Integrative analysis of the metabolome and transcriptome reveals the potential mechanism of fruit flavor formation in wild hawthorn(Crataegus chungtienensis)

Hawthorns are important medicinal and edible plants with a long history of health protection in China.Besides cultivated hawthorn,other wild hawthorns may also have excellent medicinal and edible value,such as Crataegus chungtienensis,an endemic species distributed in the Southwest of China.In this study,by integrating the flavor-related metabolome and transcriptome data of the ripening fruit of C.chungtienensis,we have developed an understanding of the formation of hawthorn fruit quality.The results show that a total of 849 metabolites were detected in the young and mature fruit of C.chungtienensis,of which flavonoids were the most detected metabolites.Among the differentially accumulated metabolites,stachyose,maltotetraose and cis-aconitic acid were significantly increased during fruit ripening,and these may be important metabolites affecting fruit flavor change.Moreover,several flavonoids and terpenoids were reduced after fruit ripening compared with young fruit.Therefore,using the unripe fruit of C.chungtienensis may allow us to obtain more medicinal active ingredients such as flavonoids and terpenoids.Furthermore,we screened out some differentially expressed genes(DEGs)related to fruit quality formation,which had important relationships with differentially accumulated sugars,acids,flavonoids and terpenoids.Our study provides new insights into flavor formation in wild hawthorn during fruit development and ripening,and at the same time this study lays the foundation for the improvement of hawthorn fruit flavor.

Are Polyploid Species Less Vulnerable to Climate Change? A Simulation Study in North American Crataegus

Understanding the mechanisms underlying plant responses to climate change is an important step toward developing effective mitigation strategies. Polyploidy is an important evolutionary trait that can influence the capacity of plants to adapt to climate change. The environmental flexibility of polyploids suggests their resiliency to climate change, however, such hypotheses have not yet received empirical evidence. To understand how ploidy level may influence response to climate change, we modeled the current and future distribution of 54 Crataegus species under moderate to severe environments and compared the range change between diploids and polyploids. The majority of studied species are predicted to experience considerable range expansion. We found a negative interaction between ploidy and ecoregions in determining the response to climate change. In extreme environments, polyploids are projected to experience a higher range expansion than diploids with climate change, while the opposite is true for moderate environments. The range expansion of Crataegus species can be attributed to their tolerance for a wide range of environmental conditions. Despite the higher tolerance of polyploids to extreme environments, they do not necessarily outperform diploids in moderate environments, which can be attributed to the varying nature of species interactions along a stress gradient.

Morphological and ISSR molecular markers reveal genetic diversity of wild hawthorns (Crataegus songorica K. Koch.) in Xinjiang, China

The wild hawthorn species, Crataegus songorica K. Koch., is an important wild germplasm resource in Xinjiang, China that has been endangered in recent years. The genetic diversity of C. songorica K. Koch. germplasm in five populations from Daxigou, Xinjiang, China were evaluated based on phenotypic traits and ISSR molecular markers to provide basic infor- mation on resource protection, rational utilization and genetic improvement. The F-value for the phenotypic differentiation coefficient of the 33 traits measured ranged from 0.266 to 15.128, and mean value was 13.85%. The variation among populations was found to be lower than that within population. A total of 303 loci were detected within the five populations by 12 primers. Within 298,polymorphic loci, the polymorphism was 98.35%, showing a high genetic diversity in C. songorica K. Koch. The gene diversity within population, total population genetic diversity, genetic differentiation coefficient and gene flow were 0.2779, 0.3235, 0.1408, and 3.0511, respectively. Our results showed that C. songorica K. Koch. from Xinjiang has a high level of genetic diversity at both the phenotypic and molecular levels. Significant genetic differentiation existed within population and the differentiation trend showed a regional association. And in this study, in situ and ex situ conser- vation approaches were raised for wild hawthorn protection utilization.

