Crataegus Pinnatifida Bge Research Articles

Chemical composition, antibacterial activity and action mechanism of different extracts from hawthorn (Crataegus pinnatifida Bge.)

Present study was designed to compared the total flavonoids and polyphenols contents and antibacterial activity of hawthorn extracts with different polarities as well as the underlying antibacterial mechanisms. The results showed that among all hawthorn extracts, methanol and ethanol extracts (ME and EE) exhibited high levels of total flavonoids and polyphenols contents, followed by acetone, ethyl acetate, trichloromethane and petroleum ether extracts. ME exhibited the strongest antibacterial activity against tested bacteria, especially Staphylococcus aureus with a 1.25 μg/mL of the minimum inhibitory concentration (MIC) and minimum bactericide concentration (MBC). Further analysis revealed that the main phenolic compounds from ME were epicatechin (281.6 mg/100 g DW), procyanidin B2 (243.5 mg/100 g DW), chlorogenic acid (84.2 mg/100 g DW) and quercetin (78.4 mg/100 g DW). The action mechanism of ME against S. aureus could be ascribed to ME damaging cell wall and cell membrane integrity, inhibiting intracellular enzyme activity, increasing reactive oxygen species (ROS), also changing expression of associated genes and then inducing apoptosis of S. aureus. In addition, the antimicrobial activity of ME against S. aureus has also been demonstrated to be efficient in the food matrix (whole milk).

Self-compatibility of Crataegus pinnatifida Bge. ‘Zizhenzhu’ is associated with the mutation of a pistil-part non-S factor

‘Zizhenzhu’ is a spontaneous, self-compatible hawthorn (Crataegus pinnatifida Bge.) mutant of unknown parentage. Although ‘Zinzhenzhu’ possesses typical S-RNase-based gametophytic self-incompatibility, the molecular mechanism underlying its self-compatible phenotype is still unclear. In this study, two full-length RNase T2 gene family S-RNases were cloned from ‘Zizhenzhu’, and their deduced amino acid sequences were found to have a conserved structure similar to S-RNases from Pyrus and Malus. In a phylogenetic tree, the two S-RNases clustered with Malus and Pyrus, thereby suggesting that the self-incompatibility recognition system in hawthorn is the same as in Malus and Pyrus. Genetic analysis of selfed and crossed progeny revealed that the switch to self-compatibility in ‘Zizhenzhu’ is due to the mutation of a non-S factor in the style. In addition, evidence for apomixis, an interesting phenomenon, was detected in the crossed progeny of ‘Zizhenzhu’ for the first time.

The Spectrum–Effect Relationship Between HPLC Fingerprint and the Invigorating Blood and Dissolving Stasis Effect of Hawthorn Leaves

Hawthorn leaves, officially listed in the 2015 Chinese Pharmacopoeia, comes from Crataegus pinnatifida Bge. var. major N. E. Br. and Crataegus pinnatifida Bge. It has the effect of invigorating blood and dissolving stasis, regulating qi and dredging veins, turbid, and lipid-lowering. This work was designed to explore the spectrum–effect relationship between high-performance liquid chromatography (HPLC) fingerprint and the invigorating blood and dissolving stasis effect of hawthorn leaves. Four extracts from hawthorn leaves, 50% ethanol extract (EE), macroporous resin extract (MRE), ethyl acetate extract (EAE), and n-butanol extract (BAE) were administered to rats at three doses of 0.4 g·kg−1, 0.6 g·kg−1, and 0.8 g·kg−1 (crude medicine weight/rat weight), respectively. Twenty-seven indexes were obtained and analyzed by principal component analysis (PCA) and one-way ANOVA. The spectrum–effect relationship was constructed by multiple linear regression (MLR) analysis. With the above separation method, HPLC fingerprints of 12 batches (three concentrations of four extracts) of hawthorn leaves extracts were obtained, and 32 common peaks were marked by similarity analysis. Additionally, 9 indexes of fibrinogen (FIB), erythrocyte total (ET), platelet count (PC), mean platelet volume (MPV), plateletcrit (PCT), whole blood viscosity-WBV(LSR-20S−1), WBV(LSR-60S−1), WBV(LSR-150S−1), and plasma viscosity (PV) were selected as the relevant indexes with pertinence and statistical significance (p < 0.05) on invigorating blood and dissolving stasis, and taken for the subsequent spectrum–effect experiments. Finally, the results of spectrum–effect relationships between 32 common peaks and 9 indexes showed the main peaks responsible for invigorating blood and dissolving stasis were 1–6, 8–11, 13–15, 17, 19, 21, 23, and 27–32. Compared with the standards and other references, peaks 3, 14, 17, and 19 were, respectively, identified as chlorogenic acid, vitexin glycoside, vitexin, and rutin. This work was helpful to the discovery of the active substance and quality control of hawthorn leaves.

