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European Journal of Plant Pathology

, Volume 155, Issue 4, pp 1367–1371 | Cite as

Development of specific molecular markers to distinguish and quantify broomrape species in a soil sample

  • Radi AlyEmail author
  • Vinay Kumar Bari
  • Avishai Londner
  • Jackline Abu Nassar
  • Ran Lati
  • Leena Taha-Salaime
  • Hanan Eizenberg
Article
  • 124 Downloads

Abstract

Broomrapes (Orobanche and Phelipanche spp.) are destructive obligate plant parasites in Israel and in the Mediterranean basin. Conventional methods for parasitic weeds detection are difficult, since the parasite seeds are extremely small (dust-like seeds) and survive in the soil for several decades. Here, we report the development of specific molecular markers rbcL1 based on rbcL (large subunit of the ribulose-bisphosphate carboxylase) gene from Orobanche crenata and ITS100 based upon unique sequences in the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA of Phelipanche aegyptiaca. Genomic DNA was extracted from soil samples artificially infested with broomrape seeds or tissue of P. aegyptiaca, O. cumana and O. crenata and subjected to PCR analysis. rbcL1 marker, successfully differentiate between O. crenata and O. cumana, amplified a specific PCR products (1300 bp with O. crenata and 1000 bp with O. cumana). However, the rbcL1 marker failed to amplify soil samples with seeds or tissues of P. aegyptiaca or any soil-borne DNA. ITS100 marker and Real-Time PCR, allowed quantitative diagnostic of the parasite O. cumana in a soil sample; amplified a specific PCR products (100 bp). As expected the universal control primer (UCP-555) amplified a PCR product (555 bp), when genomic DNA extracted from soil samples with or without broomrape tissues. The development of an efficient, simple and robust molecular marker to detect and distinguish between broomrape species, has a significant insights on the assessment level of infestation and planning eradication program of the parasite in a field crop.

Keywords

Plant parasite Phelipanche aegyptiaca Orobanche spp. ribulose-1,5-bisphosphate carboxylase (rbcLInternal transcribed spacer (ITS

Notes

Acknowledgements

Results of this research were supported by the Chief Scientist of the Ministry of Agriculture and Rural Development - Israel, grant No.132-1499-10 (MEZAMALEKET). V.K.B is grateful to the ARO-Volcani Center, Agricultural Ministry of Israel for providing the Postdoctoral fellowship.

Compliance with ethical standards

Ethical responsibility

Our manuscript is original research and it is not submitted to full or in parts to other journal for publication.

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal studies

Our study did not involve human participants and/or animal as experimental model.

Informed consent

All authors consent to this submission.

References

  1. Agarwal, M., Shrivastava, N., & Padh, H. (2008). Advances in molecular marker techniques and their applications in plant sciences. Plant Cell Reports, 27, 617–631.CrossRefGoogle Scholar
  2. Aly, R. (2007). Conventional and biotechnological approaches for control of parasitic weeds. In Vitro Cell Dev-Pl, 43, 304–317.CrossRefGoogle Scholar
  3. Aly, R., Eizenberg, H., Kocherman, M., Abu-Nassar, J., Taha, L., & Saadi, I. (2012). Use of ITS nuclear sequences from Phelipanche aegyptiaca as a direct tool to detect single seeds of broomrape species in the soil. European Journal of Plant Pathology, 133, 523–526.CrossRefGoogle Scholar
  4. Aly, R., Lati, R., Abu-Nassar, J., Ziadna, H., Achdari, G., Münchow, C. S.v., Wicke, S., Bari, V. K., & Eizenberg, H. (2019). The weedy parasite Phelipanche aegyptiaca attacks Brassica rapa var. rapa L. for the first time in Israel. Plant Disease Note, 103, 1796.Google Scholar
  5. Cusimano, N., & Wicke, S. (2016). Massive intracellular gene transfer during plastid genome reduction in nongreen Orobanchaceae. The New Phytologist, 210, 680–693.CrossRefGoogle Scholar
  6. Delavault, P., Sakanyan, V., & Thalouarn, P. (1995). Divergent evolution of two plastid genes, rbcL and atpB, in a non-photosynthetic parasitic plant. Plant Molecular Biology, 29, 1071–1079.CrossRefGoogle Scholar
  7. Dongo, A., Leflon, M., Simier, P., & Delavault, P. (2012). Development of a high-throughput real-time quantitative PCR method to detect and quantify contaminating seeds of Phelipanche ramosa and Orobanche cumana in crop seed lots. Weed Research, 52, 34–41.CrossRefGoogle Scholar
  8. Habimana, S., Nduwumuremyi, A., & Chinama, R. J. D. (2014). Management of orobanche in field crops- a review. Journal of Soil Science and Plant Nutrition, 14, 43–62.Google Scholar
  9. Joel, D. M., Hershenhorn, Y., Eizenberg, H., Aly, R., Ejeta, G., Rich, P. J., Ransom, J. K., Sauerborn, J., & Rubiales, D. (2006). Biology and management of weedy root parasites. In Horticultural reviews (pp. 267–349). New Jersey: Wiley.Google Scholar
  10. Leebens-Mack, J., & de Pamphilis, C. (2002). Power analysis of tests for loss of selective constraint in cave crayfish and nonphotosynthetic plant lineages. Molecular Biology and Evolution, 19, 1292–1302.CrossRefGoogle Scholar
  11. McNeal, J. R., Bennett, J. R., Wolfe, A. D., & Mathews, S. (2013). Phylogeny and origins of holoparasitism in Orobanchaceae. American Journal of Botany, 100, 971–983.CrossRefGoogle Scholar
  12. Park, J. M., Manen, J. F., Colwell, A. E., & Schneeweiss, G. M. (2008). A plastid gene phylogeny of the non-photosynthetic parasitic Orobanche (Orobanchaceae) and related genera. Journal of Plant Research, 121, 365–376.CrossRefGoogle Scholar
  13. Parker, C., & Riches, C. R. (1993). Parasitic weeds of the world: Biology and control. Wallingford: CAB International.Google Scholar
  14. Schneeweiss, G. M., Colwell, A., Park, J. M., Jang, C. G., & Stuessy, T. F. (2004). Phylogeny of holoparasitic Orobanche (Orobanchaceae) inferred from nuclear ITS sequences. Molecular Phylogenetics and Evolution, 30, 465–478.CrossRefGoogle Scholar
  15. Wicke, S., Muller, K. F., de Pamphilis, C. W., Quandt, D., Wickett, N. J., Zhang, Y., Renner, S. S., & Schneeweiss, G. M. (2013). Mechanisms of functional and physical genome reduction in photosynthetic and nonphotosynthetic parasitic plants of the broomrape family. Plant Cell, 25, 3711–3725.CrossRefGoogle Scholar

Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2019

Authors and Affiliations

  1. 1.Department of Plant Pathology and Weed Research, Newe Ya’ar Research CenterAgricultural Research Organization (ARO)Ramat YishayIsrael

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