Genome-Wide Characterization and Functional Validation of the ACS Gene Family in the Chestnut Reveals Its Regulatory Role in Ovule Development.

Ovule abortion significantly contributes to a reduction in chestnut yield. Therefore, an examination of the mechanisms underlying ovule abortion is crucial for increasing chestnut yield. In our previous study, we conducted a comprehensive multiomic analysis of fertile and abortive ovules and found that ACS genes in chestnuts (CmACS) play a crucial role in ovule development. Therefore, to further study the function of ACS genes, a total of seven CmACS members were identified, their gene structures, conserved structural domains, evolutionary trees, chromosomal localization, and promoter cis-acting elements were analyzed, and their subcellular localization was predicted and verified. The spatiotemporal specificity of the expression of the seven CmACS genes was confirmed via qRT-PCR analysis. Notably, CmACS7 was exclusively expressed in the floral organs, and its expression peaked during fertilization and decreased after fertilization. The ACC levels remained consistently greater in fertile ovules than in abortive ovules. The ACSase activity of CmACS7 was identified using the genetic transformation of chestnut healing tissue. Micro Solanum lycopersicum plants overexpressing CmACS7 had a significantly greater rate of seed failure than did wild-type plants. Our results suggest that ovule fertilization activates CmACS7 and increases ACC levels, whereas an overexpression of CmACS7 leads to an increase in ACC content in the ovule prior to fertilization, which can lead to abortion. In conclusion, the present study demonstrated that chestnut ovule abortion is caused by poor fertilization and not by nutritional competition. Optimization of the pollination and fertilization of female flowers is essential for increasing chestnut yield and reducing ovule abortion.

Short-term responses of soil enzyme activities and stoichiometry to litter input in Castanopsis carlesii and Cunninghamia lanceolata plantations.

Litter input triggers the secretion of soil extracellular enzymes and facilitates the release of carbon (C), nitrogen (N), and phosphorus (P) from decomposing litter. However, how soil extracellular enzyme activities were controlled by litter input with various substrates is not fully understood. We examined the activities and stoichiometry of five enzymes including ß-1,4-glucosidase, ß-D-cellobiosidase, ß-1,4-N-acetyl-glucosaminidase, leucine aminopeptidase and acidic phosphatase (AP) with and without litter input in 10-year-old Castanopsis carlesii and Cunninghamia lanceolata plantations monthly during April to August, in October, and in December 2021 by using an in situ microcosm experiment. The results showed that: 1) There was no significant effect of short-term litter input on soil enzyme activity, stoichiometry, and vector properties in C. carlesii plantation. In contrast, short-term litter input significantly increased the AP activity by 1.7% in May and decreased the enzymatic C/N ratio by 3.8% in August, and decreased enzymatic C/P and N/P ratios by 11.7% and 10.3%, respectively, in October in C. lanceolata plantation. Meanwhile, litter input increased the soil enzymatic vector angle to 53.8° in October in C. lanceolata plantations, suggesting a significant P limitation for soil microorganisms. 2) Results from partial least squares regression analyses showed that soil dissolved organic matter and microbial biomass C and N were the primary factors in explaining the responses of soil enzymatic activity to short-term litter input in both plantations. Overall, input of low-quality (high C/N) litter stimulates the secretion of soil extracellular enzymes and accelerates litter decomposition. There is a P limitation for soil microorganisms in the study area.

Royal jelly from different floral sources possesses distinct wound-healing mechanisms and ingredient profiles.

