Grain physical characteristics and milling behavior of a durum wheat line in which both wildtype puroindoline genes were translocated and stabilized after backcrossing (Svevo-Pin) were compared with the parent line (Svevo). The only observed differences between grain characteristics were the mechanical resistance and starchy endosperm porosity revealed through vitreosity measurement. A significant increase of flour and a decrease of semolina yield and break milling energy were observed from Svevo-Pin in comparison with the nonrecombinant parent line in accordance to the lower grain mechanical resistance and higher porosity measurements. Moreover, the particle size distribution shown for Svevo-Pin flour appeared consistent with a lower adhesion between starch granules and the protein matrix attributed to the presence of wild-type puroindolines. Coarse bran yield was conversely increased. This appeared to be due to a lower starchy endosperm recovery as a higher proportion of grain starch was found in this bran fraction. Flour from the durum parent line was inversely enriched in phytic acid, a cellular marker of the aleurone layer. Starch damage was also lower in Svevo-Pin flours in comparison with Svevo. All of the observed differences between translocation and parent lines were confirmed independent of the culture growth conditions (n=12).

Self-assembling protein nanocontainers are promising candidates for an increasingly wide scope of purposes. Their applications range from drug delivery vehicles and imaging agents to nanocompartments for controlled enzymatic activity. In order to exploit their full potential in these different fields, characterization of their properties is vital. For example, their mechanical properties give insight into the stability of a particle as a function of their internal content. The mechanics can be probed by atomic force microscopy nanoindentation, and while this single particle method is increasingly used to probe material properties of viral nanocages, it has hardly been used to characterize nonviral nanocages. Here we report nanoindentation studies on two types of nonviral nanocontainers: (i) lumazine synthase from Aquifex aeolicus (AaLS), which naturally self-assembles into icosahedral cages, and (ii) the artificial protein cage O3-33 originating from a computational design approach. In addition, we tested particles that had been engineered toward improved cargo loading capacity and compared these nanocages in empty and loaded states. We found that the thermostable AaLS cages are stiffer and resist higher forces before breaking than the O3-33 particles, but that mutations affecting the size of AaLS particles have a dramatic effect on their structural stability. Furthermore, we show that cargo packaging can occur while maintaining the cage's mechanical properties.

Contact-resonance atomic force microscopy (CR-AFM) is of great interest and very valuable for a deeper understanding of the mechanics of biological materials with moduli of at least a few GPa. However, sample surfaces can present a high topography range with significant slopes, where the local angle can be as large as  ± 50°. The non-trivial correlation between surface slope and CR-frequency hinders a straight-forward interpretation of CR-AFM indentation modulus measurements on such samples. We aim to demonstrate the significant influence of the surface slope on the CR-frequency that is caused by the local angle between sample surface and the AFM cantilever and present a practical method to correct the measurements. Based on existing analytical models of the effect of the AFM set-up's intrinsic cantilever tilt on CR-frequencies, we compute the non-linear variation of the first two (eigen)modes CR-frequency for a large range of surface angles. The computations are confirmed by CR-AFM experiments performed on a curved surface. Finally, the model is applied to directly correct contact modulus measurements on a durum wheat starch granule as an exemplary sample.