Insight into the Efficacy and Potential Mechanism of Fructus Aurantii on Skin Based on Network Pharmacology

[Objectives]This study was conducted to explore the mechanism and pharmacological activity of Fructus Aurantii on human skin through network pharmacology.[Methods]The active components and targets of Fructus Aurantii were screened byTCMSPand SymMap data-bases,and the targets were humanized after de-duplication by UniProt database.Relevant therapeutic targets were searched in Gencards data-base with"skin"as the key word,and those with higher weights were retained.An effective component-human skin action target network of Fructus Aurantii was established by Cytoscape software.Then,the topological analysis of protein-protein interaction(PPI)network was made by using String database and Cytoscape software.Gene Ontology(GO)funetional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were carried out by david database,and the analysis results were visualized by Hiplot soft-ware.[Results]Thirteen active compounds of Fructus Aurantii were obtained by screening,and 99 gene targets of Fructus Aurantii which might have pharmacological activity on human skin were obtained by intersection analysis.PTGS2,PTGS1,HSP90AB1,HSP90AAI,NCOA2 and PIK3CG might be core action targets of Fructus Aurantii on human skin according to the topological analysis of the active component-target network.The analysis of PPI network showed that IL-1β,INS,TNF,TP53 and ESR1 might be core action proteins of Fructus Auranti.GO enrichment analysis showed that Fructus Aurantii might balance the imbalance of skin microenvironment caused by various in-vitro stimuli by participating in biological processes such as response to xenobiotic stimulus and regulation of small molecule metabolic process.KEGG pathway enrichment analysis showed that the targets of Fructus Aurantii acting on human skin were enriched in pathways in cancer,Kaposi sarcoma-associated herpesvirus infection,pathways in measles,AGE-RAGE signaling pathway in diabetic complications,IL.-17 signaling pathway,etc.[Conclusions]Fructus Aurantii may act on human skin targets through a variety of active components,and play a regulatory role in skin-related diseases.

茶二叉蚜（Toxoptera aurantii Fonscolombe）在不同茶树品种上生存能力的差异

茶二叉蚜（ＴｏｘｏｐｔｅｒａａｕｒａｎｔｉｉＦｏｎｓｃｏｌｏｍｂｅ）在不同茶树品种上生存能力的差异朱建国，况荣平，刘胜，扈克明，陶滔关键词茶二叉蚜，生态学特性，生殖特性ＴＨＥＤＩＦＦＥＲＥＮＣＥＯＦＳＵＲＶＩＶＡＬＡＢＩＬＩＴＹＯＦＢＬＡＣＫＣＩＴＲ...

茶蚜Toxoptera aurantii Boyer在西藏林芝的发生

本文阐明茶蚜的生活史,生物学习性。揭示茶蚜的发生与温度、湿度、降雨、茶田管理、生态环境的关系。提出了以农业防治、生物防治、物理防治为主的绿色综合治理措施。

Mechanism of aqueous fructus aurantii immaturus extracts in neuroplexus of cathartic colons

AIM: To examine the effect of aqueous fructus aurantii immaturus(FAI) extracts on the intestinal plexus of cathartic colons.METHODS: Cathartic colons were induced in rats with dahuang,a laxative used in traditional Chinese medicine. Once the model was established(after approximately 12 wk),rats were administered mosapride(1.54 mg/kg) or various doses of aqueous FAI extracts(1-4 g/kg) for 14 d. Transit function was assessed using an ink propulsion test. Rats were then sacrificed,and the ultramicrostructure of colonic tissue was examined using transmission electron microscopy. The expression of the 5-hydroxytryptamine receptor 4(5-HTR4) and neurofilament-H was assessed in colon tissues using real-time PCR,Western blot,and immunohistochemistry.RESULTS: Mosapride and high dose(4 g/kg) of aqueous FAI extracts significantly improved the bowel movement in cathartic colons compared to untreated model colons as measured by the intestinal transit rate(70.06 ± 7.25 and 72.02 ± 8.74,respectively,vs 64.12 ± 5.19; P < 0.05 for both). Compared to controls,the ultramicrostructure of cathartic colons showed signsof neural degeneration. Treatment with mosapride and aqueous FAI extracts resulted in recovery of ultrastructural pathology. Treatment with mosapride alone upregulated the gene and protein expression of 5-HTR4 compared to untreated controls(P < 0.05 for both). Treatment with aqueous FAI extracts(≥ 2 g/kg) increased 5-HTR4 m RNA levels(P < 0.05),but no change in protein level was observed by Western blot or immunohistochemistry. The m RNA and protein levels of neurofilament-H were significantly increased with mosapride and ≥ 2 g/kg aqueous FAI extracts compared to controls(P < 0.05 for all).CONCLUSION: Aqueous FAI extracts and mosapride strengthen bowel movement in cathartic colons via increasing the expression of 5-HTR4 and neurofilament-H.