The first tripyrrolic chlorophyll catabolites isolated from Crataegus pinnatifida Bge. var. major brown leaves

Two diastereoisomers, namely, Crataegus pinnatifida tripyrrolic Chl catabolites or Cp-TCC-1 (1) and Cp-TCC-2 (2), were isolated from the leaves of Crataegus pinnatifida Bge. var. major. Compounds 1 and 2 were identified using spectroscopic methods including HR-ESI-TOF-MS, 1D-NMR, 2D-NMR. A probable biosynthetic pathway to Cp-TCC-1 (1) and Cp-TCC-2 (2) is expounded.

A new dineolignan with anti-β-amyloid aggregation activity from the fruits of crataegus pinnatifida bge

A new dineolignan, crataeguslignan A (1), along with one known dineolignan (2) were isolated from the fruits of Crataegus pinnatifida Bge. Its chemical structure was identified by comprehensive spectroscopic analyses. All the isolated compounds were investigated with regard to their Aβ 1–42 inhibition activity. Among them, 1 displayed the most potent Aβ 1–42 inhibitory ability with the inhibition rate of 85.2% at the concentration of 20 μM.

Chemical constituents from the leaves of Crataegus pinnatifida Bge

The phytochemical investigations on the leaves of Crataegus pinnatifida led to the isolation of 20 compounds, including seven triterpenoids (1-7), three hydroxycinnamic acids (8-10), three lignans (11-13) and seven flavonoids (14-20). All chemical structures were established on the basis of NMR (1H NMR, 13C NMR) spectroscopic data. Meanwhile, compounds 3-12 are reported for the first time from Crataegus genus. In addition, compounds 10-11 are isolated from the family Rosaceae for the first time. On the basis of chemical research, the chemotaxonomic significance of the isolated compounds has been discussed.

Chiral resolution and neuroprotective activities of enantiomeric 8-O-4′ neolignans from the fruits of Crataegus pinnatifida Bge

Five pairs of enantiomeric 8-O-4′ neolignans (1a/1b-5a/5b), including seven new compounds (1a/1b, 2a, 3a, 4a/4b and 5b), were obtained from the fruits of Crataegus pinnatifida Bge. Their structures were determined by comprehensive spectroscopic analyses. The absolute configurations of the enantiomers were determined by comparison of the experimental ECD with the calculated data. Additionally, all the enantiomeric neolignans were evaluated for their neuroprotective activity against H2O2-induced cell injury in human neuroblastoma SH-SY5Y cells, and most of them showed potent selective neuroprotective activity. Especially, 3b (72.98%) showed the best protective effect, better than 3a (63.49%) and trolox (62.86%) at 25 μM.

Pharmacological and transcriptome profiling analyses of Fufang E'jiao Jiang during chemotherapy-induced myelosuppression in mice

