A Highly Sensitive Nested-PCR Method Using a Single Closed Tube for the Detection of Colletotrichum gloeosporioides causing Greater Yam Anthracnose

Authors

  • Mithun Raj Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala
  • Jeeva ML Principal Scientist Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala
  • Vishnu S Nath Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala
  • Senthil Sankar Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala
  • Pravi Vidhyadharan Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala
  • Archana PV Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala
  • Vinayaka Hegde Div. of Crop protection, Central Tuber Crops Research Institute , Sreekariyam Trivandrum 695 017, Kerala

Keywords:

Greater yam, anthracnose, diagnosis, nested PCR

Abstract

Greater yam is an important species of yam grown in different parts of India. Anthracnose or die back  disease caused by Colletotrichum gloeosporioides reduces the yield up to 90 per cent. The pathogen  survives in the soil debris and transmits the disease to the next season through tubers. In this study,a sensitive method for the specific diagnosis of C. gloeosporioides in soil and planting material was  developed. The standard nested PCR previously described was re-standardized to run a single nested  PCR in a closed tube. The problems of cross contamination and the increased risk of PCR product  contamination, while handling the product of first PCR could be avoided. This technique could beachieved by adjusting the concentration of primers and the annealing temperature along with the  standardization of PCR cycles so as to produce a single amplicon without compromising the limit of  detection and specificity. The lowest amount of DNA that could be determined by this method was  200 pg ml-1.

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Published

2015-01-01

How to Cite

Raj, M., ML, J., Nath, V. S., Sankar, S., Vidhyadharan, P., PV, A., & Hegde, V. (2015). A Highly Sensitive Nested-PCR Method Using a Single Closed Tube for the Detection of Colletotrichum gloeosporioides causing Greater Yam Anthracnose. JOURNAL OF ROOT CROPS, 39(2), 163–167. Retrieved from https://ojs338.isrc.in/index.php/jrc/article/view/105

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