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Botryosphaeriaceae species are the major causal agents of walnut dieback worldwide, along with Diaporthe species. Botryosphaeria dothidea and Neofusicoccum parvum are the only two Botryosphaeriaceae species associated with this recently emergent disease in France, and little is known about their diversity, structure, origin and dispersion in French walnut orchards. A total of 381 isolates of both species were genetically typed using a sequence-based microsatellite genotyping (SSR-seq) method. This analysis revealed a low genetic diversity and a high clonality of these populations, in agreement with their clonal mode of reproduction. The genetic similarity among populations, regardless of the tissue type and the presence of symptoms, supports the hypothesis that these pathogens can move between fruits and twigs and display latent pathogen lifestyles. Contrasting genetic patterns between N. parvum populations from Californian and Spanish walnut orchards and the French ones suggested no conclusive evidence for pathogen transmission from infected materials. The high genetic similarity with French vineyards populations suggested instead putative transmission between these hosts, which was also observed with B. dothidea populations. Overall, this study provides critical insight into the epidemiology of two important pathogens involved in the emerging dieback of French walnut orchards, including their distribution, potential to mate, putative origin and disease pathways.

Background: Constipation is one of the most common gastrointestinal complaints during pregnancy. Consuming fruits and vegetables is often the first line of treatment due to their fiber content. Therefore, the purpose of the present study was to determine the effect of combined fig-walnut syrup on functional constipation (FC) and quality of life (QoL) in pregnant women. Methods: In this double-blind, randomized controlled clinical trial, 90 pregnant women with FC were randomly assigned to receive combined fig-walnut syrup, fig syrup, or placebo (n = 30 in each group) using block randomization. Participants received 15 ml of syrup once daily at night, half an hour before bedtime, for 14 days and were followed up for 2 weeks after the end of the intervention. The FC, quality of life-gravidity (QOL-GRAV), and food frequency questionnaires (FFQ) were used to collect data. The questionnaires were completed once before the intervention and then the FC questionnaire was completed once a week for four weeks and the QoL questionnaire was completed at the end of the fourth week. To compare the outcomes among the study groups, one-way AONOVA, ANCOVA, Mann- Whitney U test, and Kruskal- Wallis test were used. Results: Following the intervention, although there was no statistically significant difference in the number of defecations between the combined fig-walnut syrup group and the fig syrup group (P > 0.05) at every four weeks, there was a statistically significant difference compared to the placebo (P < 0.05). Combined fig-walnut syrup (adjusted mean difference (AMD): -3.4; 95% confidence interval: -0.7 to -6.1; P = 0.008) and fig syrup (AMD: -5.8; 95% CI: -3.1 to -8.6; P < 0.001) improved QoL compared to the placebo group and there was no statistically significant difference between the combined fig-walnut and fig syrup (AMD: 2.4; 95% CI: 5.2 to -0.3; P = 0.104). Conclusion: The consumption of fig-walnut syrup and fig syrup may help improve constipation symptoms and enhance QoL during pregnancy. Further studies are needed to reach a conclusive determination. Trial registration: Iranian Registry of Clinical Trials (IRCT): IRCT2012071801032N79. Date of registration: 07.12.2023. [ABSTRACT FROM AUTHOR]

Crown gall disease (Agrobacterium tumefaciens), crown/root rot disease (Phytophthora spp.), root lesion disease (Pratylenchus vulnus) and tree vigor are key traits affecting the productivity and quality of walnuts in California. Unchallenged hybrid rootstocks were analyzed by RNA-seq to examine pre-formed factors affecting these traits. Enrichment analysis of the differentially expressed genes revealed that the increased expression of cell wall biogenesis-related genes plays a key role in susceptibility to A. tumefaciens, susceptibility to Phytophthora spp. and increased vigor. Analysis of the predicted subcellular loci of the encoded proteins revealed that many gene products associated with vigor and susceptibility were targeted to the plasma membrane and extracellular space, connecting these traits to sustaining barrier function. We observed that RNA processing and splicing, along with predicted nuclear targeting, were associated with resistance to A. tumefaciens, resistance to Phytophthora spp. and low vigor. Four genes within the J. microcarpa QTL region for resistance to A. tumefaciens and Phytophthora spp. were represented among our transcripts, with two of the genes being differentially expressed in association with resistance to A. tumefaciens and decreased vigor. No differential expression related to Phytophthora spp. or P. vulnus resistance was observed in this region. Additionally, the J. microcarpa haplotype expressed more transcripts associated with resistance to A. tumefaciens, Phytophthora spp. and low vigor, but not P. vulnus, than the J. regia haplotype. We also report unique and shared hormone and defense responses associated with each trait. This research suggests a link between cell wall biogenesis, vigor and critical root diseases of walnut.

Walnut (Juglans spp.) trees hold immense significance in both economic and ecological contexts within agri-horticultural ecosystems. The comprehension of the intricate mechanisms underpinning walnut growth and development stands as a pivotal endeavor, essential for advancing sustainable yield practices. This comprehensive review delves into the multifaceted factors that contribute to the growth and development of walnuts, encompassing hormonal, biochemical, and genetic dimensions. Notably, hormones such as gibberellic acids (GAs) and sugars assume pivotal roles in the initiation and maturation of walnut flowers, with specific investigations demonstrating that the application of GAs has the capacity to augment male flower counts. The levels of endogenous auxins and gibberellins exhibit variations across distinct phases of walnut development, with the highest concentrations observed in young tissues. The molecular underpinnings of walnut growth and development involve a complex interplay of genetic regulation, hormonal dynamics, and environmental factors. Distinct sets of genes exhibit activation at discrete developmental stages, thereby influencing fundamental processes such as cell division, differentiation, and food reserve metabolism. Several key regulatory genes, including ACC, ASMT, SAD, FAD, SOC, and TFL1, emerge as pivotal orchestrators, steering essential processes encompassing cell division, differentiation, flowering, and fruit development. Conclusively, this article provides a detailed exploration of the diverse aspects of walnut growth and development, from genetic regulation to hormonal and biochemical processes. This will provide a valuable resource for researchers, horticulturalists, and biotechnologists aiming to improve walnut productivity and resilience in the face of changing environmental conditions. [ABSTRACT FROM AUTHOR]

Soil fungal communities are critical for forest ecosystem functions in the Central Hardwood Region (CHR) of the USA. This evaluation, which took place in 2022–2023, investigates the influence of Juglans nigra (BW, black walnut) and Quercus rubra (NRO, Northern red oak) on soil properties and fungal community structures across three CHR sites. The objectives of this study are to investigate how the fungal communities identified beneath J. nigra and Q. rubra serve to influence biodiversity and soil health within hardwood plantations. Soils from two locations in Indiana and one in Michigan were examined and assessed for variations in fungal composition and diversity. Soil fungal communities were characterized using Illumina high-throughput sequencing while multivariate analysis was applied to analyze patterns in these fungal communities. These data provided insights into how environment, location, and tree species affect fungal community structure. Results indicate that J. nigra soils exhibited higher carbon (0.36%, 1.02%, 0.72%), nitrogen (25%, 29%, 56%), and pH (0.46, 1.08, 1.54) levels than Q. rubra soils across all three sites and foster greater fungal diversity. Specifically, J. nigra was associated with increased Ascomycota diversity, whereas Q. rubra supported a higher prevalence of Basidiomycota. Basidiomycota were negatively correlated with carbon and pH, while Ascomycota showed positive correlations with these variables. These findings highlight how crucial it is to understand how different tree species influence fungal communities and, consequently, how they influence forest soil health. Our findings serve to improve forest management practices by emphasizing the importance of fungal communities in maintaining the function and resilience of an ecosystem. Our study underscores that grasping these specific interactions is essential for effective forest management, especially when considering how to use fungal communities to boost plant growth. This work focuses on hardwood plantations rather than either agricultural ecosystems, monocultures, or native forests, thus filling a gap in the current literature where many studies are limited to specific fungal groups such as mycorrhizae. In future research, it is important to examine a wider range of tree species. This will deepen our understanding of fungal community dynamics and their impact on maintaining healthy forest ecosystems. Our hardwood plantation focus also notes the potential for adaptive forest management as environmental conditions change. [ABSTRACT FROM AUTHOR]