In recent years, population aging together with the increased prevalence of diabetes and obesity has fuelled a surge in the instances of cutaneous non-healing wounds. Royal jelly (RJ) is a traditional remedy for wound repair; however, the subjacent mechanisms and ingredient profiles are still largely unknown. Our previous study found that Castanea mollissima Bl. RJ (CmRJ-Zj) possessed superior wound healing-promoting effects on both the in vivo and in vitro models than Brassica napus L. RJ (BnRJ-Zj). This study conducted an in-depth investigation on the wound-repairing mechanisms of CmRJ-Zj and BnRJ-Zj to explain the previously observed phenomenon and also comprehensively characterized their constituents. It was found that chestnut RJ could enhance cutaneous wound healing by boosting the growth and mobility of keratinocytes, modulating the expression of aquaporin 3 (AQP3), regulating MAPK and calcium pathways, and mediating inflammatory responses. By employing LC-MS/MS-based proteomic and metabolomic techniques, the comprehensive molecules present in CmRJ-Zj and BnRJ-Zj were elucidated, resulting in a clear discrimination from each other. A total of 15 and 631 differential proteins and compounds were identified, and 217 proteins were newly found in RJ proteome. With bioinformatic functional analysis, we speculated that some differential components were responsible for the wound-healing properties of CmRJ-Zj. Therefore, this study provides an insight into the wound-healing mechanisms of RJ and is the first to explore the compositions of RJ from different nectar plants. It will facilitate the development of therapeutic agents from RJ to treat difficult-to-heal wounds and the distinction of different RJ categories.

Influence of forest stand characteristics on physical, mechanical properties and chemistry of chestnut wood.

Site conditions and forest management affect dendrometric parameters of chestnut (Castanea sativa Mill.) coppices, but there is modest knowledge on the effect of stand dendrometric characters on physical and mechanical wood characteristics. The aim of this study was to verify these relationships in chestnut coppices that were 12-14 years old. Wood density, compression and bending strength, shrinkages were measured on shoots of five different stand in a vulcanic site in Monte Amiata (Central - Italy). Investigated stands differ in number of stools/ha and dominant height, diameter/basal area of the shoots. The main difference in the physical characters among the stands is density. The initial results of the study showed that physical, mechanical wood characters are more dependent by the shoot than by the site. There is a positive relationships between the number of stools/ha and density and a negative one among shoot dominant height and basal area with wood density. Spectroscopic profile by FTIR has not showed relevant differences among the stands. Wood anatomy has showed the breakpoint at cellular level.

The relationship between anthocyanin accumulation and photoprotection in young leaves of two dominant tree species in subtropical forests in different seasons.

Increasing amounts of experimental evidence show that anthocyanins provide physiological protection to plants under stress. However, the difference in photoprotection mediated by anthocyanins and other photoprotective substances in different seasons is still uncertain. To determine the relationship between anthocyanin accumulation and the photoprotective effects in different seasons, Castanopsis chinensis and Acmena acuminatissima, whose anthocyanin accumulation patterns differ in different seasons, were used as materials to explain how plants adapt to different seasons; as such, their physiological and biochemical responses were analyzed. Young leaves of C. chinensis and A. acuminatissima presented different colors in the different seasons. In summer, the young leaves of C. chinensis were purplish red, while those of A. acuminatissima were light green. In winter, the young leaves of C. chinensis were light green, while those of A. acuminatissima were red. Compared with the young red leaves, the young light green leaves that did not accumulate anthocyanins had higher flavonoid and phenolics contents, total antioxidant capacity, non-photochemical quenching (NPQ), and relative membrane leakage, and a slower recovery rate in the maximum photochemical efficiency (Fv/Fm) after high-light treatment. In addition, the net photosynthesis rate (Pn), transpiration rate (Tr), stomatal conductance (gs), and the effective quantum yield of PSII (ΦPSII) of the young leaves in winter were significantly lower than those in summer, while the activities of catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7), and superoxide dismutase (SOD, EC 1.15.1.1) were significantly higher than those in summer. These data indicate that to adapt to seasonal changes anthocyanins, other antioxidative substances and antioxidative enzymes, as well as components involved in the safe dissipation of excitation energy as heat need to cooperate with one another.

Integrated Proteomic and Metabolomic Profiling of Phytophthora cinnamomi Attack on Sweet Chestnut (Castanea sativa) Reveals Distinct Molecular Reprogramming Proximal to the Infection Site and Away from It.

