Visible light-responsive polylactic acid@pullulan-chitosan/homojunction g-C3N4 bilayer antimicrobial films for fruit preservation.

The development of novel packaging materials with antimicrobial properties is crucial in preventing the microbial-induced spoilage of fruits, vegetables, and foodborne illnesses. In this study, homojunction g-C3N4 (HCN) photocatalysts with excellent photocatalytic performance were incorporated into a matrix consisting of pullulan/chitosan (Pul/CS). These photocatalysts were then electrostatically spun onto polylactic acid (PLA) films to fabricate PLA@Pul/CS/HCN nanofibrous composite films. The design of the bilayer films aimed to combine the physical properties of PLA film with the excellent antibacterial properties of nanofiber films, thereby achieving synergistic advantages. The incorporation of the HCN photocatalysts resulted in enhanced hydrophobicity, barrier function, and mechanical properties of the composite films. Under visible light irradiation, the PLA@Pul/CS/HCN films exhibited approximately 3.43 log and 3.11 log reductions of Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA), respectively, within 2 h. The excellent antimicrobial performance could be attributed to the synergistic effect of CS and the release of reactive oxygen species (ROS) from HCN. Moreover, the strawberries packaged in the PLA@Pul/CS/HCN film demonstrated diminished quality degradation and a prolonged shelf life following visible light irradiation treatment. This study will provide new insights into the exploration of safe and efficient antimicrobial food packaging.

Antimicrobial carbon dots/pectin-based hydrogel for promoting healing processes in multidrug-resistant bacteria-infected wounds.

Multidrug-resistant (MDR) bacterial infections have become a significant threat to global healthcare systems. Here, we developed a highly efficient antimicrobial hydrogel using environmentally friendly garlic carbon dots, pectin, and acrylic acid. The hydrogel had a porous three-dimensional network structure, which endowed it with good mechanical properties and compression recovery performance. The hydrogel could adhere closely to skin tissues and had an equilibrium swelling ratio of 6.21, indicating its potential as a wound dressing. In particular, the bactericidal efficacy following 24-h contact against two MDR bacteria could exceed 99.99 %. When the hydrogel was applied to epidermal wounds infected with methicillin-resistant Staphylococcus aureus (MRSA) on mice, a remarkable healing rate of 93.29 % was observed after 10 days. This was better than the effectiveness of the traditionally used antibiotic kanamycin, which resulted in a healing rate of 70.36 %. In vitro cytotoxicity testing and hemolysis assay demonstrated a high biocompatibility. This was further proved by the in vivo assay where no toxic side effects were observed on the heart, liver, spleen, lung, or kidney of mice. This eco-friendly and easy-to-prepare food-inspired hydrogel provides an idea for the rational use of food and food by-products as a wound dressing to control MDR bacterial infections.

Gelatin-based multifunctional composite films integrated with dialdehyde carboxymethyl cellulose and coffee leaf extract for active food packaging.

In this study, dialdehyde carboxymethyl cellulose (DCMC, 10 wt% based on gelatin) and varying contents of coffee leaf extract (CLE, 1, 3, 5 and 7 wt% based on gelatin) were incorporated into gelatin (GEL) matrix to develop multifunctional food packaging films. DCMC acted as a physical reinforcing filler through crosslinking with GEL matrix by Schiff-base reaction, CLE served as an active filler to confer film functional properties. The micro-morphology, micro-structure, physicochemical and functional properties of the GEL/DCMC/CLE composite film were investigated. The results demonstrated that mechanical, barrier properties and thermal stability of films were significantly improved by incorporation of CLE. Compared with pure GEL film, the GEL/DCMC/5%CLE film exhibited excellent UV light blocking while kept enough transparency, the best mechanical property, water resistance, water vapor and oxygen barrier, as well as thermal stability. GEL/DCMC/5%CLE film also possessed strong antioxidant activity and some antibacterial activity against E. coli and S. aureus. Packaging application testing demonstrated that the resultant GEL/DCMC/5%CLE film effectively delayed the lipid oxidation of walnut oil and preserved the postharvest freshness of fresh walnut kernels under ambient conditions.

Oleocanthal alleviated lipopolysaccharide-induced acute lung injury in chickens by inhibiting TLR4/NF-κB pathway activation.

