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Ipomoea batatas: papain propeptide inhibits cysteine protease in main plant parasites and enhances resistance of transgenic tomato to parasites


Different parasites cause severe lose in quantity and quality of crops. Many parasites develop haustorial cells and stylets that penetrate the host using secreted enzymes and mechanical pressure. Cysteine proteases are pre-pro-enzyme produced by parasites that are essential for normal parasitism. Papain is also a kind of cysteine proteases such that its propeptide segment has inhibitory properties and limits the protease activity of papain. To investigate the inhibitory effects of papain propeptide on some parasite proteases, we cloned inhibitory propeptide of papain of Ipomoea batatas, and enzymatic fragments of Diabrotica virgifera cathepsin L-like protease-1, Meloidogyne incognita cathepsin L-like protease 1, Heterodera glycines cysteine protease-1, Cuscuta chinesis cysteine protease and Orobanche cernua cysteine protease. After purification of recombinant inhibitory propeptide and enzymatic fragments, the inhibition activity of propeptide on cysteine proteases was measured. Finally inhibitory propeptide was transformed into tomato and transgenic plants resistance to parasites (bioassay) were examined. We demonstrated papain-propeptide inhibits cysteine protease of mentioned parasites. In transgenic tomato plants, papain-inhibitory propeptide effectively interrupted haustoria development. Haustoria-digitate cells of dodder could not differentiate and develop into the phloem and xylem hyphae on transgenic tomatoes. Parasites grown on transgenic tomatoes showed reduction in vigor and productivity due to defective connection of haustoria. Lower ratio of female nematodes and a decrease of nematode egg mass per transgenic line indicated biocontrol of nematode. The changes in growth factors of parasite challenged transgenic lines relative to controls, indicates the efficacy of papain propeptide in control of parasitism.

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  1. Albert M, Kaiser B, van der Krol S, Kaldenhoff R (2010) Calcium signaling during the plant-plant interaction of parasitic Cuscuta reflexa with its hosts. Plant Signal Behav 59:1144–1146

    Article  Google Scholar 

  2. Amini M, Saify Nabiabad H, Deljou A (2017) Host-synthesized cysteine protease-specific inhibitor disrupts Cuscuta campestris parasitism in tomato. Plant Biotechnol Rep 11:289–298

    Article  Google Scholar 

  3. Amini M, Saify Nabiabad H, Deljou A (2018) The role of cuscutain-propeptide inhibitor in haustoria parasitism and enhanced resistance to dodder in transgenic alfalfa expressing this propeptide. Plant Biotechnol Rep 58:144–146

    Google Scholar 

  4. Ashigh J, Marquez E (2010) Dodder (Cuscuta spp.) biology and management. NM State University, Guide A-615. vol 54. pp 31–34

  5. Birschwilks M, Sauer N, Scheel D, Neumann S (2007) Arabidopsis thaliana is a susceptible host plant for the holoparasite Cuscuta spec. Planta 226:1231–1241

    Article  CAS  PubMed  Google Scholar 

  6. Bleischwitz M, Albert M, Fuchsbauer HL, Kaldenhoff R (2010) Significance of cuscutain, a cysteine protease from Cuscuta reflexa, in host-parasite interactions. PMC Plant Biol 10:227

    Google Scholar 

  7. Borsics T, Lados M (2002) Dodder infection induces the expression of a pathogenesis-related gene of the family PR-10 in alfalfa. J Exp Bot 53:1831–1832

    Article  CAS  PubMed  Google Scholar 

  8. Dennis C (2013) Alfalfa management guide for Ningxia. United Nations Food and Agriculture Organization, pp 1–2. Retrieved 3 August 2013

  9. Dinh PTY, Zhang L, Brown CR, Elling AA (2014) Plant- mediated RNA interference of effector gene Mc16D10L confers resistance against Meloidogyne chitwoodi in diverse genetic backgrounds of potato and reduces pathogenicity of nematode offspring. Nematology 16:669–682.