Phytophthora cinnamomi is one of the most invasive tree pathogens that devastates wild and cultivated forests. Due to its wide host range, knowledge of the infection process at the molecular level is lacking for most of its tree hosts. To expand the repertoire of studied Phytophthora-woody plant interactions and identify molecular mechanisms that can facilitate discovery of novel ways to control its spread and damaging effects, we focused on the interaction between P. cinnamomi and sweet chestnut (Castanea sativa), an economically important tree for the wood processing industry. By using a combination of proteomics, metabolomics, and targeted hormonal analysis, we mapped the effects of P. cinnamomi attack on stem tissues immediately bordering the infection site and away from it. P. cinnamomi led to a massive reprogramming of the chestnut proteome and accumulation of the stress-related hormones salicylic acid (SA) and jasmonic acid (JA), indicating that stem inoculation can be used as an easily accessible model system to identify novel molecular players in P. cinnamomi pathogenicity.

Comparison of Biochemical Constituents and Contents in Floral Nectar of Castanea spp.

Pollination is essential for efficient reproduction in pollinator-dependent crops that rely on the attraction of pollinators to flowers. Especially, floral nectar is considered to be an important factor attracting pollinator like honey bees, but differences among major chestnut species (Castanea crenata, C. mollissima, C. dentata, and C. sativa) are still little explored. This study aims to evaluate the value of honey source by analyzing floral nectar characteristics and comparing the composition of volatile organic compounds (VOCs) that mediate plant-pollinator interaction. In this study, we analyzed nectar samples obtained from male flowers using HPLC and HS-SPME/GC-MS. The five chestnuts showed significant differences between the volume of secreted nectar, free sugar composition, amino acid content and VOCs composition. Furthermore, C. crenata (Japanese cultivar 'Ungi') was revealed to emit the highest total amounts of VOCs and high levels of benzenoid compounds that are generally associated with flower-visiting insects. The sugar content per catkin, which is used to determine the honey yield, was the highest in C. crenata, suggesting that C. crenata 'Ungi' can be highly valued as a honey tree. Therefore, a better understanding of the relationship between pollinator and nectar characteristics of C. crenara could contribute to a prospective honey plant.

Allelopathic Effects of Castanea henryi Aqueous Extracts on the Growth and Physiology of Brassica pekinensis and Zea mays.

The present study investigated the allelopathic effects of aqueous extracts of Castanea henryi litter on the growth and physiological responses of Brassica pekinensis and Zea mays. Treatment with high concentrations of leaf extract (0.05 g/ml for B. pekinensis and 0.10 g/ml for Z. mays) significantly increased malonaldehyde content and reduced seed germination, seedling growth, chlorophyll content, and the activity levels of antioxidant enzymes. These effects generally increased with increasing extract concentration. However, in Z. mays, low extract concentrations actually promoted seed germination, shoot growth, chlorophyll content, and antioxidant enzyme activity. The allelopathic effects of the various C. henryi extracts decreased as follows: leaf extract > twig extract > shell extract. Eleven potential allelochemicals including rutin, quercetin, luteolin, procyanidin A2, kaempferol, allantoin, propionic acid, salicylic acid, jasmonic acid, methylmalonic acid, and gentisic acid were identified in the leaves of C. henryi which were linked to the strongest allelopathic effects. These findings suggest that the allelopathic effects of C. henryi differ depending on receptor plant species, and that leaves are the most allelopathic litter in C. henryi.

Transcriptome Analysis and Identification of Genes Associated with Starch Metabolism in Castanea henryi Seed (Fagaceae).

