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

Abstract

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

Molecular Analysis and Effectiveness Assay of AV1 Gene in Transgenic Tobacco for Resistance to Begomovirus

“…In previous experiments, genetic transformation of tobacco plants with Begomovirus-AV1 gene through Agrobacterium tumefaciens vector has produced about 1399 transformant plants (Santoso et al, 2011). Transgenic tobacco plants has brought kanamyssin antibiotic resistance gene (nptII).…”