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The Chinese chestnut (Castanea mollissima Blume) is a dioecious plant in which the number of male inflorescences far exceeds that of female inflorescences. Therefore, it is of interest to study the differentiation of male flowers in C. mollissima. In this study, the male inflorescences of "Yanshanhongli" C. mollissima were observed. Then, the main components of the cell wall were stained by fluorescence and immunofluorescence labeling techniques to examine the changes in cell wall components. The results showed that the development of the male inflorescence of C. mollissima had different transverse elongation and longitudinal elongation. In addition, the development of male inflorescences and anthers were divided into five and four major stages, respectively, according to the microstructural changes. Immunostaining indicated that the fluorescence intensity of cellulose increased with the development of male inflorescences of C. mollissima. The fluorescence intensity of low-esterified homogalacturonan (HG) was stronger in the early stage, while the fluorescence intensity of high-esterified HG was stronger thereafter. This research provides new insights into the changes in cell wall components during the growth and development of male inflorescences of C. mollissima and important clues for exploring the growth and development mechanism of male inflorescences of C. mollissima. [ABSTRACT FROM AUTHOR]

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Copyright of Journal of Agricultural Science & Technology (1008-0864) is the property of Journal of Agricultural Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

• The abortive rate of ovules could go as high as 94 % in an ovary of Castanea mollissima , the occurrence of abortive ovules with abnormal structure was related to the obstruction of nutrient transport. • Abortive ovules exhibit the presence of starch grains at an earlier stage than fertile ovules. • Cytological features proved that the selective death of ovules was dependent on programmed cell death. Chinese chestnut (Castanea mollissima Blume.) ovary contains 12–18 ovules, but only one can develop and grow normally, which indicates that the abortive rate of ovules can go as high as 94 %. In this study, we observed the developmental characteristics and cytological changes of abortive ovules and fertile ovules during development. Morphological analysis showed it would take about 15∼20 days from globular to cotyledon embryo in fertile ovules, which is accompanied with the formation and apoptosis of endosperm. But there were stark differences in the nutrient transport, microstructure and ultrastructure of fertile and abortive ovules. The fluorescence disodium fluorescein and FDA indicated that all ovules were viable at early stages, but some ovules became inactive at different times as the ovary developed. Fluorescence labelling and ultrastructure showed cell nuclei of abortive ovules were disintegrated at 25 days after anthesis (DAA). Compared with the fertile ovules, starch grains were synthesized at earlier periods, but disappeared immediately, and the cells were usually irregular in shape, and had folded cell membranes in the abortive ovules. Furthermore, the Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was carried out to assess programmed cell death in fertile and abortive ovules, results showed that DNA fragmentation was occurred in the cells of abortive ovules. In short, these results provide new insights into ovule abortion in the angiosperm. [ABSTRACT FROM AUTHOR]

Boron (B) is essential to plant development. However, excessive B is toxic to plants. This research was performed to evaluate the effects of B toxicity on cell wall architecture of Chinese chestnut (Castanea mollissima Blume) pollen tubes with emphasis on the relationship among pectins, cellulose, and callose. Results showed that 0.8 mM H3BO3 inhibited pollen germination and led to abnormal morphology of the pollen tubes. B toxicity also affected the distribution of cell wall components of the pollen tube. In control pollen tube, esterified and acid pectins were distributed unevenly, with the former mainly at the tip and the latter on the distal region. Cellulose was distributed uniformly on the surface with less at the tip; callose reduced gradually from base to sub-tip of the pollen tubes and no callose at the tip of the tube was detected. B toxicity led to the deposition of esterified and acid pectins, cellulose, and callose at the tip of the pollen tube. Results from scanning electron microscopy and transmission electron microscopy showed that B toxicity also altered pollen tube wall ultrastructure. The results from enzymatic treatment illustrated that there existed a close relationship among pectins, cellulose, and callose. B toxicity also altered the relationship. In a word, B toxicity altered deposition and relationship of pectins, cellulose, and callose of pollen tube wall.

