Identification and Characterization of Wall-Associated Kinase (WAK) and WAK-like (WAKL) Gene Family in Juglans regia and Its Wild Related Species Juglans mandshurica

Wall-associated kinase (WAK) and WAK-like kinase (WAKL) are receptor-like kinases (RLKs), which play important roles in signal transduction between the cell wall and the cytoplasm in plants. WAK/WAKLs have been studied in many plants, but were rarely studied in the important economic walnut tree. In this study, 27 and 14 WAK/WAKL genes were identified in Juglans regia and its wild related species Juglans mandshurica, respectively. We found tandem duplication might play a critical role in the expansion of WAK/WAKL gene family in J. regia, and most of the WAK/WAKL homologous pairs underwent purified selection during evolution. All WAK/WAKL proteins have the extracellular WAK domain and the cytoplasmic protein kinase domain, and the latter was more conserved than the former. Cis-acting elements analysis showed that WAK/WAKL might be involved in plant growth and development, plant response to abiotic stress and hormones. Gene expression pattern analysis further indicated that most WAK/WAKL genes in J. regia might play a role in the development of leaves and be involved in plant response to biotic stress. Our study provides a new perspective for the evolutionary analysis of gene families in tree species and also provides potential candidate genes for studying WAK/WAKL gene function in walnuts.

Impact of Hydro-Alcoholic Solvents on the Oil and Phenolics Extraction from Walnut (Juglans regia L.) Press-Cake and the Self-Emulsification of Extracts

The objective was to evaluate the performance of four hydro-alcoholic solvents to simultaneously extract oil and more polar molecules as phenolics, among others, to produce complex extracts that eventually could self-emulsify after solvent removal. Walnut press-cake was selected as the sourcing material. Extractions were performed as a semi-continuous operation up to a solvent-to-solid ratio of 28, with a fractional collection of the effluent. Among the solvents, labelled by their alcohol content EtOH 58, EtOH 86, iPro 60 and iPro 90 for ethanol (EtOH) and isopropanol (iPro), iPro 90 allowed to reach an oil extraction efficiency of 97% while the recovery for the other solvents was in the range of 30–40%. For both alcohols, the increase of the solvent hydration negatively influenced the oil extraction but positively increased the recovery of phenolics that reached 17.6 mg GAE/gcake when EtOH 58 was used. Several fractions contained enough surface-active material and oil to self-assemble as emulsions. IPro 90 and EtOH 86 showed better performances in the sense that most extracts were able to emulsify, though extraction kinetics pointed out differences. The most hydrated solvents behaved equally, with extraction yields in the same range and a similar but limited emulsifying capacity of only few fractions.

Physical and Chemical Changes in Hydrothermally Modified Wood

In this study, the simulation of artificial conditions, containing various chemical additives similar to a bog environment, was hypothesised as a relatively short way to develop a more sustainable wood product with better properties. This study aimed to determine how hydrothermal treatment with different iron additives with and without tannic acid, performed at 100 °C for 75 h, changed the mass and density of wood of different tree species (Pinus sylvestris L., Picea abies (L.) H. Karst., Pseudotsuga menziesii (Mirb.) Franco, Juglans regia L., and Acer platanoides L.); total phenolic compounds in biobased solvents after the hydrothermal processing andiron concentration in different wood layers. We also aimed to determine the wood chemical composition using Fourier-transform infrared technique. The study results showed that the softwood samples were more susceptible to hydrothermal modification than the hardwood samples, indicated by minorly different intensities on infrared spectra. The FTIR results showed bands of hardwood and softwood samples which along with the spectral characterisation of the wood constituents, which can further be utilized for creating a larger FTIR spectral database of wood samples for their identification. The factors with the most significant effect on the hydrothermal modification were tannic acid and inorganic Fe3O2 salt, used as an additive. The results indicated the suitability of inorganic Fe3O2 salt and tannic acid combination applied in this type of modification, because a higher concentration of iron was distributed on the inner and intermediate sublayers of wood samples. The hydrothermal modification approach to anchor tannic acid–Fe macromolecules in wood shown in this study has promising capabilities and may attract interest in the field of wood protection against weathering.

Introduction of a New Interesting Walnut Cultivar “Leto”

Ιn an effort to create walnut cultivars (Juglans regia) with high productivity, fruit quality and lateral bearing, a new cultivar, named “Leto”, was created by the cross “Gustine” × “Pedro”. Its main phenological and pomological characteristics were assessed according to the criteria of IPGR (1994) and UPOV-TG/125/6 (1999), for 10 consecutive years and compared with its maternal cultivars and “Chandler”. Observations showed that “Leto” has high lateral bearing habit (90%) and presents satisfactory yield at the full production age. The tree size is smaller than that of its parents and “Chandler”, female flowers bloom from 11 to 22 of April and male from 3 to 6 of April. “Leto” nuts are harvested at the end of September, present easy hull dehiscence and high kernel percentage. Other positive nut characteristics of “Leto” are light kernel color, well kernel filling and easy removal of the kernel halves. “Leto” is a mid-early cultivar of great interest due to its high- quality nuts, suitable for dense plantings, in regions where the last spring frosts occur in late March to early April, thus, making it a promising cultivar for Greece, but also for other regions with similar geomorphological and climatic conditions.

Green Husk of Walnuts (Juglans regia L.) from Southern Italy as a Valuable Source for the Recovery of Glucans and Pectins

Walnut green husk is an agricultural waste produced during the walnut (Juglans regia L.) harvest, that could be valued as a source of high-value compounds. In this respect, walnut green husks from two areas of Southern Italy (Montalto Uffugo and Zumpano), with different soil conditions, were investigated. Glucans and pectins were isolated from dry walnut husks by carrying out alkaline and acidic extractions, respectively, and then they were characterized by FT-IR, scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). The colorimetric method for the enzymatic measurement of α- and β-glucans was performed. The maximum total glucan yield was recovered from Montalto walnut husks (4.6 ± 0.2 g/100 g DM) with a β-glucan percentage (6.3 ± 0.4) higher than that calculated for Zumpano walnut husks (3.6 ± 0.5). Thermal analysis (DSC) confirmed the higher degree of crystallinity of glucans from Zumpano. The pectin content for Montalto husks was found to be 2.6 times that of Zumpano husks, and the esterification degree was more than 65%. The results suggested that J. regia L. green husks could be a source of glucans and pectins, whose content and morphological and thermal characteristics were influenced by different soil and climate conditions.