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Using Real time PCR Technique for the Detection of Xanthomonas citri subsp. citri, Causal Agent of Citrus Canker for Certification of Canker Free Orchards |
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| รหัสดีโอไอ | |
| Creator | 1. Nuttima Kositcharoenkul 2. Tippawan Kanhayart 3. Buranee Puawongphat 4. Rungnapha Thong kreng 5. Boonpiyatida Klongklew |
| Title | Using Real time PCR Technique for the Detection of Xanthomonas citri subsp. citri, Causal Agent of Citrus Canker for Certification of Canker Free Orchards |
| Publisher | Department of Agriculture |
| Publication Year | 2560 |
| Journal Title | Thai Agricultural Research Journal |
| Journal Vol. | 35 |
| Journal No. | 1 |
| Page no. | 74-90 |
| Keyword | โรคแคงเคอร์, Xanthomonas axonopodic PV. citri, ส้มโอปลอดโรคแคงเคอร์, การตรวจรับรอง, Canker disease, Xanthomonas axonopodis pv. citri, detection, Real time PCR |
| URL Website | http://at.doa.go.th/journal |
| Website title | Thai Agricultural Research Journal |
| ISSN | 0125-8389 |
| Abstract | To export pomelo (Citrus maxima Merr.) to the European Union (EU), exporteris obliged to follow the International Standards for Phytosanitary Measures No.6 andNo.10 (ISPM No. 6 and ISPM No.10) Beside, Guidelines for Surveillance Requirementsfor the Establishment of Pest Free Places of Production and Pest Free ProductionSites must be followed. Canker disease of pomelo caused by Xanthomonas citrisubsp. citri is a quarantine pest of the EU. Therefore, Pomelo orchards certification forcanker free is necessary and the effective detection method for X. citri subsp. citrineeds to be developed. This work was to utilize the Real time PCR, the rapid andhigh accuracy technique, to detect X. citri subsp. citri for the canker-free certificationof pomelo orchards. The primers 2/3 and D1/D2 were designed from avirulence/pathogenicity gene (pthA gene) of X. citri subsp. citri which had high specificity andcould detect as minimum as 5 picrograms of DNA or 81 CFU/ml of the bacterial cells.The method showed high accuracy of detection of X. citri subsp. citri in bothsymptom and symptomless samples. In 2014, this Real time PCR method was usedfor the establishment of canker-free production sites in pomelo orchards at WiangKaen Districts, Chiang Rai Province. From 130 orchards, 84 canker-free productionorchards were successfully established.The Real time PCR method developedin this study was eventually accepted bythe EU for the pomelo production inThailand for export to the EU andrepresenting as a model for the cankerfreecertification of others crops forcountries where X. citri subsp. citri is aquarantine pests |
