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
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Abstract
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.
| Item Type: | Article |
|---|---|
| Divisions: | Research Program : Asia |
| CRP: | UNSPECIFIED |
| Uncontrolled Keywords: | Chickpea, Fusarium wilt, Genetics, Molecular biology |
| Subjects: | Others > Molecular Biology Mandate crops > Chickpea Others > Genetics and Genomics |
| Depositing User: | Mr Nagaraju T |
| Date Deposited: | 14 Nov 2023 05:32 |
| Last Modified: | 14 Nov 2023 05:32 |
| URI: | http://oar.icrisat.org/id/eprint/12292 |
| Official URL: | https://www.nature.com/articles/s41598-023-29730-6... |
| Projects: | Boosting Diagnostics project |
| Funders: | Australian Government Department of Agriculture, Fisheries and Forestry |
| Acknowledgement: | We are grateful to so many people who played a significant role in ensuring the success of this project. Firstly, we would like to thank Professor Brendan Rodoni for seeking funding for this project and Dr. Fiona Contable for being the Agriculture Victoria project leader. Secondly, a heartfelt appreciation to Drs Linda Zheng, Yuhong Liu, Arati Agarwal and Peter Mee on sharing their knowledge and experiences on designing the LAMP assays. Finally, without the isolates, the project could not have been carried out this comprehensively, hence a heartful appreciation to Drs Yu Pei Tan, Jordan Bailey, Edward Liew, Paul Taylor, Sophia Callaghan and Duy Le for providing Fusarium oxysporum cultures from their collections. |
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