Advertisement

Molecular Biotechnology

, Volume 61, Issue 12, pp 929–937 | Cite as

Host-Induced Silencing of Some Important Genes Involved in Osmoregulation of Parasitic Plant Phelipanche aegyptiaca

  • Zahra Farrokhi
  • Hassan AlizadehEmail author
  • Houshang Alizadeh
  • Fariba Abooei Mehrizi
Original paper
  • 70 Downloads

Abstract

Broomrape is an obligate root-parasitic weed that acts as a competitive sink for host photoassimilates. Disruption of essential processes for growth of broomrape using host plant-mediated systemic signals can help to implement more specific and effective management plans of this parasite. Accordingly, we tested the possibility of transient silencing three involved genes (PaM6PR, PaCWI, and PaSUS1) in osmoregulation process of broomrape using syringe agroinfiltration of dsRNA constructs in tomato. The highest decrease in mRNA levels, enzyme activity, and amount of total reducing sugars was observed in Phelipanche aegyptiaca when grown on agroinfiltrated tomato plants by PaM6PR dsRNA construct than control. In addition, PaSUS1 dsRNA construct showed high reduction in mRNA abundance (32-fold fewer than control). The lowest decrease in mRNA levels was observed after infiltration of PaCWI dsRNA construct (eightfold fewer than control). While the highest reduction in PaM6PR and PaSUS1 expression levels was detected in the parasite at 3 days post-infiltration (dpi), the maximum reduction in both of the total reducing sugars amount and M6PR and SUS1 activities was observed at 8 dpi. On the contrary, CWI activity, PaCWI expression level, and amount of total reducing sugars in broomrape shoots simultaneously decreased at the day 3 after the dsRNA construct infiltration against PaCWI. On the whole, our results indicated that the three studied genes especially PaM6PR may constitute appropriate targets for the development of transgenic resistance in host plants using silencing strategy.

Keywords

Down-regulation Invertase Mannose 6-phosphate reductase Sucrose synthase Syringe agroinfiltration 

Notes

Acknowledgements

The authors would like to thank the Iran National Science Foundation (INSF) for funding this research under Grant Number 93033866.

Author Contributions

HA and HA conceived and designed research. ZF conducted experiments and analyzed the data. ZF wrote the first version of the manuscript. All authors read and approved the manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no competing interests.