Eastern black walnut, Juglans nigra L., is an economically important tree species valued for its high-quality timber and edible nuts. A regional industry for the species' nut and kernel products resides in Missouri, where over 9 million kg of hulled in-shell nuts are purchased in masting years. The crop is primarily based upon nuts harvested from wild trees, placing a ceiling on nut volume and quality (e.g. small nut size, dark pellicle color, and 10–14% kernel by weight). Orchards of named cultivars, like 'Kwik Krop' and 'Sparrow', supply up to 22,000 kg of nuts with a higher kernel percentage (> 26%) and improved quality. Such cultivars often represent chance wild or on-farm seedlings, clonally propagated since the late 1800's by enthusiasts. Today, continued improvement of eastern black walnut as an orchard crop is limited by a long generation time, delayed expression of important traits, and space requirements – creating a strong need for marker-trait association studies that inform progeny selection. The first linkage map for J. nigra was recently created using the 'Sparrow' × 'Schessler' F1 population and loci for phenology traits discovered. The objective of this study is to utilize these genetic resources to detect quantitative trait loci (QTL) and report associated DNA markers for the spur-bearing habit, which promotes precocity and high yield. Using single-year data from the 11-year-old population, we observe that segregation for the spur-bearing habit appears to be recessive and multigenic. Three QTLs (p > 0.99) were identified on linkage group (LG) 8, LG11, and LG16 that explain 7.2%, 8.7%, and 10% of trait variation, respectively. Regions between flanking DNA markers were 3.16 cM, 4.32 cM, and 9.69 cM, respectively. This study is the first to examine the genetic control of bearing habit and yield in eastern black walnut and informs breeders' approach for their future genetic improvement. [ABSTRACT FROM AUTHOR]

East Asia constitutes one of the largest and most complex areas of plant diversity globally. This complexity is attributable to the geological history and climatic diversity of East Asia. However, controversy persists over the biogeographical hypotheses and different studies on this topic. The demographical complexity of temperate deciduous woody plants and heterogeneity of geological and climatic fluctuations in East Asia remain poorly understood. We aimed to assemble the chloroplast and nuclear genomes of 360 individuals from five walnut (Juglans) species in East Asia and comprehensively analyze the phylogenetic inference, genetic structure, population demographic, and selection pressure. The results showed that East Asian walnuts could be divided into two major groups, that is, section Juglans/Dioscaryon and sect. Cardiocaryon. Within sect. Cardiocaryon, the samples were categorized into the northern clade and southern clade, both of which were affected by Quaternary glaciation; however, the southern clade was affected to a lesser extent. The population expansion events observed in sect. Cardiocaryon and sect. Dioscaryon since the late Miocene indicated that Neogene climate cooling substantially affected the distribution of Tertiary relict plants in East Asia. Biogeographic analysis results showed that the uplift of the Qinghai–Tibetan Plateau (QTP) might have contributed to the divergence within sect. Dioscaryon. Our findings highlight the heterogeneity of climatic fluctuations in the northern and southern regions of East Asia during the late Neogene and Quaternary periods and suggest that the uplift of QTP could have facilitated the divergence of temperate deciduous woody plants. [ABSTRACT FROM AUTHOR]

Juglans californica, California walnut, is a vulnerable small tree that is locally abundant but restricted to woodland and chaparral habitats of Southern California threatened by urbanization and land use change. This species is the dominant species in a unique woodland ecosystem in California. It is one of 2 endemic California walnut species (family Juglandaceae). The other species, Northern California black walnut (J. hindsii), has been suggested controversially to be a variety of J. californica. Here, we report a new, chromosome-level assembly of J. californica as part of the California Conservation Genomics Project (CCGP). Consistent with the CCGP common methodology across ~150 genomes, we used Pacific Biosciences HiFi long reads and Omni-C chromatin-proximity sequencing technology to produce a de novo assembled genome. The assembly comprises 137 scaffolds spanning 551,065,703 bp, has a contig N50 of 30 Mb, a scaffold N50 of 37 Mb, and BUSCO complete score of 98.9%. Additionally, the mitochondrial genome has 701,569 bp. In addition, we compare this genome with other existing high-quality Juglans and Quercus genomes, which are in the same order (Fagales) and show relatively high synteny within the Juglans genomes. Future work will utilize the J. californica genome to determine its relationship with the Northern California walnut and assess the extent to which these 2 endemic trees might be at risk from fragmentation and/or climate warming.

Persian walnut is a drought-sensitive species with considerable genetic variation in the photosynthesis and water use efficiency of its populations, which is largely unexplored. Here, we aimed to elucidate changes in the efficiency of photosynthesis and water content using a diverse panel of 60 walnut families which were submitted to a progressive drought for 24 days, followed by two weeks of re-watering. Severe water-withholding reduced leaf relative water content (RWC) by 20%, net photosynthetic rate (Pn) by 50%, stomatal conductance (gs) by 60%, intercellular CO2 concentration (Ci) by 30%, and transpiration rate (Tr) by 50%, but improved water use efficiency (WUE) by 25%. Severe water-withholding also inhibited photosystem II functionality as indicated by reduced quantum yield of intersystem electron transport (φEo) and transfer of electrons per reaction center (ET0/RC), also enhanced accumulation of QA (VJ) resulted in the reduction of the photosynthetic performance (PIABS) and maximal quantum yield of PSII (FV/FM); while elevated quantum yield of energy dissipation (φDo), energy fluxes for absorption (ABS/RC) and dissipated energy flux (DI0/RC) in walnut families. Cluster analysis classified families into three main groups (tolerant, moderately tolerant, and sensitive), with the tolerant group from dry climates exhibiting lesser alterations in assessed parameters than the other groups. Multivariate analysis of phenotypic data demonstrated that RWC and biophysical parameters related to the chlorophyll fluorescence such as FV/FM, φEo, φDo, PIABS, ABS/RC, ET0/RC, and DI0/RC represent fast, robust and non-destructive biomarkers for walnut performance under drought stress. Finally, phenotype-environment association analysis showed significant correlation of some photosynthetic traits with geoclimatic factors, suggesting a key role of climate and geography in the adaptation of walnut to its habitat conditions.

Soil salinization has become one of the major problems that threaten the ecological environment. The aim of this study is to explore the mechanism of salt tolerance of hybrid walnuts (Juglans major × Juglans regia) under long-term salt stress through the dynamic changes of growth, physiological and biochemical characteristics, and anatomical structure. Our findings indicate that (1) salt stress inhibited seedling height and ground diameter increase, and (2) with increasing salt concentration, relative water content (RWC) decreased, and proline (Pro) and soluble sugar (SS) content increased. The Pro content reached a maximum of 549.64 μg/g on the 42nd day. The increase in superoxide dismutase (SOD) activity (46.80–117.16%), ascorbate peroxidase (APX) activity, total flavonoid content (TFC), and total phenol content (TPC) under salt stress reduced the accumulation of malondialdehyde (MDA). (3) Increasing salt concentration led to increases and subsequent decreases in the thickness of palisade tissues, spongy tissues, leaves, and leaf vascular bundle diameter. Upper and lower skin thickness, root periderm thickness, root diameter, root cortex thickness, and root vascular bundle diameter showed different patterns of change at varying stress concentrations and durations. Overall, the study concluded that salt stress enhanced the antireactive oxygen system, increased levels of osmotic regulators, and low salt concentrations promoted leaf and root anatomy, but that under long-term exposure to high salt levels, leaf anatomy was severely damaged. For the first time, this study combined the anatomical structure of the vegetative organ of hybrid walnut with physiology and biochemistry, which is of great significance for addressing the challenge of walnut salt stress and expanding the planting area. [ABSTRACT FROM AUTHOR]

Abstract Persian walnut (Juglans regia) and Manchurian walnut (Juglans mandshurica) belong to Juglandaceae, which are vulnerable, temperate deciduous perennial trees with high economical, ecological, and industrial values. 4-Coumarate: CoA ligase (4CL) plays an essential function in plant development, growth, and stress. Walnut production is challenged by diverse stresses, such as salinity, drought, and diseases. However, the characteristics and expression levels of 4CL gene family in Juglans species resistance and under salt stress are unknown. Here, we identified 36 Jr4CL genes and 31 Jm4CL genes, respectively. Based on phylogenetic relationship analysis, all 4CL genes were divided into three branches. WGD was the major duplication mode for 4CLs in two Juglans species. The phylogenic and collinearity analyses showed that the 4CLs were relatively conserved during evolution, but the gene structures varied widely. 4CLs promoter region contained multiply cis-acting elements related to phytohormones and stress responses. We found that Jr4CLs may be participated in the regulation of resistance to anthracnose. The expression level and some physiological of 4CLs were changed significantly after salt treatment. According to qRT-PCR results, positive regulation was found to be the main mode of regulation of 4CL genes after salt stress. Overall, J. mandshurica outperformed J. regia. Therefore, J. mandshurica can be used as a walnut rootstock to improve salt tolerance. Our results provide new understanding the potential functions of 4CL genes in stress tolerance, offer the theoretical genetic basis of walnut varieties adapted to salt stress, and provide an important reference for breeding cultivated walnuts for stress tolerance.

