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Indian Phytopathology

, Volume 71, Issue 3, pp 317–323 | Cite as

Early detection of Phytophthora infestans in potato using carbohydrate binding module 1 protein (CBD1) based antiserum

  • S. SundareshaEmail author
  • Sanjeev Sharma
  • A. Jeevalatha
  • Sadhana Sharma
  • Vandana Thakur
  • Shubhangi Sharma
  • Bir Pal Singh
  • S. K. Chakrabarti
Research Article
  • 11 Downloads

Abstract

Phytophthora infestans is a global challenging pathogen among the biotic stresses which affect healthy and sustainable potato production. Desirable and successful management strategies depend on understanding of its biology, disease cycle and more importantly rapid and accurate detection of the pathogen during early stage of infection. Here, we report production of recombinant antibodies against whole cellulose binding domain (CBD-1) protein of P. infestans and a synthetic signal peptide of CBD1 and their effectiveness in detection of P. infestans both in infected potato leaf and tuber. The recombinant whole CBD1 protein and synthetic signal peptide of CBD1 protein was used to immunize the rabbits and blood serum was collected, purified and measured by SDS–PAGE. Among the two antibodies, synthetic peptide based antibody showed high specificity for mycelia, sporangia and zoospores. It also showed strong signal intensity for P. infestans infected leaf and tuber tissues. The data indicated that the produced synthetic peptide antiserum was efficient and accurate in discrimination of negative and positive samples in ELISA. Reaction of antiserum with early stage of infected leaf and tuber tissue confirmed the expression of the CBD protein, which in turn confirmed the presence of the pathogen in the plant tissues. Therefore it will enable to detect the pathogen in the potato seed before planting as well as in the standing crop in the field thus, would be helpful to ensure production of healthy seed potatoes and for effective schedule for management practices.

Keywords

Phytophthora infestans Diagnosis Seed potato CBD1 

Abbreviations

CBD1

Carbohydrate binding module1

ELISA

Enzyme linked immune sorbent assay

DAS

Double antibody sandwich

RT-PCR

Reverse transcriptase polymerase chain reaction

Notes

Acknowledgements

The authors acknowledge the financial grant received from Phytofura project ICAR, New Delhi, India.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Indian Phytopathological Society 2018

Authors and Affiliations

  1. 1.ICAR-Central Potato Research InstituteShimlaIndia
  2. 2.ICAR-Indian Institute of Spices ResearchCalicutIndia

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