In the last decade, the availability of gene sequences of many plant species, including tomato, has encouraged the development of strategies that do not rely on genetic transformation techniques (GMOs) for imparting desired traits in crops. One of these new emerging technology is TILLING (Targeting Induced Local Lesions In Genomes), a reverse genetics tool, which is proving to be very valuable in creating new traits in different crop species.

Row crops represent the most important crops in terms of global cultivated area. Such crops include soybean, corn, wheat, rice, rapeseed, sunflower, and cotton. Row crops agriculture is generally an intensive system of farming used to obtain high yields by employing elevated quantities of organic and mineral fertilizers. Considering this, and the decrease in area of arable land, it becomes crucial to ensure high yield and quality using alternative strategies, such as the use of plant biostimulants. These compounds are increasingly recognized as sustainable solution to optimize nutrient uptake, crop yield, quality, and tolerance to abiotic stresses. In this work, by means of high-throughput plant phenotyping, we evaluated the effectiveness of a set of three new foliar biostimulant prototypes (coded as 52096, 52097, 52113) applied on corn and soybean at application rates 2.5 and 5 mL/L (corresponding to 1 and 2 L/ha respectively). This allowed us to select the most effective prototype (52097, commercial name “YieldOn^®”) in increasing digital biovolume (DB) and greener area (GGA) either in soybean (both application rates) or corn (rate 5 mL/L) and decreasing Stress Index (SI) in soybean (both application rates). Molecular mechanism of action of selected prototype 52097 was subsequently characterized through Next Generation Sequencing (NGS). In corn, genes involved in hormone (cytokinin and auxin) metabolism/catabolism, maltose biosynthesis, sugar transport and phloem loading were upregulated after application of prototype 52097. In soybean, genes involved in nitrogen metabolism, metal ion transport (mainly zinc and iron), sulfate reduction, and amino acid biosynthesis were induced. The proposed approach supports the integration of multiple omics to open new perspectives in the discovery, evaluation, and development of innovative and sustainable solutions to meet the increasing needs of row-crops agriculture.

The identification of heat stress (HS)-resilient germplasm is important to ensure food security under less favorable environmental conditions. For that, germplasm with an altered activity of factors regulating the HS response is an important genetic tool for crop improvement. Heat shock binding protein (HSBP) is one of the main negative regulators of HS response, acting as a repressor of the activity of HS transcription factors. We identified a TILLING allele of Solanum lycopersicum (tomato) HSBP1. We examined the effects of the mutation on the functionality of the protein in tomato protoplasts, and compared the thermotolerance capacity of lines carrying the wild-type and mutant alleles of HSBP1. The methionine-to-isoleucine mutation in the central heptad repeats of HSBP1 leads to a partial loss of protein function, thereby reducing the inhibitory effect on Hsf activity. Mutant seedlings show enhanced basal thermotolerance, while mature plants exhibit increased resilience in repeated HS treatments, as shown by several physiological parameters. Importantly, plants that are homozygous for the wild-type or mutant HSBP1 alleles showed no significant differences under non-stressed conditions. Altogether, these results indicate that the identified mutant HSBP1 allele can be used as a genetic tool in breeding, aiming to improve the thermotolerance of tomato varieties.