Walnut (Juglans regia L.) is an important woody oilseed species cultivated throughout the world. In this study, comparative transcript profiling was performed using high-throughput RNA sequencing technology at the following three stages of walnut fat synthesis in the “Lvling” walnut cultivar: the initial developmental stage (L1), the fast developing stage (L2), and the last developing stage (L3). A total of 68.18 GB of data were obtained on the three developmental stages, and 92% to 94% of clean data were able to be located to the reference genome. Further comparisons of the transcripts in the three libraries revealed that 724, 2027, and 4817 genes were differentially expressed between the L2 and L1 (L2vsL1), L3 and L2 (L3vsL2), and L3 and L1 (L3vsL1) samples, respectively. Through the GO gene enrichment analysis, differentially expressed genes (DEGs) in L2vsL1, L3vsL2, and L3vsL1 were enriched into 3, 0, and 2 functional categories, respectively. According to the KEGG enrichment analysis, DEGs in L2vsL1, L3vsL2, and L3vsL1 were annotated into 77, 110, and 3717 taxonomic metabolic pathways in the KEGG database, respectively. Next, we analyzed expression levels of genes related to fat synthesis. Our results indicated that ACCase, LACS, and FAD7 were the key genes related to fat synthesis. The high-throughput transcriptome sequencing of walnut in different developmental stages has greatly enriched the current genomic available resources. The comparison of DEGs under different developmental stages identified a wealth of candidate genes involved in fat synthesis, which will facilitate further genetic improvement and molecular studies of the walnut.

Understanding how the soil environment impacts root water uptake location and magnitude is important for better management of plant irrigation. In this study, stable hydrogen and oxygen isotope composition were used to determine seasonal variations in the depth of water taken up by walnut trees during different phenological stages in an irrigated arid hilly area in the Taihang Mountains in China. The contributions of soil water at different depths to the water taken up were quantified by the MixSIAR Bayesian isotope mixing model. The results indicated that water taken up by the walnut trees was sourced mainly from soil water in the 0–20 cm soil layer at the sprouting and leaf expansion stages (62.95%), and the 20–40 cm soil layer at blossoming and fruit-bearing (43.45%), fruit expansion (41.8%), and fruit maturity (39.15%) stages. The mean soil depth of the water taken up by the walnut trees gradually decreased as the phenological stages advanced. The proportions of various soil layer water contributions to the walnut trees differed throughout the phenological stages, and the proportion of deeper soil water contributions gradually increased as the phenological stages of walnut trees advanced. The results of the present study indicated that water sources for walnut trees varied by depth during different phenological stages. In addition to soil moisture, soil temperature may also be an important factor affecting the depth of water taken up by walnut trees. The results also provided scientific implications for water-saving irrigation management.

Knowledge of the spatial–temporal variations of soil organic carbon (SOC) quantity and quality and its microbial regulation mechanisms is essential for long-term SOC sequestration in agroecosystems; nevertheless, this information is lacking in the process of walnut plantations. Here, we used the modified Walkley-Black method, phospholipid fatty acid analysis, and micro-plate enzyme technique to analyze the evolution of SOC stocks and quality/lability as well as microbial communities and enzyme activities at different soil depths in walnut plantations with a chronosequence of 0-, 7-, 14-, and 21-years in the Eastern Taihang Mountains, China. The results indicated that long-term walnut plantations (14-and 21-years) enhanced SOC stocks, improved SOC quality/lability (as indicated by the lability index), and promoted microbial growth and activities (i.e., hydrolase and oxidase activities) in the 0–40 cm soil layers. Besides, these above-mentioned SOC-and microbial-related indices (except for oxidase activities) decreased with increasing soil depths, while oxidase activities were higher in deeper soils (40–60 cm) than in other soils (0–40 cm). The partial least squares path model also revealed that walnut plantation ages and soil depths had positive and negative effects on microbial attributes (e.g., enzyme activities, fungal and bacterial communities), respectively. Meanwhile, the SOC stocks were closely related to the fungal community; meanwhile, the bacterial community affected SOC quality/liability by regulating enzyme activities. Comprehensively, long-term walnut plantations were conducive to increasing SOC stocks and quality through altering microbial communities and activities in the East Taihang Mountains in Hebei, China.

To reveal the regular patterns of water transport in fruit-bearing shoots of walnut (Juglans regia), water transport pathways for various organs were observed using dye-tracing technique; water potential, water status and water transport rate were determined for each of these organs. Water potential in the pedicel, petiole, lateral shoot and main shoot was -2.59, -2.85, -1.68 and -0.37 MPa, respectively. Significant differences were noted among various organs in terms of water status. The ratio of bound water to free water in the petiole was the highest (1.93 RBC), followed by the main shoot, lateral shoot and pedicel. Water transport speed in the petiole was the fastest (3.13 cm/min), followed by the lateral shoot, pedicel and main shoot. However, the water transport rate of the main shoot was the highest, and higher than the sum of the other three organs. The xylem in the petiole was separated from the main shoot; at the top of the main shoot, the xylem was divided into two parts, one of which was connected to the xylem of the pedicel and the other to the lateral shoot. During the shell-hardening period of fruits, the dye was found at the edge of the fruit vascular bundle, although, it was not at the central vascular bundle. No dye was found in the main and accessory buds, instead, only a small part of the xylem was dyed between the accessory bud and the main shoot. Water transport pathways were different for various organs. Thus, the transport and distribution of water in various organs was affected by their water potential, tissue structure and water requirement.