Two Carya Species, Carya hunanensis and Carya illinoinensis, Used as Rootstocks Point to Improvements in the Heat Resistance of Carya cathayensis.

Grafting as a crucial horticultural technique has been widely used in the cultivation of Carya cathayensis (Chinese hickory), which is a unique and important economic tree in the northeast of Zhejiang Province and the south of Anhui Province. However, the existing literature lacks research on the potential impact of various rootstocks on the thermal tolerance of Chinese hickory. The objectives of this study were to evaluate heat tolerance in four distinct groups of Chinese hickory, including C. cathayensis grafted onto Carya hunanensis and Carya illinoinensis, one self-grafted group (C. cathayensis grafted onto C. cathayensis), and one non-grafted group (C. cathayensis). We examined photosynthesis parameters, phytohormones, and differentially expressed genes in the four various hickory groups subjected to 25 °C, 35 °C, and 40 °C heat stress (HS). The results demonstrated that grafting onto C. hunanensis and C. illinoinensis exhibited a higher net photosynthetic rate and stomatal conductance, lower intercellular CO2 concentration, and smaller changes in plant hormone content compared to self-grafted and non-grafted group under HS. The transcriptome results revealed that the majority of differentially expressed genes (DEGs) associated with photosynthetic pathways exhibited downregulation under HS, while the degree of variation in grafted groups using C. hunanensis and C. illinoinensis as rootstocks was comparatively lower than that observed in self-grafted and non-grafted groups. The alteration in the expression patterns of DEGs involved in plant hormone synthesis and metabolism under HS corresponded to changes in plant hormone contents. Overall, Chinese hickory grafted onto C. hunanensis and C. illinoinensis exhibited enhanced resistance to high-temperature stress at the juvenile stage.

Fractionation of phenolic compounds from hickory by-products using solid phase extraction-sonication: Chemical composition, antioxidant and antimicrobial activity.

Hickory is an abundant source of phenolic compounds that exhibit a diverse range of bioactivities. In this study, phenolic compounds were extracted and purified from hickory green husk (HG), hickory nutshell (HN), and hickory seed coat (HS) using solid-phase extraction and ultrasonication (SPE-US). The effects of the SPE-US treatment on the structure and properties of the phenolic compounds were then investigated, including their composition, antioxidant activity, and antimicrobial activity. The dominant phenolic substances in the different extracts after SPE-US treatment were: ellagic acid and trans ferulic acid (HS); ellagic acid and sinapic acid (HN); and rutin (HG). The HS-SPE-US1 extract exhibited the highest total polyphenol content (416 ± 11 mg GAE/g DW), total flavonoid content (47.51 ± 0.68 mg RE/g DW), Fe3+ reduction ability (74.2 ± 1.0 mmol Fe2+/g DW), radical (DPPH and ABTS) scavenging ability, and antimicrobial activity against Staphylococcus aureus.

The Comparative Genomics of Botryosphaeriaceae Suggests Gene Families of Botryosphaeria dothidea Related to Pathogenicity on Chinese Hickory Tree.

Trunk canker poses a major threat to the production of Chinese hickory tree (Carya cathayensis Sarg.), which is primarily determined by Botryosphaeriaceae. In our previous work, we identified Botryosphaeria dothidea as the predominant pathogen of this disease. However, it is still unclear about corresponding gene families and mechanisms associated with B. dothidea's pathogenicity on Chinese hickory tree. Here, we present a comparative analysis of high-quality genome assemblies of Botryosphaeria dothidea and other isolated pathogens, showing highly syntenic relationships between B. dothidea and its closely related species and the conservative evolution of the Botryosphaeriaceae family. Higher GC contents were found in the genomes of B. dothidea and three other isolated pathogens (Botryshaeria cortices, Botryshaeria fabicerciana, and Botryshaeria qingyuanensis) compared to Macrophomina phaseolina, Neofusicoccum parvum, Diplodia corticola, and Lasiodiplodia theobromae. An investigation of genes specific to or expanded in B. dothidea revealed that one secreted glucanase, one orsellinic acid biosynthesis enzyme, and two MFS transporters positively regulated B. dothidea's pathogenicity. We also observed an overrepresentation of viral integrase like gene and heterokaryon incompatibility proteins in the B. dothidea's genome. In addition, we observed one LRR-domain-containing protein and two Sec-domain-containing proteins (Sec_1 and Sec_7) that underwent positive selection. This study will help to understand B. dothidea's pathogenicity and potential influence on the infection of Chinese hickory, which will help in the development of disease control and ensure the security of Chinese hickory production.