This document provides information about a new walnut cultivar called 'Xin Xiong' that was selected from indigenous individuals in Hotan County, China. The cultivar has unique characteristics, including long secondary male inflorescences that have ornamental value and can be used for studying walnut flower organs. 'Xin Xiong' has moderate tree vigor, a spreading growth habit, and yellowish-green branches. The document also includes detailed descriptions of the nuts, their weight, and the percentage of kernels. 'Xin Xiong' has high and stable yields and is suitable for cultivation in the oasis at the margin of Tarim Basin. The document highlights the importance of walnut genetic resources for future breeding approaches. [Extracted from the article]

The R2R3-MYB gene family, encoding plant transcriptional regulators, participates in many metabolic pathways of plant physiology and development, including flavonoid metabolism and anthocyanin synthesis. This study proceeded as follows: the JrR2R3-MYB gene family was analyzed genome-wide, and the family members were identified and characterized using the high-quality walnut reference genome "Chandler 2.0". All 204 JrR2R3-MYBs were established and categorized into 30 subgroups via phylogenetic analysis. JrR2R3-MYBs were unevenly distributed over 16 chromosomes. Most JrR2R3-MYBs had similar structures and conservative motifs. The cis-acting elements exhibit multiple functions of JrR2R3-MYBs such as light response, metabolite response, and stress response. We found that the expansion of JrR2R3-MYBs was mainly caused by WGD or segmental duplication events. Ka/Ks analysis indicated that these genes were in a state of negative purifying selection. Transcriptome results suggested that JrR2R3-MYBs were widely entangled in the process of walnut organ development and differentially expressed in different colored varieties of walnuts. Subsequently, we identified 17 differentially expressed JrR2R3-MYBs, 9 of which may regulate anthocyanin biosynthesis based on the results of a phylogenetic analysis. These genes were present in greater expression levels in 'Zijing' leaves than in 'Lvling' leaves, as revealed by the results of qRT-PCR experiments. These results contributed to the elucidation of the functions of JrR2R3-MYBs in walnut coloration. Collectively, this work provides a foundation for exploring the functional characteristics of the JrR2R3-MYBs in walnuts and improving the nutritional value and appearance quality of walnuts. [ABSTRACT FROM AUTHOR]

Persian walnut (Juglans regia) and Manchurian walnut (Juglans mandshurica) belong to Juglandaceae, which are vulnerable, temperate deciduous perennial trees with high economical, ecological, and industrial values. 4-Coumarate: CoA ligase (4CL) plays an essential function in plant development, growth, and stress. Walnut production is challenged by diverse stresses, such as salinity, drought, and diseases. However, the characteristics and expression levels of 4CL gene family in Juglans species resistance and under salt stress are unknown. Here, we identified 36 Jr4CL genes and 31 Jm4CL genes, respectively. Based on phylogenetic relationship analysis, all 4CL genes were divided into three branches. WGD was the major duplication mode for 4CLs in two Juglans species. The phylogenic and collinearity analyses showed that the 4CLs were relatively conserved during evolution, but the gene structures varied widely. 4CLs promoter region contained multiply cis-acting elements related to phytohormones and stress responses. We found that Jr4CLs may be participated in the regulation of resistance to anthracnose. The expression level and some physiological of 4CLs were changed significantly after salt treatment. According to qRT-PCR results, positive regulation was found to be the main mode of regulation of 4CL genes after salt stress. Overall, J. mandshurica outperformed J. regia. Therefore, J. mandshurica can be used as a walnut rootstock to improve salt tolerance. Our results provide new understanding the potential functions of 4CL genes in stress tolerance, offer the theoretical genetic basis of walnut varieties adapted to salt stress, and provide an important reference for breeding cultivated walnuts for stress tolerance. [ABSTRACT FROM AUTHOR]

Similar to other cropping systems, few walnut cultivars are used as scion in commercial production. Germplasm collections can be used to diversify cultivar options and hold potential for improving crop productivity, disease resistance and stress tolerance. In this study, we explored the anatomical and biochemical bases of photosynthetic capacity and response to water stress in 11 Juglans regia accessions in the U.S. department of agriculture, agricultural research service (USDA-ARS) National Clonal Germplasm. Net assimilation rate (An ) differed significantly among accessions and was greater in lower latitudes coincident with higher stomatal and mesophyll conductances, leaf thickness, mesophyll porosity, gas-phase diffusion, leaf nitrogen and lower leaf mass and stomatal density. High CO2 -saturated assimilation rates led to increases in An under diffusional and biochemical limitations. Greater An was found in lower-latitude accessions native to climates with more frost-free days, greater precipitation seasonality and lower temperature seasonality. As expected, water stress consistently impaired photosynthesis with the highest % reductions in lower-latitude accessions (A3, A5 and A9), which had the highest An under well-watered conditions. However, An for A3 and A5 remained among the highest under dehydration. J. regia accessions, which have leaf structural traits and biochemistry that enhance photosynthesis, could be used as commercial scions or breeding parents to enhance productivity.

Soil fungal communities are critical for forest ecosystem functions in the Central Hardwood Region (CHR) of the USA. This evaluation, which took place in 2022–2023, investigates the influence of Juglans nigra (BW, black walnut) and Quercus rubra (NRO, Northern red oak) on soil properties and fungal community structures across three CHR sites. The objectives of this study are to investigate how the fungal communities identified beneath J. nigra and Q. rubra serve to influence biodiversity and soil health within hardwood plantations. Soils from two locations in Indiana and one in Michigan were examined and assessed for variations in fungal composition and diversity. Soil fungal communities were characterized using Illumina high-throughput sequencing while multivariate analysis was applied to analyze patterns in these fungal communities. These data provided insights into how environment, location, and tree species affect fungal community structure. Results indicate that J. nigra soils exhibited higher carbon (0.36%, 1.02%, 0.72%), nitrogen (25%, 29%, 56%), and pH (0.46, 1.08, 1.54) levels than Q. rubra soils across all three sites and foster greater fungal diversity. Specifically, J. nigra was associated with increased Ascomycota diversity, whereas Q. rubra supported a higher prevalence of Basidiomycota. Basidiomycota were negatively correlated with carbon and pH, while Ascomycota showed positive correlations with these variables. These findings highlight how crucial it is to understand how different tree species influence fungal communities and, consequently, how they influence forest soil health. Our findings serve to improve forest management practices by emphasizing the importance of fungal communities in maintaining the function and resilience of an ecosystem. Our study underscores that grasping these specific interactions is essential for effective forest management, especially when considering how to use fungal communities to boost plant growth. This work focuses on hardwood plantations rather than either agricultural ecosystems, monocultures, or native forests, thus filling a gap in the current literature where many studies are limited to specific fungal groups such as mycorrhizae. In future research, it is important to examine a wider range of tree species. This will deepen our understanding of fungal community dynamics and their impact on maintaining healthy forest ecosystems. Our hardwood plantation focus also notes the potential for adaptive forest management as environmental conditions change.