Phytochemical and biological activities of Crataegus sinaica growing in Egypt

Objective:To evaluate the cardiac activity and hepatoprotection of Crataegus sinaica (C.sinaica).Methods:All the isolated compounds were isolated by open-column liquid chromatography(CC) using sephadex LH-20 as stationary phase.Elution of the column was performed with EtOH or MeOH.The phytochemical investigation of the young stem of C.sinaica for the first time together with the leaves and flowers lead to the isolation and identification of quercetin,hyperoside,vitexin-2"-0-rhamnoside,epicatechin,procyanidin B2 and procyanidins CI.Results:Rats treated with the low and high dose of C.sinaica leaves with flowers extract showed 15%and 17%reduction in the heart rate,and reduction in the STsegment by 107%and 57%;respectively.The T-amplitude was decreased by 59%of the high dose extract.On the other hand,the young steins and leaves with flowers extracts of C.sinaica on primary culture of rat hepatocytes monolayer indicated a hepatoprotection for the total extract,ethyl acetate,butanol,and chlor of orm fractions at 100μg/mL,75μg/mL,50μg/mL,and 25μg/mL; respectively.Conclusions:The results of these chemical and biological studies suggest the use of C.sinaica growing in Egypt as a preventive drug against cardiovascular and hepatic diseases. The chemical studies suggest the use of woody young stems as a newly investigated bioactive organ.The extraction of unsaturated fatty acids from the seeds of the plant would serve as a good health and nutritive product.

Two New C-glucoside Flavonoids from Leaves of Crataegus pinnatifida Bge.var.major N.E.Br.

Two new C-glucoside flavonoids, namely 8-C-b-D-(2-O-acetyl) glucofuranosyl apigenin and 3-O-acetylvitexin, were isolated from leaves of Crataegus pinnatifida Bge. var. major N. E. Br.. Their structures were elucidated by the spectroscopic means and chemical evidence.

A New Flavonoid Ketohexofuranoside from Leaves of Crataegus pinnatifida Bge.var. major N.E.Br.

A new flavonoid, namely pinnatifine I (1), was isolated from the leaves of Crataegus pinnatifida Bge. var. major N.E.Br.. Its structure was elucidated by spectroscopic analysis and chemical evidence.

Inhibitory Activity of Prunus mume,Copitis chinensis and Crataegus pinnatifida on Vibrio harveyi and Its Biofilm

[Objective] This study was conducted to determine the in-vitro inhibitory effects of Prunus mume,Coptis chinensis and Crataegus pinnatifida on Vibrio harveyi and its biofilm. [Method]The inhibitory zone diameters of the three Chinese herbal medicines against V. harveyi were determined by agar diffusion method; the minimal inhibitory concentration( MIC) and minimal bactericidal concentration( MBC) values of the three Chinese herbal medicines against V. harveyi were determined by doubling dilution method; and the effects of the three Chinese herbal medicines on the formation of V. harveyi biofilm were determined by methyl thiazolyl tetrazolium( MTT) method. [Result]The three Chinese herbal medicines all inhibited V. harveyi to different degrees. C. chinensis and C. pinnatifida and P. mume exhibited the inhibitory zone diameters of( 17. 62 ± 0. 04),( 20. 16 ± 0. 08) and( 30. 76 ± 0. 26) mm against V. harveyi,respectively. P. mume and C. pinnatifida had strong inhibitory effects on V. harveyi. The MIC and MBC values of P. mume against V. harveyi were 7. 812 5 mg/ml; the MIC and MBC values of C. pinnatifida against V. harveyi were 31. 25 mg/ml; and the MIC and MBC values of C. chinensis against V. harveyi were 62. 5 mg/ml. P. mume had the strongest antibacterial and bactericidal ability. The MIC values of C. pinnatifida,C. chinensis and P. mume against V. harveyi were 7. 81,7. 81 and 1. 96 mg/ml,respectively,i. e.,P. mume exhibited the lowest MIC. [Conclusion] P. mume,C. pinnatifida and C. chinensis all have inhibitory effects on V. harveyi and its biofilm,and P. mume has the strongest bactericidal ability.

Optimization of PCR-SSCP Reaction System and Reaction Conditions in Crataegus spp.