Polyphenols from hickory nut reduce the occurrence of atherosclerosis in mice by improving intestinal microbiota and inhibiting trimethylamine N-oxide production.

BACKGROUND: Trimethylamine N-oxide (TMAO), a metabolite produced by intestinal microbiota through metabolizing phosphatidylcholine, choline, l-carnitine and betaine in the diet, has been implicated in the pathogenesis of atherosclerosis (AS). Concurrently, dietary polyphenols have garnered attention for their potential to ameliorate obesity, diabetes and atherosclerosis primarily by modulating the intestinal microbial structure. Hickory (Carya cathayensis) nut, a polyphenol-rich food product favored for its palatability, emerges as a candidate for exploration. HYPOTHESIS/PURPOSE: The relationship between polyphenol of hickory nut and atherosclerosis prevention will be firstly clarified, providing theoretical basis for the discovery of natural products counteracting TMAO-induced AS process in hickory nut. STUDY DESIGN AND METHODS: Employing Enzyme-linked Immunosorbent Assay (ELISA) and histological examination of aortic samples, the effects of total polyphenol extract on obesity index, inflammatory index and pathological changes of atherosclerosis in C57BL/6 J mice fed with high-fat and high choline diet were evaluated. Further, the composition, abundance, and function of mouse gut microbiota were analyzed through 16srDNA sequencing. Concurrently, the levels of TMAO and the expression of key enzymes (CutC and FMO3) involved in its synthesis are quantified using ELISA, Western Blot and Real-Time Quantitative PCR (RT-qPCR). Additionally, targeted metabolomic profiling of the hickory nut polyphenol extract was conducted, accompanied by molecular docking simulations to predict interactions between candidate polyphenols and the CutC/FMO3 using Autodock Vina. Finally, the docking prediction were verified by microscale thermophoresis (MST) . RESULTS: Polyphenol extracts of hickory nut improved the index of obesity and inflammation, and alleviated the pathological changes of atherosclerosis in C57BL/6 J mice fed with high-fat and high-choline diet. Meanwhile, these polyphenol extracts also changed the composition and function of intestinal microbiota, and increased the abundance of microorganisms in mice. Notably, the abundance of intestinal microbiota endowed with CutC gene was significantly reduced, coherent with expression of CutC catalyzing TMA production. Moreover, polyphenol extracts also decreased the expression of FMO3 in the liver, contributing to the reduction of TMAO levels in serum. Furthermore, metabonomic profile analysis of these polyphenol extracts identified 647 kinds of polyphenols. Molecular docking predication further demonstrated that Casuariin and Cinnamtannin B2 had the most potential inhibition on the enzymatic activities of CutC or FMO3, respectively. Notably, MST analysis corroborated the potential for direct interaction between CutC enzyme and available polyphenols such as Corilagin, (-)-Gallocatechin gallate and Epigallocatechin gallate. CONCLUSION: Hickory polyphenol extract can mitigate HFD-induced AS by regulating intestinal microflora in murine models. In addition, TMA-FMO3-TMAO pathway may play a key role in this process. This research unveils, for the inaugural time, the complex interaction between hickory nut-derived polyphenols and gut microbial, providing novel insights into the role of dietary polyphenols in AS prevention.

High-quality genome assembly and genetic transformation system of Lasiodiplodia theobromae strain LTTK16-3, a fungal pathogen of Chinese hickory.