球孢白僵菌Beauveria bassiana对茶蚜Toxoptera aurantii致病力筛选及毒力活性研究

茶蚜Toxoptera aurantii(Boyer de Fonscolombe, 1841)是茶园重要害虫之一,开展茶蚜的绿色防控是茶叶安全优质生产的重要保障。研究以无翅茶蚜为研究对象,通过喷雾塔喷雾接菌方法,对供试的9株球孢白僵菌菌株对无翅茶蚜的致病力活性进行筛选,获得了3株致死中时LT50在4.5 d以内,7 d校正死亡率在90%以上的潜在高致病菌株。进一步通过活性评价试验,优化出高致病球孢白僵菌Bb2032菌株,并对该菌株开展了侵染无翅茶蚜毒力学活性评价研究,其4 d、8 d致死中浓度LC50分别为2.881×107孢子/mL和2.102×104孢子/mL。最后通过显微技术观察高致病球孢白僵菌株Bb2032侵染茶蚜的症状,表明菌丝主要从无翅茶蚜的腹管和头部两个位置浸染。该研究将为白僵菌应用在茶园有害生物综合治理中提供理论基础,为茶叶的安全生产提供技术支撑,具有广阔的应用前景。

Chromatographic fingerprint analysis of Fructus Aurantii Immaturus by HPLC-DAD and chemometric methods

An efficient method for quality control of Fructus Aurantii Immaturus (FAI),a famous traditional Chinese medicine (TCM) was established. A simple and reliable high-performance liquid chromatography-photodiode array detector (HPLC-DAD) procedure coupled with chemometric methods was developed for fingerprint analysis,qualitative analysis and quantitative determination of this herb. In qualitative and quantitative analyses,heuristic evolving latent projection (HELP) method was employed to resolve the overlapping peaks of the tested samples. Two bioactive components,namely hesperidin and naringin,are confirmed and determined,together with four flavonoids compounds tentatively identified including two new ones. From fingerprint analysis,the fingerprint data were processed with correlation coefficients for quantitative expression of their similarity and dissimilarity. The developed method based on an integration of chromatographic fingerprint and quantitative analysis is scientific,and the obtained results can be applied to the quality control of herb medicine.

Essential oil extracted from Quzhou Aurantii Fructus prevents acute liver failure through inhibiting lipopolysaccharide-mediated inflammatory response

