Biotechnological intervention in Betelvine(Piper betle L.):A review on recent advances and future prospects

Betelvine(Piper betle L.) is cultivated for its deep green heart shaped leaf for(15-20) million Indian and 2 billion foreign consumers annually. The crop provides Rs(6 000-7 000) million of national income per year and at the same time leaves worth Rs(30-40) million is exported to other countries. The leaves are not only used directly for chewing purposes but also possesses antioxidant, anti-inflammatory, anti-apoptotic, anti-cancer and anti-microbial properties. Besides, the leaves also contain eugenol rich essential oil(1%-3%) which is the source for medicine, stimulant, antiseptic, tonic and other ayurvedic formulations. The essential oil also contains chavibetol, caryophyllene and methyl eugenol which are the potent source for preparation in ayurvedic medicine and herbal products. Cost of betelvine essential oil is 10$ per 5 mL. In spite of its great economical and medicinal importance betelvine is still neglected by the researchers for proper characterization and authentication for selection of elite landraces. Lack of awareness among people, use of same planting material for many generations, existing of many synonyms for a single landraces, no proper characterization of available landraces are some of the significant constraints for its commercialization. Our review endeavours a complete advance in the research on betelvine, existing lacunae for its proper characterization and commercial cultivation. It also attempts to provide a comprehensive account on biotechnological interventions made in betelvine aimed at complementing conventional programmes for improvement of this nutraceutically important cash crop.

Fusarium solani Upregulated Sesquiterpene Synthase Expression, Sesquiterpene Production and Allelopathic Activity in Piper betle

Activators of sesquiterpene synthase(STS) gene expression and sesquiterpene production in Piper betle L. were examined using quantitative real time PCR and gas chromatography mass spectrometry methods, and the allelopathic activity of untreated and Fusarium solani-treated betel extracts was tested on seed germination and on the shoot and root growth of Thai rice variety PSL2(Oryza sativa cv. Phitsanulok 2) and three dominant paddy weeds(Eclipta prostrata, Echinochloa crus-galli and Chloris barbata). The results demonstrated that F. solani dramatically upregulated STS expression and productions of β-cubebene, β-caryophyllene and germacrene D sesquiterpene when compared with the untreated control, and that betel extracts had a greater inhibitory effect on weeds than on rice. The effects were more clearly detected on seed germination and root growth than on shoot growth, and they were found to be dose-dependent. It is also noted that F. solani-treated extract had stronger effects than the untreated extract. The species most sensitive to the allelopathic effects was C. barbata, germination of which was completely inhibited even at a dose of 0.1 mg/mL untreated extract. With regards to rice, although betel extract at 1.0 mg/mL showed no inhibition on germination, it affected the elongation of rice roots, in addition to those of the tested weeds. The obtained data suggested that F. solani has potential as an activator of sesquiterpene allelochemical production via STS expression, the latter leading to the treated betel extract having a stronger phytotoxic effect. These results were beneficial in the promotion of natural herbicide production using biotechnology.

Research progress on the main chemical constituents and pharmacological effects of Piper betle L.

The Piper betle,belonging to Piper genus of the Piperaceae family,is an evergreen and perennial creeper used in several traditional medicines.This plant has been known to possess antioxidant,antitumor,antibacterial,anti-inflammatory and antimalarial activities.A wide range of chemical compounds including alkaloids,simple phenylpropanoids,lignans,flavonoids,steroids,triterpenoids,sesquiterpenes,monoterpenes have been isolated from this plant.The present review summarized the information concerning phytochemistry and biological activity of P.betle,providing references for the future research of this plant.

Progress on chemical constituents and pharmacological effects of Piper nigrum L.

Piper nigrum L.,belonging to Piper genus of the Piperaceae family,is a medicinal and edible plant.Studies have shown that there are many chemical constituents in this plant,including alkaloids,lignans,steroids and other compounds.In addition,some chemical components of P.nigrum have potential values to treat cancer,inflammation and other diseases.Research progress on the chemical constituents and pharmacological effects of P.nigrum was reviewed in this study to better explore its potential medicinal value.

