Crystal structure and interconversion of monomers and domain-swapped dimers of the walnut tree phytocystatin.

Biotechnological applications of phytocystatins have garnered significant interest due to their potential applications in crop protection and improve crop resistance to abiotic stress factors. Cof1 and Wal1 are phytocystatins derived from Coffea arabica and Juglans regia, respectively. These plants hold significant economic value due to coffee's global demand and the walnut tree's production of valuable timber and widely consumed walnuts with culinary and nutritional benefits. The study involved the heterologous expression in E. coli Lemo 21(DE3), purification by immobilized metal ion affinity and size exclusion chromatography, and biophysical characterization of both phytocystatins, focusing on isolating and interconverting their monomers and dimers. The crystal structure of the domain-swapped dimer of Wal1 was determined revealing two domain-swapped dimers in the asymmetric unit, an arrangement reminiscent of the human cystatin C structure. Alphafold models of monomers and Alphafold-Multimer models of domain-swapped dimers of Cof1 and Wal1 were analyzed in the context of the crystal structure. The methodology and data presented here contribute to a deeper understanding of the oligomerization mechanisms of phytocystatins and their potential biotechnological applications in agriculture.

Development and characterization of new microsatellites for walnut (Juglans regia).

The expressed sequence tag (EST) database represents a potentially valuable resource for the development of simple sequence repeat (SSR) markers for use in evolutionary studies. EST-SSRs reveal polymorphisms not only within the source taxon, but in related taxa as well. In this paper, we describe a case study in which the publicly available walnut (Juglans regia) EST database was used to develop SSR markers for use in the genetic analysis of the widespread Juglans nigra and Carya cathayensis and an endangered species Annamocarya sinensis. A total of 7262 unigenes, including 1911 contigs and 5351 singletons, were obtained from 13,559 ESTs retrieved from the NCBI database. The 7262 unigenes were further reduced to 706 EST-SSR sequences containing 805 SSR loci. Then, 309 EST-SSR primers were randomly designed, and 77 were identified with five high across-species transferability cross-species: namely, J. regia, J. nigra, C. cathayensis, Carya dabieshanensis, and A. sinensis. Thirteen highly polymorphic EST-SSRs were further used for genetic analyses in these above five species.

Walnut (Juglans regia L.): genetic resources, chemistry, by-products.

Walnut (Juglans regia L.) is the most widespread tree nut in the world. There is a great diversity of genotypes differing in forestry, productivity, physical and chemical nut traits. Some of them have been evaluated as promising and may serve as germplasm sources for breeding. The nutritional importance of the nut is related to the seed (kernel). It is a nutrient-dense food mainly owing to its oil content (up to 740 g kg(-1) in some commercial varieties), which can be extracted easily by screw pressing and consumed without refining. Walnut oil composition is dominated largely by unsaturated fatty acids (mainly linoleic together with lesser amounts of oleic and linolenic acids). Minor components of walnut oil include tocopherols, phospholipids, sphingolipids, sterols, hydrocarbons and volatile compounds. Phenolic compounds, present at high levels in the seed coat but poorly extracted with the oil, have been extensively characterised and found to possess strong antioxidant properties. The oil extraction residue is rich in proteins (unusually high in arginine, glutamic and aspartic acids) and has been employed in the formulation of various functional food products. This review describes current scientific knowledge concerning walnut genetic resources and composition as well as by-product obtainment and characteristics.