Quzhou Aurantii Fructus(QAF)has a long history as a folk medicine and food for the treatment of liver diseases.While our earlier study provided evidence of hepatoprotective properties contained within the flavonoids and limonins constituents in QAF,the potential preventative effects afforded by essential oil components present within QAF remains enigmatic.In this study,we prepared Quzhou Aurantii Fructus essential oil(QAFEO)and confirmed its anti-inflammatory effects on liver inflammation through experimentation on lipopolysaccharide and D-galactosamine(LPS/D-GalN)induced acute liver failure(ALF)mouse models.Using RNA-sequence(RNA-seq)analysis,we found that QAFEO prevented ALF by systematically blunting the pathways involved in response to LPS and toll-like receptor signaling pathways.QAFEO effectively suppressed the phosphorylation of tank-binding kinase 1(TBK1),TGF-beta activated kinase 1(TAK1),interferon regulatory factor 3(IRF3),and the activation of mitogen activated kinase-like protein(MAPK)and nuclear factor-kappa B(NF-κB)pathways in vivo and in vitro.Importantly,QAFEO substantially reduced myeloid differentiation primary response gene 88(MyD88)-toll-like receptor 4(TLR4)interaction levels.Moreover,8 compounds from QAFEO could directly bind to REAL,TAK1,MyD88,TBK1,and IRF3.Taken together,the results of our study support the notion that QAFEO exerts a hepatoprotective effect through inhibiting LPS-mediated inflammatory response.

GC-MS Analysis and Antibacterial Activity Study of Volatile Oil from 11 Kinds of Aurantii Fructus Immaturus Processed Products

[Objectives]To study the volatile components and antibacterial effects of 11 kinds of Aurantii Fructus Immaturus processed products.[Methods]11 kinds of Aurantii Fructus Immaturus processed products were obtained according to the traditional processing method,the volatile oil was extracted by steam distillation,and the composition of volatile oil in the 11 kinds of processed products was analyzed by gas chromatography-mass spectrometry(GC-MS).the relative percentage content of each component in these 11 kinds of processed products was determined using the peak area normalization.The drug sensitivity activity of the volatile oil of these 11 kinds of processed products was tested using the K-B paper diffusion method and the minimum inhibition volume fraction of volatile oil of these 11 kinds of processed products was tested using the microdilution method.[Results]The highest yield of volatile oil of 11 kinds of these processed products was baking(5.193%),and the lowest was stir-bake to scorch(1.998%).A total of 36 chemical components were identified from the volatile oils of these 11 kinds of processed products.The components with the most volatile oil were stir-bake to scorch(24 kinds),and the components with the least volatile oil were the method of processing with rice-washed water(15 kinds).They contain 8 kinds of common chemical components,such as limonene,linalool,myrcene,α-pinene.The highest content of limonene came from processing with honey(60.93%),the lowest came from processing with rice-washed water(55.25%);the highest content of linalool came from processing with rice-washed water(7.139%),the lowest came from processing with honey(5.436%);the highest content of myrcene came from processing with honey(1.899%),the lowest came from stir-bake to scorch(1.632%);the highest content ofα-pinene came from raw Aurantii Fructus Immaturus(2.355%),and the lowest came from stir-bake to scorch(1.618%).The volatile oil of these 11 kinds of Aurantii Fructus Immaturus processed products has good antibacterial effect on Escherichia coli and Staphylococcus aureus.[Conclusions]The oil yields of volatile oils of 11 kinds of Aurantii Fructus Immaturus processed products are different,the content of limonene is significantly different,and the changes of other chemical components and their contents are not significantly different.The volatile oil of 11 kinds of Aurantii Fructus Immaturus processed products has certain antibacterial effect.

多成分含量测定结合网络药理学识别湘枳壳种质资源品质评价的质量标志物

建立多指标成分的色谱定量分析方法,结合网络药理学、分子对接技术和多变量统计分析从“谱-效”关系找出用于湘枳壳种质资源品质评价的质量标志物(quality marker, Q-Marker)。采用高效液相色谱法、顶空固相微萃取-气相色谱法对湖南省多个县市枳壳种质资源样本进行含量测定;对多指标成分进行靶点搜集和网络药理学分析,构建“成分-靶点-通路”网络,并预测候选Q-Marker和核心作用靶点;通过分子对接技术对配体与受体结合活性进行理论模拟找出关键Q-Marker,借助主成分荷载因子分子和聚类分析挖掘出区分枳壳种质资源样本的Q-Marker。结果表明种质资源样本以柚皮苷、新橙皮苷和D-柠檬烯含量最高,各目标化合物的含量差异波动较大;橘皮素等7个候选Q-Marker通过PI3K/AKT和RAS/MAPK通路的PIK3CD等7个核心靶点发挥抗氧化、抗炎等作用,它们之间的分子对接活性良好;多变量统计分析表明柠檬苦素等5个Q-Marker可用于区分种质资源样本,它们有类群特异性,但样本间差异与地域无关。该方法可对湘枳壳种质资源进行品质评价,为品种选育提供参考。