References

  1. 1.
    Abbes, Z., Kharrat, M., Delavault, P., Chaïbi, W., & Simier, P. (2009). Nitrogen and carbon relationships between the parasitic weed Orobanche foetida and susceptible and tolerant faba bean lines. Plant Physiology and Biochemistry, 47, 153–159.CrossRefGoogle Scholar
  2. 2.
    Aly, R., et al. (2009). Gene silencing of mannose 6-phosphate reductase in the parasitic weed Orobanche aegyptiaca through the production of homologous dsRNA sequences in the host plant. Plant Biotechnology Journal, 7, 487–498.CrossRefGoogle Scholar
  3. 3.
    Aly, R., Plakhin, D., & Achdari, G. (2006). Expression of sarcotoxin IA gene via a root-specific tob promoter enhanced host resistance against parasitic weeds in tomato plants. Plant Cell Reports, 25, 297–303.CrossRefGoogle Scholar
  4. 4.
    Bhaskar, P. B., Venkateshwaran, M., Wu, L., Ané, J.-M., & Jiang, J. (2009). Agrobacterium-mediated transient gene expression and silencing: A rapid tool for functional gene assay in potato. PLoS ONE, 4, e5812.CrossRefGoogle Scholar
  5. 5.
    Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.  https://doi.org/10.1016/0003-2697(76)90527-3.CrossRefPubMedGoogle Scholar
  6. 6.
    Broghammer, A., et al. (2012). Legume receptors perceive the rhizobial lipochitin oligosaccharide signal molecules by direct binding. Proceedings of the National Academy of Sciences United States of America, 109, 13859–13864.CrossRefGoogle Scholar
  7. 7.
    Chauhan, B. S., & Mahajan, G. (2014). Recent advances in weed management. New York: Springer.CrossRefGoogle Scholar
  8. 8.
    Delavault, P., Simier, P., Thoiron, S., Véronési, C., Fer, A., & Thalouarn, P. (2002). Isolation of mannose 6-phosphate reductase cDNA, changes in enzyme activity and mannitol content in broomrape (Orobanche ramosa) parasitic on tomato roots. Physiologia Plantarum, 115, 48–55.  https://doi.org/10.1034/j.1399-3054.2002.1150105.x.CrossRefPubMedGoogle Scholar
  9. 9.
    Dellaporta, S. L., Wood, J., & Hicks, J. B. (1983). A plant DNA minipreparation: Version II. Plant Molecular Biology Reporter, 1, 19–21.CrossRefGoogle Scholar
  10. 10.
    Doyle, J. J. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemistry, 19, 11–15.Google Scholar
  11. 11.
    Draie, R., et al. (2011). Invertases involved in the development of the parasitic plant Phelipanche ramosa: Characterization of the dominant soluble acid isoform, PrSAI1. Molecular Plant Pathology, 12, 638–652.  https://doi.org/10.1111/j.1364-3703.2010.00702.x.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Dubey, N. K., et al. (2017). Enhanced host-parasite resistance based on down-regulation of Phelipanche aegyptiaca target genes is likely by mobile small RNA Frontiers in plant science, 8, 1574.Google Scholar
  13. 13.
    Everard, J. D., Cantini, C., Grumet, R., Plummer, J., & Loescher, W. H. (1997). Molecular cloning of mannose-6-phosphate reductase and its developmental expression in celery. Plant Physiology, 113, 1427–1435.CrossRefGoogle Scholar
  14. 14.
    Farrokhi, Z., Alizadeh, H., & Alizadeh, H. (2019). Developmental patterns of enzyme activity, gene expression, and sugar content in sucrose metabolism of two broomrape species. Plant Physiology and Biochemistry, 142, 8–14.  https://doi.org/10.1016/j.plaphy.2019.06.014.CrossRefPubMedGoogle Scholar
  15. 15.
    Feder, M., & Walser, J. C. (2005). The biological limitations of transcriptomics in elucidating stress and stress responses. Journal of Evolutionary Biology, 18, 901–910.CrossRefGoogle Scholar
  16. 16.
    Fernández-Aparicio, M., Reboud, X., & Gibot-Leclerc, S. (2016). Broomrape weeds. Underground mechanisms of parasitism and associated strategies for their control: A review. Frontiers in Plant Science, 7, 135.  https://doi.org/10.3389/fpls.2016.00135.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391, 806–811.CrossRefGoogle Scholar
  18. 18.
    Foy, C. L., Jain, R., & Jacobsohn, R. (1989). Recent approaches for chemical control of broomrape (Orobanche spp.). Reviews of Weed Science, 4, 123–152.Google Scholar
  19. 19.
    Gleave, A. P. (1992). A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Molecular Biology, 20, 1203–1207.  https://doi.org/10.1007/bf00028910.CrossRefPubMedGoogle Scholar
  20. 20.
    Grunberg-Manago, M. (1999). Messenger RNA stability and its role in control of gene expression in bacteria and phages. Annual Review of Genetics, 33, 193–227.CrossRefGoogle Scholar
  21. 21.