Successful yield in orchards is the culmination of a series of events that start with plants entering dormancy with adequate energy reserves (non-structural carbohydrates; NSC). These NSC are responsible for the maintenance of activities during dormancy and extending onto the period of activeness. Using multi-year yield information and monthly NSC content in twigs, we show that high levels of carbohydrate in Prunus dulcis, Pistachio vera, and Juglans regia during the winter months are indeed associated with high yield, while high levels of the NSC in late summer often correlate with low yield. An evaluation of monthly NSC level importance on yield revealed that for P. dulcis high levels in February were a good predictor of yield and that low levels throughout summer were associated with high yield. In P. vera, high levels of NSC in December were best predictors of yield. J. regia exhibited peculiar patterns; while high pre-budbreak reserves were associated with high yields they only played a minor role in explaining crop, the most important months for predicting yields were June and July. Results suggest that NSC levels can serve as good predictors of orchard yield potential and should be monitored to inform orchard management.

Walnut, a woody plant, is regarded as having difficulty rooting when propagated by vegetative methods, such as cutting and layering. A layering experiment was conducted in 2018 and 2022. In 2018, some Juglans species, including J. regia L. seedling (JR), J. regia cv. Liaoning 1 (JR LN1), J. hopeinesis Hu seedling (JH), J. mandshurica Maxim seedling (JM), and J. nigra L. seedling (JN), were the mother plants. The specific research hypotheses were that own-rooted walnut propagule could be obtained through layering. the rooting capacity of different Juglans species would be different, and the rooting ability of JN would be the highest among the samplings. The results indicated that all of these species in the experiment could be rooted by etiolation and indole-3-butyric acid (IBA) treatment and that root occurrence was found 6 to 7 weeks after IBA treatment. The layers (shoots from the mother plant) on the seedlings of JR, JH, and JM obtained rooting percentages (RP) of 75.55%, 84.45%, and 86.67%, respectively, and root numbers (RNs) of 21.8, 42.8, and 38.8, respectively, after 20 days of etiolation and 1% IBA treatment. JR LN1 had difficulty rooting in equal conditions and had a RP of 31.11%. In 2022, JR LN1 was the only mother plant and the IBA concentration was increased to obtain satisfactory RP and RN. With the 4% and 8% IBA treatments, RPs of 88.9% and 93.3% and RNs of 40.3 and 27.7, respectively, were achieved. During the experiment, the RP, RN, root length (RL), and root diameter (RD), as well as the layer height (LH) and layer diameter (LD), were investigated and evaluated. Layers with low vigor were more likely to root, as shown by a nonparametric test conducted for the height and diameter of the layers of the rooting and nonrooting groups. A significantly negative correlation (r 5 20.548) was observed between RN and LH. Moreover, the quality of the best results of JR LN1 layering propagule and that with 'liaoning 1' 1-yearold seedling were compared. Our results provide more support for the possibility of vegetative propagation of walnut by layering and more information regarding the clonal cultivation of walnut trees and the own-rooted seedling establishment of walnut cultivars. [ABSTRACT FROM AUTHOR]

Multiplex polymerase chain reaction (PCR) of microsatellite loci allows for simultaneous amplification of two or more pairs of primers in a single PCR reaction; hence, it is cost and time effective. However, very few attempts have been reported in non-model species. In this study, by combining a genome-based de novo development and cross-species application approach, a multiplex PCR system comprising 5 PCR reactions of 33 microsatellites consisting of 26 novel genomic and 7 literature-sourced loci was tested for polymorphisms, cross-species transferability, and the ability to assess genetic diversity and population structure of three walnut species (Juglans spp.). We found that the genome-based approach is more efficient than other methods. An allelic ladder was developed for each locus to enhance consistent genotyping among laboratories. The population genetic analysis results showed that all 33 loci were successfully transferred across the three species, showing high polymorphism and a strong genetic structure. Hence, the multiplex PCR system is highly applicable in walnut species. Furthermore, we propose an efficient pipeline to characterize and genotype polymorphic microsatellite loci. The novel toolbox developed here will aid future ecology and evolution studies in walnut and could serve as a model for other plant species. [ABSTRACT FROM AUTHOR]

Walnut blight is a significant above-ground disease of walnuts caused by Xanthomonas arboricola pv. juglandis (Xaj). The secreted form of chorismate mutase (CM), a key enzyme of the shikimate pathway regulating plant immunity, is highly conserved between plant-associated beta and gamma proteobacteria including phytopathogens belonging to the Xanthomonadaceae family. To define its role in walnut blight disease, a dysfunctional mutant of chorismate mutase was created in a copper resistant strain Xaj417 (XajCM). Infections of immature walnut Juglans regia (Jr) fruit with XajCM were hypervirulent compared with infections with the wildtype Xaj417 strain. The in vitro growth rate, size and cellular morphology were similar between the wild-type and XajCM mutant strains, however the quantification of bacterial cells by dPCR within walnut hull tissues showed a 27% increase in XajCM seven days post-infection. To define the mechanism of hypervirulence, proteome analysis was conducted to compare walnut hull tissues inoculated with the wild type to those inoculated with the XajCM mutant strain. Proteome analysis revealed 3296 Jr proteins (five decreased and ten increased with FDR ≤ 0.05) and 676 Xaj417 proteins (235 increased in XajCM with FDR ≤ 0.05). Interestingly, the most abundant protein in Xaj was a polygalacturonase, while in Jr it was a polygalacturonase inhibitor. These results suggest that this secreted chorismate mutase may be an important virulence suppressor gene that regulates Xaj417 virulence response, allowing for improved bacterial survival in the plant tissues.

The Asian walnut moth, Garella musculana (Erschov, 1874) (Lepidoptera, Nolidae) is an alien pest originating from Central Asia and is now spreading in Europe, attacking walnut trees. In this study, we updated the current distribution of G. musculana, focusing on the Italian range, where it was reported for the first time in 2021. Field surveys showed an extensive distribution of G. musculana in northern Italy, particularly in the Veneto Region. In this area, the Asian walnut moth developed on English and black walnut, attacking almost exclusively tree shoots. Based on current distribution data, further investigations are required in the nearby regions as well as in those that were less surveyed. Lastly, it is imperative to conduct more studies on insect biology and the impact on walnut production.

Soil salinization has become one of the major problems that threaten the ecological environment. The aim of this study is to explore the mechanism of salt tolerance of hybrid walnuts (Juglans major × Juglans regia) under long-term salt stress through the dynamic changes of growth, physiological and biochemical characteristics, and anatomical structure. Our findings indicate that (1) salt stress inhibited seedling height and ground diameter increase, and (2) with increasing salt concentration, relative water content (RWC) decreased, and proline (Pro) and soluble sugar (SS) content increased. The Pro content reached a maximum of 549.64 μg/g on the 42nd day. The increase in superoxide dismutase (SOD) activity (46.80–117.16%), ascorbate peroxidase (APX) activity, total flavonoid content (TFC), and total phenol content (TPC) under salt stress reduced the accumulation of malondialdehyde (MDA). (3) Increasing salt concentration led to increases and subsequent decreases in the thickness of palisade tissues, spongy tissues, leaves, and leaf vascular bundle diameter. Upper and lower skin thickness, root periderm thickness, root diameter, root cortex thickness, and root vascular bundle diameter showed different patterns of change at varying stress concentrations and durations. Overall, the study concluded that salt stress enhanced the antireactive oxygen system, increased levels of osmotic regulators, and low salt concentrations promoted leaf and root anatomy, but that under long-term exposure to high salt levels, leaf anatomy was severely damaged. For the first time, this study combined the anatomical structure of the vegetative organ of hybrid walnut with physiology and biochemistry, which is of great significance for addressing the challenge of walnut salt stress and expanding the planting area.

Walnut blight is a significant above-ground disease of walnuts caused by Xanthomonas arboricola pv. juglandis (Xaj). The secreted form of chorismate mutase (CM), a key enzyme of the shikimate pathway regulating plant immunity, is highly conserved between plant-associated beta and gamma proteobacteria including phytopathogens belonging to the Xanthomonadaceae family. To define its role in walnut blight disease, a dysfunctional mutant of chorismate mutase was created in a copper resistant strain Xaj417 (XajCM). Infections of immature walnut Juglans regia (Jr) fruit with XajCM were hypervirulent compared with infections with the wildtype Xaj417 strain. The in vitro growth rate, size and cellular morphology were similar between the wild-type and XajCM mutant strains, however the quantification of bacterial cells by dPCR within walnut hull tissues showed a 27% increase in XajCM seven days post-infection. To define the mechanism of hypervirulence, proteome analysis was conducted to compare walnut hull tissues inoculated with the wild type to those inoculated with the XajCM mutant strain. Proteome analysis revealed 3296 Jr proteins (five decreased and ten increased with FDR ≤ 0.05) and 676 Xaj417 proteins (235 increased in XajCM with FDR ≤ 0.05). Interestingly, the most abundant protein in Xaj was a polygalacturonase, while in Jr it was a polygalacturonase inhibitor. These results suggest that this secreted chorismate mutase may be an important virulence suppressor gene that regulates Xaj417 virulence response, allowing for improved bacterial survival in the plant tissues.