Network pharmacology-based virtual screening of active alkaloid ingredients from Piper nigrum L.and the potential targets against Parkinson’s disease

Parkinson’s disease(PD)is a common neurodegenerative disease.Despite a great deal of research on this disease,there is still no definitive treatment plan.Literature investigations have shown that Piper nigrum L.has a protective effect on the nerve center.Therefore,network pharmacology was used in this study to investigate the therapeutic potential of P.nigrum for PD.As a result,six active alkaloid ingredients and four potential targets were obtained.This study laid the foundation for finding the lead compounds from P.nigrum to treat PD.

Isolation of Bulk Amount of Piperine as Active Pharmaceutical Ingredient (API) from Black Pepper and White Pepper (<i>Piper nigrum</i>L.)

In the pharmaceutical world the majority of the active pharmaceutical ingredients (API) have been obtained from the natural products. Piperine is such naturally occurring alkaloid which can be considered as major bioactive phytochemical having broad spectrum of pharmacological activities. It is obtained from the most valuable ethnomedicinal spices peppercorns i.e. black pepper and white pepper, which are the fruits of the Asian vine Piper nigrum L. Because of the widespread traditional uses of this medicinal compound, present article reveals a simple and effective isolation method of bulk piperine. The novelty of this investigation is to provide an idea for utilizing such natural method of large scale commercial piperine production as API drug in spite of chemical synthesis. Piperine was isolated in a pure crystal form and characterized by its melting point, X-Ray diffraction (XRD) studies and spectral data, including two-dimensional nuclear magnetic resonance (2D-NMR) spectroscopy. Chromatographic techniques like Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC) were applied to determine the purity of the yielded piperine. It was found that piperine yield from black pepper was within 2.5% - 3.0% and from white pepper within 4.0% - 4.5% and the purity of the yielded piperine was found to be up to 98.5% for black pepper and 98.2% for white pepper. Considering this yield value and purity it is indicated that, such effective isolation method can be successfully utilized for industrial large-scale production commercially. According to the result, it can be claimed that, as a natural product the isolated piperine can also be utilized as API drug like other expensive chemically synthesized piperine in different drug formulation.

荜茇提取物的抗氧化活性及改善LPS所致大鼠听力损伤的作用研究

目的 探讨荜茇提取物的抗氧化活性及改善LPS所致大鼠听力损伤的作用。方法 测试浓度为0.05~1.0 mg/mL的荜茇提取物对DPPH自由基的清除率;测试浓度为25~500μg/mL荜茇提取物对LPS诱导RAW 264.7细胞分泌NO的抑制作用。随机将24只SD大鼠分成空白对照组、模型组、氢化可的松组和荜茇提取物组,每组6只,空白对照组不造模处理,模型组、氢化可的松组及荜茇提取物组连续鼓室内注射5 mg/mL的LPS 5 d后,模型组生理盐水灌胃,氢化可的松组1.81 mg/kg氢化可的松灌胃,荜茇提取物组按生药量1 g/mL荜茇提取物灌胃,连续灌胃10 d后,测试各组大鼠8、16 kHz ABR反应阈值、耳蜗组织中谷氨酸、MDA、TNF-α、NO因子的表达量,并行耳蜗基底膜铺片观察。结果 荜茇提取物对DPPH自由基清除的IC50为0.505 mg/mL;浓度为25~500μg/mL的荜茇提取物对LPS诱导的RAW264.7细胞过度分泌NO有显著的抑制作用,抑制率的IC50为280.55μg/mL。连续灌胃10 d后,与空白对照组相比,模型组8、16 kHz ABR阈值显著升高,耳蜗组织中谷氨酸、MDA、TNF-α、NO的表达显著升高,SOD含量显著降低;与模型组相比,氢化可的松组和荜茇提取物组8、16 kHz的ABR反应阈值降低(P<0.01),大鼠耳蜗组织谷氨酸、MDA、TNF-α、NO因子表达也显著降低(分别为P<0.05、P<0.01、P<0.001),SOD活力提高(P<0.05)。基底膜铺片结果示,空白组毛细胞无明显缺失,三排外毛细胞纤毛排列整齐,结构完整,无缺失;模型组耳蜗毛细胞大部分明显缺失,并且细胞肿胀明显、结构严重不完整,氢化可的松组及荜茇提取物组耳蜗毛细胞排列较为整齐,毛细胞肿胀缓解。结论 荜茇提取物可能通过抑制耳蜗组织中的谷氨酸、自由基的过度表达以及炎症反应,提高超氧化歧化酶SOD活力,产生良好的改善听力作用。

