PJB-2022-384
Detection of Phytophthora nicotianae induced citrus gummosis by the loop-mediated isothermal amplification
Hadeed Ahmad, Nasir Ahmed Rajput, Muhammad Atiq, Ghalib Ayaz Kachelo, Muhammad Usman, Hamza Tariq and Muhammad Wahab
Abstract
Citrus fruits are indeed one of the prime fruits of Pakistan. The decline of citrus orchards due to gummosis became a potential threat to successful production of fruit during the last few years and is now continuously hammering farmer’s as well as the economy of Pakistan. In field, control of gummosis is significantly depending on quick and exact detection of Phytophthora nicotianae. The current study used a highly sensitive loop-mediated isothermal amplification (LAMP) test with hydroxynaphthol blue (HNB) as a fluorescent indicator to detect P. nicotianae. On the basis of translation elongation factor 1-alpha (EF1α) sequence of P. nicotianae, four different specific LAMP primers were designed. The LAMP assay showed positive results on different 40 isolates of P. nicotianae that were collected from different districts of Punjab, Pakistan. While none of the cross reaction was seen with other different pathogens. From 25 μL of reaction mixture, the detection frontier of P. nicotianae was 10 pg genomic DNA. It is concluded that the detection of P. nicotianae can be performed from infected tissues by the LAMP assay, which is simple, fast, sensitive and specific
To Cite this article:
Ahmad, H., N.A. Rajput, M. Atiq, G.A. Kachelo, M. Usman, H. Tariq and M. Wahab. 2024. Detection of Phytophthora nicotianae induced citrus gummosis by the loop-mediated isothermal amplification. Pak. J. Bot., 56(5): DOI: http://dx.doi.org/10.30848/PJB2024-5(5)
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