Lasiodiplodia theobromae, as one of the causative agents associated with Chinese hickory trunk cankers, has caused huge economic losses to the Chinese hickory industry. Although the biological characteristics of this pathogen and the occurrence pattern of this disease have been well studied, few studies have addressed the related mechanisms due to the poor molecular and genetic study basis of this fungus. In this study, we sequenced and assembled L. theobromae strain LTTK16-3, isolated from a Chinese hickory tree (cultivar of Linan) in Linan, Zhejiang province, China. Phylogenetic analysis and comparative genomics analysis presented crucial cues in the prediction of LTTK16-3, which shared similar regulatory mechanisms of transcription, DNA replication, and DNA damage response with the other four Chinese hickory trunk canker-associated Botryosphaeria strains including, Botryosphaeria dothidea, Botryosphaeria fabicerciana, Botryosphaeria qingyuanensis, and Botryosphaeria corticis. Moreover, it contained 18 strain-specific protein clusters (not conserved in the other L. theobromae strains, AM2As and CITRA15), with potential roles in specific host-pathogen interactions during the Chinese hickory infection. Additionally, an efficient system for L. theobromae protoplast preparation and polyethylene glycol (PEG) -mediated genetic transformation was firstly established as the foundation for its future mechanisms study. Collectively, the high-quality genome data and the efficient transformation system of L. theobromae here set up the possibility of targeted molecular improvements for Chinese hickory canker control.IMPORTANCEFungi with disparate genomic features are physiologically diverse, possessing species-specific survival strategies and environmental adaptation mechanisms. The high-quality genome data and related molecular genetic studies are the basis for revealing the mechanisms behind the physiological traits that are responsible for their environmental fitness. In this study, we sequenced and assembled the LTTK16-3 strain, the genome of Lasiodiplodia theobromae first obtained from a diseased Chinese hickory tree (cultivar of Linan) in Linan, Zhejiang province, China. Further phylogenetic analysis and comparative genomics analysis provide crucial cues in the prediction of the proteins with potential roles in specific host-pathogen interactions during the Chinese hickory infection. An efficient PEG-mediated genetic transformation system of L. theobromae was established as the foundation for the future mechanisms exploration. The above genetic information and tools set up valuable clues to study L. theobromae pathogenesis and assist in Chinese hickory canker control.

Analysis of Delta(9) fatty acid desaturase gene family and their role in oleic acid accumulation in Carya cathayensis kernel.

Carya cathayensis, commonly referred to as Chinese hickory, produces nuts that contain high-quality edible oils, particularly oleic acid (18:1). It is known that stearoyl-ACP desaturase (SAD) is the first key step converting stearic acid (C18:0, SA) to oleic acid (C18:1, OA) in the aminolevulinic acid (ALA) biosynthetic pathway and play an important role in OA accumulation. Thus far, there is little information about SAD gene family in C. cathayensis and the role of individual members in OA accumulation. This study searched the Chinese Hickory Genome Database and identified five members of SAD genes, designated as CcSADs, at the whole genome level through the comparison with the homologous genes from Arabidopsis. RNA-Seq analysis showed that CcSSI2-1, CcSSI2-2, and CcSAD6 were highly expressed in kernels. The expression pattern of CcSADs was significantly correlated with fatty acid accumulation during the kernel development. In addition, five full-length cDNAs encoding SADs were isolated from the developing kernel of C. cathayensis. CcSADs-green fluorescent protein (GFP) fusion construct was infiltrated into tobacco epidermal cells, and results indicated their chloroplast localization. The catalytic function of these CcSADs was further analyzed by heterologous expression in Saccharomyces cerevisiae, Nicotiana benthamiana, and walnut. Functional analysis demonstrated that all CcSADs had fatty acid desaturase activity to catalyze oleic acid biosynthesis. Some members of CcSADs also have strong substrate specificity for 16:0-ACP to synthesize palmitoleic acid (C16:1, PA). Our study documented SAD gene family in C. cathayensis and the role of CcSSI2-1, CcSSI2-2, and CcSAD6 in OA accumulation, which could be important for future improvement of OA content in this species via genetic manipulation.

Melatonin alleviates aluminum toxicity by regulating aluminum-responsive and nonresponsive pathways in hickory.