Ethnopharmacological relevanceFufang E'jiao Jiang (FEJ), a famous traditional Chinese medicine formula from “Liangyi Ointment”, consists of five crude drugs, Colla corii asini, Radix Ginseng Rubra, Radix Rehmanniae Preparata, Codonopsis pilosula, and Crataegus pinnatifida Bge. It has pronounced functions of qi-nourishing and blood-activating. Recently, it has been widely used in China as a medication against myelosuppression in cancer treatment. Aim of the studyWe aimed to investigate the complex mode of action and underlying mechanisms of Fufang E'jiao Jiang (FEJ) regarding its hematopoietic effect. Main methodsMice were divided into 5 groups of control, model, high dose FEJ (HFEJ), medium dose FEJ (MFEJ) and low dose FEJ (LFEJ). After 10 days from the administration, bone marrow cells (BMCs) were extracted for nucleated cells counts, flow cytometry analysis of hematopoietic stem cells (HSCs) population, as well as hematopoietic progenitor cells (HPCs) colony-forming unit (CFU) assay. A portion of bone marrow nucleated cells (BMNCs) of MFEJ group were prepared for RNA sequencing (RNA-Seq). The transcriptome data were analyzed based on the differentially expressed genes (DEGs). The molecular mechanisms of FEJ were deducted based on the biological processes and protein-protein interaction (PPI) network. ResultsFEJ could significantly increase the percentage of HSCs and the quantities of BFU-E and CFU-GM in BMSCs. FEJ could stimulate the proliferation of HSC and the differentiation of HPC to all lineages, which may thereby accelerate the recovery of hematopoietic function in myelosuppressive mice. By providing transcriptome profile we highlighted several genes and biological processes that might be applicable for FEJ to treat chemotherapy-induced myelosuppression. GO analysis showed that the co-expressed DEGs in FEJ vs model and model vs control group were involved in biological processes including ossification, osteoblast differentiation, bone mineralization and bone development. The KEGG pathway analysis pointed out ECM-receptor interaction and PI3K-AKT signaling pathway as the most relevant pathways to the function of FEJ on myelosuppression. PPI network showed MMP2 and COL1A1 were the relatively large nodes. ConclusionFEJ has the hematopoietic effect in chemotherapy-induced myelosuppression mice. It might be achieved by improving the proliferative capacity of HSCs and the differentiation ability of HPCs. The molecular mode of action of FEJ might be the improvement of the bone marrow microenvironment via ECM-receptor interaction, the promoted proliferation of HSC through regulation of PI3K-AKT signaling pathway, and the involvement of osteoblasts and osteoclasts. MMP2 and COL1A1 appear to be the key relevant regulatory molecules. These results provide significant insight into the hematopoietic effects of FEJ in myelosuppression and point out novel targets for future validating analyses.

Pectic polysaccharides from hawthorn: Physicochemical and partial structural characterization

The fruit of hawthorn (Crataegus pinnatifida Bge. var. major) is rich in pectic polysaccharides and has been used for centuries in China as food and herb. Hawthorn pectin (HP) recently has attracted rising attention due to the reported functional properties (e.g. gelling, emulsification) and health benefits towards the cardiovascular system. In this study, using hawthorn wine residue as raw material, a high methoxylated pectic polysaccharide HSP (degree of esterification up to 70%) was obtained by mild acid extraction (yield up to 20 wt%). Results from HPSEC coupled with triple detector revealed a widely dispersed (polydispersity index = 2) pectic polysaccharide with weight average molecular weight (Mw) of 111.7 kDa. Compared to pectin extracted from the hawthorn fruit, HSP showed a relatively lower molecular weight and higher polydispersity index (PDI), which suggested that the degradation of HP molecular chain during the wine-making processing (fermentation). Functionally, HSP demonstrated relatively high apparent viscosity but did not form gel in aqueous solution at concentrations up to 8 wt%. Structurally, HSP molecule was composed of over 70 mol% of 4-GalpA, with a small amount of glucose, galactose, xylose and arabinose. Under pectinase hydrolysis, over 95 wt% of the HSP was degraded into small fragments and monosaccharides; a small portion of relatively higher molecular weight rhamnose rich fragments were obtained from the hydrolytes. Combining both methylation analysis and 1D NMR spectroscopy, structural features of HSP were partially elucidated.

