Auxin controls multiple aspects of plant growth and development. However, its role in stress responses remains poorly understood. Auxin acts on the transcriptional regulation of target genes, mainly through Auxin Response Factors (ARF). This study focuses on the involvement of SlARF4 in tomato tolerance to salinity and osmotic stress. Using a reverse genetic approach, we found that the antisense down-regulation of SlARF4 promotes root development and density, increases soluble sugars content and maintains chlorophyll content at high levels under stress conditions. Furthermore, ARF4-as displayed higher tolerance to salt and osmotic stress through reduced stomatal conductance coupled with increased leaf relative water content and Abscisic acid (ABA) content under normal and stressful conditions. This increase in ABA content was correlated with the activation of ABA biosynthesis genes and the repression of ABA catabolism genes. Cu/ZnSOD and mdhar genes were up-regulated in ARF4-as plants which can result in a better tolerance to salt and osmotic stress. A CRISPR/Cas9 induced SlARF4 mutant showed similar growth and stomatal responses as ARF4-as plants, which suggest that arf4-cr can tolerate salt and osmotic stresses. Our data support the involvement of ARF4 as a key factor in tomato tolerance to salt and osmotic stresses and confirm the use of CRISPR technology as an efficient tool for functional reverse genetics studies.

Background and aims: Trichomes are epidermal structures with an enormous variety of ecological functions and economic applications. Glandular trichomes produce a rich repertoire of secondary metabolites, whereas non-glandular trichomes create a physical barrier against biotic and abiotic stressors. Intense research is underway to understand trichome development and function and enable breeding of more resilient crops. However, little is known on how enhanced trichome density would impinge on leaf photosynthesis, gas exchange and energy balance. Methods: Previous work has compared multiple species differing in trichome density, instead here we analyzed monogenic trichome mutants in a single tomato genetic background (cv. Micro-Tom). We determined growth parameters, leaf spectral properties, gas exchange and leaf temperature in the hairs absent ( h ), Lanata ( Ln ) and Woolly ( Wo ) trichome mutants. Key results: Shoot dry mass, leaf area, leaf spectral properties and cuticular conductance were not affected by the mutations. However, the Ln mutant showed increased carbon assimilation ( A ) possibly associated with higher stomatal conductance ( g _s ), since there were no differences in stomatal density or stomatal index between genotypes. Leaf temperature was furthermore reduced in Ln in the early hours of the afternoon. Conclusions: We show that a single monogenic mutation can increase glandular trichome density, a desirable trait for crop breeding, whilst concomitantly improving leaf gas exchange and reducing leaf temperature. HIGHLIGHT: A monogenic mutation in tomato increases trichome density and optimizes gas exchange and leaf temperature