Molecular Biotechnology

, Volume 52, Issue 3, pp 277–284 | Cite as

Polymerase Chain Reaction Assay for Rapid, Sensitive Detection, and Identification of Colletotrichum gloeosporioides causing Greater Yam Anthracnose

  • Mithun Raj
  • M. L. Jeeva
  • V. Hegde
  • Pravi Vidyadharan
  • P. V. Archana
  • M. Senthil alias Sankar
  • S. Vishnu Nath
Research
First Online:

Abstract

Anthracnose caused by Colletotrichum gloeosporioides is an economically important disease which affects greater yam (Dioscorea alata L.) worldwide. Apart from airborne conidia, the pathogen propagules surviving in soil and planting material are the major sources of inoculum. A nested PCR assay has been developed for specific detection of C. gloeosporioides in soil and planting material. In conventional (single-round) PCR, the limit of detection was 20 pg, whereas in nested PCR the detection limit increased to 0.2 pg of DNA. The primers designed were found to be highly specific and could be used for accurate identification of the pathogen up to species level. The protocol was standardized for detection of the pathogen in artificially and naturally infected field samples.

Keywords

Dioscorea alata Molecular diagnosis rDNA Nested PCR Glomerella cingulata Phytopathogen Die back 
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Notes

Acknowledgments

The funding provided for research study by the National Fund for Basic, Strategic, and Frontier Application Research in Agriculture (NFBSFARA), ICAR, New Delhi, was gratefully acknowledged. The authors thanked the Director, Central Tuber Crops Research Institute, Thiruvananthapuram for providing the infrastructure facilities.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Mithun Raj
    • 1
  • M. L. Jeeva
    • 1
  • V. Hegde
    • 1
  • Pravi Vidyadharan
    • 1
  • P. V. Archana
    • 1
  • M. Senthil alias Sankar
    • 1
  • S. Vishnu Nath
    • 1
  1. 1.Division of Crop ProtectionCentral Tuber Crops Research Institute, SreekariyamThiruvananthapuramIndia

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