The Chinese chestnut (Castanea mollissima Bl.) is a woody nut crop with a high ecological value. Although many cultivars have been selected from natural seedlings, elite lines with comprehensive agronomic traits and characters remain rare. To explore genetic resources with aid of whole genome sequence will play important roles in modern breeding programs for chestnut. In this study, we generated a high-quality C. mollissima genome assembly by combining 90× Pacific Biosciences long read and 170× high-throughput chromosome conformation capture data. The assembly was 688.93 Mb in total, with a contig N50 of 2.83 Mb. Most of the assembled sequences (99.75%) were anchored onto 12 chromosomes, and 97.07% of the assemblies were accurately anchored and oriented. A total of 33,638 protein-coding genes were predicted in the C. mollissima genome. Comparative genomic and transcriptomic analyses provided insights into the genes expressed in specific tissues, as well as those associated with burr development in the Chinese chestnut. This highly contiguous assembly of the C. mollissima genome provides a valuable resource for studies aiming at identifying and characterizing agronomical-important traits, and will aid the design of breeding strategies to develop more focused, faster, and predictable improvement programs.

Chinese chestnuts (Castanea mollissima Blume.) contain 12–18 ovules in one ovary, but only one ovule develops into a seed, indicating a high ovule abortion rate. In this study, the Chinese chestnut ‘Huaihuang’ was used to explore the possible mechanisms of ovule abortion with respect to morphology and proteomics. The morphology and microstructure of abortive ovules were found to be considerably different from those of fertile ovules at 20 days after anthesis (20 DAA). The fertile ovules had completely formed tissues, such as the embryo sac, embryo and endosperm. By contrast, in the abortive ovules, there were no embryo sacs, and wide spaces between the integuments were observed, with few nucelli. Fluorescence labelling of the nuclei and transmission electron microscopy (TEM) observations showed that cells of abortive ovules were abnormally shaped and had thickened cell walls, folded cell membranes, condensed cytoplasm, ruptured nuclear membranes, degraded nucleoli and reduced mitochondria. The iTRAQ (isobaric tag for relative and absolute quantitation) results showed that in the abortive ovules, low levels of soluble protein with small molecular weights were found, and most of differently expressed proteins (DEPs) were related to protein synthesis, accumulation of active oxygen free radical, energy synthesis and so on. These DEPs might be associated with abnormal ovules formation.

CMBP was an alcohol-soluble polysaccharide extracted from Castanea mollissima blume. Our study aimed to investigate the preliminary structural characteristics and in vivo antitumor activity of CMBP. The contents of total sugar, protein and uronic acid in CMBP was 93.68%, 0.89% and 1.78%, respectively. GC and FTIR results suggested galactose and glucose were detected for CMBP (2.86 × 103 Da), while glucose was the dominant monosaccharide in CMBP and pyranose form and α- and β-type glycosidic linkages were founded for CMBP. Meanwhile, CMBP could exhibite strong antitumor effects on S180 tumor-bearing mice. The antitumor effects of CMBP were related to improving the immune response, which result from improving the levels of serum cytokines (TNF-α, IL-2 and IFN-γ), activating immune cells (macrophages, lymphocytes and NK cells), adjusting the distribution of lymphocyte subsets. The H&E staining and cell cycle results indicated CMBP could induce tumor cells apoptosis and extensive injuries.

A galactoglucan (HCPA) was extracted from C. mollissima Blume fruits using hot water, which was composed of glucose and galactose, with a molecular weight of 5.5 kDa. Methylation and 1 D, 2 D NMR spectroscopy analysis showed that HCPA contained [→4)-α-D-Glcp(1→], [→4,6)-α-D-Glcp(1→], [→4)-β-D-Galp(1→] and [α-D-Glcp(1→] motifs in a molar ratio of approximately 6.11:1.08:1.00:1.03. HCPA showed a spherical-like structure as detected by scanning electron microscope (SEM). HCPA biological activity was investigated in vitro. The results showed that HCPA inhibited the proliferation of HeLa, MCF-7 and MGC-803 cells, induced HeLa cells apoptosis and arrested cell cycle in G2/M phase of HeLa cells. HCPA also enhanced the viability and phagocytic ability of RAW 264.7 cells and stimulated the production of NO, TNF-α and IL-6. The results demonstrated that HCPA had a potent antitumor and immunomodulating effects in vitro, suggesting its potential use as functional food and drug products. [ABSTRACT FROM AUTHOR]