Effects and mechanisms of Danshen-Shanzha herb-pair for atherosclerosis treatment using network pharmacology and experimental pharmacology

Ethnopharmacological relevanceThe danshen (the root of Salvia miltiorrhiza Bge.)-shanzha (the fruit of Crataegus pinnatifida Bge. var. major N.E.Br.) (DS) herb combination is a commonly used traditional Chinese medicine with cardiovascular disease (CVD) treatment potential. Materials and methodsIn this study, we investigated the anti-atherosclerotic effects and mechanisms of DS by the integration of network pharmacology and polypharmacology. Eight main components were selected for target fishing by PharmMapper. ResultsThe network pharmacological study indicated that DS may target 41 proteins and 16 pathways associated with inflammation, lipid metabolism and endothelial protection, which indicates that DS probably adjusts these processes as part of its anti-atherosclerotic activities. Furthermore, this hypothesis was verified by polypharmacology using an atherosclerotic model. Histopathological examination showed that DS inhibited pathological changes in the arteries of atherosclerotic rats and reduced the intima-media thickness (IMT). DS significantly reduced the levels of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein-cholesterol (LDL-C) and increased the high-density lipoprotein-cholesterol (HDL-C) level in the blood. DS also decreased the concentrations of interleukin (IL)-1β and IL-18, indicating anti-inflammation activity. In addition, DS increased the serum levels of nitric oxide (NO) and 6-keto-prostaglandin F1α (6-keto-PGF1α) and decreased the serum levels of endothelin (ET) and thromboxane B2 (TXB2), indicating an endothelial protective effect. ConclusionsIn conclusion, DS has an anti-atherosclerotic ability to lower lipid concentrations and to protect endothelial function, and it also has anti-inflammatory activity.

Aqueous Two-phase System for the Extraction of Flavonoids from Hawthorn (Crataegus pinnatifida Bge.) Leaves

in the paper, the effect of aqueous two-phase system (ATPS) on the extraction of flavonoids from hawthorn (Crataegus pinnatifida Bge.) leaves was investigated. Based on the yield of flavonoids and partition coefficient (K), PEG 11.5% and (NH4)2SO4 14% was selected as the optimum solvent for extracting flavonoids. When the ratio of material to solvent was 1:35, the yield of flavonoids was 2.79%.The present study demonstrated that the aqueous two-phase system (ATPS) was an efficient method to extract flavonoids from hawthorn (Crataegus pinnatifida Bge.) leaves.

Near-infrared Spectroscopy as a Process Analytical Technology Tool for Monitoring the Parching Process of Traditional Chinese Medicine Based on Two Kinds of Chemical Indicators.

Background:The active ingredients and thus pharmacological efficacy of traditional Chinese medicine (TCM) at different degrees of parching process vary greatly.Objective:Near-infrared spectroscopy (NIR) was used to develop a new method for rapid online analysis of TCM parching process, using two kinds of chemical indicators (5-(hydroxymethyl) furfural [5-HMF] content and 420 nm absorbance) as reference values which were obviously observed and changed in most TCM parching process.Materials and Methods:Three representative TCMs, Areca (Areca catechu L.), Malt (Hordeum Vulgare L.), and Hawthorn (Crataegus pinnatifida Bge.), were used in this study. With partial least squares regression, calibration models of NIR were generated based on two kinds of reference values, i.e. 5-HMF contents measured by high-performance liquid chromatography (HPLC) and 420 nm absorbance measured by ultraviolet–visible spectroscopy (UV/Vis), respectively.Results:In the optimized models for 5-HMF, the root mean square errors of prediction (RMSEP) for Areca, Malt, and Hawthorn was 0.0192, 0.0301, and 0.2600 and correlation coefficients (Rcal) were 99.86%, 99.88%, and 99.88%, respectively. Moreover, in the optimized models using 420 nm absorbance as reference values, the RMSEP for Areca, Malt, and Hawthorn was 0.0229, 0.0096, and 0.0409 and Rcal were 99.69%, 99.81%, and 99.62%, respectively.Conclusions:NIR models with 5-HMF content and 420 nm absorbance as reference values can rapidly and effectively identify three kinds of TCM in different parching processes. This method has great promise to replace current subjective color judgment and time-consuming HPLC or UV/Vis methods and is suitable for rapid online analysis and quality control in TCM industrial manufacturing process.SUMMARY Near-infrared spectroscopy.(NIR) was used to develop a new method for online analysis of traditional Chinese medicine.(TCM) parching processCalibration and validation models of Areca, Malt, and Hawthorn were generated by partial least squares regression using 5.(hydroxymethyl) furfural contents and 420.nm absorbance as reference values, respectively, which were main indicator components during parching process of most TCMThe established NIR models of three TCMs had low root mean square errors of prediction and high correlation coefficientsThe NIR method has great promise for use in TCM industrial manufacturing processes for rapid online analysis and quality control. Abbreviations used: NIR: Near-infrared Spectroscopy; TCM: Traditional Chinese medicine; Areca: Areca catechu L.; Hawthorn: Crataegus pinnatifida Bge.; Malt: Hordeum vulgare L.; 5-HMF: 5-(hydroxymethyl) furfural; PLS: Partial least squares; D: Dimension faction; SLS: Straight line subtraction, MSC: Multiplicative scatter correction; VN: Vector normalization; RMSECV: Root mean square errors of cross-validation; RMSEP: Root mean square errors of validation; Rcal: Correlation coefficients; RPD: Residual predictive deviation; PAT: Process analytical technology; FDA: Food and Drug Administration; ICH: International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use.