Walnut pellicle color is a key quality attribute that drives consumer preference and walnut sales. For the first time a high-throughput, computer vision-based phenotyping platform using a custom algorithm to quantitatively score each walnut pellicle in L* a* b* color space was deployed at large-scale. This was compared to traditional qualitative scoring by eye and was used to dissect the genetics of pellicle pigmentation. Progeny from both a bi-parental population of 168 trees ('Chandler' × 'Idaho') and a genome-wide association (GWAS) with 528 trees of the UC Davis Walnut Improvement Program were analyzed. Color phenotypes were found to have overlapping regions in the 'Chandler' genetic map on Chr01 suggesting complex genetic control. In the GWAS population, multiple, small effect QTL across Chr01, Chr07, Chr08, Chr09, Chr10, Chr12 and Chr13 were discovered. Marker trait associations were co-localized with QTL mapping on Chr01, Chr10, Chr14, and Chr16. Putative candidate genes controlling walnut pellicle pigmentation were postulated.

BACKGROUND:Unravelling the genetic architecture of agronomic traits in walnut such as budbreak date and bearing habit, is crucial for climate change adaptation and yield improvement. A Genome-Wide Association Study (GWAS) using multi-locus models was conducted in a panel of 170 walnut accessions genotyped using the Axiom™ J. regia 700 K SNP array, with phenological data from 2018, 2019 and legacy data. These accessions come from the INRAE walnut germplasm collection which is the result of important prospecting work performed in many countries around the world. In parallel, an F1 progeny of 78 individuals segregating for phenology-related traits, was genotyped with the same array and phenotyped for the same traits, to construct linkage maps and perform Quantitative Trait Loci (QTLs) detection. RESULTS:Using GWAS, we found strong associations of SNPs located at the beginning of chromosome 1 with both budbreak and female flowering dates. These findings were supported by QTLs detected in the same genomic region. Highly significant associated SNPs were also detected using GWAS for heterodichogamy and lateral bearing habit, both on chromosome 11. We developed a Kompetitive Allele Specific PCR (KASP) marker for budbreak date in walnut, and validated it using plant material from the Walnut Improvement Program of the University of California, Davis, demonstrating its effectiveness for marker-assisted selection in Persian walnut. We found several candidate genes involved in flowering events in walnut, including a gene related to heterodichogamy encoding a sugar catabolism enzyme and a cell division related gene linked to female flowering date. CONCLUSIONS:This study enhances knowledge of the genetic architecture of important agronomic traits related to male and female flowering processes and lateral bearing in walnut. The new marker available for budbreak date, one of the most important traits for good fruiting, will facilitate the selection and development of new walnut cultivars suitable for specific climates.

The interaction between the plant host, walnut (Juglans regia; Jr), and a deadly pathogen (Xanthomonas arboricola pv. juglandis 417; Xaj) can lead to walnut bacterial blight (WB), which depletes walnut productivity by degrading the nut quality. Here, we dissect this pathosystem using tandem mass tag quantitative proteomics. Walnut hull tissues inoculated with Xaj were compared to mock-inoculated tissues, and 3972 proteins were identified, of which 3296 are from Jr and 676 from Xaj. Proteins with differential abundance include oxidoreductases, proteases, and enzymes involved in energy metabolism and amino acid interconversion pathways. Defense responses and plant hormone biosynthesis were also increased. Xaj proteins detected in infected tissues demonstrate its ability to adapt to the host microenvironment, limiting iron availability, coping with copper toxicity, and maintaining energy and intermediary metabolism. Secreted proteases and extracellular secretion apparatus such as type IV pilus for twitching motility and type III secretion effectors indicate putative factors recognized by the host. Taken together, these results suggest intense degradation processes, oxidative stress, and general arrest of the biosynthetic metabolism in infected nuts. Our results provide insights into molecular mechanisms and highlight potential molecular tools for early detection and disease control strategies.

The release of the first reference genome of walnut (Juglans regia L.) enabled many achievements in the characterization of walnut genetic and functional variation. However, it is highly fragmented, preventing the integration of genetic, transcriptomic, and proteomic information to fully elucidate walnut biological processes. Here, we report the new chromosome-scale assembly of the walnut reference genome (Chandler v2.0) obtained by combining Oxford Nanopore long-read sequencing with chromosome conformation capture (Hi-C) technology. Relative to the previous reference genome, the new assembly features an 84.4-fold increase in N50 size, with the 16 chromosomal pseudomolecules assembled and representing 95% of its total length. Using full-length transcripts from single-molecule real-time sequencing, we predicted 37,554 gene models, with a mean gene length higher than the previous gene annotations. Most of the new protein-coding genes (90%) present both start and stop codons, which represents a significant improvement compared with Chandler v1.0 (only 48%). We then tested the potential impact of the new chromosome-level genome on different areas of walnut research. By studying the proteome changes occurring during male flower development, we observed that the virtual proteome obtained from Chandler v2.0 presents fewer artifacts than the previous reference genome, enabling the identification of a new potential pollen allergen in walnut. Also, the new chromosome-scale genome facilitates in-depth studies of intraspecies genetic diversity by revealing previously undetected autozygous regions in Chandler, likely resulting from inbreeding, and 195 genomic regions highly differentiated between Western and Eastern walnut cultivars. Overall, Chandler v2.0 will serve as a valuable resource to better understand and explore walnut biology.

Abstract Background Late Embryogenesis Abundant (LEA) proteins are a class of proteins associated with plant stress resistance. Two Juglans species, Juglans regia and J. mandshurica, are both diploid (2n = 32), monoecious perennial economic tree species with high edible, pharmaceutical, and timber value. The identification, characterization, and expression patterns of LEA proteins in J. regia and its wild relative, J. mandshurica, would not only provide the genetic basis of this gene family, but it would also supply clues for further studies of the evolution and regulating mechanisms of LEA proteins in other tree species. Results In this study, we identified 25 and 20 members of the LEA gene family in Juglans regia and its wild relative, Juglans mandshurica, respectively. The results of phylogenetic analysis showed that the LEA members were divided into eight main subgroups. Predictions of their physicochemical properties showed the variable characteristics of LEA proteins, and the subcellular localization analysis indicated that most LEA proteins are localized in the nucleus. Chromosomal localization analysis and gene replication pattern prediction indicated that WGD is the predominant duplication mode of LEA genes. The results of the comparative analysis indicated a high level of collinearity between the two Juglans species. Analysis of cis-acting elements indicated that LEA genes had a relatively wide range of responses to abiotic stresses and phytohormonal processes, particularly in two phytohormones, methyl jasmonate and abscisic acid. Transcriptome profiling and qRT-PCR experiments showed that JrLEAs are commonly expressed in leaves, green husks, and male and female flowers, and most JmLEAs are more highly expressed in male flowers. We also hypothesized that JrLEAs are involved in the process of anthracnose resistance. Anthracnose-resistant varieties of JrLEAs presented relatively high expression levels at later stages. Conclusion In this study, we provide a theoretical basis for the functional study of LEA genes in J. regia and J. mandshurica. Analysis of cis-acting elements and gene expression indicated that JrLEAs and JmLEAs play important roles in resistance to biotic stresses in these species.