Aluminum (Al) toxicity is a significant constraint on agricultural productivity worldwide. Melatonin (MT) has been shown to alleviate Al toxicity in plants; however, the underlying mechanisms remain largely unknown. Here, we employed a combination of physiological and molecular biology techniques to examine the role of MT in mitigating Al toxicity of hickory. We found that MT decreased the contents of cell wall pectin, hemicellulose, Al, and Al-induced massive reactive oxygen species accumulation in the roots of hickory. Transcriptomic analysis revealed that MT may alleviate root tip Al stress by regulating Al-responsive and nonresponsive pathways. Co-expression regulatory network and dual-luciferase receptor assays revealed that transcription factors, CcC3H12 and CcAZF2, responded to MT and significantly activated the expression of two cell wall pectin-related genes, CcPME61 and CcGAE6, respectively. Further, yeast one-hybrid and electrophoretic mobility shift assay (EMSA) assays verified that CcC3H12 and CcAZF2 regulated CcPME61 and CcGAE6, respectively, by directly binding to their promoters. Overexpression of CcPME61 enhanced the Al sensitivity of Arabidopsis thaliana. Our results indicate that MT can improve Al tolerance of hickory via multiple pathways, which provides a new perspective for the study of the mechanism of MT in alleviating abiotic stress.

Lipid droplets proteome reveals dynamic changes of lipid droplets protein during embryonic development of Carya cathayensis nuts.

Lipid droplets (LD) is an important intracellular organelle for triacylglycerols (TAGs) storage. A variety of proteins on the surface of LD coordinately control the contents, size, stability and biogenesis of LD. However, the LD proteins in Chinese hickory (Carya cathayensis) nuts, which is rich in oil and composed of unsaturated fatty acids, have not been identified and their roles in LD formation still remain largely unknown. In present study, LD fractions from three developmental stages of Chinese hickory seed were enriched and the LD fraction accumulated proteins were then isolated and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Protein compositions throughout the various developmental phases were calculated using label-free intensity-based absolute quantification (iBAQ) algorithm. The dynamic proportion of high abundance lipid droplets proteins such as oleosins 2 (OLE2), caleosins 1 (CLO1) and steroleosin 5 (HSD5) increased parallelly with the embryo development. For low abundance lipid droplets proteins, seed LD protein 2 (SLDP2), sterol methyltransferase 1 (SMT1) and LD-associated protein 1 (LDAP1) were the predominant proteins. Moreover, 14 low abundance OB proteins such as oil body-associated protein 2 A (OBAP2A) were selected for future investigation that may associate with embryo development. Overall, 62 differentially expressed proteins (DEPs) were determined by label free quantification (LFQ) algorithms and may involve in LD biogenesis. Furthermore, the subcellular localization validation indicated that selected LD proteins were targeted to the lipid droplets, confirming the promising of proteome data. Taken together, this comparative study may shed light on further study to understand the lipid droplets function in the seed, which contains high oil content.

Microencapsulation with Different Starch-Based Polymers for Improving Oxidative Stability of Cold-Pressed Hickory (Carya cathayensis Sarg.) Oil.

Hickory (Carya cathayensis Sarg.) oil is a nutrient-dense edible woody oil, with its unsaturated fatty acids accounting for more than 90% of total ones, and liable to oxidation spoilage. To efficiently improve its stability and expand its application fields, the microencapsulation of cold-pressed hickory oil (CHO) by the molecular embedding method and freeze-drying technique was performed using malt dextrin (MD), hydroxylpropyl-ß-cyclodextrin (HP-ß-CD), ß-cyclodextrin (ß-CD), or porous starch (PS) as a wall material. Two wall materials and/or their CHO microcapsulates (CHOM) with higher encapsulation efficiencies (EE) were selected to carry out physical and chemical characterizations using laser particle size diffractometer, scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, derivative thermogravimetry, and oxidative stability tests. Results indicated ß-CDCHOM and PSCHOM had significantly higher EE values (80.40% and 75.52%) than MDCHOM and HP-ß-CDCHOM (39.36% and 48.32%). The particle sizes of the two microcapsules selected were both widely distributed with their spans being more than 1 µm and a certain degree of polydispersity. Microstructural and chemical characterizations indicated that ß-CDCHOM had comparatively stable structure and good thermal stability compared with PSCHOM. Storage performances under light, oxygen, and temperature showed that ß-CDCHOM was superior to PSCHOM, especially in terms of thermal and oxidative stability. This study demonstrates that ß-CD embedding can be applied to improve the oxidative stability of vegetable oils such as hickory oil and act as a means of preparing functional supplementary material.

Comparative plastomes of Carya species provide new insights into the plastomes evolution and maternal phylogeny of the genus.