This study aimed to investigate the ameliorative effect of oleocanthal (OC) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in chickens and its possible mechanisms. In total, 20 chickens were randomly divided into 4 groups: control (CON) group, LPS group, LPS + OC group, and OC group. LPS + OC and OC groups were intragastrically administered a 5 mg/kg·d OC dose for 7 d. On d 8, the LPS group and LPS + OC group were intratracheally administered 2 mg/kg LPS for 12 h. It was found that OC ameliorated the pathological morphology and significantly suppressed apoptosis after OC treatment in LPS-induced ALI chicken (P < 0.01). Antioxidant capacity was higher in the LPS + OC group compared with the LPS group (P < 0.01). OC downregulated the related genes and proteins expression of toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) pathway in LPS group (P < 0.01). In conclusion, OC supplementation can alleviate LPS-induced ALI in chickens by suppressing apoptosis, enhancing lung antioxidant capacities and inhibiting TLR4/NF-κB pathway activation.

The De Novo Genome Assembly of Olea europaea subsp. cuspidate, a Widely Distributed Olive Close Relative.

The olive complex, comprising six subspecies, is a valuable plant for global trade, human health, and food safety. However, only one subspecies (Olea europaea subsp. europaea, OE) and its wild relative (Olea europaea subsp. europaea var. sylvestris, OS) have genomic references, hindering our understanding of the evolution of this species. Using a hybrid approach by incorporating Illumina, MGI, Nanopore, and Hi-C technologies, we obtained a 1.20-Gb genome assembly for the olive subspecies, Olea europaea subsp. cuspidate (OC), with contig and scaffold N50 values of 5.33 and 50.46 Mb, respectively. A total of 43,511 protein-coding genes were predicted from the genome. Interestingly, we observed a large region (37.5 Mb) of "gene-desert" also called "LTR-hotspot" on chromosome 17. The gene origination analyses revealed a substantial outburst (19.5%) of gene transposition events in the common ancestor of olive subspecies, suggesting the importance of olive speciation in shaping the new gene evolution of OC subspecies. The divergence time between OC and the last common ancestor of OE and OS was estimated to be 4.39 Mya (95% CI: 2.58-6.23 Mya). The pathways of positively selected genes of OC are related to the metabolism of cofactors and vitamins, indicating the potential medical and economic values of OC for further research and utilization. In summary, we constructed the de novo genome assembly and protein-coding gene pool for Olea europaea subsp. cuspidate (OC) in this study, which may facilitate breeding applications of improved olive varieties from this widely distributed olive close relative.

Ellagic acid Alleviates hepatic ischemia-reperfusion injury in C57 mice via the Caspase-1-GSDMD pathway.

BACKGROUND: Ellagic acid (EA) has improving function against oxidative damage and inflammatory reaction in many disorders. Hepatic ischemia-reperfusion injury (IRI) is a common pathophysiological phenomenon in the veterinary clinic. In the present study, the protective effects of EA pretreatment against hepatic IRI-induced injury and the underlying mechanisms were investigated. RESULTS: We found that pyroptosis is involved in hepatic IRI, which is manifested in increasing the expression of pyroptosis-related genes and promoting the expression of active caspase-1, thereby cleaving GSDMD-N to cause pyroptosis, and caspase-1-/- mice were used to verify this conclusion. In addition, we found that EA protects against hepatic IRI by inhibiting pyroptosis, including reducing the activity of caspase-1 and its expression in the liver, inhibiting the lysis of GSDMD-N, and reducing the levels of IL-18 and IL-1ß. CONCLUSIONS: The present results have demonstrated that prophylactic administration of EA ameliorated hepatic IRI by inhibiting pyroptosis induced in hepatic ischemia-reperfusion in vivo through the caspase-1-GSDMD axis, providing a potential therapeutic option prevent hepatic IRI in pets.

Walnut oil alleviates LPS-induced intestinal epithelial cells injury by inhibiting TLR4/MyD88/NF-κB pathway activation.