Starch is the most important form of carbohydrate storage and is the major energy reserve in some seeds, especially Castanea henryi. Seed germination is the beginning of the plant's life cycle, and starch metabolism is important for seed germination. As a complex metabolic pathway, the regulation of starch metabolism in C. henryi is still poorly understood. To explore the mechanism of starch metabolism during the germination of C. henryi, we conducted a comparative gene expression analysis at the transcriptional level using RNA-seq across four different germination stages, and analyzed the changes in the starch and soluble sugar contents. The results showed that the starch content increased in 0-10 days and decreased in 10-35 days, while the soluble sugar content continuously decreased in 0-30 days and increased in 30-35 days. We identified 49 candidate genes that may be associated with starch and sucrose metabolism. Three ADP-glucose pyrophosphorylase (AGPase) genes, two nucleotide pyrophosphatase/phosphodiesterases (NPPS) genes and three starch synthases (SS) genes may be related to starch accumulation. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate the expression levels of these genes. Our study combined transcriptome data with physiological and biochemical data, revealing potential candidate genes that affect starch metabolism during seed germination, and provides important data about starch metabolism and seed germination in seed plants.

An atmospheric pollutant (inorganic nitrogen) alters the response of evergreen broad-leaved tree species to extreme drought.

Drought and nitrogen (N) deposition are important components of global climate and environmental change. In this greenhouse study, we investigated the ecophysiological responses of the seedlings of three subtropical forest plant species (Schima superba, Castanopsis fissa, and Michelia macclurei) to short-term experimental drought stress, N addition, and their interaction. The results showed that drought stress reduced the activities of antioxidant enzymes [superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)] and total antioxidant capacity (T-AOC), but increased the malondialdehyde (MDA), abscisic acid (ABA), and proline (PRO) contents in plants. The PRO content, T-AOC, and antioxidant enzyme activities were increased, and ABA and MDA contents were decreased by N addition alone. Furthermore, N addition under drought stress increased antioxidant enzymes activities, PRO content, and T-AOC. The treatments, however, did not significantly affect the chlorophyll fluorescence parameters of the species. T-AOC was positively correlated with antioxidant enzyme activities in each species, indicating that antioxidant enzymes were important for plant resistance to oxidative stress. MDA content increased with the increase of ABA content, indicating that ABA may help regulate stomatal movement and drought-induced oxidative injury in plants. T-AOC was positively correlated with PRO content, probably because PRO participated in osmotic regulation of cells and increased osmotic stress resistance. These results indicate that N addition can reduce drought stress of subtropical forest plants and will help researchers predict how evergreen broad-leaved forests will respond to global change in the future.

Complete chloroplast genome of Castanopsis sclerophylla (Lindl.) Schott: Genome structure and comparative and phylogenetic analysis.

Castanopsis sclerophylla (Lindl.) Schott is an important species of evergreen broad-leaved tree in subtropical areas and has high ecological and economic value. However, there are few studies on its chloroplast genome. In this study, the complete chloroplast genome sequence of C. sclerophylla was determined using the Illumina HiSeq 2500 platform. The complete chloroplast genome of C. sclerophylla is 160,497 bp long, including a pair of inverted repeat (IR) regions (25,675 bp) separated by a large single-copy (LSC) region of 90,255 bp and a small single-copy (SSC) region of 18,892 bp. The overall GC content of the chloroplast genome is 36.82%. A total of 131 genes were found; of these, 111 genes are unique and annotated, including 79 protein-coding genes, 27 transfer RNA genes (tRNAs), and four ribosomal RNA genes (rRNAs). Twenty-one genes were found to be duplicated in the IR regions. Comparative analysis indicated that IR contraction might be the reason for the smaller chloroplast genome of C. sclerophylla compared to three congeneric species. Sequence analysis indicated that the LSC and SSC regions are more divergent than IR regions within Castanopsis; furthermore, greater divergence was found in noncoding regions than in coding regions. The maximum likelihood phylogenetic analysis showed that four species of the genus Castanopsis form a monophyletic clade and that C. sclerophylla is closely related to Castanopsis hainanensis with strong bootstrap values. These results not only provide a basic understanding of Castanopsis chloroplast genomes, but also illuminate Castanopsis species evolution within the Fagaceae family. Furthermore, these findings will be valuable for future studies of genetic diversity and enhance our understanding of the phylogenetic evolution of Castanopsis.