Carya, in the Juglandiodeae subfamily, is to a typical temperate-subtropical forest-tree genus for studying the phylogenetic evolution and intercontinental disjunction between eastern Asia (EA) and North America (NA). Species of the genus have high economic values worldwide for their high-quality wood and the rich healthy factors of their nuts. Although previous efforts based on multiple molecular markers or genome-wide SNPs supported the monophyly of Carya and its two EA and NA major subclades, the maternal phylogeny of Carya still need to be comprehensively evaluated. The variation of Carya plastome has never been thoroughly characterized. Here, we novelly present 19 newly generated plastomes of congeneric Carya species, including the recently rediscovered critically endangered C. poilanei. The overall assessment of plastomes revealed highly conservative in the general structures. Our results indicated that remarkable differences in several plastome features are highly consistent with the EA-NA disjunction and showed the relatively diverse matrilineal sources among EA Carya compared to NA Carya. The maternal phylogenies were conducted with different plastome regions and full-length plastome datasets from 30 plastomes, representing 26 species in six genera of Juglandoideae and Myrica rubra (as root). Six out of seven phylogenetic topologies strongly supported the previously reported relationships among genera of Juglandoideae and the two subclades of EA and NA Carya, but displayed significant incongruencies between species within the EA and NA subclades. The phylogenetic tree generated from full-length plastomes demonstrated the optimal topology and revealed significant geographical maternal relationships among Carya species, especially for EA Carya within overlapping distribution areas. The full-length plastome-based phylogenetic topology also strongly supported the taxonomic status of five controversial species as separate species of Carya. Historical and recent introgressive hybridization and plastid captures might contribute to plastome geographic patterns and inconsistencies between topologies built from different datasets, while incomplete lineage sorting could account for the discordance between maternal topology and the previous nuclear genome data-based phylogeny. Our findings highlight full-length plastomes as an ideal tool for exploring maternal relationships among the subclades of Carya, and potentially in other outcrossing perennial woody plants, for resolving plastome phylogenetic relationships.

Whole-Transcriptome Analysis Reveals Long Noncoding RNAs Involved in Female Floral Development of Hickory (Carya cathayensis Sarg.).

Hickory, an endemic woody oil and fruit tree species in China, is of great economic value. However, hickory has a long juvenile period and an inconsistent flowering of males and females, thus influencing the bearing rates and further limiting fruits yield. Currently, it is reported that long noncoding RNAs (lncRNAs) play critical regulatory roles in biological processes. However, the role of lncRNAs in the development of hickory female flowers remains unclear. In this study, a total of 6,862 putative lncRNAs were identified from the female flower transcriptomes in three different growth stages of hickory. We proposed that lncRNAs might play an important role in phytohormone signaling processes for flower formation, especially in the abscisic acid and jasmonic acid pathways, according to the results of our Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Moreover, we predicted the interactions among four microRNAs (miRNAs), three lncRNAs, and four genes. We proposed that facing the changing environment, LNC_002115 competes with PHOSPHATE2 (PHO2) for the binding sites on cca-miR399f, and protects PHO2 from suppression. In addition, cis-acting LNC_002115 regulates the expression of the SHORT VEGETATIVE PHASE (SVP) by influencing ABRE-binding factor (ABF). In brief, LNC_002115 regulates hickory female floral development by impacting both PHO2 and SVP. This study was the first to identify lncRNAs involved in hickory female floral development, and provided new insight to elucidate how lncRNAs and their targets play a role in female floral development in hickory, thus unfolding the opportunities for functional characterization of blossom-related lncRNAs in further studies.

Flavonoid Synthesis and Metabolism During the Fruit Development in Hickory (Carya cathayensis).

Hickory (Carya cathayensis) kernel is rich in powerful bioactive flavonoids, which can remove excess free radicals in the human body and play an important role in regulating the physiological metabolism of the plant. This study investigated the changes of flavonoids in hickory exocarp and embryo during development. In this study, 72 DEGs involved in the regulation of flavonoid biosynthesis in fruits were identified, and TT4, CCoAOMT1, UGT71D1, C4H, F3H, TT8, FLS1, and LDOX were identified as the core genes of flavonoid biosynthesis. A total of 144 flavonoid-related metabolites were detected by metabolite analysis. Transcriptome and metabolome analysis combined to construct the flavonoid biosynthesis regulatory pathway in the development stage of hickory fruit. Our results provide a theoretical basis for the exploration and regulation of functional genes related to flavonoid biosynthesis and metabolism in hickory and other plants and the breeding of new walnut varieties.