The Asian walnut moth, Garella musculana (Erschov, 1874) (Lepidoptera, Nolidae) is an alien pest originating from Central Asia and is now spreading in Europe, attacking walnut trees. In this study, we updated the current distribution of G. musculana, focusing on the Italian range, where it was reported for the first time in 2021. Field surveys showed an extensive distribution of G. musculana in northern Italy, particularly in the Veneto Region. In this area, the Asian walnut moth developed on English and black walnut, attacking almost exclusively tree shoots. Based on current distribution data, further investigations are required in the nearby regions as well as in those that were less surveyed. Lastly, it is imperative to conduct more studies on insect biology and the impact on walnut production. [ABSTRACT FROM AUTHOR]

Despite non-structural carbohydrate (NSC) importance for tree productivity and resilience, little is known about their seasonal regulations and trade-off with growth and reproduction. We characterize the seasonal dynamics of NSC in relation to the aboveground phenology and temporal growth patterns of three deciduous Mediterranean species: almond (Prunus dulcis (Mill.) D. A. Webb), walnut (Juglans regia L.) and pistachio (Pistacia vera L.). Seasonal dynamics of NSC were synchronous between wood tissues from trunk, branches and twigs. Almond had almost identical levels and patterns of NSC variation in twigs, branches and trunks whereas pistachio and walnut exhibited clear concentration differences among plant parts whereby twigs had the highest and most variable NSC concentration, followed by branches and then trunk. While phenology had a significant influence on NSC seasonal trends, there was no clear trade-off between NSC storage and growth suggesting that both were similarly strong sinks for NSC. A temporal trade-off observed at the seasonal scale was influenced by the phenology of the species. We propose that late senescing species experience C allocation trade-off at the end of the growing season because of C-limiting thermal conditions and priority allocation to storage in order to survive winter.

Walnut shell suture strength directly impacts the ability to maintain shell integrity during harvest and processing, susceptibility to insect damage and other contamination, and the proportion of kernel halves recovered during cracking. Suture strength is therefore an important breeding objective. Here, two methods of phenotyping this trait were investigated: 1) traditional, qualitative and rather subjective scoring on an interval scale by human observers, and; 2) quantitative and continuous measurements captured by a texturometer. The aim of this work was to increase the accuracy of suture strength phenotyping and to then apply two mapping approaches, quantitative trait loci (QTL) mapping and genome wide association (GWAS) models, in order to dissect the genetic basis of the walnut suture trait. Using data collected on trees within the UC Davis Walnut Improvement Program (n = 464), the genetic correlation between the texturometer method and qualitatively scored method was high (0.826). Narrow sense heritability calculated using quantitative measurements was 0.82. A major QTL for suture strength was detected on LG05, explaining 34% of the phenotypic variation; additionally, two minor QTLs were identified on LG01 and LG11. All three QTLs were confirmed with GWAS on corresponding chromosomes. The findings reported in this study are relevant for application towards a molecular breeding program in walnut.

The interaction between the plant host, walnut (Juglans regia; Jr), and a deadly pathogen (Xanthomonas arboricola pv. juglandis 417; Xaj) can lead to walnut bacterial blight (WB), which depletes walnut productivity by degrading the nut quality. Here, we dissect this pathosystem using tandem mass tag quantitative proteomics. Walnut hull tissues inoculated with Xaj were compared to mock-inoculated tissues, and 3972 proteins were identified, of which 3296 are from Jr and 676 from Xaj. Proteins with differential abundance include oxidoreductases, proteases, and enzymes involved in energy metabolism and amino acid interconversion pathways. Defense responses and plant hormone biosynthesis were also increased. Xaj proteins detected in infected tissues demonstrate its ability to adapt to the host microenvironment, limiting iron availability, coping with copper toxicity, and maintaining energy and intermediary metabolism. Secreted proteases and extracellular secretion apparatus such as type IV pilus for twitching motility and type III secretion effectors indicate putative factors recognized by the host. Taken together, these results suggest intense degradation processes, oxidative stress, and general arrest of the biosynthetic metabolism in infected nuts. Our results provide insights into molecular mechanisms and highlight potential molecular tools for early detection and disease control strategies.

Following photosynthesis, sucrose is translocated to sink organs, where it provides the primary source of carbon and energy to sustain plant growth and development. Sugar transporters from the SWEET (sugar will eventually be exported transporter) family are rate-limiting factors that mediate sucrose transport across concentration gradients, sustain yields, and participate in reproductive development, plant senescence, stress responses, as well as support plant-pathogen interaction, the focus of this study. We identified 25 SWEET genes in the walnut genome and distinguished each by its individual gene structure and pattern of expression in different walnut tissues. Their chromosomal locations, cis-acting motifs within their 5' regulatory elements, and phylogenetic relationship patterns provided the first comprehensive analysis of the SWEET gene family of sugar transporters in walnut. This family is divided into four clades, the analysis of which suggests duplication and expansion of the SWEET gene family in Juglans regia. In addition, tissue-specific gene expression signatures suggest diverse possible functions for JrSWEET genes. Although these are commonly used by pathogens to harness sugar products from their plant hosts, little was known about their role during Xanthomonasarboricola pv. juglandis (Xaj) infection. We monitored the expression profiles of the JrSWEET genes in different tissues of "Chandler" walnuts when challenged with pathogen Xaj417 and concluded that SWEET-mediated sugar translocation from the host is not a trigger for walnut blight disease development. This may be directly related to the absence of type III secretion system-dependent transcription activator-like effectors (TALEs) in Xaj417, which suggests different strategies are employed by this pathogen to promote susceptibility to this major aboveground disease of walnuts.

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Walnut consumption can provide both vascular and metabolic health benefits, and walnut-induced changes in lipoprotein particle chemical payloads may be responsible for these health benefits. To explore this possibility with a focus on metabolic health, this study investigated the impact of walnut consumption on lipoprotein lipid composition and changes in LDL anti-inflammatory properties, as reported by inflamed adipocyte. Hypercholesterolemic, postmenopausal females were treated with 40 g/day (i.e., 1.6 servings/day; n=15) of walnuts for 4 weeks. Fatty acids and their oxygenated metabolites, i.e., oxylipins, were quantified in isolated lipoproteins. Human primary adipocytes were exposed to LDL and TNFα-stimulated adipokine production was measured. Walnut treatment elevated α-linolenic acid and its epoxides in all lipoproteins and depleted mid-chain alcohols in VLDL and LDL, but not HDL. Walnuts also reduced TNFα-induced diabetic adipocyte production of IL-6 (-48%, P=.0006) and IL-8 (-30%, P=.01), changes inversely correlated with levels of α-linolenic acid-derived epoxides but not α-linolenic acid itself. In conclusion, modest walnut consumption can alter lipoprotein lipid profiles and enhance their ability to inhibit TNFα-dependent pro-inflammatory responses in human diabetic primary adipocytes. Moreover, this study suggests the oxylipins, rather than the parent fatty acids, mediate LDL action of adipocytes.

Over the last 20 years, global production of Persian walnut (Juglans regia L.) has grown enormously, likely reflecting increased consumption due to its numerous benefits to human health. However, advances in genome-wide association (GWA) studies and genomic selection (GS) for agronomically important traits in walnut remain limited due to the lack of powerful genomic tools. Here, we present the development and validation of a high-density 700K single nucleotide polymorphism (SNP) array in Persian walnut. Over 609K high-quality SNPs have been thoroughly selected from a set of 9.6 m genome-wide variants, previously identified from the high-depth re-sequencing of 27 founders of the Walnut Improvement Program (WIP) of University of California, Davis. To validate the effectiveness of the array, we genotyped a collection of 1284 walnut trees, including 1167 progeny of 48 WIP families and 26 walnut cultivars. More than half of the SNPs (55.7%) fell in the highest quality class of 'Poly High Resolution' (PHR) polymorphisms, which were used to assess the WIP pedigree integrity. We identified 151 new parent-offspring relationships, all confirmed with the Mendelian inheritance test. In addition, we explored the genetic variability among cultivars of different origin, revealing how the varieties from Europe and California were differentiated from Asian accessions. Both the reconstruction of the WIP pedigree and population structure analysis confirmed the effectiveness of the Applied Biosystems™ Axiom™ J. regia 700K SNP array, which initiates a novel genomic and advanced phase in walnut genetics and breeding.