Major triterpenoids in Chinese hawthorn “Crataegus pinnatifida” and their effects on cell proliferation and apoptosis induction in MDA-MB-231 cancer cells

The cytotoxicity and antiproliferative effect of phytochemicals presenting in the fruits of Chinese hawthorn (Crataegus pinnatifida) were evaluated. Shanlihong (Crataegus pinnatifida Bge. var. major N.E.Br.) variety possessed significant levels of flavonoids and triterpenoids, and showed potent antiproliferative effect against HepG2, MCF-7 and MDA-MB- 231 human cancer cells lines. Triterpenoids-enriched fraction (S9) prepared by Semi-preparative HPLC, and its predominant ingredient ursolic acid (UA) demonstrated remarkably antiproliferative activities for all the tested cancer cell lines. DNA flow cytometric analysis showed that S9 fraction and UA significantly induced G1 arrest in MDA-MB-231 cells in a dose-dependent manner. Western blotting analysis revealed that S9 fraction and UA significantly induced PCNA, CDK4, and Cyclin D1 downregulation in MDA-MB-231 cells, followed by p21Waf1/Cip1 up-regulation. Additionally, flow cytometer and DNA ladder assays indicated that S9 fraction and UA significantly induced MDA-MB-231 cells apoptosis. Mitochondrial death pathway was involved in this apoptosis as significantly induced caspase-9 and caspase-3 activation. These results suggested that triterpenoids-enriched fraction and UA exhibited antiproliferative activity through the cell cycle arrest and apoptosis induction, and was majorly responsible for the potent anticancer activity of Chinese hawthorn.

In Vitro Study on Anti-Hepatitis C Virus Activity of Spatholobus suberectus Dunn.

Hepatitis C virus (HCV) infects 200 million people worldwide, and 75% of HCV cases progress into chronic infections, which consequently cause cirrhosis and hepatocellular carcinoma. HCV infection is treated with currently considered standard drugs, including direct anti-viral agents (DAAs), alone or in combination with peginterferon-α plus ribavirin. However, sustained viral responses vary in different cohorts, and high costs limit the broad use of DAAs. In this study, the ethanol and water extracts of 12 herbs from Lingnan in China were examined in terms of their inhibitory effect on HCV replication. Among the examined extracts, Spatholobus suberectus ethanol extracts suppressed HCV replication. By comparison, Extracts from Fructus lycii, Radix astragali (root), Rubus chingii Hu (fruit), Flos chrysanthemi Indici (flower), Cassia obtusifolia (seed), Lonicera japonica Thunb (flower), Forsythia suspense Thunb (fruit), Poria cocos (sclerotia), Carthamus tinctorius L. (flower), Crataegus pinnatifida Bge. (fruit), and Leonurus japonicas Houtt. (leaf) extracts failed to show a similar activity. Active S. suberectus fractions containing tannins as the major component also inhibited the in vitro translation of HCV RNA. The combination treatments of single compounds, such as epigallocatechin gallate and epicatechin gallate, were not as potent as crude S. suberectus fractions; therefore, crude S. suberectus extract may be a potential alternative treatment against HCV either alone or in combination with other agents.