[Objective] This study was conducted to optimize PCR-SSCP reaction system and conditions for hawthorn （Crataegus spp.）. [Method] The chloroplast DNA and nuclear DNA of hawthorn leaf were extracted with improved CTAB method. Pdmers for the PCR amplification of chloroplast DNA and nuclear DNA were select- ed from eight pairs of candidate pdmers, and the PCR-SSCP reaction system and reaction conditions were optimized. The PCR products were detected by agarose gel electrophoresis, and the denatured PCR-SSCP products were analyzed by native polyacrylamide gel. [Result] Five pairs primers （psbA-tmH, ropB, ropL, rpoC1 and ITS2） were proved to be suitable for PCR-SSCP in hawthorn, including four for chloroplast DNA and one for nuclear DNA. The clear electrophoretogram of PCR- SSCP in hawthorn was obtained by performing electrophoresis in 0.5×TBE buffer, at 4 ℃ and 200 V for 3-4 h, using 6% native polyacrylamide gel （crosslinking ratio at 29：1）, and the PCR product had been mixed with an equal volume of loading buffer containing 1% NaOH （without glycerol） and denatured at 98℃ for 15 min. [Conclu- sion] The results may lay the foundation of SSCP analysis of hawthom.

Anti-Obesity and Antihyperglycemic Effects of <i>Crataegus aronia</i>Extracts: <i>In Vitro</i>and <i>in Vivo</i>Evaluations

Hypocholesterolemic activity of Crataegus aronia L. (Rosaceae) is therapeutically praised. Its potent antiobesity (P < 0.001, n = 6 - 8) as well as marked triacylglycerol-reducing efficacies (P < 0.001, n = 6 - 8) in 10 weeks-high cholesterol diet (HCD) fed rats are demonstrated. Pancreatic triacylglycerol lipase (PL), α-amylase and α-glucosidase are an interesting pharmacological target for the management of dyslipidemia, atherosclerosis, diabetes and obesity. Comparable to acarbose, acute starch induced postprandial hyperglycaemia as well glycemic excursions in normoglycemic overnight fasting rats was highly significantly (P < 0.001) dampened by C. aronia 100, 200 and 400 mg/Kg b.wt aqueous extracts (AE), but not acute glucose evoked postprandial hyperglycaemia increments, unlike diabetes pharmaco-therapeutics metformin and glipizide. C. aronia aerial parts as well as fruits AEs (0.1 - 10 mg/mL) were identified as in vitro dual inhibitors of α-amylase and α-glucosidase with respective IC50 (mg/mL) of 2.1 ± 0.3 and 3.5 ± 0.7. Still, it lacked on in vitro hindrance of glucose movement, dissimilar to guar gum. Equivalent to orlistat (PL IC50 of 0.1 ± 0.0 μg/mL), C. aronia tested AEs and its purified bioactive phytoconstituents;quercetin and rutin, inhibited highly substantially in a dose dependent trend PL in vitro (n = 3), in an ascending order of obtained PL-IC50 (μg/mL): quercetin;30.1 ± 2.8, rutin;77.3 ± 11.7, C. aronia aerial parts;225.2 ± 33.4 and C. aronia fruits;286.1 ± 37.4. Flavonoid-rich C. aronia, as a functional food and a nutraceutical, modulating gastrointestinal carbohydrate and lipid digestion and absorption, maybe be advocated as an exquisite and potential candidate for combinatorial obesity-diabetes prevention and phytotherapy.

Determination of Nine Phenolic Components in Leaves of Crataegus pinnatifida Bge.

[Objectives] To determine the nine phenolic components in the leaves of Crataegus pinnatifida Bge. [Methods] The reversedphase high-performance liquid chromatography( RP-HPLC) was applied. [Results] Nine phenolic components showed a good linear relationship in the range of 2-500 μg/m L with r in the range of 0. 999 5-0. 999 9. The recovery rate of spiked samples ranged from 93. 7% to110. 2%,and the relative standard deviation was in the range of 0. 69%-4. 58%. The leaves of 29 cultivars of C. pinnatifida Bge. were measured,and the average content of the nine phenolic components was as follows: isoquercitrin,hyperoside,procyanidin C1,procyanidin D1,epicatechin,procyanidin B2,chlorogenic acid,eucomic acid,and vitexin 2 "-O-rhamnoside. The contents of flavonoids and phenolic acids were high,up to 15 mg/g D. W,and the content of procyanidins was up to 6 mg/g D. W. [Conclusions]This method is easy and accurate in determination of phenolic components in the leaves of C. pinnatifida Bge.