Influence of amylose on the pasting and gel texture properties of chestnut starch during thermal processing.

The quality of starchy foods is highly correlated with their amylose content. After boiling or roasting, the amylose content in cooked chestnut starch decreased to 16.6-26.8%, which was significantly lower than that of raw chestnut starch (26.6-28.1%). After cooking, the pasting temperature and peak time increased, while the breakdown, peak viscosity, final viscosity and setback of starches decreased. The amylose content showed a negative correlation with the pasting characteristics of the starch gel of cooked chestnut. The amylose content negatively affected the springiness and chewiness of starch gel, and positively correlated with its cohesiveness and stringiness. The freeze-thaw stability of starch from cooked chestnut was lower than that of raw chestnut. The amylose content was negatively correlated with freeze-thaw stability and positively correlated with the solubility of starch. The results demonstrated that amylose plays an important role in the pasting and gel texture properties of starch during cooking.

Spatiotemporal Coupling of Vessel Cavitation and Discharge of Stored Xylem Water in a Tree Sapling.

Water discharge from stem internal storage compartments is thought to minimize the risk of vessel cavitation. Based on this concept, one would expect that water storage compartments involved in the buffering of xylem tensions empty before the onset of vessel cavitation under drought stress, and potentially refill after soil saturation. However, scant in vivo data exist that elucidate this localized spatiotemporal coupling. In this study on intact saplings of American chestnut (Castanea dentata), x-ray computed microtomography (microCT) showed that the xylem matrix surrounding vessels releases stored water and becomes air-filled either concurrent to or after vessel cavitation under progressive drought stress. Among annual growth rings, the xylem matrix of the current year remained largely water-filled even under severe drought stress. In comparison, microtomography images collected on excised stems showed that applied pressures of much greater than 0 MPa were required to induce water release from the xylem matrix. Viability staining highlighted that water release from the xylem matrix was associated primarily with emptying of dead fibers. Refilling of the xylem matrix and vessels was detected in intact saplings when the canopy was bagged and stem water potential was close to 0 MPa, and in leafless saplings over the winter period. In conclusion, this study indicates that the bulk of water stored in the xylem matrix is released after the onset of vessel cavitation, and suggests that capillary water contributes to overall stem water storage under drought but is not used primarily for the prevention of drought-induced vessel cavitation in this species.

Chestnut (Castanea sativa Miller.) Burs Extracts and Functional Compounds: UHPLC-UV-HRMS Profiling, Antioxidant Activity, and Inhibitory Effects on Phytopathogenic Fungi.

Chestnut (Castanea sativa Miller.) burs (CSB) represent a solid waste produced during the edible fruit harvesting. Their usual disposal in the field increases the environmental and economic impact of the agricultural process. HPLC-UV-HRMS profiling revealed that CSB organic and aqueous extracts (CSB-M, CSB-H, CSB-A) contain several hydrolyzable tannins, mainly ellagitannins, and glycoside flavonols. Ellagic acid (EA) and chestanin are predominant components (5⁻79 and 1⁻13 mg/g dry extract, respectively). NMR analysis confirmed the chemical structures of the major constituents from CSB-M. The extracts displayed a significant scavenging activity against DPPH (EC50 12.64⁻24.94 µg/mL) and ABTS⁺ radicals (TEAC value 2.71⁻3.52 mM Trolox/mg extract). They were effective in inhibiting the mycelial growth (EC50 6.04⁻15.51 mg/mL) and spore germination (EC50 2.22⁻11.17 mg/mL) of Alternaria alternata and Fusarium solani. At the highest concentration, CSB-M was also active against Botrytis cinerea both in mycelium and spore form (EC50 64.98 and 16.33 mg/mL). The EA contributed to the antifungal activity of extracts (EC50 on spore germination 13.33⁻112.64 µg/mL). Our results can support the upgrading of chestnut burs from agricultural wastes to a resource of natural fungicides for managing fruit and vegetable diseases.

