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Establishment and evaluation of a TEF1-α based loop-mediated isothermal amplification assay for detection of Phomopsis longicolla

Establishment and evaluation of a TEF1-α based loop-mediated isothermal amplification assay for... Phomopsis longicolla is an important seed-borne pathogen affecting soybean. In this research, transcription elongation factor 1-α (TEF1-α) was identified as a suitable target for detecting P. longicolla using Loop-mediated isothermal amplification (LAMP) method. The specificity of the method was tested against 54 P. longicolla isolates, 13 oomycetes isolates and 28 other fungal isolates. With the addition of hydroxynaphthol blue (HNB) prior to amplification, a sky-blue color was only observed in the presence of P. longicolla, whereas other isolates showed no color change. The method was sensitive enough to detect as little as 100 pg/uL fungal DNA. In addition, the assay also detected P. longicolla from diseased soybean tissues and residues from different origins. The results demonstrated that the LAMP assay provides a rapid and sensitive tool for detecting P. longicolla from plant tissues. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Australasian Plant Pathology Springer Journals

Establishment and evaluation of a TEF1-α based loop-mediated isothermal amplification assay for detection of Phomopsis longicolla

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References (11)

Publisher
Springer Journals
Copyright
Copyright © 2016 by Australasian Plant Pathology Society Inc.
Subject
Life Sciences; Plant Pathology; Plant Sciences; Agriculture; Entomology; Ecology
ISSN
0815-3191
eISSN
1448-6032
DOI
10.1007/s13313-016-0415-6
Publisher site
See Article on Publisher Site

Abstract

Phomopsis longicolla is an important seed-borne pathogen affecting soybean. In this research, transcription elongation factor 1-α (TEF1-α) was identified as a suitable target for detecting P. longicolla using Loop-mediated isothermal amplification (LAMP) method. The specificity of the method was tested against 54 P. longicolla isolates, 13 oomycetes isolates and 28 other fungal isolates. With the addition of hydroxynaphthol blue (HNB) prior to amplification, a sky-blue color was only observed in the presence of P. longicolla, whereas other isolates showed no color change. The method was sensitive enough to detect as little as 100 pg/uL fungal DNA. In addition, the assay also detected P. longicolla from diseased soybean tissues and residues from different origins. The results demonstrated that the LAMP assay provides a rapid and sensitive tool for detecting P. longicolla from plant tissues.

Journal

Australasian Plant PathologySpringer Journals

Published: May 6, 2016

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