Abstract A 1,6-α-D-glucan (CPA) with a high molecular weight of 2.0 × 103 kDa was isolated from the fruits of Castanea mollissima Blume. CPA showed good water solubility, low side effects on normal cells, and moderate cytotoxicity on cancer cells. Selenium nanoparticles (SeNPs) possess significant anticancer effects, but poor water solubility and dispersity. Therefore, an effective nanocomposite was designed in order to enhance the stability and anticancer effects of SeNPs decorated with CPA. Water soluble and dispersed selenium nanoparticles (CPA-SeNPs) were fabricated using CPA and Na 2 SeO 3 by redox reaction with ascorbic acid. The average diameter of CPA-SeNPs was 53.7 ± 4.0 with a Se content of approximately 171.4 mg/g. CPA-SeNPs showed a significant anti-proliferative effect on HeLa cells in vitro. Since these NPs could induce apoptosis and S phase arrest. The apoptotic pathway triggered by CPA-SeNPs on HeLa cells was also characterized as induced by reactive oxygen species (ROS) overproduction, mitochondrial dysfunction and activation of caspase-3. This study suggested that CPA-SeNPs could be developed as a potential candidate in cancer treatment, especially against human cervical cancer. Highlights • CPA-SeNPs was fabricated using CPA and Na 2 SeO 3 by reducing with ascorbic acid. • CPA-SeNPs was approximately 50 nm with a Se content about 171.4 mg/g. • CPA-SeNPs showed a significant anti-proliferative effect on HeLa cells in vitro. • CPA-SeNPs induced HeLa cells apoptosis in an intrinsic apoptosis pathway. [ABSTRACT FROM AUTHOR]

Polyphenol extracts from the burs of Castanea mollissima Blume (CMPE) exhibited potential antioxidant and hypoglycemic activities. The α-glucosidase inhibitory activities of CMPE were assessed as a means of elucidating the mechanism behind its hypoglycemic activities. In vitro studies showed that CMPE significantly inhibited both yeast α-glucosidase, through a noncompetitive mode with an IC50 of 0.33 μg/mL, and rat intestinal α-glucosidase. In vivo studies revealed that oral administration of CMPE at doses of 600 mg/kg significantly reduced postprandial blood glucose levels by 27.2% in normal rats following sucrose challenges. Gel permeation chromatography revealed that CMPE exhibited typical characteristics of high-molecular-mass polymers with mean (Mn) and weight (Mw) average molecular weights of 35.4 and 50.7 kDa, respectively, and a polydispersity (Mw/Mn) of 1.432. Acid hydrolysis analysis indicated the presence of ellagitannins. These data suggest that CMPE, enriched with ellagitannins, would be an efficacious dietary supplement for diabetes management through the inhibition of alpha-glucosidase.

The present study aimed to characterize the glucan from C. mollissima Blume fruits and its selenium derivative, then investigate their antitumor activity in vitro. A glucan, designated as CPA, was firstly isolated from the fruits of C. mollissima Blume. Structure analysis indicated that CPA was a linear 1,6-α-D-glucan with the average molecular weight about 2.0 × 10 kDa. The selenylation modification derivative of CPA (sCPA), exhibited a stronger antiproliferative effect on tumor cells than CPA in vitro. CPA and sCPA could induce HeLa cells apoptosis and decrease mitochondrial membrane potential. sCPA could also arrest HeLa cells in S phase, promote reactive oxygen species generation and activate caspase-3 activity in HeLa cells. These results manifest that CPA and sCPA inhibit the proliferation of HeLa cells via different mechanisms, which is meaningful for their potential use as antitumor drugs. [ABSTRACT FROM AUTHOR]

