## 2 Materials and Methods
### Sampling Procedures, Measurements, and Methods  
To obtain oilseed cake samples, amelina seeds weighing a total of 300 g were harvested from each experimental plot. The seeds were cleaned from foreign materials and then cold-pressed using a cold mechanical screw pressing (TabyPress Type 20, Orebro, Sweden) operated at a temperature below 50 \({}^{\circ}\)C. The press cake was left to dry at room conditions for 24 h before chemical composition analysis. The seed cake extraction rate (%) was determined by dividing the mass of cake extracted with initial weight of amelina seeds and multiplying by 100. The seed cake yield was calculated by multiplying the seed cake extraction rate with the seed yield. As regards chemical analysis, the oilseed cake samples were crushed to pass through a 1 mm Wiley mill screen (Thomas T4274.E15 Steel Model 4 Wiley Mill; [PERSON], Philadelphia, PA, USA). The samples were assessed for dry matter (DM) analysis (method 943.01), crude ash (CA; method 924.05), and crude protein (CP; method 984.13) with a fully automated Kjeldahl analyzer (Kjeldlec 8400; Foss Tecator AB, Hoganas, Sweden), as well as crude fat (CF; method 920.39) and crude fiber (CFr; method 978.10) [35]. The crude protein (CP) content was estimated by multiplying the Kjeldahl nitrogen content by 6.25. Concerning the carbohydrate content (CHO), it was calculated as the remainder up to 100% of the sample dry matter (DM) after subtraction of the sum of crude fat (CF), crude protein (CP), and crude ash (CA). The residual oil from the oilseed cake was extracted using the Soxhlet n-hexane method. Specifically, 250 mL of solvent and 10 g of amelina oilseed cake sample were used in order to maintain a solvent to solid ratio of 25:1. The total extraction process was completed within one 1 h. The extracted phase was then distilled to separate the oil from the solvent. The residual oil content (%) was determined by dividing the mass of oil extracted by the initial weight of amelina cake and multiplying by 100. The quantification and determination of fatty acids (FAs) were also performed in the oilseed cake samples using an Agilent 6890 GC chromatograph (Agilent Technologies, Santa Clara, CA, USA) equipped with a flame ionization detector (FID) and a single capillary column HP-88 GC (60 m \(\times\) 0.25 mm i.d. \(\times\) 0.20 um film thickness; Hewlett-Packard Corporation, Palo Alto, CA, USA) after the transesterification procedure described by [PERSON] et al. [36]. The temperatures in the initial and final columns were 170 \({}^{\circ}\)C and 230 \({}^{\circ}\)C, respectively, and the temperature increased at a rate of 4 \({}^{\circ}\)C min\({}^{-1}\). The temperatures of the injector and the detector were both 230 \({}^{\circ}\)C. The FID temperature was set to 260 \({}^{\circ}\)C, and the chromatographic analysis included a temperature-programmed run beginning at 120 \({}^{\circ}\)C and lasting 1 min. Then, there were two step-ramps, one at 1.25 \({}^{\circ}\)C min\({}^{-1}\) to 230 \({}^{\circ}\)C and another at 10 \({}^{\circ}\)C min\({}^{-1}\) to 240 \({}^{\circ}\)C, each for 3 min. The flame gases were hydrogen (at least \(>\)99% purity) and synthetic air. The carrier gas was helium with a linear velocity set at 30 cm s\({}^{-1}\). Fatty acid identification was made by comparing the relative retention times from samples with a mix standard of 37 component FA methyl esters (FAME) (Supelco, Sigma-Aldrich, St. Louis, MO, USA). Extra FA standards were also utilized for cis-9, trans-11, C18:2, trans-10, cis-12 C18:2, and trans-11 C18:1 FA (Sigma-Aldrich, St. Louis, MO, USA). The results were processed using a Hewlett-Packard 3365 Chemstation data analysis software (Hewlett-Packard Corporation, Palo Alto, CA, USA). In the present study, the seven major fatty acids (\(\sim\)99%) identified in amelina oilseed cake were studied and expressed as the relative fraction of each individual fatty acid contained in the sample.