Recently, there has been an increasing interest in wheat landraces and historical varieties mainly cultivated at the beginning of the 19th century, defined as conservation varieties. Most of the Italian tetraploid conservation varieties originate from Sicily, where they are yet boosting the local economy. One of the main reasons for the growing interest in these varieties is the consumers’ perception that they are healthier and safer than their modern counterparts. It is thought that their consumption could ameliorate symptoms of non-celiac wheat sensitivity (NCWS) and irritable bowel syndrome (IBS). Amylase/trypsin-inhibitors (ATIs) and fructans are among the wheat grain components linked to these conditions. In this study we quantified ATI and fructans in five Sicilian conservation varieties (Bidì, Perciasacchi, Russello, Timilia reste bianche, Timilia reste nere) and three modern cultivars (Iride, Orizzonte, Simeto) in a multi-environment trial. Moreover, we calculated the percentage of unextractable polymetric proteins (%UPP) to predict their technological quality. Our findings revealed no significant differences between conservation varieties and modern cultivars for total ATI content, although variations were observed in the composition of individual ATIs. The fructan content was lower in conservation varieties, especially in Bidì and Russello. Overall, %UPP was generally higher in modern cultivars, although Bidì often displayed comparable values. The environmental influence was significant for all analysed traits; however, high heritability was yet observed for all, except for total ATI content. Our results indicate that only minor differences in grain composition exist between tetraploid conservation varieties and modern cultivars.

Thirteen ATIs were extracted according to Geisslitz et al., (2020) and quantified using the same targeted LC-MS/MS method based on SIDA reported in Jahn et al., (2025). The calculation of the content of individual ATI was based on Geisslitz et al., (2020). Peptide separation was achieved with a Vanquish UHPLC system (Thermo Fisher Scientific, Waltham, MA, USA) equipped with an Aeris 1.7 µm PEPTIDE XB-C18 (100 Å, 150 × 2,1 mm, Phenomenex, Aschaffenburg, Germany) using formic acid (0.1%, v/v) in water as solvent A and formic acid (0.1%, v/v) in acetonitrile as solvent B. Gradient, injection volume and column temperature were the same of Jahn et al., (2025). Calibration was performed exactly as reported by Geisslitz et al., (2020). Scheduled parallel reaction monitoring was performed with a Q Exactive Plus Orbitrap (Thermo Fisher Scientific) and it was operated as described in Jahn et al., (2025).

Five tetraploid wheat conservation varieties (Bidì, Perciasacchi, Russello, Timilia reste bianche, Timilia reste nere) and three modern cultivars (Iride, Orizzonte, Simeto) were cultivated for three years (2020-2021, 2021-2022, 2022-2023) across two different locations in Sicily: Caltavuturo (37°47'43"N 13°54'20"E) and Selinunte (37°54'20"N 12°54'07"E). Kernels of the different conservation varieties were collected from Cinozoo 3R srl (Bisacquino, Palermo, Italy). All the varieties belong to T. turgidum ssp. durum, except for Perciasacchi, which belongs to T. turgidum ssp. turanicum. A randomized complete block design with three replications was implemented at each site. Wheat samples were harvested, and grains were milled in a Cyclotec 1093 (FOSS, Denmark) equipped with a 0.5 mm sieve.