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Diagnosis of Fusarium oxysporum f. sp. ciceris causing Fusarium wilt of chickpea using loop‑mediated isothermal amplification (LAMP) and conventional end‑point PCR

OAR@ICRISAT

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Relation http://oar.icrisat.org/12292/
https://www.nature.com/articles/s41598-023-29730-6#citeas
https://doi.org/10.1038/s41598-023-29730-6
 
Title Diagnosis of Fusarium oxysporum f. sp. ciceris causing Fusarium wilt of chickpea using loop‑mediated isothermal amplification (LAMP) and conventional end‑point PCR
 
Creator Achari, S R
Mann, R C
Sharma, M
Edwards, J
 
Subject Molecular Biology
Chickpea
Genetics and Genomics
 
Description Fusarium oxysporum (Fo) is ubiquitous in soil and forms a species complex of pathogenic and putatively non-pathogenic strains. Pathogenic strains cause disease in over 150 plant species. Fusarium oxysporum f. sp. ciceris (Foc) is a major fungal pathogen causing Fusarium wilt in chickpeas (Cicer arietinum). In some countries such as Australia, Foc is a high-priority pest of biosecurity concern. Specific, sensitive, robust and rapid diagnostic assays are essential for effective disease management on the farm and serve as an effective biosecurity control measure. We developed and validated a novel and highly specific PCR and a LAMP assay for detecting the Indian Foc race 1 based on a putative effector gene uniquely present in its genome. These assays were assessed against 39 Fo formae speciales and found to be specific, only amplifying the target species, in a portable real-time fluorometer (Genie III) and qPCR machine in under 13 min with an anneal derivative temperature ranging from 87.7 to 88.3 °C. The LAMP assay is sensitive to low levels of target DNA (> 0.009 ng/µl). The expected PCR product size is 143 bp. The LAMP assay developed in this study was simple, fast, sensitive and specific and could be explored for other Foc races due to the uniqueness of this marker to the Foc genome.
 
Publisher Nature Publishing Group
 
Date 2023-02-14
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
Rights cc_attribution
 
Identifier http://oar.icrisat.org/12292/1/Scientific%20Reports_13_1-10_2023.pdf
Achari, S R and Mann, R C and Sharma, M and Edwards, J (2023) Diagnosis of Fusarium oxysporum f. sp. ciceris causing Fusarium wilt of chickpea using loop‑mediated isothermal amplification (LAMP) and conventional end‑point PCR. Scientific Reports (TSI), 13. pp. 1-10. ISSN 2045-2322