrid strategy”. Nucleic Acids Research 45 (2017): 365-373.
  23. Laskowski RA and Swindells MB. “LigPlot+: multiple ligand-protein interaction diagrams for drug discovery”. Journal of Chemical Information and Modeling 51 (2011): 2778-2786.
  24. DeLano WL. “Pymol: An open-source molecular graphics tool”. CCP4 Newsletter on Protein Crystallography 40 (2002): 82-92.
  25. Kim SG., et al. “Evaluation of Resistance to Ralstonia solanacearum in Tomato Genetic Resources at Seedling Stage”. The Plant Pathology Journal 32 (2016): 58-64.
  26. I Arulpandi and R Sangeetha. “Antibacterial Activity of Fistulin: A Protease Inhibitor Purified from the Leaves of Cassia fistula”. International Scholarly Research Network ISRN Pharmaceutics (2012): 584073.
  27. Patrícia MGP., et al. “Protease inhibitors from plants: Biotechnological insights with emphasis on their effects on microbial pathogens” (2013).
  28. Kim JY., et al. “Protease inhibitors from plants with antimicrobial activity”. International Journal of Molecular Sciences 10 (2009): 2860-2872.
  29. Ngai P and Ng, T. “A napin-like polypeptide from dwarf Chinese white cabbage seeds with translation-inhibitory, trypsin-inhibitory, and antibacterial activities”. Peptides 25 (2004): 171-176.
  30. Senthilkumar R., et al. “Genetically pyramiding protease-inhibitor genes for dual broad-spectrum resistance against insect and phytopathogens in transgenic tobacco”. Plant Biotechnology Journal 8 (2010): 65-75.
  31. Brogden Kim. “Antimicrobial peptides: Pore formers or metabolic inhibitors in bacteria?” Nature Reviews Microbiology 3 (2005): 238-250.
  32. Ekiert DC., et al. “Architectures of Lipid Transport Systems for the Bacterial Outer Membrane”. Cell 169 (2017): 273-285.
  33. Kamischke C., et al. “The Acinetobacter baumannii Mla system and glycerophospholipid transport to the outer membrane”. Elife 8 (2019): e40171.
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Citation

Citation: Theertha Prasad D., et al. “Cocculus hirsutus Trypsin Inhibitor Confers Resistance to Ralstonia solanacearum: In silico Analysis". Acta Scientific Microbiology 3.6 (2020): 63-68.




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