一测多评法同时测定不同配伍比例枳实-厚朴药对中11种成分的含量

目的建立一测多评法同时测定不同配伍比例枳实-厚朴药对中辛弗林、紫丁香苷、木兰花碱、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、柠檬苦素、柚皮素、和厚朴酚、厚朴酚的含量。方法枳实、厚朴分别按1∶1、1∶2、2∶1比例配伍。分析采用Agilent 5TC-C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相甲醇-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长280 nm。以橙皮苷峰为内标,计算其他10种成分相对校正因子,测定其含量。结果11种成分在各自范围内线性关系良好(r≥0.9990),平均加样回收率97.04%~99.45%,RSD 1.22%~2.48%,一测多评法所得结果与外标法接近,配伍比例为1∶1各成分含量最高。结论该方法简便准确,重复性好,可用于枳实-厚朴药对质量控制。2种药材配伍比例为1∶1时,更有利于各成分提取。

枳实化学成分和药理作用研究进展及其质量标志物预测分析

枳实作为临床常用理气中药,其化学成分主要包括黄酮类、生物碱类、香豆素类和挥发油类,药理作用主要为调节胃肠运动、抗氧化、抗炎、保肝等。本文在综述了枳实化学成分和药理作用的基础上,从植物亲缘、传统药性、药理作用和成分可测性方面对枳实的质量标志物进行预测分析,建议将柚皮苷、橙皮苷、新橙皮苷、川陈皮素、野漆树苷、柠檬烯、芳樟醇等化学成分纳入枳实质量标志物的考虑范畴,以期为完善枳实质量评价体系和提高质量标准提供科学依据。

食蚜瘿蚊对茶蚜的捕食功能反应

茶蚜Toxoptera aurantii(Boyer)是茶树的重要害虫,评估食蚜瘿蚊Aphidoletes aphidimyza(Rondani)对茶蚜的防控潜能,为大田释放天敌食蚜瘿蚊防治茶蚜提供理论依据。在室内条件下,探索了饥饿处理24 h的食蚜瘿蚊3龄幼虫对3龄茶蚜的捕食功能反应、搜寻效应和自身的种内干扰作用,试验完成后继续饲喂,直至食蚜瘿蚊羽化,观察其性别。结果表明:食蚜瘿蚊雌、雄幼虫对茶蚜的捕食功能反应均符合HollingⅡ模型,雌、雄幼虫的最大日捕食量分别为19.1、24.5头,瞬时攻击率(a′)分别为0.780、0.459,处理时间(T_(h))分别为0.052、0.041 d;食蚜瘿蚊捕食量随着茶蚜密度的增加而增加,搜寻效应随着茶蚜密度的增加而下降,自身密度干扰方程为E=0.421 P^(-0.892),相同的猎物密度下,雌虫的搜寻效应高于雄虫。食蚜瘿蚊对茶蚜具有较好的控害潜能,且雌虫对茶蚜的控害能力优于雄虫。