In this study, we investigated the protective effects of walnut oil (WO) on mouse intestinal epithelial cells using used MODE-K cells as a model and explored the underlying mechanisms. Our data suggested that WO attenuated lipopolysaccharide (LPS)-induced pathological changes and inhibited the rate of LPS-induced apoptosis in MODE-K cells. Furthermore, WO down-regulated LPS-induced gene and protein expression of toll-like receptor 4 (TLR4), myeloid differentiation primary response gene 88 (MyD88), nuclear factor-κB (NF-κB), tumor necrosis factor-α, and interleukin-6. In conclusion, this study shows that WO exerts an anti-inflammatory effect on LPS-induced MODE-K cells injury by inhibiting the TLR4/MyD88/NF-κB pathway activation. Based on our data, a prominent functional food candidate can be provided for inflammatory bowel disease treatment. PRACTICAL APPLICATIONS: Walnut oil (WO) has excellent anti-inflammatory properties and is widely used in traditional dietary supplements. However, whether WO causes anti-lipopolysaccharide (LPS)-induced intestinal injury remains unclear. In this study, we investigated the protective effects of WO on mouse intestinal epithelial cells using MODE-K cells as a model and explored their potential mechanisms. Our data showed that WO ameliorated the pathological morphology, inhibited the apoptosis of LPS-induced MODE-K cell injury, decreased the release of pro-inflammatory cytokines, and down-regulated the related genes and proteins expression of the LPS-TLR4/MyD88/NF-κB inflammatory pathway. The results of this study would enhance the utilization of WO in the prevention of gastrointestinal diseases in animals and humans inflammatory bowel disease as well as in functional foods formulations.

Walnut oil alleviates DSS-induced colitis in mice by inhibiting NLRP3 inflammasome activation and regulating gut microbiota.

Ulcerative colitis (UC) has become a global disease and closely related to changes in intestinal oxidative stress, inflammatory factors and gut microbiota. Furthermore, the NLRP3 inflammasome activation is a key cause in the pathogenesis of dextran sulfate sodium (DSS)-induced colitis. Recent data showed the potential antioxidative and anti-inflammatory advantage of walnut oil, which widely used in traditional medicine and has become a dietary supplement for some patients. Therefore, we investigated whether walnut oil exerts an anti-inflammatory effect on DSS-induced colitis mice by targeting NLRP3 inflammasome and gut microbiota. Our data showed that walnut oil ameliorated the pathological morphology, decreased the reactive oxygen species (ROS) production and pro-inflammatory cytokines release, down-regulated the related gene proteins expression of NLRP3/ASC/Caspase-1 inflammatory pathway, inhibited apoptosis, shifted from more pathogens towards probiotics, and increased the levels of short-chain fatty acids (SCFAs) in DSS-induced damaging process. Collectively, our study concludes that walnut oil exerts anti-inflammatory effect on DSS-induced colitis in mice by inhibiting the NLRP3 inflammasome activation and modulating gut microbiota, and may be a prominent functional food candidate for UC treatment.

The protective effect of walnut oil on lipopolysaccharide-induced acute intestinal injury in mice.

Walnut oil (WO) is widely used in traditional medicine, and it has become a dietary supplement in many countries. We isolated walnut oil from Juglans sigillata and evaluated its protective effects on acute intestinal injury, and Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway in lipopolysaccharide (LPS)-induced mice was studied. The results showed that the LPS + WO group significantly decreased serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1ß levels and increased the jejunum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels compared with the LPS group. Walnut oil ameliorated the pathological morphology of the LPS-induced acute jejunum injury and decreased jejunum cells apoptosis rate and TLR4/NF-κB protein expression. Furthermore, the expression of the TLR4/NF-κB pathway key gene mRNA significantly reduced after treatment with walnut oil. This study concludes that walnut oil can exert the protective effect on LPS-induced acute intestinal injury in mice by inhibiting the TLR4/NF-κB signaling pathway.

Gamma-aminobutyric acid facilitates the simultaneous production of biomass, astaxanthin and lipids in Haematococcus pluvialis under salinity and high-light stress conditions.

The effects of γ-aminobutyric acid (GABA) on the biomass and astaxanthin and lipids production in Haematococcus pluvialis under combined salinity stress and high-light stresses were investigated. The results showed that the highest biomass (1.65 g L-1), astaxanthin production (3.86 mg L-1 d-1) and lipids content (55.11%) in H. pluvialis LUGU were observed under the 0.25 mM GABA treatment. Moreover, compared with salinity and high-light stress, GABA treatment also increased the transcript levels of biosynthesis genes, the contents of endogenous GABA and carbohydrates but decreased reactive oxygen species (ROS) levels. Further evidence revealed that intracellular GABA could regulate cell growth, astaxanthin production and lipids synthesis by mediating carotenogenesis, lipogenesis and ROS signalling. Collectively, this study provides a combined strategy for promoting the coproduction of astaxanthin and lipids and sheds light on the regulatory mechanism through which GABA affects cell growth, astaxanthin production and lipids biosynthesis in H. pluvialis under unfavourable conditions.

