This paper describes the application of the wide-band non-contact ultrasonic resonant spectroscopy technique to layered plant tissues (leaves), a method to extract the properties of main component tissues: palisade parenchyma and spongy mesophyll, a verification of the obtained properties, and a discussion of the implications of the observed elastic anisotropy. Transmission coefficient spectra of Ligustrum lucidum leaves with the thickness in the range of 250–850 μm revealing several order thickness resonances have been measured. A leaf acoustic model based on a two-layered structure and a metaheuristic (simulated annealing algorithm) is used to solve the inverse problem. The extracted parameters of these two layers of tissue are consistent with cross-sectional cryo-SEM images and other independent measurements. The extracted resonant frequency and the impedance of each layer explain the origin of the observed resonances. Finally, the elastic modulus of each layer is extracted and analyzed. The presented technique is a unique tool to study (in vivo and in a completely non-invasive way) the ultrasonic, elastic, and viscoelastic properties of layered plant tissues which could lead to a better understanding of the relationship between the tissue microstructure and the tissue function with macroscopic properties and how this may affect water relations.
The support from the Spanish State Research Agency and European Regional Development Fund (ERDF/FEDER), Grant No. DPI2016-78876-R, is acknowledged.