Genome-Wide Identification of TLP Gene Family and Their Roles in Carya cathayensis Sarg in Response to Botryosphaeria dothidea.

Hickory (Carya cathayensis) is a critical tree species of the genus Carya from the Juglandaceae family that contains nutrient-rich nuts. Due to large-scale soil degradation, the pests and diseases of hickory are becoming more and more serious. Thaumatin-like proteins (TLPs) are vital proteins involved in the complex defense process of plant pathogens. In this study, 40 CcTLP genes were identified genome-widely and phylogenetically grouped into three subfamilies. The sequence of CcTLPs had a conservative pattern, such as eight stable disulfide bonds, REDDD, and G-X-[GF]-X-C-X-T-[GA]-D-C-X(1,2)-G-X-(2,3)-C structure. In total, 57 cis-elements related to stress-responsive, light-responsive, phytohormone-responsive, and plant-responsive were discovered. Under salicylate (SA), methyl jasmonate (MeJA), and ethephon (ETH) treatments, the expressions of CcTLP28, CcTLP29, CcTLP30, CcTLP31, CcTLP32, CcTLP33, CcTLP37, CcTLP38, and CcTLP39 had different patterns. This is an indication that most of the TLP genes were upregulated by SA and downregulated by MeJA. Notably, seven TLP genes were significantly upregulated under the Botryosphaeria dothidea inoculation, especially CcTLP31, with an over 20-fold change. Nine genes were shown by subcellular localization analysis to be located at the plasma membrane and cytoplasm. The knowledge of the disease-resistant function of the CcTLP family in hickory is promoted by these results. A foundation reference for the molecular breeding of this plant in the future is provided by our findings.

Multi-nutrient stoichiometry of Chinese hickory (Carya cathayensis) saplings: plant organs vary in their response to nitrogen fertilization.

Nitrogen (N) enrichment from excessive fertilization in managed forests affects biogeochemical cycles on multiple scales, but our knowledge of how N availability shifts multi-nutrient stoichiometries (including macronutrients: N, phosphorus, potassium, calcium, magnesium and micronutrients: manganese, iron and zinc) within and among organs (root, stem and leaf) remains limited. To understand the difference among organs in terms of multi-nutrient stoichiometric homeostasis responding to N fertilization, a six-level N supply experiment was conducted through a hydroponic system to examine stem growth, multi-nutrient concentrations and stoichiometric ratios in roots, stems and leaves of 2-year-old Chinese hickory (Carya cathayensis Sarg.) saplings. Results showed that N supply significantly enhanced leaf length, width, basal diameter and sapling height. Increasing the rates of N also significantly altered multi-nutrient concentrations in roots, stems and leaves. Macronutrients generally respond more positively than micronutrients within organs. Among organs, leaves and stems generally responded more actively to N supply than roots. The stoichiometric ratios of nutrients within different organs changed significantly with N supply, but their direction and degree of change varied by organ. Specifically, increased N supply reduced the ratios of both macronutrients and micronutrients to N in plant organs, while increased N supply elevated the ratios of P to other nutrients. With N fertilization, ratios of micronutrients decreased in leaves and stems and increased in roots. In particular, leaf N and stem Mn stoichiometries responded strongly to N availability, indicating stimulated N uptake but a decreased risk of Mn2+ accumulation to excessive N. Overall, Chinese hickory saplings responded positively to increasing N availability in terms of stem growth, but the multi-nutrient stoichiometric homeostasis was distinctively organ-dependent. These results are expected to enhance our understanding of N-induced changes in homeostasis of multiple nutrients at the organ level and may offer new insights into how plants adapt to increasing N fertilization.

The rediscovery of Caryapoilanei (Juglandaceae) after 63 years reveals a new record from China.