Fermented Castanea crenata Inner Shell Extract Increases Fat Metabolism and Decreases Obesity in High-Fat Diet-Induced Obese Mice.

The anti-obesity effects of fermented Castanea crenata inner shell extract (FCCE) were investigated using high-fat diet (HFD)-induced obese mice. In the FCCE intake groups, body weight gain and adipocyte area were significantly reduced, especially body weight gain in the 250 mg/kg FCCE group (G4) decreased by 37%, respectively, compared with negative control group (G2, HFD group). After oral administration of the FCCE, the increase of serum low-density lipoprotein (LDL)-cholesterol induced by HFD was suppressed significantly, as well as the level of aspartate aminotransferase, and alanine aminotransferase, which are markers of hepatitis induced by obesity. Serum leptin in G4 group was significantly decreased to less than that of G2 group. Also, in G4 and 500 mg/kg FCCE group (G5), enzymes-related lipogenesis, citrate synthase, and ATP citrate lyase were decreased, whereas the level of enoyl-CoA hydratase used for ß-oxidation was significantly increased in comparison with normal diet group. Furthermore, the FCCE stimulated the expression of lipolytic regulators, especially AMP-activated protein kinase. In conclusion, we suggest that the FCCE may ameliorate in diet-induced obesity by regulating lipid metabolism.

Alterations in leaf nitrogen metabolism indicated the structural changes of subtropical forest by canopy addition of nitrogen.

Globally, nitrogen deposition increment has caused forest structural changes due to imbalanced plant nitrogen metabolism and subsequent carbon assimilation. Here, a 2 consecutive-year experiment was conducted to reveal the effects of canopy addition of nitrogen (CAN) on nitrogen absorption, assimilation, and allocation in leaves of three subtropical forest woody species (Castanea henryi, Ardisia quinquegona, and Blastus cochinchinensis). We hypothesized that CAN altered leaf nitrogen absorption, assimilation and partitioning of different plants in different ways in subtropical forest. It shows that CAN increased maximum photosynthetic rate (Amax), photosynthetic nitrogen use efficiency (PNUE), and metabolic protein content of the two understory species A. quinquegona and B. cochinchinensis. By contrary, for the overstory species, C. henryi, Amax, PNUE, and metabolic protein content were significantly reduced in response to CAN. We found that changes in leaf nitrogen metabolism were mainly due to the differences in enzyme (e.g. Ribulose-1,5-bisphosphate carboxylase, nitrate reductase, nitrite reductase and glutamine synthetase) activities under CAN treatment. Our results indicated that C. henryi may be more susceptible to CAN treatment, and both A. quinquegona and B. cochinchinensis could better adapt to CAN treatment but in different ways. Our findings may partially explain the ongoing degradation of subtropical forest into a community dominated by small trees and shrubs in recent decades. It is possible that persistent high levels of atmospheric nitrogen deposition will lead to the steady replacement of dominant woody species in this subtropical forest.

Hydrothermal treatment of chestnut shells (Castanea sativa) to produce oligosaccharides and antioxidant compounds.

Hydrothermal treatment is an environmentally friendly technology that allows the solubilisation of hemicellulosic oligosaccharides with potential for their use as prebiotics. The purpose of this study was to solubilize oligosaccharides and antioxidant compounds from chestnut shells by a hydrothermal processing. The highest content of oligosaccharides (18.3 g/L), with a relatively low level of monosaccharides (2.4 g/L) and degradation products (0.5 g/L) was obtained at 180 °C (severity of 3.08). In addition, the liquors presented a high content of phenolic and flavonoid compounds with good antioxidant properties. The GC-MS revealed that the most abundant phenolic compound was pyrogallol (13.2%). The molecular weight distribution of the solubilization products showed that a 26.5% presented an apparent Mw of 6077 g/mol and a 73.5% presented an apparent Mw of 586 g/mol with a high polydispersity index. MALDI-TOF, FTIR, and TGA analyses revealed structural information of these compounds and their thermal stability.