    Hibberd, J. M., Quick, W. P., Press, M. C., Scholes, J. D., & Jeschke, W. D. (1999). Solute fluxes from tobacco to the parasitic angiosperm Orobanche cernua and the influence of infection on host carbon and nitrogen relations. Plant, Cell & Environment, 22, 937–947.  https://doi.org/10.1046/j.1365-3040.1999.00462.x.CrossRefGoogle Scholar
  22. 22.
    Huang, G., Allen, R., Davis, E. L., Baum, T. J., & Hussey, R. S. (2006). Engineering broad root-knot resistance in transgenic plants by RNAi silencing of a conserved and essential root-knot nematode parasitism gene. Proceedings of the National Academy of Sciences Unites States of America, 103, 14302–14306.CrossRefGoogle Scholar
  23. 23.
    McCarthy, J. E., & Gualerzi, C. (1990). Translational control of prokaryotic gene expression. Trends in Genetics, 6, 78–85.CrossRefGoogle Scholar
  24. 24.
    McDonald, J. P., Tissier, A., Frank, E. G., Iwai, S., Hanaoka, F., & Woodgate, R. (2001). DNA polymerase iota and related Rad30–like enzymes. Philosophical Transactions of the Royal Society B, 356, 53–60.CrossRefGoogle Scholar
  25. 25.
    Miller, G. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428.CrossRefGoogle Scholar
  26. 26.
    Mlotshwa, S., et al. (2002). RNA silencing and the mobile silencing signal. The Plant Cell, 14, S289–S301.CrossRefGoogle Scholar
  27. 27.
    Parker, C., & Riches, C. R. (1993). Parasitic weeds of the world: Biology and control. Wallingford: Cab Intl.Google Scholar
  28. 28.
    Péron, T., et al. (2012). Role of the sucrose synthase encoding PrSus1 gene in the development of the parasitic plant Phelipanche ramosa L. (Pomel). Molecular Plant-Microbe Interactions, 25, 402–411.CrossRefGoogle Scholar
  29. 29.
    Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Research, 29, e45.CrossRefGoogle Scholar
  30. 30.
    Price, D. R., & Gatehouse, J. A. (2008). RNAi-mediated crop protection against insects. Trends in Biotechnology, 26, 393–400.CrossRefGoogle Scholar
  31. 31.
    Prins, M., Laimer, M., Noris, E., Schubert, J., Wassenegger, M., & Tepfer, M. (2008). Strategies for antiviral resistance in transgenic plants. Molecular Plant Pathology, 9, 73–83.  https://doi.org/10.1111/j.1364-3703.2007.00447.x.CrossRefPubMedGoogle Scholar
  32. 32.
    Robert, S., Simier, P., & Fer, A. (1999). Purification and characterization of mannose 6-phosphate reductase, a potential target for the control of Striga hermonthica and Orobanche ramosa. Functional Plant Biology, 26, 233–237.  https://doi.org/10.1071/PP98138.CrossRefGoogle Scholar
  33. 33.
    Ruan, Y.-L., Jin, Y., Yang, Y.-J., Li, G.-J., & Boyer, J. S. (2010). Sugar input, metabolism, and signaling mediated by invertase: Roles in development, yield potential, and response to drought and heat. Molecular Plant, 3, 942–955.CrossRefGoogle Scholar
  34. 34.
    Sambrook, J., & Russell, W. (2001). Molecular cloning: A laboratory manual. New York: Cold pring Harbor Laboratory Press.Google Scholar
  35. 35.
    Smith, N. A., Singh, S. P., Wang, M.-B., Stoutjesdijk, P. A., Green, A. G., & Waterhouse, P. M. (2000). Gene expression: Total silencing by intron-spliced hairpin RNAs. Nature, 407, 319–320.CrossRefGoogle Scholar
  36. 36.
    Varshavsky, A. (1996). The N-end rule: Functions, mysteries, uses. Proceedings of the National Academy of Sciences United States of America, 93, 12142–12149.CrossRefGoogle Scholar
  37. 37.
    Voinnet, O. (2008). Post-transcriptional RNA silencing in plant–microbe interactions: A touch of robustness and versatility. Current Opinion in Plant Biology, 11, 464–470.CrossRefGoogle Scholar
  38. 38.
    Wang, K. (2006). Agrobacterium protocols (Vol. 1). New York: Springer.Google Scholar
  39. 39.
    Wesley, S. V., et al. (2001). Construct design for efficient, effective and high-throughput gene silencing in plants. The Plant Journal, 27, 581–590.CrossRefGoogle Scholar
  40. 40.
    Wiesen, J. L., & Tomasi, T. B. (2009). Dicer is regulated by cellular stresses and interferons. Molecular Immunology, 46, 1222–1228.CrossRefGoogle Scholar
  41. 41.
    Yang, Z., Wang, T., Wang, H., Huang, X., Qin, Y., & Hu, G. (2013). Patterns of enzyme activities and gene expressions in sucrose metabolism in relation to sugar accumulation and composition in the aril of Litchi chinensis Sonn. Journal of Plant Physiology, 170, 731–740.  https://doi.org/10.1016/j.jplph.2012.12.021.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Agronomy & Plant Breeding, College of Agriculture & Natural ResourcesUniversity of TehranKarajIran

Personalised recommendations