Identification of virus causing the yellow leaf curl diseases on chili pepper in Lombok Island by PCR-RFLP technique

Abstract

Yellow leaf curl virus causes pepper yellow leaf curl diseases that caused by Geminiviruses (Begomovirus) became epidemic in Indonesia on 2000. This disease is widespread at several chili pepper production centers in Java, Sumatera, Bali, Kalimantan, and Sulawesi. There is no report about existence of it in Lombok island, West Nusa Tenggara Province, one of chili pepper production centre in Indonesia. The aim of this research is to identify Begomovirus which infect chili pepper with molecular approach and to find out the variabilities genetic of Begomovirus from Lombok island, Central Jawa and DIY with DNA fingerprint pattern based on Polymerase Chain Reaction (PCR) and Reaction Fragment Length Polymorphisme (RFLP). This research conducted with this following step: field observation of yellow leaf curl symptoms, DNA extraction, amplification the coat protein with spesific primer PYLCV, and RFLP analyses. Field result that conducted in three regency of Lombok island (East Lombok, West Lombok and Central Lombok) shown that infection symptoms in chili pepper such as various yellow-green spot in leaves, leaf curl, become small and wrapped on top, and in plant that have severe infection the plant become stunted. Results of molecular identification using PCR in sample plant aredetected the Begomovirus existence with amplification products sized 840 bp. The analysis results of PCR-RFLP using restriction enzymes HaeIII have shown the genetic variability between isolate from Lombok island, Centre of Java, and DIY. The result of this research has proved the ability of PYLCV primer in PCR to detect Begomovirus obtained from different places. This research proved that the universal primer ability in PCR for Begomovirus detection from different area and as the first report there are infections of Pepper Yellow Leaf Curl Virus in Lombok island NTB.Yellow leaf curl virus causes pepper yellow leaf curl diseases that caused by Geminiviruses (Begomovirus) became epidemic in Indonesia on 2000. This disease is widespread at several chili pepper production centers in Java, Sumatera, Bali, Kalimantan, and Sulawesi. There is no report about existence of it in Lombok island, West Nusa Tenggara Province, one of chili pepper production centre in Indonesia. The aim of this research is to identify Begomovirus which infect chili pepper with molecular approach and to find out the variabilities genetic of Begomovirus from Lombok island, Central Jawa and DIY with DNA fingerprint pattern based on Polymerase Chain Reaction (PCR) and Reaction Fragment Length Polymorphisme (RFLP). This research conducted with this following step: field observation of yellow leaf curl symptoms, DNA extraction, amplification the coat protein with spesific primer PYLCV, and RFLP analyses. Field result that conducted in three regency of Lombok island (East Lombok, West Lombok and Central Lo...