Multiplex PCR assay for simultaneous detection of Phytophthora, Pythium and Fusarium associated with foot rot and yellowing diseases of black pepper

Multiple PCR for the detection of pathogens infecting black pepper

Authors

  • A Jeevalatha ICAR-Indian Institute of Spices Research, Kozhikode-673 012, Kerala, India
  • Fathimath Zumaila ICAR-Indian Institute of Spices Research, Kozhikode-673 012, Kerala, India
  • C N Biju ICAR-Indian Institute of Spices Research, Kozhikode-673 012, Kerala, India
  • Mohammed Faisal Peeran ICAR- Indian Institute of Spices Research Regional Station, Appangala, Madikeri-571201, Karnataka, India

DOI:

https://doi.org/10.25081/josac.2022.v31.i1.7739

Keywords:

black pepper, detection, Foot rot, multiplex PCR, yellowing

Abstract

In this study, a multiplex PCR assay was developed to detect Phytophthora, Pythium and Fusarium infecting black pepper. Genus-specific primers were designed from the conserved region of ITS and a multiplex PCR was optimized by manipulating the annealing temperature and primer as well as MgCl2 concentrations. The black pepper 18S rRNA gene-specific primers were also included in the multiplex PCR assay as internal control. The assay successfully detected the pathogens from artificially inoculated black pepper roots and did not show any cross amplification with other fungal pathogens of black pepper such as Rhizoctonia solani, Sclerotium rolfsii and Colletotrichum sp. Hence, the developed multiplex PCR assay will help in early diagnosis of the cause of black pepper yellowing leading to timely adoption of management strategies.

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References

Aravind R, Kumar A, Dinu A & Eapen S J 2011 Single tube duplex PCR for simultaneous detection of Phytophthora capsici and Radopholus similis infecting black pepper (Piper nigrum). Indian Phytopath. 64(4): 353–357.

Costa S S, Moreira G M & Pfenning L H 2017 Development of a PCR protocol for the identification and detection of Fusarium solani f. sp. piperis from soil and roots of black pepper (Piper nigrum). Trop. plant pathol. 42(1): 55–59.

Jeevalatha A, Biju C N & Bhai R S 2021 Ypt1 gene-based recombinase polymerase amplification assay for Phytophthora capsici and P. tropicalis detection in black pepper. Eur. J. Plant Pathol. 159(4): 863–875.

Pandian R T P, Bhat A I, Biju C N & Sasi S 2018 Development of diagnostic assays for rapid and sensitive detection of Phytophthora infecting major spices and plantation crops. J. Spices Arom Crops. 27(2): 119–130.

Sheji C, Renu S G, Balaji S & Anandaraj M 2009 Ribosomal DNA analysis of three Phytophthora species occurring in India. Indian Phytopathol. 62(2): 155–162.

Silvar C, Duncan J M, Cooke D E L, Williams N A, Díaz J & Merino F 2005 Development of specific PCR primers for identification and detection of Phytophthora capsici Leon. Eur. J. Plant Pathol. 112: 43–52.

Subila K P & Bhai R S 2020 Pythium deliense, a pathogen causing yellowing and wilt of black pepper in India. New Dis. Rep. 42: 6.

Published

19-07-2022

How to Cite

Jeevalatha, A., Zumaila, F., Biju, C. N., & Peeran, M. F. (2022). Multiplex PCR assay for simultaneous detection of Phytophthora, Pythium and Fusarium associated with foot rot and yellowing diseases of black pepper: Multiple PCR for the detection of pathogens infecting black pepper. Journal of Spices and Aromatic Crops, 31(1), 92–96. https://doi.org/10.25081/josac.2022.v31.i1.7739