<i>Agrobacterium</i>-mediated transformation of tomato (<i>Lycopersicon esculentum</i>L. cv. Hezuo 908) with improved efficiency

Abstract

We developed an efficient and rapid Agrobacterium-mediated tomato transformation protocol by using cotyledon and hypocotyl as explants. The transgenic nature of the regenerants was confirmed through β-glucuronidase activity staining, polymerase chain reaction (PCR), quantitative real-time PCR (qPCR) analysis and northern blot analysis. In the used protocol, the optimized experimental conditions of the tomato genetic transformation were Agrobacterium liquid concentration, equivalent to optical density (OD600) of 1.0, 20 min (cotyledon) or 30 min (hypocotyl) infection time and 2 d of co-culturing period. The optimized medium for shoot induction was Murashige & Skoog (MS) medium + 0.5 mg/L indole-3-acetic acid (IAA) + 0.5 mg/L 6-benzylaminopurine (BAP) for cotyledon and MS + 0.5 mg/L IAA + 2.0 mg/L BAP for hypocotyl. The root induction rate was the highest on the MS medium, supplemented with 0.1 mg/L IAA. The transformation frequency reached 40% per explant. The received results are a base for a larger scale development of transformation procedure.