Despite having been first published in 1941, Caryapoilanei (A.Chev.) J.-F.Leroy is only known from three collections in Vietnam, Laos and Thailand. It has not been recollected since then and was long suspected to have become extinct through repeated deforestation events. Here, we report the rediscovery, and meanwhile the first new record in China, of this extremely rare gigantic hickory species at Yunnan province 63 years after its last collection in 1958. Three small patchy subpopulations were found with a total of about 50 adult trees having diameter at breast height (DBH) larger than 60 cm, together with some seedlings and saplings, but the fruit set was low. Based on new and fresh material, we present a revised morphological description of C.poilanei, and an updated distribution map for the species. In addition, we also provide a key for the hickories in China. Lastly, we suggest C.poilanei should be listed as a Critically Endangered (CR) species according to the latest IUCN Red List Categories and Criteria.

Comprehensive identification and analysis of circRNAs during hickory (Carya cathayensis Sarg.) flower bud differentiation.

Flower bud differentiation represents a crucial transition from vegetative growth to reproductive development. Carya cathayensis (hickory) is an important economic species in China, with a long juvenile period that hinders its commercial development. In recent years, circular RNAs (circRNAs) have been widely studied and identified as sponges for miRNA regulation of mRNA expression. However, little is known regarding the role of circRNAs in flower buds. In this study, we sequenced circRNAs at three developmental stages (undifferentiated, differentiating, and fully differentiated) in both female and male buds. A total of 6,931 circRNAs were identified in the three developmental stages and 4,449 and 2,209 circRNAs were differentially expressed in female and male buds, respectively. Gene ontology demonstrated that many circRNA host genes participated in various processes, for example, cellular and intracellular pH regulation. Function annotation identified 46 differentially expressed circRNAs involved in flowering regulation, with 28 circRNAs found only in female buds, 4 found only in male buds, and 11 found in both female and male buds. A circRNA-miRNA-mRNA network was predicted based on 13 flowering-related circRNAs and their seven putative interacting miRNAs to describe the regulatory mechanism. Our preliminary results demonstrated a potential involvement of circRNA in bud differentiation. They provided a preliminary theoretical basis for how circRNA might participate in flower development in hickory, perhaps in woody plants.

Comparison analysis of widely-targeted metabolomics revealed the variation of potential astringent ingredients and their dynamic accumulation in the seed coats of both Carya cathayensis and Carya illinoinensis.

Pecan and hickory nuts are two of consumers' favorite ones. Pecan seeds can be eaten fresh, while hickory ones must remove astringency before eating. Here, we reported that total phenols, flavonoids and condensed tannins of hickory seeds were reduced after de-astringent treatments. They gradually increased with development, showing higher levels in hickory seed coat at mid-late periods than that in pecan's. Widely-targeted metabonomics analysis of developing testa identified 424 kinds of components, including 101, 38, 58, 27 classes of flavonoids, tannins, phenolic acids, organic acids and others, showing 16 different changing trends. Notably, most kinds of flavonoids, hydrolysable tannins and phenolic acids at maturity were more than that of pecan's, while oligomeric condensed tannins were opposite. Gene expression analysis provided further explanations for their dynamic accumulation. These results unraveled potential astringent components in hickory testa and preliminary molecular mechanisms of their dynamic changes, offering theoretical basis for the targeted de-astringency.

Analysis of lipidomics profile of Carya cathayensis nuts and lipid dynamic changes during embryonic development.

Hickory (Carya cathayensis) nuts contain higher amount of lipids, and possess high nutritional value and substantial health benefits. However, their lipid composition and dynamic changes during embryogenesis have not been thoroughly investigated. Therefore, lipidomics profile and lipid dynamic changes during embryonic development were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Totally, 544 kinds of lipids were identified in mature hickory nuts with higher proportions of glycerolipids (59.94%) and glycerophospholipids (38.66%). Notably, diacylglycerols showed gradual uptrends, which corresponded with total glycerolipid and glycerophospholipid at middle and late stage of embryogenesis, suggesting the pivotal role of diacylglycerols in the accumulation of glycerolipids and glycerophospholipids. Moreover, triacylglycerols, diacylglycerols, phosphatidylethanolamines and phosphatidylcholines had high relative content with abundance of unsaturated fatty acids, specifically oleic acid, linoleic acid and linolenic acid, localized mainly at sn-2 lipid position. Together, our study provides innovative perspectives for studying the nutritional benefits of hickory nut lipids.