Identitas dan Keragaman Genetik Begomovirus yang Berasosiasi dengan Penyakit Keriting pada Tomat Berdasarkan Teknik Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP)

Abstract

<p>Begomoviruses, members of the Geminivirus,<br />are considered as emerging plant viruses. This was due to<br />the increasing incidences and severities of the diseases in a<br />number of economically important crops, including tomato.<br />Genetic diversities of the Begomovirus isolates infecting<br />tomato (Lycopersicon esculentum) of several areas in Indonesia<br />were analyzed by using Polymerase Chain Reaction-<br />Restriction Fragment Length Polymorphism (PCR-RFLP)<br />technique. A 1500 base pairs of PCR fragments amplified by<br />using degenerate primers for Begomovirus was digested<br />using four restriction enzymes, i.e., DraI, EcoRI, RsaI, and<br />PstI. The pattern of RE digested fragments of 8 Begomovirus<br />isolates and the predicted RFLP fragments of the Begomovirus<br />isolates in the GeneBank database were used to determine<br />the genetic identities and diversities among the isolates.<br />Positive results of the PCR amplifications proved that<br />diseased tomato plant samples collected from 8 locations in<br />Java and Sumatra were infected with at least one Begomovirus<br />isolate. The PCR amplification products, which were<br />digested using the four restriction enzymes indicated the<br />presence of polimorfisms among the DNA fragments of the<br />Begomovirus isolates. Identifications of the Begomovirus<br />indicated that the Brastagi, Bogor, Sragen, Ketep, and Boyolali<br />isolates were Tomato Leaf Curl Virus (ToLCV); the<br />isolates from Malang and Blitar isolates were Ageratum<br />Yellow Vein Virus (AYVV), while one isolate from Kaliurang<br />was Tomato Yellow Leaf Curl Virus (TYLCV). Results of the<br />phylogenetic analysis of the 8 Begomovirus isolates based<br />on Begomoviruses from the DNA database indicated that<br />they belonged to three different groups.</p>