Defence-related gene expression in transgenic lemon plants producing an antimicrobial Trichoderma harzianum endochitinase during fungal infection

Constitutive over-expression of antifungal genes from microorganisms involved in plant defence mechanisms represents a promising strategy for conferring genetic resistance against a broad range of plant pathogenic fungi. In the present work, two transgenic lemon clones with the chit42 gene from Trichoderma harzianum were tested for resistance to fungal disease and expression level of defence-related genes was evaluated. Different resistance-related processes, such as production of reactive oxygen species (ROS), systemic acquired resistance (SAR) and induced systemic resistance (ISR), were monitored in transgenic and wild type lemon clones inoculated with Botrytis cinerea, the causal agent of grey mould in citrus. Expression of genes that encode gluthatione peroxidase (GPX), a producer of ROS, chitinases, glucanases (SAR), PAL, HPL, and AOS (ISR) was measured by quantitative PCR during the first 24 h after leaf inoculation. Leaves of transgenic lemon plants inoculated with B. cinerea showed significantly less lesion development than wild type leaves. Tissues from detached leaves of different transgenic lemon clones showed a significant correlation between resistance and transgene expression. On the other hand, the over-expression of the transgenic fungal gene enhanced by two-three folds transcript levels of genes associated with enhanced ROS production and ISR establishment, while the expression of native chitinase and glucanase genes involved in SAR was down-regulated.