Effect of walnut (Juglans sigillata) oil on intestinal antioxidant, anti-inflammatory, immunity, and gut microbiota modulation in mice.

The study investigated the anti-oxidant, anti-inflammatory, immunity, and gut microbiota modulation in mice (n = 60; 15 mice/group) after intragastric administration of walnut oil (WO; three groups (low (LD), medium (MD), and high doses (HD): 2.5, 5, and 10 ml/kg, respectively) and normal control (NC, saline). WO significantly increased the median villous height/crypt depth (VH/CD) ratio, the activities of superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in intestinal mucosa. WO exerted the anti-inflammatory effects by decreasing the expression of tumor necrosis factor-α (TNF-α) in the duodenal mucosa. All groups shared 157 operational taxonomic units (OTUs; 97% similarity) representing nine phyla. The relative abundance in gut microbiota shifted from more pathogenic bacteria-Helicobacter (NC: 22% versus MD: 3%) toward probiotic-Lactobacillus (NC: 19% versus MD: 40%). The immune organ index (spleen) and contents of secretory immunoglobulin A (S-IgA) were increased from small intestine. In conclusion, WO decreased the oxidative stress, inflammation, and improved the immunity and beneficial gut microbiota in the mice. PRACTICAL APPLICATIONS: Walnut oil (WO) is widely used in traditional medicine around the world and is prescribed as beneficial food oil in agro-industry. However, the intestinal benefits of WO have not been explored extensively, and even its therapeutic mechanism still remains unknown in modern medicine. In this study, WO from Juglans sigillata was investigated for its preventive and protective effects on the intestinal mucosa in mice including anti-oxidant, anti-inflammatory, immunity, and gut microbiota modulation. WO decreased the oxidative stress, inflammation, and improved immunity and beneficial gut microbiota in the mice. WO has shown strong probiotic effect on the gut, and thus, can be considered as a potential candidate in food. The study outcome would enhance utilization of WO for the prevention of gastrointestinal diseases (e.g., Helicobacter, etc.) both in animals and human (inflammatory bowel diseases, IBD) and the formulation of functional foods.

Integration of physiological and metabolomic profiles to elucidate the regulatory mechanisms underlying the stimulatory effect of melatonin on astaxanthin and lipids coproduction in Haematococcus pluvialis under inductive stress conditions.

The effect of melatonin (MT) on the coproduction of astaxanthin and lipids was studied in Haematococcus pluvialis under inductive stress conditions. The contents of astaxanthin and lipids were enhanced by 1.78- and 1.3-fold, respectively. MT treatment upregulated the transcription levels of carotenogenic, lipogenic and antioxidant system-related genes and decreased the levels of abiotic stress-induced reactive oxidative species (ROS). Further metabolomic analysis suggested that the intermediates in glycolysis and TCA cycle facilitate the accumulation of astaxanthin and lipids in algae treated with MT. Meanwhile, MT treatment upregulated the metabolite levels of the γ-aminobutyric acid (GABA) shunt, which might regulate the carbon-nitrogen balance and the antioxidant system. After MT treatment, exogenous linoleic acid, succinate, and GABA further increased the astaxanthin content. This study may help to elucidate the specific responses to MT induction in H. pluvialis and to identify novel biomarkers that may be employed to further promote astaxanthin and lipids coproduction.

Chromosomal-level assembly of Juglans sigillata genome using Nanopore, BioNano, and Hi-C analysis.