In Vitro Efficacy of Crataegus oxycantha L. (Hawthorn) and Its Major Components against ATCC and Clinical Strains of Ureaplasma urealyticum

Crataegus oxycantha L., commonly known as hawthorn, has traditionally been used for its beneficial effect on cardiovascular health, which is related to its flavonoid content. The aim of the present study was to evaluate the antibacterial properties of a fluid extract and a hydro-ethanolic macerate from buds of Crataegus oxycantha against clinical isolates of Ureaplasma urealyticum. The major purified flavonoids present in the extracts were also tested against ATCC strains and clinical isolates. Both the fluid extract and the hydro-ethanolic macerate were active against thirty-due clinical strains of U. urealyticum, with MIC ranges between 15.6 and 250 μg/ml and 15.6 and 62.5 μg/ml, respectively. All pure organic compounds, with the exception of rutin, showed activity against the strains tested, luteolin 3,7-diglucoside being the most active compound (MICs in the range of 0.48 and 1.95 μg/ml), followed by apigenin-7-O-glucoside (MICs in the range of 0.48 and 3.9 μg/ml). The activity of the pure flavonoids was greater against the clinical isolates compared to the ATCC strains. The data presented here demonstrate that flavonoids present in Crataegus oxycantha are effective against clinical isolates of U. urealyticum and could be used in combination with antibiotics in order to combat resistance.

Phenolic compounds obtained from the fruit of Crataegus pinnatifida with neuroprotective activities

Ten phenolic compounds(1a/1b,2–9)including a pair of enantiomers(1a/1b),along with eight analogues(2–9)were isolated from the fruit of Crataegus pinnatifida.Enantiomers 1a/1b were separated successfully by chiral chromatographic column.Their structures were established by comprehensive spectroscopic analyses,and the absolute configurations of enantiomers were determined by comparison between the experimental and calculated electronic circular dichroism(ECD)spectra.In addition,all isolates were investigated for their neuroprotective effects against H2O2–induced oxidative stress in human neuroblastoma SH-SY5Y cells.It was found that enantiomers 1a and 1b displayed significant neuroprotective activities but no enantioselectivity.In addition,compounds 3–6 showed obvious neuroprotective effects at different concentrations,while compound 8 exhibited potential neuroprotective effect at higher concentration(50μmol/L).

The chemistry,stability and health effects of phenolic compounds in cultivated hawthorn(Crataegus pinnatifida var.major):a review

Cultivated hawthorn(Crataegus pinnatifida var.major),the principal cultivated hawthorn species in China,is rich in phenolic compounds.These phytochemicals are considered essential bioactive ingredients in different organs of cultivated hawthorn,contributing to its health effects.However,there is lack of a comprehensive review that systematically summarizes the chemistry,health effects,and stability of phenolic substances.This review summarizes recent progress in the characterization,classification,and distribution of phenolic compounds in C.pinnatifida var.major,including procyanidins,flavonoids,phenolic acids,lignans,and others.Importantly,the stability of these compounds was found to be influenced by factors such as the hawthorn development process,postharvest storage,postharvest processing,and the gastrointestinal tract environment.Additionally,the health effects of phenolic compounds in cultivated hawthorn and their underlying mechanisms were discussed.This review provides valuable insights into the utilization of hawthorn resources and the development of the hawthorn industry.