种植年限对枳壳根际微生物群落和土壤性质的影响

根际微生物影响土壤养分的有效性与转化,和植物的生长及健康密切相关。探讨不同种植年限枳壳根际微生物和土壤性质的变化,可为枳壳的田间管理提供科学依据。采集重庆市种植4、20和40a的枳壳根际土壤,高通量测序技术分析不同种植年限土壤细菌和真菌多样性和群落结构变化规律,进一步研究微生物群落与土壤性质的相关性。结果表明,连续种植40 a枳壳的土壤有机质、全氮、碱解氮、有效磷和速效钾含量显著高于4 a和20 a,而土壤pH则显著降低(P<0.05)。随着种植年限增加,细菌的多样性指数无明显变化,而真菌的多样性和均匀度指数显著增加(P<0.05)。主坐标分析显示,种植40 a的枳壳根际土壤细菌和真菌群落结构和组成与其他年限显著不同。在门水平上,优势细菌门为变形菌门、酸杆菌门、放线菌门和芽单胞菌门,占总丰度的61.07%~87.79%;优势真菌门为子囊菌门和担子菌门,占总丰度的64.70%~85.75%。在属水平上,随种植年限的增加,40年枳壳根际土壤有益细菌和真菌属丰度较第4年的显著减少(P<0.05),如鞘氨醇单胞菌属、苍白杆菌属、类诺卡氏菌属、假单胞菌属、毛孢子菌属和毛壳菌属,分别下降了51.21%、91.32%、78.60%、81.87%、70.58%和60.74%。此外,致病菌属丰度显著增加,如镰刀属、Plectosphaerella属和Ilyonectria属分别增加了117.6%、323.9%和1631%。Mantel分析表明,土壤pH和营养因素是影响细菌和真菌群落组成的重要环境因子。枳壳长期种植会改变根际微生物群落结构和组成,随着种植年限增加,真菌的多样性指数显著增加,有益的根际细菌和真菌属丰度减少,而潜在的致病菌属丰度增加,这可能是枳壳长期栽培长势变差和病害逐渐加重的主要原因。

发酵枳实残渣对仔猪生长性能、抗氧化能力和肠道健康的影响

本试验旨在探究发酵枳实残渣对仔猪生长性能、抗氧化能力和肠道健康的影响。选取32头体重为(7.65±0.18)kg杜×长×大三元杂交仔猪,随机分为4组,每组8个重复,每个重复1头猪。各组分别饲喂发酵枳实残渣添加水平为0(对照组)、4%(4%FF组)、8%(8%FF组)、12%(12%FF组)的试验饲粮。试验期30 d。结果表明:1)与对照组相比,8%FF组的平均日增重和平均日采食量显著提高(P<0.05)。2)与对照组相比,8%FF组的血清总蛋白(TP)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)含量和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性显著提高(P<0.05),4%FF、8%FF、12%FF组的血清总抗氧化能力(T-AOC)显著提高(P<0.05),4%FF和8%FF组的血清丙二醛(MDA)含量显著降低(P<0.05)。3)与对照组相比,8%FF组的空肠绒毛高度、杯状细胞数量和绒毛高度/隐窝深度显著提高(P<0.05),空肠隐窝深度显著降低(P<0.05)。4)与对照组相比,8%FF和12%FF组的空肠闭锁小带蛋白-1(ZO-1)、超氧化物歧化酶1(SOD1)和核因子E2相关因子2(Nrf2)的mRNA相对表达水平显著提高(P<0.05),8%FF组的空肠封闭蛋白(Occludin)的mRNA相对表达水平显著提高(P<0.05)。5)与对照组相比,8%FF和12%FF组的结肠拟杆菌门(Bacteroidetes)相对丰度显著提高(P<0.05),结肠厚壁菌门(Firmicutes)相对丰度显著降低(P<0.05);4%FF和8%FF组的结肠乳杆菌属(Lactobacillus)相对丰度显著提高(P<0.05),12%FF组的结肠普氏菌属(Prevotella)相对丰度显著提高(P<0.05),4%FF、8%FF和12%FF组的结肠链球菌属(Streptococcus)相对丰度显著降低(P<0.05)。综上所述,饲粮中添加发酵枳实残渣对仔猪生长性能无负面影响,并且能提高机体抗氧化能力,改善肠道形态,增加肠道微生物多样性,最终改善仔猪的生长性能。本试验条件下,仔猪饲粮中发酵枳实残渣添加水平以8%为宜。

