Konstruksi Kandidat Gen AV1 Begomovirus pada pBI121 dan Introduksinya ke dalam Tembakau Menggunakan Vektor Agrobacterium tumefaciens

Tri Joko Santoso, Muhammad Herman, Sri H Hidayat, Hajrial Aswidinnoor, Sudarsono Sudarsono

Abstract


Construction of Begomovirus AV1 Gene Candidate into
pBI121 and Its Introduction into Tobacco by using
Agrobacterium tumefaciens Vector. Tri J. Santoso,
Muhammad Herman, Sri H. Hidayat, Hajrial
Aswidinnoor, and Sudarsono. Infection of Begomovirus
has caused leaf curl disease in tomato. This infection has
significantly impact on yield losses of tomato production.
Recently, in Indonesia there was no effectively way to
control this disease. The use of resistant tomato variety is
one of strategies to control this virus. Genetic engineering
technology gives an opportunity to develop the transgenic
tomato resistant to Begomovirus through pathogen derived
resistance (PDR) approach. The objectives of this study
were to construct the Begomovirus AV1 candidate gene in
the pBI121 and to introduce the construct into tobacco plant
genome through Agrobacterium tumefaciens vector. A series
activites in gene construct have been conducted include
PCR amplification of AV1 gene using a pair of specific
primer, cloning the gene into pGEM-T easy, transformation of
the clone into Escherichia coli DH5α competent cell,
construct the gene into pBI121, and transform the construct
into A. tumefaciens. Leaf segments of in vitro tobacco plant
were transformed by co-cultivation with A. tumefaciens
containing ToLCV-AV1 construct. In the research activitiy,
Indonesian Begomovirus AV1 gene was successfully
amplified and inserted in expression vector plasmid pBI121.
Tobacco transformants carrying kanamycin-resistant gene
(nptII gene) were regenerated and established in the
glasshouse. Those transformant plants are expected
containing the AV1 gene.


Keywords


AV1 gene; Begomovirus; Nicotiana tabaccum; Agrobacterium tumefaciens; coat protein

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DOI: http://dx.doi.org/10.21082/jbio.v7n1.2011.p9-18

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