Classification and phylogenetic analysis of Chinese hawthorn assessed by plant and pollen morphology.

The Chinese hawthorn (Crataegus pinnatifida Bge. var. major N.E.Br.) is uniquely originated in northern China. The ecological and horticultural importance of Chinese hawthorn is considerable and some varieties are valued for their fruit or medicine extracts. Its taxonomy and phylogeny remain poorly understood. Apart from general plant morphological traits, pollen is an important trait for the classification of plants and their evolutionary origin. However, few studies have investigated the pollen of Chinese hawthorn. Here, an analysis of plant and pollen morphological characteristics was conducted in 57 cultivars from the Shenyang region. Thirty plant morphological characters and nine pollen grain characters were investigated. The plant morphological analysis revealed that the coefficient of variation for 13 traits was >20%, which indicates a high degree of variability. We also found that the pollen grains varied greatly in size, shape (from prolate to perprolate), and exine pattern (striate-perforate predominantly). The number of apertures was typically three. Based on these findings, we suggest that pollen morphology associated with plant morphological traits can be used for classification and phylogenetic analysis of Chinese hawthorn cultivars. In sum, our results provide new insights and constitute a scientific basis for future studies on the classification and evolution of Chinese hawthorn.

English

Hawthorn (Crataegus pinnatifida Bge.) flavonoids constituents have recently received greater attention for their health and medical properties. In order to relgulate the flavonoids constituents, one-year-old hawthorn plants were subjected to foliar application of L-glutamic acid (GLA, 800 mg L–1 ), glycine (GLY, 20 mg L–1), L-phenylalanine (PHE, 20 mg L–1), L-arginine (ARG, 20 mg L–1). The levels of rhamnosylvitexin, rutin and hyperoside in both leaves and root exudates were significantly (p<0.05) increased in GLA and PHE treatments. Application of GLY and PHE increased the levels of rutin and hyperoside in hawthorn roots, especially for PHE treatment, and the level of hyperoside in PHE treatment was significantly (p<0.05) higher than in other treatments. No positive effects were observed for quercetin under exogenous amino acids treatments in both leaves and roots. The total flavonoids content of leaves, roots and root exudates under GLA and PHE treatments were significantly increased compared to the control. In addition, the phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activities of leaves and roots were also increased. These results suggest that some exogenous amino acids did affect the flavonoids content, which may have significance in regulation of flavonoids in hawthorn. Key words: Amino acids, flavonoids, phenylalanine ammonia-lyase.

Antioxidant effects of 14 Chinese traditional medicinal herbs against human low-density lipoprotein oxidation

The relationship between the antioxidant activities and inhibitory effect of 14 Chinese medicinal herbs against oxidized low-density lipoprotein (LDL) formation was evaluated. Prolongation of the lag phase of LDL oxidation depended on the concentration of the herbs. The concentration of each herb that was able to prolong the lag time by about two-fold was calculated and expressed as doubling-time concentration. The lower the doubling-time concentration, the stronger the inhibitory effect exhibited toward LDL oxidation. Among them, Chrysanthemi Flos (Chrysanthemum morifolium ramat; 甘菊花 gān jú huā), Crataegi Fructus (Crataegus pinnatifida Bge. var. major N.E.Br.; 山楂 shān zhā), and Roselle (Hibiscus sabdariffa Linn.; 洛神 luò shén) showed significant inhibitory effects. Correlation coefficients between doubling-time concentration and radical-scavenging activities were high; the total phenolic content was also high. In conclusion, phenolic compounds contributed not only to antioxidant activities, but also to the inhibitory effect against LDL oxidation. Chrysanthemi Flos, Crataegi Fructus, and H. sabdariffa, with lower doubling-time concentrations, could be potent phytochemical agents to reduce LDL oxidation and prevent the progression of atherosclerosis.