Effects of Condensed and Hydrolyzable Tannins on Rumen Metabolism with Emphasis on the Biohydrogenation of Unsaturated Fatty Acids.

The hypothesis that condensed tannins have higher inhibitory effect on ruminal biohydrogenation than hydrolyzable tannins was tested. Condensed tannin extract from mimosa (CT) and hydrolyzable tannin extract from chestnut (HT) or their mixture (MIX) were incorporated (10%) into oil supplemented diets and fed to rumen fistulated sheep. Fatty acid and dimethyl acetal composition of rumen contents and bacterial biomass were determined. Selected rumen bacteria were analyzed by quantitative real time PCR. Lower ( P < 0.05) rumen volatile fatty acids concentrations were observed with CT compared to HT. Moreover, lower concentration ( P < 0.05) of Fibrobacter succinogenes, Ruminococcus flavefaciens, Ruminococcus albus, and Butyrivibrio proteoclasticus were observed with CT compared to HT. The extension of biohydrogenation of 18:2n-6 and 18:3n-3 did not differ among treatments but was much more variable with CT and MIX than with HT. The trans-/ cis-18:1 ratio in bacterial biomass was higher ( P < 0.05) with HT than CT. Thus, mimosa condensed tannins had a higher inhibitory effect on ruminal metabolism and biohydrogenation than chestnut hydrolyzable tannins.

Comprehensive Transcriptome Analysis of Phytohormone Biosynthesis and Signaling Genes in the Flowers of Chinese Chinquapin (Castanea henryi).

Chinese chinquapin (Castanea henryi) nut provides a rich source of starch and nutrients as food and feed, but its yield is restricted by a low ratio of female to male flowers. Little is known about the developmental programs underlying sex differentiation of the flowers. To investigate the involvement of phytohormones during sex differentiation, we described the morphology of male and female floral organs and the cytology of flower sex differentiation, analyzed endogenous levels of indole-3-acetic acid (IAA), gibberellins (GAs), cytokinins (CKs), and abscisic acid (ABA) in the flowers, investigated the effects of exogenous hormones on flower development, and evaluated the expression profiles of genes related to biosyntheses and signaling pathways of these four hormones using RNA-Seq combined with qPCR. Morphological results showed that the flowers consisted of unisexual and bisexual catkins, and could be divided into four developmental stages. HPLC results showed that CK accumulated much more in the female flowers than that in the male flowers, GA and ABA showed the opposite results, while IAA did not show a tendency. The effects of exogenous hormones on sex differentiation were consistent with those of endogenous hormones. RNA-Seq combined with qPCR analyses suggest that several genes may play key roles in hormone biosynthesis and sex differentiation. This study presents the first comprehensive report of phytohormone biosynthesis and signaling during sex differentiation of C. henryi, which should provide a foundation for further mechanistic studies of sex differentiation in Castanea Miller species and other nonmodel plants.

[Analysis of transicriptomes and exploring flavonoid biosynthetic pathway genes in Lithocarpus polystachyus].

The sweet taste and health effect of Lithocarpus polystachyus are mainly related flavonoid. To obtain Lithocarpus transcriptome database and flavonoid biosynthesis-related genes, the RNA-Seq techology (Illumina HiSeq 4000) was used to sequence its transcriptome. Six Gb database was assembled after assembly steps, and 41 043 of L. polystachyus unigenes were obtained. With blasting them with 7 data banks, all unigenes were involved in 51 GO-terms and 237 metabolic pathways. And furthermore 28 genes of the flavonoid biosynthesis-related were found. After using the MicroSatallite, 18 161 SSR were obtained, the single-nucleotide-repeated was the richest at 7 346. These data represent abundant messages about transcripts and provide valuable genome data sources in molecular biology of L. polystachyus.