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Detection of Begomoviruses Causing Pepper Yellow Leaf Curl Disease in Thailand Using Broad-Spectrum Primers |
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| รหัสดีโอไอ | |
| Creator | Srihunsa Malichan |
| Title | Detection of Begomoviruses Causing Pepper Yellow Leaf Curl Disease in Thailand Using Broad-Spectrum Primers |
| Contributor | Parin Taweechotworakul, Ratchanee Hongprayoon |
| Publisher | Department of Agriculture |
| Publication Year | 2562 |
| Journal Title | Thai Agricultural Research Journal |
| Journal Vol. | 37 |
| Journal No. | 3 |
| Page no. | 250-264 |
| Keyword | begomovirus, pepper yellow leaf curl disease, detection, PCR |
| URL Website | http://at.doa.go.th/journal |
| Website title | Thai Agricultural Research Journal |
| ISSN | 0125-8389 |
| Abstract | Begomoviruses are causal agents of yellow leaf curl disease in peppers causing serious threat for pepper's quality and quantity. Their distribution was reported throughout the country. The viruses are highly variable in nature and have evolved and adaptedrapidly due to several reported factors, and may lead to incorrect detection by thepreviously designed universal primers in polymerase chain reaction technique. A morespecific and reliable detection method will provide an effective tool for disease surveillance,control and breeding for resistance. In this research, the new broad-spectrum Unibego-Pepprimers were designed to amplify the target consensus sequence in the AV1 gene regionsof 8 pepper-infecting begomoviruses, i.e., Chili leaf curl virus (ChiLCV), Eggplant goldenmosaic virus (EGMV), Pepper leaf curl virus (PepLCV), Pepper yellow leaf curl Thailand virus(PepYLCTHV), Pepper yellow leaf curl Indonesia virus (PepYLCIV), Tobacco leaf curl virus(TbLCTHV), Tomato yellow leaf curl Kanchanaburi virus (TYLCKaV) and Tomato yellow leafcurl Thailand virus (TYLCTHV). The efficiency of the primers was investigated by testing4 reference samples, all of which gave positive results giving an approximately 380 bpfragment. The testing was also performed with 41 pepper samples showing diseasesymptoms collected from 20 provinces and the target band could be amplified fromthe samples in all regions. Analysis of the amplified products by sequence alignment withthe GenBank database showed five pepper-infecting begomoviruses were at 93-99%identity, i.e., EGMV (93.62-98.49%), PepLCV-[MY] (93.43-96.69%), PepYLCTHV (97.83-99.69%),PepYLCIV (98.25-99.40%) and TYLCKaV (97.38-99.12%). The positive samples werecategorized based on a phylogenetic tree constructed and representatives from each groupfurther characterized by full-length DNA-A rolling circle amplification and sequencing toconfirm the 380 bp results. BLASTn analysis showed good correlation of 7 out of 8 samplesexamined. This newly designed primers has very strong potential to be an effective tool forthe detection of pepper-infecting begomoviruses in Thailand. |
