Duplex PCR for Detection of Phytophthora katsurae Causing Chestnut Ink Disease
| Autor: | Jong Kyu Lee, Sang Yong Lee, Sang Hyun Lee, Hye Jeong Kim, Sun Keun Lee, Dong Hyeon Lee |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: | |
| Zdroj: | Research in Plant Disease, Vol 18, Iss 2, Pp 73-79 (2012) |
| ISSN: | 2233-9191 1598-2262 |
| Popis: | Phytophthora katsurae is a fungal pathogen responsible for chestnut ink disease. We designed two duplexprimer sets (SOPC 1F/1R+KatI 3F/5R, SOPC 1-1F/1-1R+KatI 3F/5R) to detect P. katsurae. SOPC 1F/1R andSOPC 1-1F/1-1R primer pairs were designed for sequence characteristic amplification regions (SCAR)marker, and KatI 3F/5R primer pair was used for P. katsurae-specific primer designed from internaltranscribed spacer (ITS) region. To assess the sensitivity of duplex PCR, genomic DNA was serially diluted10-fold to make the final concentrations from 1 mg/ml to 1 ng/ml. The sensitivity for two primer sets were 1μg/ml and 100 ng/ml, respectively. To find detection limits for zoospores of P. katsurae, each zoosporesuspension was serially diluted 10-fold to make the final concentrations from 1× 106 to 1 × 102 cells/ml, andthen DNA was extracted. The limits of detection for all of two primer sets were 1 × 105 cells/ml. All of twoprimer sets were specific to P. katsurae in PCR detection and did not produce any P. katsurae-specific PCRamplicons from other 16 Phytophthora species used as the control. This study shows that duplex PCR usingtwo primer sets might be a useful tool for rapid and efficient detection of P. katsurae. |
| Databáze: | OpenAIRE |
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