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.

山楂(Crataegus pinnatifida Bge．)遗传多样性的RAPD和ISSR标记分析

利用RAPD和ISSR标记对35份山楂（Crataegus pinnatifida Bge．）资源进行了DNA多态性分析。12个RAPD引物共扩增出110条清晰的谱带，其中89条显示多态性，平均每个引物扩增出7．4条多态性谱带。13个ISSR引物共扩增出110条清晰的谱带，其中94条显示多态性，平均每个引物扩增出7．2条多态性谱带。基于RAPD和ISSR标记，利用UPGMA分别构建了35份山楂资源的聚类树状图。距离系数分别为0～0．62（RAPD）和0～0．64（ISSR），表明山楂具有较高的遗传多样性。

葡萄糖对盐胁迫下山楂(Crataegus pinnatifida Bge.)幼苗光合荧光特性的影响

为了探讨外源葡萄糖对盐胁迫下山楂叶片的缓解效应,采用光合和叶绿素荧光技术,研究了葡萄糖对盐胁迫下山楂幼苗叶片光合与叶绿素荧光特性及细胞保护酶活性的影响。结果表明:葡萄糖能减轻因盐胁迫造成的叶片光合机构的破坏程度,提高叶绿素含量、净光合速率(Pn)及气孔导度(Gs),降低胞间CO2浓度(Ci),提高PSⅡ的最大光化学效率(Fv/Fm),PSⅡ潜在光化学效率(Fv/Fo),反应中心吸收的光能用于电子传递的量子产额(ΦEo)及以吸收光能为基础的光合性能指数(PIABS)。各浓度外源葡萄糖处理Pn、Fv/Fm及PIABS值比单一盐胁迫处理分别高3.4%～59.8%、5.4%～18.2%和5.6%～78.4%,表明外源葡萄糖提高了盐胁迫下山楂叶片光能利用率及光系统Ⅱ的光化学效率。同时,外源葡萄糖还能够显著提高其叶片内SOD和POD的活性,减少MDA的含量。5.0mmol.L-1外源葡萄糖处理叶片内SOD和POD活性比单一盐胁迫植株分别高21.1%和15.3%,而MDA的含量降低30.1%。

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.

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.

Inhibitory Effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio parahaemolyticus and Its Biofilm in Vitro

In order to explore the inhibitory effects of Prunus mume,Crataegus pinnatifida and Coptis chinensis on Vibrio parahaemolyticus and its biofilm in vitro,the agar diffusion method was applied. These three Chinese herbal medicines had different inhibitory effects on V. parahaemolyticus. The inhibition zone of C. pinnatifida to V. parahaemolyticus was( 15. 25 ± 0. 53) mm,and the minimum inhibitory concentration( MIC) and minimum bactericidal concentration( MBC) of C. pinnatifida on V. parahaemolyticus were both 31. 25 mg/m L; the inhibition zone of C. chinensis to V. parahaemolyticus was( 18. 08 ± 0. 10) mm,and the MIC and MBC of C. chinensis on V. parahaemolyticus were both15. 63 mg/m L; the inhibition zone of P. mume to V. parahaemolyticus was( 28. 99 ± 0. 47) mm,and the MIC and MBC of P. mume on V. parahaemolyticus were both 7. 81 mg/m L. The effects of three traditional Chinese medicines on the biofilm formation of V. parahaemolyticus were tested by MTT colorimetric method using methylthiazolyl tetrazolium( MTT). P. mume,C. pinnatifida and C. chinensis have significant inhibitory effects on V. parahaemolyticus biofilm and their MIC are 7. 81 mg/m L,3. 125 mg/m L,and 62. 5 mg/m L,respectively( P < 0. 01).The experimental results are expected to provide certain references for the development of new fishery drugs.

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.

低共熔溶剂提取山里红多糖工艺优化研究

目的优化超声辅助低共熔溶剂提取山里红总多糖工艺并考察其抗氧化活性。方法以多糖含量为指标,低共熔溶剂种类、摩尔比、料液比、含水量和超声时间为影响因素,采用超声单因素分析和响应面方法优化低共熔溶剂提取山里红总多糖的工艺参数,并以DPPH自由基和·OH自由基清除能力对其抗氧化活性进行评价。结果山里红总多糖成分的最佳提取工艺:以氯化胆碱-丙二醇为低共熔溶剂,摩尔比为1∶3,料液比为1∶20,含水量为20.12%,超声时间为38.98 min。在此工艺下,山里红总多糖含量可达到(523.49±8.79)mg/g。在抗氧化实验中,山里红多糖对DPPH自由基清除作用的IC50为1.33,对·OH自由基清除的IC50为1.23。结论超声辅助低共熔溶剂优化的多糖提取工艺提取率高且工艺稳定,山里红多糖具有较好的抗氧化作用。