    Article  CAS  Google Scholar 

  10. Ejeta G, Butler LG, Hess DE, Vogler RK (1991) Genetic and breeding strategies for Striga resistance in sorghum. In: Ransom JK, Musselman LJ, Worsham AD, Parker C (eds) Proceedings of the 5th international symposium of parasitic weeds, CIMMYT, Nairobi, Kenya, 1991, pp 539–544

  11. Furuhashi T, Furuhashi K, Weckwerth W (2011) The parasitic mechanism of the holostemparasitic plant Cuscuta. J Plant Interact 6:207–219

    Article  CAS  Google Scholar 

  12. Kim SH, Hamada T (2005) Rapid and reliable method of extracting DNA and RNA from sweetpotato, Ipomoea batatas (L). Lam. Biotechnol Lett 27:1841–1845

    Article  CAS  PubMed  Google Scholar 

  13. Marra B, Souza D, Aguiar J, Firmino A et al (2009) Protective effects of a cysteine proteinase propeptide expressed in transgenic soybean roots. Peptides 30:825–831

    Article  CAS  PubMed  Google Scholar 

  14. Mishra JS (2009) Biology and management of Cuscuta species. Indian J Weed Sci 41:1–11

    Google Scholar 

  15. Okamoto T, Shimada T, Hara-Nishimura I, Nishimura M, Minamikawa T (2003) C-terminal KDEL sequence of a KDEL-tailed cysteine-proteinase (sulfhydryl-endopeptidase) is involved in formation of KDEL vesicle in efficient vacuolar transport of sulfhydryl-endopeptidase. Plant Physiol 132:1892–1900

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Papolu PK, Gantasala NP, Kamaraju D, Banakar P, Sreevathsa R, Rao U (2013) Utility of host delivered RNAi of two FMRF amide like peptides, flp-14 and flp-18, for the management of root-knot nematode, Meloidogyne incognita. PLoS ONE 8:e80603.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Saify Nabiabad H, Yaghoobi M, Jalali-Javaran M, Hosseinkhani S (2011) Expression analysis and purification of human recombinant tissue type plasminogen activator (rtPA) from transgenic tobacco plants. Prep Biochem Biotechnol 41:175–186

    Article  CAS  Google Scholar 

  18. Sajid M, McKerrow JH (2002) Cysteine proteases of parasitic organisms. Mol Biochem Parasitol 120:1–21

    Article  CAS  PubMed  Google Scholar 

  19. Santamaria M, Arnaiz A, Mendoza M, Martinez M, Diaz I (2015) Inhibitory properties of cysteine protease pro-peptides from barley confer resistance to spider mite feeding. PLoS ONE 10(6):e0128323

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Saric-Krsmanovic M, Bozic D, Malidza G, Radivojevic L et al (2015) Chemical control of field dodder in alfalfa. Pestic Phytomedicine 30:107–114

    Article  Google Scholar 

  21. Vaughn KC (2002) Attachment of the parasitic weed dodder to the host. Protoplasma 219:227–237

    Article  CAS  PubMed  Google Scholar 

  22. Wilhite E, Elden TC, Brzin J, Smigocki AC (2000) Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil mid gut with biochemical and plant-derived proteinase inhibitors. Insect Biochem Mol Biol 30:1181–1188

    Article  CAS  PubMed  Google Scholar 

  23. Yoder JI, Scholes JD (2010) Host plant resistance to parasitic weeds; recent progress and bottlenecks. Curr Opin Plant Biol 13:478–484

    Article  CAS  PubMed  Google Scholar 

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The research is partially supported by the Bu Ali Sina University.

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Correspondence to Haidar Saify Nabiabad.

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Saify Nabiabad, H., Amini, M. & Kianersi, F. Ipomoea batatas: papain propeptide inhibits cysteine protease in main plant parasites and enhances resistance of transgenic tomato to parasites. Physiol Mol Biol Plants 25, 933–943 (2019).

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  • Cuscuta chinesis
  • Papain protease
  • Parasitism
  • Nematodes