Detection of Begomoviruses Causing Pepper Yellow Leaf Curl Disease in Thailand Using Broad-Spectrum Primers
DOI:
https://doi.org/10.14456/thaidoa-agres.2019.21Keywords:
begomovirus, universal primer, Unibego-Pep, yellow leaf curl diseaseAbstract
Begomoviruses are causal agents for yellow leaf curl diseases in peppers which are recognized as a serious threat for quality and quantity of peppers in Thailand. The disease distribution was found in every regions throughout the country. Polymerase chain reaction, a specific and efficient detection method are applied to identify the presence of begomoviruses in pepper samples. However, these viruses evolve and adapt rapidly leading to incorrect detection by the previous universal primer sets. In this research, the universal primers were designed to amplify the target sequence of the begomovirus infecting peppers in Thailand, from the consensus sequences in the regions of AV1 and AV2 genes of 8 pepper-infecting begomoviruses which were Potato yellow mosaic virus (PYMV), Tomato severe leaf curl virus (ToSLCV), Chili leaf curl virus (ChiLCV), Eggplant golden mosaic virus (EGMV), Pepper yellow virus (PepYLCV), Pepper leaf curl virus (PepLCV), Tomato leaf curl virus (ToLCV) และ Tomato yellow leaf curl virus (TYLCV). The efficiency of the designed primers were analyzed by testing 44 yellow leaf curl pepper samples collected from 20 provinces in 4 regions of Thailand. The designed primer pair designated Unibego-Pep was able to amplify 360 bp target fragment from the viral DNA fragments within approximately 1 hr and 15 min. Sequencing analysis of the PCR products from all samples showed high similarity (93-99%) to 5 pepper-infecting begomoviruses, e.g. EGMV, Pepper yellow leaf curl Thailand virus (PepYLCThV), Pepper yellow leaf curl Indonesia virus (PepYLCIV), Tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), and Pepper leaf curl virus -[Malaysia] (PepLCV-[MY]).
Keywords: pepper, begomovirus, universal primer, Unibego-Pep, yellow leaf curl disease
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