基于GC-MS/MS技术对承德地区山楂中44种农药残留分析评价

目的:对承德地区山楂中44种农药残留进行检测分析并评价。方法:利用气相色谱-质谱/质谱联用方法检测山楂样品中44种农药残留含量。结果:44种农药残留组分在0.2~0.6μg·mL^(-1)线性关系良好,相关系数均大于0.998,加标回收率在70.8%~104.5%,相对标准偏差在0.18%~5.20%。结论:承德地区山楂产品安全性较高,病虫害综合防治技术较好,能够为食品安全监管提供数据支持。

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.

指纹图谱结合一测多评法评价山楂叶质量

目的 建立不同产地山楂叶药材的指纹图谱和多成分含量测定方法,评价其质量并筛选差异标志物。方法 收集产自河北省承德市、辽宁省葫芦岛市、山西省运城市、山东省临沂市的78批山楂叶药材,采用高效液相色谱(HPLC)法,利用《中药色谱指纹图谱相似度评价系统(2012版)》绘制指纹图谱并进行相似度评价;采用SPSS 19.0、MetaboAnalyst 5.0工具、SIMCA 14.1软件等进行灰色关联分析、聚类分析(CA)、主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA),以变量重要性投影(VIP)值>1且误差线不超过原点为标准筛选影响山楂叶质量的差异标志物。采用同一HPLC法结合一测多评(QAMS)法,以牡荆素鼠李糖苷为内参物,测定78批山楂叶药材中绿原酸、牡荆素葡萄糖苷、金丝桃苷和异槲皮素的含量,并与外标法测定结果进行比较。结果 4个产地的78批山楂叶药材指纹图谱中有8个共有峰被标定,有5种成分[绿原酸(峰1)、牡荆素葡萄糖苷(峰3)、牡荆素鼠李糖苷(峰4)、金丝桃苷(峰7)和异槲皮素(峰8)]被指认,相似度为0.871~0.998。河北省承德市、辽宁省葫芦岛市、山西省运城市、山东省临沂市样品的平均相对关联度分别为0.538、0.528、0.462、0.435;CA、PCA分析均显示,河北省承德市和辽宁省葫芦岛市样品大致分为一类,山东省临沂市和山西省运城市样品大致分为一类;峰1、2、3、5的VIP值均大于1。QAMS法计算得绿原酸、牡荆素葡萄糖苷、金丝桃苷、异槲皮素的平均相对校正因子分别为0.401、0.993、1.670、1.615(RSD<2%);与外标法结果比较,除2批样品(S39、S41)中的异槲皮素外,其余批次样品中各成分的含量均无明显差异(相对偏差不高于5%)。结论 所建指纹图谱和QAMS法操作简单,可用于评价山楂叶药材的质量;河北省承德市样品质量较优;绿原酸(峰1)、牡荆素葡萄糖苷(峰3)和峰2、5对应成分可能是影响山楂叶药材质量的差异标志物。

北山楂、南山楂和广山楂性状鉴别和有机酸成分研究进展

山楂药材来源除《中华人民共和国药典》(以下简称《药典》)收载的山里红和山楂(习称“北山楂”)外,国内市场上较为常用的尚有野山楂(习称“南山楂”)、台湾林檎或光萼林檎(习称“广山楂”),三者在来源品种、化学成分和药理作用等方面都具有较大差异。有机酸作为北山楂消食健脾的主要有效成分,也是《药典》规定的质量控制的重要指标。研究表明北山楂中总有机酸含量为7.75%~14.00%,南山楂为0.75%~3.35%,广山楂约为4.12%,南山楂和广山楂中总有机酸含量均低于《药典》规定的最低限度(5%)。目前部分地区仍习惯性地将南山楂和广山楂当作北山楂使用,这就大大降低了消食健脾等有机酸类成分所发挥的功效。该文分别对北山楂、南山楂、广山楂的外观性状和所含有机酸的成分、药理作用、提取工艺、检测方法及炮制对有机酸影响等方面的研究进行汇总分析,旨在为3种药材的研究和应用提供参考。

