European Journal of Plant Pathology

, Volume 128, Issue 3, pp 343–351 | Cite as

In planta PCR-based detection of early infection of plant-parasitic nematodes in the roots: a step towards the understanding of infection and plant defence

  • Syarifah Aisyafaznim Sayed Abdul Rahman
  • Zulqarnain Mohamed
  • Rofina Yasmin Othman
  • Rony Swennen
  • Bart Panis
  • Dirk De Waele
  • Serge Remy
  • Sebastien Christian Carpentier
Article
First Online:
Accepted:

Abstract

The polyphagous obligate parasites Meloidogyne spp. devastate a wide range of crop plants including bananas and plantains. Their infestations impact agriculture worldwide. Therefore, an effective combating regime against this nematode species and an in-depth understanding of plant-nematode interaction are essential. Early detection of infection by visual inspection is not possible. This hampers early control strategy efforts and makes in-depth research of the early infection and plant defence unfeasible. A simple and robust in planta PCR-based nematode detection method is described here as the first crucial step. This PCR-based detection assay exploits the existence of the Internal Transcribed Spacer 1 (ITS 1) region of the ribosomal DNA (rDNA) gene family in the nematodes for early detection of nematode penetration into the roots. The results demonstrate that this detection assay is suitable to serve as a molecular screening tool for plant root diagnostic purposes.

Keywords

Banana DNA extraction In planta detection Meloidogyne incognita Nematode PCR 

Abbreviations

IP

Infected plant root

N

Nematode individual

ND

Nematode DNA

NP

Nematode mixed with plant roots

NPD

Nematode and plant DNA

P

Plant roots

PD

Plant DNA

TR

Transformed tomato roots infected with Meloidogyne incognita

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Notes

Acknowledgement

This work was supported by the Ministry of Science, Technology and Innovation Malaysia, through the National e-Science Fund 02-01-03-SF0088. We would like to acknowledge University of Malaya, Kuala Lumpur, Malaysia for funding the first author’s stay in the Laboratory of Tropical Crop Improvement, Leuven, Belgium. We also wish to thank Annemie Elsen, Christine Vos, Els Thiry and Isabelle Henry for their continuous assistance and valuable comments during this study.

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Copyright information

© KNPV 2010

Authors and Affiliations

  • Syarifah Aisyafaznim Sayed Abdul Rahman
    • 1
    • 2
  • Zulqarnain Mohamed
    • 1
    • 3
  • Rofina Yasmin Othman
    • 1
    • 3
  • Rony Swennen
    • 2
  • Bart Panis
    • 2
  • Dirk De Waele
    • 2
  • Serge Remy
    • 2
  • Sebastien Christian Carpentier
    • 2
  1. 1.Unit of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia
  2. 2.Laboratory of Tropical Crop Improvement, Division of Biotechnics, Faculty of Bioscience EngineeringCatholic University of Leuven (K.U.Leuven)LeuvenBelgium
  3. 3.Centre for Research in Biotechnology for Agriculture (CEBAR)University of MalayaKuala LumpurMalaysia

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