Identification of Pseudoperonospora cubensis using real-time PCR and high resolution melting (HRM) analysis

Jeong Hwan Lee; Myong Ho Park; Sanghyeob Lee

doi:10.1007/s10327-016-0646-y

Identification of Pseudoperonospora cubensis using real-time PCR and high resolution melting (HRM) analysis

Jeong Hwan Lee
, Myong Ho Park
, Sanghyeob Lee^*

^*Corresponding author for this work

Department of Biological Sciences

Sejong University

Research output: Contribution to journal › Journal article › peer-review

9 Scopus citations

Abstract

To develop a molecular method to identify Pseudoperonospora cubensis in cucumber leaves with signs of downy mildew, we compared the nucleotide sequences of ribosomal DNA-internal transcribed spacer, cytochrome oxidase II, and β-tubulin genes of P. cubensis and P. humuli isolates. Seven single nucleotide polymorphisms (SNPs) distinguished P. cubensis and P. humuli based on variations in β-tubulin sequences, and one specific primer set was designed for further analysis. Real-time PCR and high resolution melting analysis showed that the primer set can be used to specifically identify P. cubensis in cucumber leaves with downy mildew.

Original language	English
Pages (from-to)	110-115
Number of pages	6
Journal	Journal of General Plant Pathology
Volume	82
Issue number	2
DOIs	https://doi.org/10.1007/s10327-016-0646-y
State	Published - 2016.03.1

Keywords

Cucurbit downy mildew
High resolution melting analysis
Pseudoperonospora cubensis
Pseudoperonospora humuli
Real-time PCR
β-tubulin gene

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10.1007/s10327-016-0646-y

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title = "Identification of Pseudoperonospora cubensis using real-time PCR and high resolution melting (HRM) analysis",

abstract = "To develop a molecular method to identify Pseudoperonospora cubensis in cucumber leaves with signs of downy mildew, we compared the nucleotide sequences of ribosomal DNA-internal transcribed spacer, cytochrome oxidase II, and β-tubulin genes of P. cubensis and P. humuli isolates. Seven single nucleotide polymorphisms (SNPs) distinguished P. cubensis and P. humuli based on variations in β-tubulin sequences, and one specific primer set was designed for further analysis. Real-time PCR and high resolution melting analysis showed that the primer set can be used to specifically identify P. cubensis in cucumber leaves with downy mildew.",

keywords = "Cucurbit downy mildew, High resolution melting analysis, Pseudoperonospora cubensis, Pseudoperonospora humuli, Real-time PCR, β-tubulin gene",

author = "Lee, \{Jeong Hwan\} and Park, \{Myong Ho\} and Sanghyeob Lee",

note = "Publisher Copyright: {\textcopyright} 2016, The Phytopathological Society of Japan and Springer Japan.",

year = "2016",

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doi = "10.1007/s10327-016-0646-y",

language = "English",

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journal = "Journal of General Plant Pathology",

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T1 - Identification of Pseudoperonospora cubensis using real-time PCR and high resolution melting (HRM) analysis

AU - Lee, Jeong Hwan

AU - Park, Myong Ho

AU - Lee, Sanghyeob

PY - 2016/3/1

Y1 - 2016/3/1

N2 - To develop a molecular method to identify Pseudoperonospora cubensis in cucumber leaves with signs of downy mildew, we compared the nucleotide sequences of ribosomal DNA-internal transcribed spacer, cytochrome oxidase II, and β-tubulin genes of P. cubensis and P. humuli isolates. Seven single nucleotide polymorphisms (SNPs) distinguished P. cubensis and P. humuli based on variations in β-tubulin sequences, and one specific primer set was designed for further analysis. Real-time PCR and high resolution melting analysis showed that the primer set can be used to specifically identify P. cubensis in cucumber leaves with downy mildew.

AB - To develop a molecular method to identify Pseudoperonospora cubensis in cucumber leaves with signs of downy mildew, we compared the nucleotide sequences of ribosomal DNA-internal transcribed spacer, cytochrome oxidase II, and β-tubulin genes of P. cubensis and P. humuli isolates. Seven single nucleotide polymorphisms (SNPs) distinguished P. cubensis and P. humuli based on variations in β-tubulin sequences, and one specific primer set was designed for further analysis. Real-time PCR and high resolution melting analysis showed that the primer set can be used to specifically identify P. cubensis in cucumber leaves with downy mildew.

KW - Cucurbit downy mildew

KW - High resolution melting analysis

KW - Pseudoperonospora cubensis

KW - Pseudoperonospora humuli

KW - Real-time PCR

KW - β-tubulin gene

UR - https://www.scopus.com/pages/publications/84959140027

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DO - 10.1007/s10327-016-0646-y

M3 - Journal article

AN - SCOPUS:84959140027

SN - 1345-2630

VL - 82

SP - 110

EP - 115

JO - Journal of General Plant Pathology

JF - Journal of General Plant Pathology

IS - 2

ER -

Identification of Pseudoperonospora cubensis using real-time PCR and high resolution melting (HRM) analysis

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Keywords

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