A new polysaccharide was extracted from Chinese chestnut (Castanea mollissima Blume) by pressurized liquid extraction (PLE) and further successive purification through DEAE-cellulose 52 and Sephadex G-200 columns. The results showed a single and symmetrical sharp peak, which suggested that purified polysaccharide was homogeneous. The molecular weight was calculated to be 3.28 × 105 Da. Monosaccharide analysis revealed that the polysaccharide was composed of rhamnose, ribose, arabinose, xylose, mannose, glucose, and D-galactose in the molar ratio of 1.42%, 13.12%, 22.68%, 1.57%, 9.39%, 21.58%, 30.24%, respectively. There was a dose-dependent relationship with measures of antioxidant activities, viz. capabilities to scavenge free radical, superoxide anion free radical, hydrogen peroxides and reducibility. The antitumor activity of the polysaccharide was tested in a cell culture model. The polysaccharide caused a significant dosedependent inhibition on the multiplication of cancer cells. [ABSTRACT FROM AUTHOR]

In the screening of biologically active constituents from medicinal plants, the 75% EtOH extract of the testas of Castanea mollissima Blume showed potent α-glucosidase inhibitory activity. By means of various chromatographic methods, the extract gave a new dammarane-type triterpene 1 along with 17 known compounds. The structure of 1 was determined to be 3β-acetoxy-20-oxo-21-nordammaran-23-oic acid by HRMS and NMR studies including 2D NMR experiments. The new compound and some known compounds showed potent α-glucosidase inhibitory activity with acarbose as a positive control. [ABSTRACT FROM AUTHOR]

• A polysaccharide had been isolated from Castanea mollissima blume. • The polysaccharide was a water-soluble polysaccharide. • Preliminary characterization of the polysaccharide was investigated. • In vitro and in vivo antitumor activities of the polysaccharide were studied. A new water-soluble polysaccharide, CMP90, with a molecular weight of 23.9 kDa was isolated from Castanea mollissima Blume and the preliminary structural characteristics and antitumor effects of CMP90 in vitro and in vivo were investigated in the research. CMP90 consists of arabinose, galactose, glucose, xylose and mannose (molar ratio: 0.08:0.11:5.14:0.12:0.08) with α- and β-anomeric units. The results of in vitro experiments indicated that CMP90 exhibited a significant inhibitory effect on the proliferation of HL-60 cells with typical apoptotic characteristics by inducing cell cycle arrested at G1/M phase. Additionally, the results in vivo suggested CMP90 was able to inhibit the growth of S180 solid tumors via protecting immune organs, improving the levels of serum cytokines (TNF-α, IL-2 and IFN-γ), enhancing the activities of immune cells (macrophages, lymphocytes and NK cells) and inducing cell apoptosis or death. Taken together, these combined data clearly indicated that CMP90 may be used as a potential candidate agent for cancer therapy. [ABSTRACT FROM AUTHOR]

• A polysaccharide had been isolated from Castanea mollissima blume. • The polysaccharide was an alcohol-soluble polysaccharide. • Preliminary characterization of the polysaccharide was investigated. • In vivo antitumor activities of the polysaccharide were studied. CMBP was an alcohol-soluble polysaccharide extracted from Castanea mollissima blume. Our study aimed to investigate the preliminary structural characteristics and in vivo antitumor activity of CMBP. The contents of total sugar, protein and uronic acid in CMBP was 93.68%, 0.89% and 1.78%, respectively. GC and FTIR results suggested galactose and glucose were detected for CMBP (2.86 × 103 Da), while glucose was the dominant monosaccharide in CMBP and pyranose form and α- and β-type glycosidic linkages were founded for CMBP. Meanwhile, CMBP could exhibite strong antitumor effects on S180 tumor-bearing mice. The antitumor effects of CMBP were related to improving the immune response, which result from improving the levels of serum cytokines (TNF-α, IL-2 and IFN-γ), activating immune cells (macrophages, lymphocytes and NK cells), adjusting the distribution of lymphocyte subsets. The H&E staining and cell cycle results indicated CMBP could induce tumor cells apoptosis and extensive injuries. [ABSTRACT FROM AUTHOR]