BACKGROUND: Juglans sigillata, or iron walnut, belonging to the order Juglandales, is an economically important tree species in Asia, especially in the Yunnan province of China. However, little research has been conducted on J. sigillata at the molecular level, which hinders understanding of its evolution, speciation, and synthesis of secondary metabolites, as well as its wide adaptability to its plateau environment. To address these issues, a high-quality reference genome of J. sigillata would be useful. FINDINGS: To construct a high-quality reference genome for J. sigillata, we first generated 38.0 Gb short reads and 66.31 Gb long reads using Illumina and Nanopore sequencing platforms, respectively. The sequencing data were assembled into a 536.50-Mb genome assembly with a contig N50 length of 4.31 Mb. Additionally, we applied BioNano technology to identify contacts among contigs, which were then used to assemble contigs into scaffolds, resulting in a genome assembly with scaffold N50 length of 16.43 Mb and contig N50 length of 4.34 Mb. To obtain a chromosome-level genome assembly, we constructed 1 Hi-C library and sequenced 79.97 Gb raw reads using the Illumina HiSeq platform. We anchored ∼93% of the scaffold sequences into 16 chromosomes and evaluated the quality of our assembly using the high contact frequency heat map. Repetitive elements account for 50.06% of the genome, and 30,387 protein-coding genes were predicted from the genome, of which 99.8% have been functionally annotated. The genome-wide phylogenetic tree indicated an estimated divergence time between J. sigillata and Juglans regia of 49 million years ago on the basis of single-copy orthologous genes. CONCLUSIONS: We provide the first chromosome-level genome for J. sigillata. It will lay a valuable foundation for future research on the genetic improvement of J. sigillata.

The newly developed genomic-SSR markers uncover the genetic characteristics and relationships of olive accessions.

BACKGROUND: Olive (Olea europaea L.) is an important oil and fruit crop worldwide, owning a rich germplasm with a large number of cultivars. Simple sequence repeats (SSRs) are excellent markers and have been used for the identification of olive cultivars. However, the limited number of SSR markers and the occurrence of confusion on the names of cultivars, as well as the possible appearance of clonal variation make it difficult to identify cultivars and interpret relationships among olive cultivars. METHOD: SSR markers were designed based on trinucleotide repeat sequences by screening the whole genome of olive, and the polymorphic SSR markers were developed that were applied to the identification of 53 olive accessions. The genetic characteristics and relationships of these olive accessions were evaluated based on the developed SSR markers. RESULTS: Twenty-one highly polymorphic genomic-SSR markers were developed, covering most chromosomes of olive. These SSR markers could well distinguish all 53 olive accessions, confirming their effectiveness. DNA fingerprints of the 53 olive accessions were constructed based on the 21 SSR markers. The dendrogram clearly divided the tested accessions into two main groups, which was also supported by the results of principal coordinate analysis. A total of 31 private alleles were detected in 15 olive accessions, which reflected the genetic diversity within 53 olive accessions to some extent. Six homonymy cases were also clarified by genetic analysis. These results suggest that the newly developed olive SSR markers are informative for the exploitation, preservation and breeding of olive.

Physiological and Metabolomics Analyses Reveal the Roles of Fulvic Acid in Enhancing the Production of Astaxanthin and Lipids in Haematococcus pluvialis under Abiotic Stress Conditions.

In this study, it was found that fulvic acid (FA) enhanced the contents of astaxanthin and lipids in Haematococcus pluvialis under high light and nitrogen starvation conditions by 2- and 1.2-fold, respectively. Meanwhile, the carbohydrate and chlorophyll contents were decreased by FA induction, whereas the levels of reactive oxygen species (ROS) and glutathione (GSH) as well as the expression of astaxanthin and lipid biosynthetic genes were increased. To further explore the interrelation between FA and the biosynthesis of astaxanthin and lipids, a metabolomics analysis of H. pluvialis by combined FA and abiotic stress exposure was conducted by using LC-MS/MS. The contents of some cytoprotective metabolites and signal molecules, including d-maltose, succinate, malic acid, melatonin (MT), and some amino acids, were increased under FA induction and abiotic stress conditions. These metabolites are intermediates in the TCA cycle and Calvin cycle, providing more precursors for the synthesis of astaxanthin and lipids. Moreover, the signal molecules might contribute to enhancing the abiotic stress tolerance. This study provided new insights into the regulatory mechanism of FA on astaxanthin and lipid accumulation in H. pluvialis.

Comparative physiological and metabolomic analyses of the hyper-accumulation of astaxanthin and lipids in Haematococcus pluvialis upon treatment with butylated hydroxyanisole.

The major goal of this study was to explore the functions of butylated hydroxyanisole (BHA) combined with abiotic stress on the cultivation of the microalga Haematococcus pluvialis for astaxanthin and lipid production. Here, the effect of BHA on astaxanthin and lipid accumulation and physiological and metabolomic profiles was investigated. These results suggested that astaxanthin content was increased by 2.17-fold compared to the control. The lipid content was enhanced by 1.22-fold. BHA treatment simultaneously reduced carbohydrates and protein and delayed the decay of chlorophyll. Furthermore, metabolomic analysis demonstrated that BHA upregulated and activated the bioprocesses involved in cellular basal metabolism and signalling systems, such as glycolysis, the TCA cycle, amino acid metabolism and the phosphatidylinositol signalling system, thus enhancing astaxanthin and lipid accumulation. Altogether, this research shows the dramatic effects of BHA on algal metabolism in the regulation of key metabolic nodes and provides novel insights into microalgal regulation and metabolism.

