GENETIC DIVERSITY OF BEGOMOVIRUS INFECTING TOMATO

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III. GENETIC DIVERSITY OF BEGOMOVIRUS INFECTING TOMATO

1 ABSTRACT NOOR AIDAWATI. Genetic Diversity of Begomovirus Infecting Tomato. Supervised by SRI HENDRASTUTI HIDAYAT, PURNAMA HIDAYAT, RUSMILAH SUSENO, and SRIANI SUJIPRIHATI. Begomovirus is an important virus that can cause severe damage on various crops. Infection of begomoviruses, mediated through its insect vector Bemisia tabaci Genn., has been reported to cause significant losses on tomato industry in some places. Availability of detection and identification method with high accuracy and sensitivity is very critical for early warning system of the begomovirus disease. This research is conducted to develop an identification method to differentiate isolates of begomovirus based on PCR-RFLP technique. Samples of tomato leaves infected by begomovirus was collected come from D.I. Yogyakarta, Central Java and West Java. Detection and identification of begomovirus was conducted with PCR technique using universal primers of begomovirus i.e PAL 1v 1978 and PAR 1c 715. The amplified viral DNA was digested w ith restriction enzymes BamHI, Eco RI, HindIII, and PstI. Clustering of isolates were obtained using Unweight Pair Group Method Average UPGMA. Using specific primers, PAL1v 1978 and PAR 1c 715, DNA of begomovirus was successfully amplified from Central Java Magelang and Boyolali, D.I.Yogyakarta Kaliurang and Kulonprogo and West Java Barusireum. Amplified viral DNA with the size of ≈ 1.600 bp was then subjected to restriction enzyme BamHI, Eco RI, HindIII, and PstI digest. Based on the restriction pattern, begomovirus isolates from Java can be compared. Strains from Magelang were similar with those strain from Kaliurang. Both strain s were different from those of Boyolali, Kulonprogo and Barusireum, whereas the last three strains were different from each other. Using NTSYS PC 2.1 it can be concluded that the begomovirus infecting tomato in Java can be differentiated into two groups. First group consist of four different strains and second group consist of two different strains. 1 Part of the dissertation has been published in Indonesian Journal for Microbiology, vol. 10, No. 1, February 2005, with a title “ Identification of Geminivirus Infecting Tomato Based on Polymerase Chain Reaction-Restriction Fragment Length Polymorphism”.

IV. KEANEKARAGAMAN GENETIK Bemisia tabaci, SERANGGA VEKTOR BEGOMOVIRUS, BERDASARKAN