Effect of Shan Zha(Hawthorn or Crataegus)on gastrointestinal cancer:A network pharmacology and molecular docking study

Background Shan Zha(Hawthorn or Crataegus)is a traditional Chinese medicine(TCM)most commonly used for the treatment of hyperlipidemia.Gastrointestinal cancer is closely correlated with blood lipid levels.This study illustrates the potential anticancer effects of Shan Zha on gastrointestinal tumors based on network pharmacology and molecular docking.Methods Hawthorn's bioactive ingredients and drug targets were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine version 2.0(TCMIP v2.0),and Herbal Ingredients'Targets Platform(HIT 2.0)databases.Validated disease targets of gastrointestinal cancer were obtained from the Therapeutic Targets Database(TTD)and HIT 2.0 databases.Protein–protein interaction analysis of intersecting genes was performed using the Search Tool for the Retrieval of Interacting Genes(STRING)database.The functions of these genes were further analyzed by performing gene ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.Molecular docking verification was performed using Molecular Operating Environment(MOE)software.Results Four main bioactive components were identified in Shan Zha.A total of 271 potential drug targets were identified,and 393 gastrointestinal-tumor targets were obtained.Through protein interaction analysis of intersecting targets,the main components of Shan Zha were found to interact more closely with proteins such as tumor protein p53(TP53),AKT serine/threonine kinase 1(AKT1),JUN proto-oncogene(JUN),interleukin 6(IL6),epidermal growth factor receptor(EGFR),and vascular endothelial growth factor A(VEGFA).KEGG pathway enrichment analysis showed a total of 127 pathways,mainly involving pathways in multiple types of cancer,the Phosphatidylinositol 3-kinase-Akt(PI3K-Akt)signaling pathway,and EGFR tyrosine kinase inhibitor resistance.Combined with The Cancer Genome Atlas(TCGA)differential analysis,key targets,including TP53,cyclin D1(CCND1),EGFR,and VEGFA,were screened.Molecular docking results showed that quercetin and kaempferol had the good binding potential for TP53,CCND1,EGFR,and VEGFA.Conclusion These findings suggest that Shan Zha exerts its effects on gastrointestinal cancers through a multitarget,multi-component,and a multi-pathway mechanism.

Cultivated hawthorn(Crataegus pinnatifida var.major)genome sheds light on the evolution of Maleae(apple tribe)

Cultivated hawthorn(Crataegus pinnatifida var.major)is an important medicinal and edible plant with a long history of use for health protection in China.Herein,we provide a de novo chromosomelevel genome sequence of the hawthorn cultivar“Qiu Jinxing.”We assembled an 823.41 Mb genome encoding 40571 genes and further anchored the779.24 Mb sequence into 17 pseudo-chromosomes,which account for 94.64%of the assembled genome.Phylogenomic analyses revealed that cultivated hawthorn diverged from other species within the Maleae(apple tribe)at approximately 35.4 Mya.Notably,genes involved in the flavonoid and triterpenoid biosynthetic pathways have been significantly amplified in the hawthorn genome.In addition,our results indicated that the Maleae share a unique ancient tetraploidization event;however,no recent independent whole-genome duplication event was specifically detected in hawthorn.The amplification of non-specific long terminal repeat retrotransposons contributed the most to the expansion of the hawthorn genome.Furthermore,we identified two paleo-sub-genomes in extant species of Maleae and found that these two sub-genomes showed different rearrangement mechanisms.We also reconstructed the ancestral chromosomes of Rosaceae and discussed two possible paleopolyploid origin patterns(autopolyploidization or allopolyploidization)of Maleae.Overall,our study provides an improved context for understanding the evolution of Maleae species,and this new highquality reference genome provides a useful resource for the horticultural improvement of hawthorn.

New norlignan enantiomers from the fruit of Crataegus pinnatifida with neuroprotective activities