HPLC DETERMINATION OF VITEXIN-4″-O-GLUCOSIDE IN MOUSE PLASMA AND TISSUES AFTER ORAL AND INTRAVENOUS ADMINISTRATION

Vitexin-4″-O-glucoside (VOG) was herein isolated from the leaves of Crataegus pinnatifida Bge. var. major, and then a sensitive and specific high performance liquid chromatography (HPLC) method was developed and validated for studying the pharmacokinetics and tissue distribution of VOG after oral and intravenous administration at a dose of 30 mg/kg. The results indicated that VOG was rapidly and widespreadly distributed throughout the whole body after administration and the oral bioavailability was 11.97 ± 0.66%. The highest VOG level after oral administration was observed in gallbladder, followed by stomach and small intestine. Whilst, the highest VOG level after intravenous administration was found in gallbladder, followed by liver and small intestine.

Structural Analysis of Proanthocyanidins Isolated from Fruit Stone of Chinese Hawthorn with Potent Antityrosinase and Antioxidant Activity

Proanthocyanidins were isolated from fruit stone of Chinese hawthorn (Crataegus pinnatifida Bge. var. major N.E.Br.). Their structures were analyzed and elucidated by methods of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and high performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS). The results demonstrated that these compounds are complicated mixtures of homo- and heteropolymers consisting of procyanidin/procyanidin gallate and prodelphinidin. They possessed structural heterogeneity in monomer units, polymer length, and interflavan linkage (A-type and B-type). Their antityrosinase and antioxidant activity were then investigated. The results revealed that they can inhibit tyrosinase activities, including the monophenolase activity and the diphenolase activity. In addition, proanthocyanidins possessed potent antioxidant activity. Our studies revealed that proanthocyanidins isolated from fruit stone of Chinese hawthorn may be applied in food, agriculture, pharmaceutical, and cosmetic industries.

First Report of Brown Rot on Crataegus pinnatifida var. major Caused by Monilia yunnanensis in China.

Crataegus pinnatifida Bge. var. major N. is a time-honored herbal medicine and an important economic fruit that is processed into various foods. Cultivated fields are mostly situated in northern China. In August 2012, a fruit brown rot disease on mature plants was found in Fushun, Liaoning Province, China. This disease reduced the yield and the quality of fruits; the incidence of fruits affected in each tree was between 20% and 35%. The initial stage of infection was marked by regular or irregular light brown spots turning dark brown and gradually supporting many grey fluffy tufts of mycelium. At last, the lesions expanded, causing the fruits to become rotted or withered. The pathogen was isolated from infected fruits by a tissue isolation method (1) and cultured on potato dextrose agar (PDA) at 25°C under 12 h light/12 h dark for 7 days. The colonies reached 60 to 75 mm in diameter after 7 days and were pale green or pale yellow with a neat greyish white margin; the aerial mycelium sometimes formed annulations. The conidia were single, hyaline, lemon-shaped or oval, and 11.3 ± 1.4 (8.0 to 16.5) × 6.4 ± 0.8 (5.1 to 7.6) μm; sporulation was sparse and stromata that formed after 15 days were abundant, black, and spherical to elliptical in shape. Conidiophores were simple or branched, bearing a single conidium on the tip. The internal transcribed spacer (ITS) region of rDNA amplified with primers ITS1 and ITS4 was directly sequenced in both directions. The ITS1-5.8S-ITS2 sequence of rDNA was 100% identical to Monilinia yunnanensis (GenBank Accession No. HQ908788.1), which is distinguished from the closely related M. fructigena (AF150679.1) and M. polystroma (JX315717.1). The pathogen was identified as M. yunnanensis on the basis of morphological characteristics (2) and the ITS sequence of rDNA (2,3). Pathogenicity tests were performed on detached healthy fruits. Four fruits were wounded with a sterilized nail and inoculated by placing a PDA plug of mycelia on upper surfaces of the fruits. Another four fruits treated with sterile PDA plugs served as a control. Fruits were cultured in petri dishes with a 12-h photoperiod at 25°C and 90% relative humidity. The initial symptoms on inoculated fruits were observed after 5 days, while no symptoms showed on the controls. The pathogen was reisolated from the inoculated fruit and identified as M. yunnanensis by the above methods. While M. yunnanensis has been reported to cause brown rot on peach (2), to our knowledge, this is the first report of M. yunnanensis infecting C. pinnatifida Bge. var. major N. in China.