Identification and application of an exocarp-preferential promoter for genetic engineering of tomato fruit.

Abstract

Tomato (Solanum lycopersicum) is a globally cultivated crop with great economic value. The exocarp determines the appearance of tomato fruit and protects it from various biotic and abiotic challenges at both pre-harvest and post-harvest stages. However, no tomato exocarp-specific promoter is currently available, which hinders exocarp-based genetic engineering. Here, we identified by RNA sequencing and reverse transcription-quantitative PCR analyses that the tomato gene SlPR10 (PATHOGENESIS RELATED 10) was abundantly and predominantly expressed in the exocarp. A fluorescent reporter expressed by a 2087-bp SlPR10 promoter (pSlPR10) was mainly detected in the exocarp of transgenic tomato plants of both Ailsa Craig and Micro-Tom cultivars. This promoter was further utilized for transgenic expression of SlANT1 and SlMYB31 in tomato, which are master regulators of anthocyanin and cuticular wax biosynthesis, respectively. pSlPR10-driven SlANT1 expression resulted in anthocyanin accumulation in the exocarp, conferring gray mold resistance and extended shelf life to the fruit, while SlMYB31 expression led to waxy thickening in the fruit skin, delaying water loss and also extending fruit shelf life. Intriguingly, pSlPR10 and two other weaker tomato exocarp-preferential promoters exhibited coincided expression specificities in the gynophore of transgenic Arabidopsis (Arabidopsis thaliana) plants, providing not only an inkling of evolutionary homology between tomato exocarp and Arabidopsis gynophore but also useful promoters for studying gynophore biology in Arabidopsis. Collectively, this work reports a desirable promoter enabling targeted gene expression in tomato exocarp and Arabidopsis gynophore and demonstrates its usefulness in genetic improvement of tomato fruit quality.