(±)-Crataegusnorin A(la/1b) and B(2a/2b),two pairs of rare 8,9’-epoxy-type norlignan enantiomers featuring a y-butyrolactone ring,were isolated from the fruit of Crataegus pinnatifida.Their structures were determined via extensive spectroscopic analyses.Gauge-independent atomic orbital(GIAO) NMR chemical shift calculations,combined with the advanced statistical method DP4+were employed to establish the relative configurations of four compounds.Next,chiral separation was accomplished by chiral chromatographic column and the absolute configurations of the four compounds were unambiguously assigned by comparison between their experimental electronic circular dichroism curves with the quantum-mechanically calculated curves based on time-dependent density functional theory(TDDFT).All the isolates were evaluated fo r their neuroprotective activities against H2O2-induced cell injury in human neuroblastoma SH-SY5Y cells.The results showed that two pairs of enantiomers 1a/1b and 2a/2b displayed diff;erent effect on neuroprotective activity.Among them,compound 2a displayed the most potent neuroprotective effect Further flow cytometry analysis indicated that 2a could protect SH-SY5Y cells from oxidative damage through inhibiting cell apoptosis.

Opposite Modulatory Effects of Crataegus aronia Aqueous Extract on Platelet Aggregation in Rats

Objectives:To reveal the mechanisms behind the dual effects of Crataegus aronia(C.aronia)aqueous extract on platelet aggregation by focusing on function,regulation,expression,and signaling of platelets P_(2)Y_(12)receptors.Methods:Adult male Wistar rats(120±10 g)were classified as control received the vehicle,C.aronia(200 mg/kg),and C.aronia(2,000 mg/kg)-treated rats.After treatments for consecutive 7 days,hematological and molecular experiments were conducted to detect alterations in platelet aggregation,thromboxane B2(THXB2)and intracellular reactive oxygen species(ROS)content;protein levels of P_(2)Y_(12),p-Akt,cyclic adenosine monophosphate(c AMP),phosphorylated vasodilator-stimulated-phosphoprotein(p-VASP),nuclear factorκB(NF-κB),P-selectin,etc.in platelets were determined by Western blot;m RNA expressions of P_(2)Y_(12)and some inflammatory markers were determined by real-time polymerase chain reaction.Results:At a concentration of 200 mg/kg,C.aronia inhibited platelet aggregation through multiple interconnected mechanisms including downregulation P_(2)Y_(12)synthesis and expression,stimulating intracellular cAMP levels and protein levels of p-VASP,inhibiting platelets THXB2 release and protein levels of P-selectin.Also,it inhibited platelets level of ROS and of NF-κB,a major signaling pathway that stimulates the expression of P_(2)Y_(12)and THXA2 synthesis.Opposite findings were seen in platelets of rats received C.aronia at a concentration of 2,000 mg/kg.Interestingly,co-administration of N-acetylcysteine prevented all hematological and molecular alterations exerted by the high dose of the extract and inhibited platelet aggregation.Conclusion:Oral administration of C.aronia at low dose inhibits platelet aggregation by reducing THXB2 release,expression of P-selectin and activating c AMP and Akt signaling through two major mechanisms including downregulation of P_(2)Y_(12)and inhibition of ROS-induced activation of NF-κB,an effect that is observed to be in the opposite direction with its high dose.

山楂研究现状

山楂是一种常用药。本草记载其味酸甘,性微温,归脾、胃、肝经能消食积、散淤血。现代研究发现山楂具有多方面的药理作用,近年又取得了新的进展。山楂临床应用也日趋广泛。山楂资源也有所发展。

长白山稀有抗寒果树资源——山楂海棠

山楂海棠[Malus komarovii(Sarg.)Rehd]俗称山苹果,蔷薇科,苹果属。曾定名为薄叶山楂,归山楂属[Crataegus tenuifolia kom.]。为国家重点保护的野生植物。分布于长白山南坡,长白县境内,海拔1,1000—1,300米的个别地段。朝鲜也有分布。是珍贵的抗寒果树种质资源。

壳聚糖涂膜处理对山楂的保鲜效果研究

采用不同浓度(0.0%、0.5%、1.0%、1.5%、2.0%)壳聚糖对山楂进行涂膜处理后,置于常温下贮藏,定期测定其理化指标,研究壳聚糖对山楂的保鲜效果。结果表明,壳聚糖涂膜处理可以降低山楂的失重率、维生素C的损失,延缓好果率和果肉硬度的下降,保持山楂的贮藏品质,其中,浓度为1.5%的壳聚糖处理对山楂的保鲜效果好且成本较低。