Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification

Gao, Y, Tan, M and Zhu, Y 2016, 'Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification', Australasian Plant Pathology, vol. 45, no. 4, pp. 361-367.


Document type: Journal Article
Collection: Journal Articles

Title Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification
Author(s) Gao, Y
Tan, M
Zhu, Y
Year 2016
Journal name Australasian Plant Pathology
Volume number 45
Issue number 4
Start page 361
End page 367
Total pages 7
Publisher Springer
Abstract Karnal bunt in wheat is caused by the fungus Tilletia indica. It is a quarantine disease, and therefore its timely and specific detection is important. Current detection protocols involve DNA amplification by polymerase chain reaction (PCR). However, this technique has encountered specificity issues due to the high DNA homology of T. indica with other Tilletia species, in particular T. walkeri. Here, we report the specific and rapid detection of T. indica DNA using the loop-mediated isothermal amplification (LAMP) at 62 °C. Alignment of the mitochondrial DNA of T. indica and T. walkeri has revealed four major unique regions in T. indica. Six LAMP primers designed in one of these unique regions were able to amplify T. indica DNA. The amplification could be completed in 30 min and was nearly as sensitive as conventional PCR. The amplification was found to be highly specific to T. indica DNA during the screening of 17 isolates of T. indica, T. walkeri, T. horrida, T. ehrhartae and T. caries. The use of the fluorescent chemical calcein has enabled endpoint detection with the naked eye. This method, with its specificity, rapidity and simple visualization, is well suited for the detection of this important disease in wheat.
Subject Agricultural Biotechnology not elsewhere classified
Keyword(s) Detection
Karnal bunt
Lamp
Loop-mediated isothermal amplification
Tilletia indica
Wheat
DOI - identifier 10.1007/s13313-016-0422-7
Copyright notice © 2016, Australasian Plant Pathology Society Inc.
ISSN 0815-3191
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 6 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 0 times in Scopus Article
Altmetric details:
Access Statistics: 5 Abstract Views  -  Detailed Statistics
Created: Tue, 26 Mar 2019, 09:36:00 EST by Catalyst Administrator
© 2014 RMIT Research Repository • Powered by Fez SoftwareContact us