Multiplex polymerase chain reaction (PCR) of microsatellite loci allows for simultaneous amplification of two or more pairs of primers in a single PCR reaction; hence, it is cost and time effective. However, very few attempts have been reported in non-model species. In this study, by combining a genome-based de novo development and cross-species application approach, a multiplex PCR system comprising 5 PCR reactions of 33 microsatellites consisting of 26 novel genomic and 7 literature-sourced loci was tested for polymorphisms, cross-species transferability, and the ability to assess genetic diversity and population structure of three walnut species (Juglans spp.). We found that the genome-based approach is more efficient than other methods. An allelic ladder was developed for each locus to enhance consistent genotyping among laboratories. The population genetic analysis results showed that all 33 loci were successfully transferred across the three species, showing high polymorphism and a strong genetic structure. Hence, the multiplex PCR system is highly applicable in walnut species. Furthermore, we propose an efficient pipeline to characterize and genotype polymorphic microsatellite loci. The novel toolbox developed here will aid future ecology and evolution studies in walnut and could serve as a model for other plant species.

BackgroundMultiple studies have shown a Mediterranean diet, characterized by their olive oil and nut consumption, to be correlated with lower depression risk.ObjectiveTo examine whether part of this reduced risk in the United States is attributable to walnut consumption, we analyzed data on walnut consumption and depression scores from the National Health and Nutrition Examination Survey (NHANES).MethodsNHANES survey data for 2005 through 2014 was pooled for adults with 24 h recall dietary data. Depression scores were based on PHQ-9 self-report responses. A total of 26,656 participants were characterized as reporting the consumption of walnuts with high certainty, walnuts with other nuts, other nuts, or no nuts.ResultsAfter an adjustment for covariates, walnut consumers showed lower depression scores compared to non-nut consumers. The least square mean for total depression score was 26% lower for walnut with high certainty consumers than for non-nut consumers (p < 0.0001), and the association was stronger among women (32%, p < 0.0001) than men (21%, p = 0.05). The significant contributors to this difference were due to walnut consumers reporting greater interest in doing things (p = 0.003), less hopelessness (p = 0.02), and feeling more energetic (p = 0.05) than non-nut consumers. Non-nut consumers were more likely to have trouble concentrating (p = 0.02), to feel they were moving or speaking abnormally (p = 0.03), and to have thought they were better off dead (p = 0.002).ConclusionsDepression scores were significantly lower among nut consumers and particularly walnut consumers as compared to non-nut consumers. After controlling for potential covariates, walnut users had scores significantly lower than other nut consumers. The difference was strongest among women, who are more likely than men to report higher depression scores.

Genomic analysis in Juglans (walnuts) is expected to transform the breeding and agricultural production of both nuts and lumber. To that end, we report here the determination of reference sequences for six additional relatives of Juglans regia: Juglans sigillata (also from section Dioscaryon), Juglans nigra, Juglans microcarpa, Juglans hindsii (from section Rhysocaryon), Juglans cathayensis (from section Cardiocaryon), and the closely related Pterocarya stenoptera While these are 'draft' genomes, ranging in size between 640Mbp and 990Mbp, their contiguities and accuracies can support powerful annotations of genomic variation that are often the foundation of new avenues of research and breeding. We annotated nucleotide divergence and synteny by creating complete pairwise alignments of each reference genome to the remaining six. In addition, we have re-sequenced a sample of accessions from four Juglans species (including regia). The variation discovered in these surveys comprises a critical resource for experimentation and breeding, as well as a solid complementary annotation. To demonstrate the potential of these resources the structural and sequence variation in and around the polyphenol oxidase loci, PPO1 and PPO2 were investigated. As reported for other seed crops variation in this gene is implicated in the domestication of walnuts. The apparently Juglandaceae specific PPO1 duplicate shows accelerated divergence and an excess of amino acid replacement on the lineage leading to accessions of the domesticated nut crop species, Juglans regia and sigillata.

Background: Late Embryogenesis Abundant (LEA) proteins are a class of proteins associated with plant stress resistance. Two Juglans species, Juglans regia and J. mandshurica, are both diploid (2n = 32), monoecious perennial economic tree species with high edible, pharmaceutical, and timber value. The identification, characterization, and expression patterns of LEA proteins in J. regia and its wild relative, J. mandshurica, would not only provide the genetic basis of this gene family, but it would also supply clues for further studies of the evolution and regulating mechanisms of LEA proteins in other tree species. Results: In this study, we identified 25 and 20 members of the LEA gene family in Juglans regia and its wild relative, Juglans mandshurica, respectively. The results of phylogenetic analysis showed that the LEA members were divided into eight main subgroups. Predictions of their physicochemical properties showed the variable characteristics of LEA proteins, and the subcellular localization analysis indicated that most LEA proteins are localized in the nucleus. Chromosomal localization analysis and gene replication pattern prediction indicated that WGD is the predominant duplication mode of LEA genes. The results of the comparative analysis indicated a high level of collinearity between the two Juglans species. Analysis of cis-acting elements indicated that LEA genes had a relatively wide range of responses to abiotic stresses and phytohormonal processes, particularly in two phytohormones, methyl jasmonate and abscisic acid. Transcriptome profiling and qRT-PCR experiments showed that JrLEAs are commonly expressed in leaves, green husks, and male and female flowers, and most JmLEAs are more highly expressed in male flowers. We also hypothesized that JrLEAs are involved in the process of anthracnose resistance. Anthracnose-resistant varieties of JrLEAs presented relatively high expression levels at later stages. Conclusion: In this study, we provide a theoretical basis for the functional study of LEA genes in J. regia and J. mandshurica. Analysis of cis-acting elements and gene expression indicated that JrLEAs and JmLEAs play important roles in resistance to biotic stresses in these species. [ABSTRACT FROM AUTHOR]

Eastern black walnut (Juglans nigra L.), one of the most valuable timber and veneer trees in North America, provides nut shells with unique industrial uses and nut kernels with distinctive culinary attributes. A mature F1 full-sib progeny orchard of 248 individuals from the cross of two eastern black walnut cultivars provides a long-term resource for discovering genetic mechanisms controlling life history, quality traits, and stress resistance. The genetic linkage map, constructed with 356 single nucleotide polymorphism (SNP) markers and 62 expressed sequence tag simple sequence repeats (EST-SSRs), is 1645.7 cM in length, distributed across the expected 16 linkage groups. In this first application of QTL mapping in J. nigra, we report QTL for budbreak, peak pistillate bloom, peak staminate bloom, and heterodichogamy. A dominant major QTL for heterodichogamy is reported, the sequence for which is syntenic with the heterodichogamy QTL on chromosome 11 of Persian walnut (J. regia L.). The mapping population parents are both protogynous, and segregation suggests a Mendelian component, with a 3:1-like inheritance pattern from heterozygous parents. Mapping the sequenced EST-SSR markers to the J. regia "Chandler" V2.0 genome sequence revealed evidence for collinearity and structural changes on two of the sixteen chromosomes. The inclusion of sequenced EST-SSR markers enables the direct comparison of this and subsequent J. nigra maps and other Juglandaceae genetic maps. This investigation initiates long-term QTL detection studies for quality and stress resistance traits in black walnut. [ABSTRACT FROM AUTHOR]

