Heterologous Expression of the Bean Chitinase Gene (ch5B) in Lettuce (Lactuca sativa L.) Confers Enhanced Tolerance to Rhizoctonia solani and Demonstrates Agronomic Substantial Equivalence in Field Trials

Abstract

Fungal pathogens cause severe yield losses in lettuce production worldwide. We developed transgenic lettuce lines overexpressing the ch5B chitinase gene from Phaseolus vulgaris (bean) under the regulation of the rbcS1 (ribulose-1,5-bisphosphate carboxylase small subunit) promoter. Molecular characterization confirmed stable transgene integration and expression across the selected lines. In vitro assays demonstrated that transgenic leaf extracts significantly inhibited Rhizoctonia solani mycelial growth. Furthermore, inoculation assays in both seedlings and adult plants in the greenhouse showed a significant reduction in lesion areas compared to non-transgenic (NT) controls. Finally, two lines were evaluated in field trials, both showing agronomic substantial equivalence to the NT, satisfying international biosafety frameworks for commercial development. Among them, the ch5B 3-4 line appears to be the most promising candidate for further studies, as it showed slightly higher fresh weight compared to the NT under field conditions.