105(4)_str45

 

ISSN 1392-3196 / e-ISSN 2335-8947
Zemdirbyste-Agriculture, vol. 105, No. 4 (2018), p. 357–362
DOI  10.13080/z-a.2018.105.045

Virus-induced gene silencing for functional analysis of eds1 gene in tomato infected with Ralstonia solanacearum

Hamid Reza BOLOK YAZDI, Seyed Kazem SABBAGH, Mahta MAZAHERI, Mohammad SALARI, Seyed Mohammad MOSHTAGHIOUN

Abstract

The efficiency of virus-induced gene silencing (VIGS) can be identified by monitoring the level of silencing of the target genes in plants. In this study, enhanced disease susceptibility 1 (eds1) gene was silenced in tomato (Lycopersicon esculentum L.) plants by VIGS, using Tobacco rattle virus (TRV) expression vector. Tomato plants at different growth stages (fully-opened cotyledon and eight-leaf) were co-infected with Agrobacterium tumefaciens C58C1 strain containing eds1 complementary deoxyribonucleic acid (cDNA) at different (20°C and 28°C) temperatures and bacterial concentrations with 1.0 × 106 and 1.5 × 108 colony forming unit (CFU) mL-1. The reverse transcriptase-polymerase chain reaction (RT-PCR) method was used to analyse the suppression level of the eds1 gene and the VIGS efficiency assay. Data analysis showed a high eds1 gene silencing at fully-opened cotyledon stage at 20°C using a A. tumefaciens at a concentration of 1.0 × 106 CFU rather than other conditions. In addition, the percentage of wilted plants was significantly varied in different conditions and the most wilted plants were observed at the 1.0 × 106 CFU and 20°C.

Based on our results, we suggest that eds1 gene silencing can be used as a possible screening marker to select bacterial wilt-resistant species.

Key words: Agrobacterium tumefaciens, agro-infiltration, eds1, Ralstonia solanacearum, virus-induced gene silencing.

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