Early detection of Phytophthora infestans in potato using carbohydrate binding module 1 protein (CBD1) based antiserum
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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.
KeywordsPhytophthora infestans Diagnosis Seed potato CBD1
Carbohydrate binding module1
Enzyme linked immune sorbent assay
Double antibody sandwich
Reverse transcriptase polymerase chain reaction
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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