基于UPLC-MS/MS探讨枳实中黄酮类化合物与辛弗林药动学的相互影响

目的:基于超高效液相色谱-三重四级杆串联质谱(UPLC-MS/MS),探讨枳实中黄酮类化合物与辛弗林药动学的相互影响,为引入黄酮类化合物作为枳实的质量定量控制指标提供科学依据。方法:取24只SD大鼠,随机分为6组(n=4),A组为辛弗林+6种黄酮类化合物(新橙皮苷、柚皮苷、芸香柚皮苷、橙皮苷、川陈皮素和野漆树苷),B组为辛弗林单独给药,C组为辛弗林+柚皮苷,D组为辛弗林+新橙皮苷,E组为6种黄酮类化合物单独给药,F组采用等体积的0.9%氯化钠溶液处理,均灌胃给药。取给药后不同时间点的血浆,测定血浆中柚皮苷、新橙皮苷和辛弗林含量,应用DAS 2.0软件计算药动学参数。利用UPLC-MS/MS方法测定血药浓度。结果:A组大鼠新橙皮苷清除率为(9337.88±8373.94)L/h·kg,显著高于E组的(2558.22±610.87)L/h·kg,差异有统计学意义(P<0.05)。A组、D组大鼠新橙皮苷的分布体积分别为(2549.72±1381.42)、(1668.41±394.73)L/kg,均明显低于E组的(23388.87±6089.53)L/kg,差异均有统计学意义(P<0.05)。A组大鼠辛弗林的平均停留时间为(2.50±0.63)h,较B组、C组和D组[(1.62±0.11)、(1.70±0.17)和(1.57±0.08)h]更长,差异均有统计学意义(P<0.05)。结论:辛弗林对新橙皮苷的消除率和分布影响较大,枳实中的黄酮类化合物可延长辛弗林的平均停留时间。

不同基原枳实水煎液黄酮类成分及抗氧化活性差异研究

目的 比较不同基原枳实水煎液黄酮类成分和抗氧化活性差异,为枳实的质量评价和临床用药提供参考。方法 建立HPLC法同时测定枳实水煎液中8种黄酮类成分(芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、柚皮素、橙皮素、川陈皮素、桔皮素)的含量,结合多元统计分析化学成分差异;采用DPPH法和ABTS法测定不同基原枳实水煎液的抗氧化活性,以灰色关联度分析和双变量相关性分析评价枳实水煎液中8种成分含量与抗氧化活性的相关性。结果 含量测定及统计分析结果表明不同基原枳实水煎液主要差异成分为柚皮苷和新橙皮苷,其中甜橙枳实水煎液中几乎不含柚皮苷和新橙皮苷,而酸橙枳实水煎液中这两种成分含量较高;抗氧化活性结果表明酸橙枳实水煎液抗氧化活性显著优于甜橙枳实水煎液(P <0.05);灰色关联度分析和双变量相关性分析显示,酸橙枳实水煎液中抗氧化作用的核心药效组分是橙皮苷、新橙皮苷、川陈皮素;甜橙枳实水煎液抗氧化作用的核心药效组分是芸香柚皮苷、橙皮素、川陈皮素。结论 本研究建立了HPLC法同时测定枳实水煎液中8种黄酮类成分的含量;不同基原枳实水煎液黄酮类成分含量与抗氧化能力均存在差异。

基于倾向性评分匹配法评价调畅气机法治疗心肌桥回顾性队列研究

目的基于倾向性评分匹配法(PSM)探讨“调畅气机法”(柴胡-桔梗-枳壳-牛膝)治疗心肌桥的临床疗效。方法采用回顾性队列研究方法,选取158例心肌桥医案,将其分为暴露组与非暴露组,暴露组为接受中药汤剂中含有调畅气机法(包含柴胡-桔梗-枳壳-牛膝)的医案,非暴露组为基础类中药临症加减的医案。运用PSM进行匹配后得到样本35对。观察治疗前后中医证候积分和心电图。结果匹配后暴露组与非暴露组治疗后中医证候积分较治疗前有显著的改善(P<0.01),以暴露组改善更明显(P<0.01);匹配后暴露组的总有效率(94.29%,33/35)高于非暴露组(77.14%,27/35);暴露组的显效率(42.86,15/35%)高于非暴露组(20.00%,7/35),均差异具有统计学意义(P<0.05);匹配后暴露组的心电图总有效率(37.14%,13/35)高于非暴露组(14.29%,5/35),差异具有统计学意义(P<0.05)。结论“调畅气机法”治疗心肌桥疗效良好,缓解临床症状,且一定程度上减轻心电图心肌缺血改变。