Walnuts are among the commonly consumed oil nuts in the world. The nuts typically contain 40–70% oil, and are a rich source of fatty acids, phenolics, phytosterols, tocopherols and carotenoids. The levels of unsaturated fatty acids and tocopherols in walnuts were significantly higher than those of many other widely consumed oil nuts. The presence of phytochemicals in walnuts gives rise to multiple bioactivities, such as antioxidant, anti-inflammatory properties, and cholesterol lowering activity. The addition of walnut to a diet could enable delay of the onset of neurodegenerative disorders, protection against the development of certain cancers. Agronomic practices, environmental and climatic conditions, geographic locations, harvesting and postharvest handling practices have impacts upon phytochemical levels in walnuts. Abbreviations: LDL: low-density lipoprotein; HDL: high-density lipoprotein; GC-FID: gas chromatography-flame ionization detector; GC-MS: gas chromatography-mass spectrometry; HPLC-DAD: high-performance liquid chromatography-diode array detector; LC-MS/MS: liquid chromatography-tandem mass spectrometry; CE-MS: capillary electrophoresis–mass spectrometry; NMR: nuclear magnetic resonance; Aβ: amyloid beta protein; ROS: reactive oxygen species; DCF: dichlorofluorescein; DCFH-DA: dichlorofluorescin diacetate; SOD: superoxide dismutase; CAT: catalase; GR: glutathione reductase; GPx: glutathione peroxidase; GST: glutathione S-transferase; GSH: glutathione; LDH: lactate dehydrogenase; VCAM-1: vascular cell adhesion molecule: ICAM-1: intracellular adhesion molecule; TNF-α: tumor necrosis factor α; IL-6: interleukin 6; NF-κB: nuclear factor kappa light chain enhancer of activated B cells; IL-1β: interleukin 1β; IL-8: interleukin 8; IL-10: interleukin 10; MCP-1: monocyte chemoattractant protein 1; mTOR: mammalian target of rapamycin; ATG7: autophagy related 7; MAP1BLC3: microtubule-associated proteins 1A/1B light chain 3; PMA: phorbol 12-myristate 13-acetate; LPS: lipopolysaccharide; iNOS: inducible nitric oxide synthase; COX-2: cyclooxygenase 2; IGF-1: plasma insulin-like growth factor 1; GSTT2: glutathione S-transferase theta 2; GSK3β: glycogen synthase kinase 3 beta; ALA: α-linolenic acid; CD133: CD133 antigen; CD44: CD44 antigen; DLK1: Protein delta homolog 1; EPA: eicosapentaenoic acid; DHA: docosahexaenoic acid; Bcl-2: B-cell lymphoma 2; IL-1α: interleukin 1α; MAO-B: monoamine oxidase B; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; MPP+: 1-methyl-4-phenylpyridinium; BLM: bleomycin; CSE: cigarette smoke emission. [ABSTRACT FROM AUTHOR]

Biochar has been used to accelerate heating profiles during composting by increasing oxygenation, which could also reduce microbial pathogens. However, the antimicrobial inactivation of foodborne pathogens in compost, by amending with biochar without increased heating profiles, has not been evaluated. In this study, we examined the ability of biochar to inactivate E. coli O157:H7 (EC) in fresh dairy manure compost by amending with one of four types of biochar. Two slow pyrolysis biochars (high-temperature walnut hull biochar [HTWHB], and walnut hull cyclone biochar [WHCB]), and two fast pyrolysis biochars were examined. Compost with 8.1 log CFU/g of EC + 10% amended biochar was held at 22 °C and analyzed for EC weekly. The control treatment sustained ca. 8.7 log CFU of EC through week 7; however, the bacterium was not detected by direct plating in WHCB compost (below the detection limit) by day 7, through the entire 49 days (which may be attributed to increased compost alkalinity [i.e., pH 10.76]). Populations of EC in compost supplemented with 10% of the three other biochars sustained EC populations ≥9.2 log through the balance of the study. The four biochars were further tested in soil at 17% moisture to determine if concentrations as low as 3.5% could inactivate EC. When 3.5% HTWHB was added to soil, populations were 5.1 log CFU lower than when 10% of the same biochar was amended into dairy compost by week 3. This may indicate that alkaline biochar, amended into lower moisture, soil may be more biocidal than when alkaline biochar is added to high moisture manure compost. The current study demonstrates that highly alkaline walnut hull cyclone biochar is capable of reducing up to 8 log of EC in high moisture fresh compost in only 7 days, while as little as 3.5% alkaline WHCB in 17% moisture soil can reduce 6.7 log of EC in only 14 days. These results may assist farmers in amending compost, manure, cattle feedlots, or soil with biochar to reduce EC, and potentially other pathogens (e.g., Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes), with the goal of reducing the dissemination of human bacterial pathogens to meat, poultry, and fresh produce., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Inc.)

Objectives: To observe the clinical effect on perimenopausal dry eye disease treated with He's fire needle therapy combined with walnut-shell moxibustion., Methods: A total of 60 patients with perimenopausal dry eye disease were randomized into an observation group (30 cases) and a control group (30 cases). In the observation group, the pricking technique with fire needle was operated at bilateral Naokong (GB19), Feishu (BL13), Xinshu (BL15), Ganshu (BL18), Pishu (BL20), Shenshu (BL23), 3 times a week, for 4 weeks, and the walnut-shell moxibustion was combined, 30 minutes each time, 3 times a week, for 4 weeks. In the control group, hyaluronic acid sodium eye drops were used, one drop each time, 3 times a day, for 4 weeks. Before and after treatment, the scores of the questionnaire for dry eye disease in China, the result of lacrimal secretion test (Schimer Ⅰ test, SⅠT) and tear film break-up time(BUT), as well as the modified Kupperman score were evaluated in the two groups. The clinical effect was compared between the two groups., Results: After 4 weeks of treatment, the results of SⅠT and BUT increased ( P <0.001), while the scores of the questionnaire for dry eye disease in China, the modified Kupperman score decreased ( P <0.001) in the two groups when compared with the baseline. In the observation group, the result of SⅠT and BUT were higher than those of the control group ( P< 0.05), while the scores of the questionnaire for dry eye disease in China, and the modified Kupperman score was lower ( P <0.05). The marked effective rate (30/60, 50.0%) in the observation group was better than that (10/60, 16.7%) of the control group ( P <0.05)., Conclusions: In treatment of perimenopausal dry eye disease, the combined therapy of fire needling and walnut-shell moxibustion obtains the superior effect to hyaluronic acid sodium eye drops. This combined therapy can effectively relieve the eye symptoms, increase the secretion of tears, prolong BUT, and ameliorate systemic symptoms.

BackgroundDietary strategies that help patients adhere to a weight reduction diet may increase the likelihood of weight loss maintenance and improved long-term health outcomes. Regular nut consumption has been associated with better weight management and less adiposity. The objective of this study was to compare the effects of a walnut-enriched reduced-energy diet to a standard reduced-energy-density diet on weight, cardiovascular disease risk factors, and satiety.MethodsOverweight and obese men and women (n = 100) were randomly assigned to a standard reduced-energy-density diet or a walnut-enriched (15% of energy) reduced-energy diet in the context of a behavioral weight loss intervention. Measurements were obtained at baseline and 3- and 6-month clinic visits. Participants rated hunger, fullness and anticipated prospective consumption at 3 time points during the intervention. Body measurements, blood pressure, physical activity, lipids, tocopherols and fatty acids were analyzed using repeated measures mixed models.ResultsBoth study groups reduced body weight, body mass index and waist circumference (time effect p

Regular nut consumption is associated with lower adiposity and reduced weight gain in adulthood. Walnut feeding studies have observed minimal effect on body weight despite potential additional energy intake. Several mechanisms may explain why consuming nuts promotes weight control, including increased early phase satiety, possibly reflected in postprandial response of gastrointestinal and pancreatic peptides hypothesized to affect appetite. The purpose of this study was to compare postprandial insulin, glucagon and gastrointestinal peptide response and satiety following a meal with ∼54% of energy from walnuts or cream cheese, using a within-subject crossover study design in overweight/obese adults (N = 28). Sixty minutes after the walnut-containing meal, glucagon-like peptide-1 was lower than after the reference meal (p=0.0433), and peptide YY, cholecystokinin and ghrelin did not differ after the two meals. Sixty and 120 min after the walnut-containing meal, pancreatic polypeptide (p = 0.0014 and p = 0.0002) and glucose-dependent insulinotropic peptide (p

BackgroundObesity is a risk factor for postmenopausal breast cancer incidence and premenopausal and postmenopausal breast cancer mortality, which may be explained by several metabolic and hormonal factors (sex hormones, insulin resistance, and inflammation) that are biologically related. Differential effects of dietary composition on weight loss and these metabolic factors may occur in insulin-sensitive vs. insulin-resistant obese women.ObjectiveTo examine the effect of diet composition on weight loss and metabolic, hormonal and inflammatory factors in overweight/obese women stratified by insulin resistance status in a 1-year weight loss intervention.Methods and resultsNondiabetic women who were overweight/obese (n=245) were randomly assigned to a lower fat (20% energy), higher carbohydrate (65% energy) diet; a lower carbohydrate (45% energy), higher fat (35% energy) diet; or a walnut-rich (18% energy), higher fat (35% energy), lower carbohydrate (45% energy) diet. All groups lost weight at follow-up (P