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Pest Manag Sci. 2016 Nov;72(11):2128-2135. doi: 10.1002/ps.4243. Epub 2016 Mar 3.

Development of a rapid and high-throughput molecular method for detecting the F200Y mutant genotype in benzimidazole-resistant isolates of Fusarium asiaticum.

Author information

1
College of Plant Protection, State and Local Joint Engineering Research Centre of Green Pesticide Invention and Application, Nanjing Agricultural University, Nanjing, China.
2
College of Plant Protection, State and Local Joint Engineering Research Centre of Green Pesticide Invention and Application, Nanjing Agricultural University, Nanjing, China. mgzhou@njau.edu.cn.

Abstract

BACKGROUND:

The point mutation at codon 200 (TTC→TAC, F200Y) of the β2 -tubulin gene confers resistance to benzimidazole fungicide in Fusarium asiaticum. These isolates with this mutation have been detected mainly by determining the minimum inhibitory concentration (MIC) of fungicides, which is always time consuming, tedious and inefficient.

RESULTS:

A visual, rapid and efficient method with high specificity was developed, based on loop-mediated isothermal amplification (LAMP). Six sets of LAMP primers were designed, and one set was optimised specifically to distinguish the F200Y mutant genotype. With the optimal LAMP primers, concentrations of LAMP components were optimised. The optimal reaction conditions were 57-64 °C for 75 min. The feasibility of the LAMP assay for detection of the F200Y mutant genotype of F. asiaticum was demonstrated by assaying diseased wheat spikelets that were artificially inoculated in the field.

CONCLUSION:

The new LAMP assay had good specificity, sensitivity, stability and repeatability. It will be useful for assessing the risk of F. asiaticum populations with carbendazim resistance developing in the field, and will also provide important reference data for integrated control of Fusarium head blight caused by F. asiaticum. © 2016 Society of Chemical Industry.

KEYWORDS:

Fusarium asiaticum; carbendazim; detection; loop-mediated isothermal amplification; resistance

PMID:
26823005
DOI:
10.1002/ps.4243
[Indexed for MEDLINE]

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