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

Tri Joko Santoso, Sri H. Hidayat, M. Herman, H. Aswidinnoor, Sudarsono Sudarsono

Abstract


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


Keywords


Begomovirus; tomato leaf curl; genetic diversity; PCR-RFLP technique.

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DOI: http://dx.doi.org/10.21082/jbio.v4n1.2008.p9-17

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P-ISSN : 1907-1094
E-ISSN : 2549-1547


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