黄芪、山药和山楂对幼猫生长性能、 毛发、抗氧化、免疫指标及肠道菌群的影响

试验旨在研究黄芪、山药和山楂对幼猫生长性能、毛发质量、粪便性状、血清抗氧化、免疫指标,及肠道菌群的影响。选取18只45~60日龄的幼猫,随机分为3组,每组6个重复,每个重复1只猫。空白对照组(H组)幼猫饲喂基础幼猫粮,试验对照组(R组)幼猫饲喂市售幼猫粮,试验组(T组)在基础幼猫粮中添加0.2%黄芪、0.2%山药、0.2%山楂。试验期30 d。结果显示,T组幼猫平均日增重高于H组和R组,料重比低于H组(P>0.05)。T组幼猫毛鳞片厚度和贴服度更高。T组幼猫血清谷胱甘肽过氧化物酶(GSH-Px)活性显著高于R组(P<0.05)。T组幼猫粪便评分高于H组和R组(P>0.05),肠道菌群丰富度更高,菌群结构更相似。研究表明,幼猫粮中添加黄芪、山药和山楂可改善幼猫生长性能、粪便性状、毛发质量、抗氧化能力和免疫能力,优化肠道菌群结构,更利于幼猫健康成长。

山楂叶质量评价研究

目的 对山楂Crataegus pinnatifida Bge.var.major N.E.Br叶质量进行评价。方法 建立该药材UPLC指纹图谱,测定其对α-淀粉酶、α-葡萄糖苷酶活性的抑制作用,双变量相关分析筛选与2种酶IC_(50)值显著相关的共有峰,绘制热图,进行聚类分析、主成分分析。结果 15批样品指纹图谱中有23个共有峰,相似度0.849~0.998。与α-淀粉酶、α-葡萄糖苷酶IC_(50)值显著相关的共有峰分别为13、14个。各批样品聚为3类,2种酶累积方差贡献率分别为85.886%、87.187%。结论 该方法简便可靠,可用于不同产地山楂叶区分及质量评价。

纤维素纳米晶-肉桂精油-壳聚糖复合涂膜制备及其对黄山楂金如意的保鲜效果

【目的】优化涂膜性能,提高其保鲜效果,用于改善黄山楂金如意贮藏品质。【方法】利用肉桂精油与咖啡酸修饰的纤维素纳米晶合成皮克林乳液,选取壳聚糖为成膜剂,制备纤维素纳米晶-肉桂精油-壳聚糖复合涂膜用于黄山楂保鲜。对涂膜力学性能、气体阻隔能力、抗菌、抗氧化性能进行测试。测定贮藏过程中黄山楂果实硬度、失重率、腐烂率,以及可溶性固形物、维生素C和可滴定酸含量等生理指标的变化。【结果】皮克林乳液的添加可以极大改善涂膜材料的力学性能、抗菌和抗氧化能力,提高其气体阻隔性能。随着乳液添加量的提高,涂膜对黄山楂金如意保鲜效果逐渐增强。当乳液体积分数为5%时制备的涂膜保鲜性能最优,在贮藏40 d后,山楂果实失重率和腐烂率显著降低,硬度可达5.4 kg·cm^(-2),可溶性固形物、维生素C和可滴定酸消耗量显著降低。【结论】向壳聚糖中加入皮克林乳液制备复合涂膜,可以极大改善涂膜保鲜性能,延缓黄山楂营养物质流失,最终提高黄山楂金如意采后贮藏品质。

茯苓配方浸膏对高脂饮食小鼠肥胖及脂质沉积的影响

为了探究茯苓配方(茯苓、山楂、黄大茶)浸膏对高脂饮食小鼠肥胖及脂质沉积的影响,将C57/BL6N小鼠随机分为对照组,模型组,奥利司他药物对照组,茯苓配方(Poria cocos Wolf formula,PWF)浸膏低、高剂量组。除对照组给予低脂饲料,其他组均以高脂饲料喂养12周。每周记录小鼠的体重、能量摄入。给药结束后,计算各组小鼠Lee’s指数、脏器系数,采用苏木精-伊红(hematoxylin-eosin,HE)染色法观察各组小鼠肝细胞与脂肪细胞形态变化,全自动生化仪测定各组小鼠血清中TG、TC、HDL-C、LDL-C、AST及ALT水平变化。结果显示,与对照组相比,模型组小鼠体重、Lee’s指数、脏器系数、TG、TC、LDL-C、AST及ALT水平均显著升高(P<0.01或P<0.001),肝脏脂滴数量与脂肪细胞直径增加;与模型组相比,茯苓配方浸膏高剂量组小鼠体重、Lee’s指数和脏器系数均显著降低(P<0.05或P<0.01),低剂量组小鼠体重、Lee’s指数和脏器系数均没有显著变化,茯苓配方浸膏高剂量组小鼠肝脏脂滴数量与脂肪细胞直径减少,TG、TC、LDL-C水平、AST及ALT水平均显著降低(P<0.05或P<0.01或P<0.001)。综上所述,茯苓配方浸膏能够改善高脂饮食小鼠的体重及脂质沉积,为开发改善肥胖的天然物质提供理论基础。