衢枳壳主要化学成分及药效学作用和机制研究进展

衢枳壳作为浙江地区道地中药材,在2018年入选新“浙八味”中药材培育品种后,其药理作用得到愈来愈多的关注。研究发现,衢枳壳及其活性成分具有抗炎、平喘、降血糖、保肝、抗氧化、降血脂、调节肠道菌群、抑菌和免疫调节等多种药理作用,具有良好的药用开发前景。本研究归纳总结了衢枳壳及其主要化学成分的药理作用和机制研究进展,为衢枳壳的进一步药用开发利用提供参考。

高效液相色谱法同时检测枳实及枳壳药材中12种黄酮苷类成分

目的建立同时检测枳实、枳壳药材中12种黄酮苷类成分的高效液相色谱法。方法色谱柱为Agilent Zorbax SB-C18柱(250 mm×4.6 mm,5μm),流动相为0.1%甲酸水溶液-乙腈(梯度洗脱),流速为1.0 mL/min,检测波长为284 nm(圣草次苷、新北美圣草苷、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、枸橘苷、柚皮素、橙皮素)、330 nm(橙皮内酯、川陈皮素、橘红素),柱温为35℃,进样量为10μL。结果上述成分质量浓度分别在0.19~46.61μg/mL、0.17~43.27μg/mL、0.20~51.07μg/mL、11.34~756.01μg/mL、0.41~101.28μg/mL、20.58~1028.79μg/mL、0.20~50.49μg/mL、0.10~25.25μg/mL、0.10~26.63μg/mL、0.24~11.96μg/mL、0.21~52.64μg/mL、0.55~27.51μg/mL范围内与峰面积线性关系良好;检测限均不大于0.10μg/mL,定量限均不大于0.41μg/mL;精密度、稳定性、重复性试验结果的RSD均小于2.0%;平均加样回收率为92.90%~114.33%,RSD为0.30%~1.98%(n=9)。结论所建立的方法简便、可靠,可用于同时检测枳实、枳壳药材中12种黄酮苷类成分。

望谟紫芽茶树优良品系‘W016’与黄金芽抗茶蚜性能比较

茶蚜Toxoptera aurantii Boyer是茶树上的重要害虫之一,深入研究抗蚜特异资源的抗性机理对培育茶树抗蚜品种具有重要意义。课题组前期收集的望谟紫芽茶树优良品系‘W016’对茶蚜抗性强,为初步探明该种质的抗蚜特性,利用刺吸电位技术测定了茶蚜在‘W016’和黄金芽(感虫品种)上的取食行为,并探明影响茶蚜刺探取食叶片的主要物理性状及生化成分因子。结果表明,茶蚜在两种供试种质上均产生9种EPG波形,依次为非刺探波(np波)、路径波(A波、B波、C波、pd波)、机械阻力波(F波)、木质部吸食波(G波)、韧皮部唾液分泌波(El波)和韧皮部汁液吸食波(E2波)。刺探取食过程中,茶蚜在‘W016’上的非刺探时间(np波)和C波总持续时间均显著长于黄金芽。二者口针遇到阻力的次数(F波)和总持续时间无差异,但韧皮部吸食汁液(E2波)时间‘W016’显著短于黄金芽。茸毛密度、下表皮厚度和栅栏组织厚度是‘W016’抗茶蚜取食胁迫的主要物理屏障因子,而咖啡碱和可溶性糖含量为主要生化因子。表明‘W016’对茶蚜的取食抗性与其叶片理化性质密切相关。