Protective effect of walnut on d-galactose-induced aging mouse model.

OBJECTIVE S: Accumulating evidence has suggested that oxidative stress and apoptosis are involved in the aging process. d-galactose (gal) has been reported to cause symptoms of aging in mice, accompanied by liver and brain injuries. Our present work was to study the potential antioxidative and anti-apoptotic effects of walnut and to explore how these effects act on mice in a d-gal-induced aging model. MATERIALS AND METHODS: Aging mice were induced by subcutaneous injection of d-gal (200 mg kg-1 d-1 for 8 weeks). Walnut samples were simultaneously administered to the d-gal-induced aging mice once daily by intragastric gavage. Finally, body weight, organ index, cognitive function, levels of antioxidative enzymes, and liver function were monitored. RESULTS: The kernel pellicles of walnut could not only improve the learning and memory ability, and the organ index, but also significantly decrease body weight and normalize the levels of activity of antioxidative enzymes in aging mice. Further, the walnut seed coat would protect damages of hippocampus and liver in aging mice. HIGHLIGHTS: In the current study, we investigated the effects of walnut kernels and walnut seed coats (WSCs) on d-galactose-induced aging mice. WSC was firstly found to have beneficial effects on d-gal-treated mouse's brain with learning and memory impairment, which probably through the underlying mechanism reduces oxidative damage and limits neuroinflammation. In addition, WSC had a protective effect on liver damage in d-galactose sensing mice.

Role of media composition in biomass and astaxanthin production of Haematococcus pluvialis under two-stage cultivation.

In the present study, the effects of four different culture media on the growth, astaxanthin production and morphology of Haematococcus pluvialis LUGU were studied under two-step cultivation. The interactions between astaxanthin synthesis and secondary messengers, reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPK) were also investigated. In the first green vegetative cell stage, maximal biomass productivity (86.54 mg L-1 day-1) was obtained in BBM medium. In the induction stage, the highest astaxanthin content (21.5 mg g-1) occurred in BG-11 medium, which was higher than in any other media. The expressions of MAPK and astaxanthin biosynthetic genes in BG-11 were higher than in any other media, whereas the ROS content was lower. Biochemical and physiological analyses suggested that the ROS, MAPK and astaxanthin biosynthetic gene expression was involved in astaxanthin biosynthesis in H. pluvialis under different culture media conditions. This study proposes a two-step cultivation strategy to efficiently produce astaxanthin using microalgae.

Precocious genotypes and homozygous tendency generated by self-pollination in walnut.

BACKGROUND: Observations of precocious (early bearing) genotypes of walnut (Juglans regia L.) under natural conditions encouraged us to study the origin and genetic control of these fascinating traits. RESULTS: In this study, the self-fertility, progeny performance, and simple sequence repeat (SSR) locus variation of iron walnut (Juglans sigillata Dode), an ecotype of J. regia, were investigated. The average self-pollinated fruit set rate of J. sigillata cv. 'Dapao' (DP) was 7.0% annually from 1979 to 1982. The average germination rate of self-pollinated seeds was 45.2% during the 4-year period. Most progeny had inbreeding depression. Nine representative self-pollinated progeny (SP1-SP9), with special or typical traits of DP, were selected. SP1-SP4 were precocious because they initiated flowers as early as 2 years after germination, compared to the 7-10-yr period that is typical of DP. SP9 had not flowered since 1980. Twelve SSR markers were used to analyze the SP and DP. The genome of SP had a tendency toward high levels of homozygosity. The high levels of homozygosity reported in 18 additional precocious walnut genotypes complemented the results of this study. CONCLUSIONS: These results provide evidence of precocious phenotypes and genomes with high levels of homozygosity that might be generated from self-pollinating walnut. This suggests that self-pollination might facilitate the generation of unique homozygous parents for subsequent use in walnut-breeding programs. The results also indicate that more attention should be focused on adequate management of precocious walnut to avoid early depression in the production of nuts.