Optimum temperature for detection of TrAP and Rep genes using SPG1/SPG2 primer design on red chili (Capsicum annuum L.)

Red chili (Capsicum annuum L.) is a vegetable plant much cultivated in Indonesia because it has a high economic value as a seasonings. In the cultivation of this plant, pathogenic infection still the main limited. One of the pathogens that could cause a disease in red chili is Begomovirus. This virus is the largest genus of the Geminiviridae family and requires whitefly vector insects (Bemisia tabaci Genn.) in its spread. Scientific information regarding disease detection is very limited in Indonesia and has never been done in Lampung. This research was conducted using the Polymerase Chain Reaction (PCR) method to amplify the Transcriptional Activator Protein (TrAP) and Replication Associated Protein (Rep) genes using SPG1/SPG2 primers. SPG1 acts as a forward primer that will amplify the TrAP and SPG2 acts as a reverse primer that will amplify the Rep of Begomovirus. This research aims to determine the temperature optimization of Polymerase Chain Reaction in the detection of TrAP and Rep genes in red chili in Lampung. The results showed that the temperature to amplify during a 30 cycle is predenaturation 95°C for 1 minute, denaturation 95°C for 15 seconds, annealing 52°C for 15 seconds, and extention 72°C for 10 seconds. The amplification results produced a specific band measuring ±912 bp which indicated the TrAP and Rep Begomovirus genes. The results of this research can be used as an early stage of virus identification to study the pattern of spread and control of this disease.