HPLC法测定胡椒根中胡椒碱的含量

目的建立胡椒根药材中胡椒碱的含量测定方法。方法采用反相高效液相色谱法,色谱柱:Luna C18柱(250mm×4.60mm,5μm);流动相:甲醇-水(77∶23);流速:1.0mL·min-1;检测波长为343nm;柱温:35℃。结果胡椒碱在0.164μg～0.984μg范围内,具有良好的线性关系,r=0.9996;平均回收率为98.09%,RSD=2.67%(n=9)。三批胡椒根药材胡椒碱含量为6.67～6.77mg·g-1。结论胡椒根含有胡椒碱,该含量测定法可用于控制胡椒根药材的质量。

HPLC测定海南产胡椒中胡椒碱的含量

测定海南不同地区产的胡椒果实及胡椒根和胡椒叶中胡椒碱的含量。运用索氏提取法对胡椒中胡椒碱进行提取,并采用HPLC对胡椒碱的含量进行测定。海南万宁产胡椒果实中胡椒碱的含量为3.17%,胡椒根中胡椒碱含量为0.43%,叶中胡椒碱含量为0.01%,万宁产的胡椒果实中胡椒碱的含量比其它地区的含量高。不同的生长环境对胡椒果实中胡椒碱的含量有一定的影响。胡椒果实中胡椒碱含量高于根部和叶子中胡椒碱含量。

FTIR方法和HPLC指纹图谱鉴别不同产地荜茇

荜茇是药食同源性中药,也是蒙、藏、维医的习用药材,临床用于治疗胃痛、腹泻等,也被用作香料或调味剂,因此其质量控制非常重要。已报道采用高效液相色谱法检测胡椒碱或荜茇宁,该方法缺乏整体性。傅里叶红外光谱法和HPLC指纹图谱联合应用考察4个产地6批次荜茇,基于共有峰率和变异峰率双指标序列分析荜茇的红外光谱,并建立荜茇的HPLC指纹图谱。结果显示两种方法对荜茇的评价结果相互印证,各批次荜茇的化学成分基本相同,质量比较稳定,其中云南产荜茇(S1)与其余5批次荜茇有一定差异。红外光谱共有峰率和变异峰率与HPLC指纹图谱均能从化学成分的角度评价药材的质量,具有整体性和综合性。荜茇的红外光谱由药材中多种化学成分的红外光谱叠加产生,而指纹图谱仅显示在波长254 nm条件下有紫外吸收的化合物,因此红外光谱在分析药材品质中更具有优势。同时考察了胡椒碱和荜茇宁的含量,云南(S1)和海南(S5, S6)较安徽亳州(S2)和广西(S3, S4)低,结果与红外和HPLC指纹图谱的结果有一定的差异,提示中药材的质量评价应更注重整体性。将FTIR和HPLC指纹图谱联合应用于荜茇的质量控制研究,两种方法互为补充,操作简便,样品使用量少,可做为药材快速鉴别的新方法,为荜茇的化学成分、质量控制和开发应用提供研究基础和依据。

HPLC法同时测定荜茇-大黄中6种成分的含量

目的建立HPLC法同时测定荜茇-大黄药对中胡椒碱、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的含量。方法采用Thermo Hypersil BDS C_(18)色谱柱(4.6 mm×250 mm,5μm);以甲醇(A)-0.1%磷酸(B)为流动相,梯度洗脱;体积流量1.0 ml/min;检测波长254 nm;柱温30℃。结果各成分色谱峰分离度良好,胡椒碱、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的质量浓度分别在6.02~240.72μg/ml(r=0.9996)、1.85~74.18μg/ml(r=0.9993)、1.79~71.78μg/ml(r=0.9996)、1.59~63.55μg/ml(r=0.9995)、5.42~216.72μg/ml(r=0.9992)、1.62~64.64μg/ml(r=0.9997)内呈良好的线性关系,平均回收率分别为99.61%(RSD=1.50%)、99.48%(RSD=1.11%)、99.10%(RSD=1.31%)、99.72%(RSD=1.29%)、99.88%(RSD=1.25%)、99.06%(RSD=0.97%)。结论该方法准确稳定,重复性好,可用于荜茇-大黄药对的质量控制。

超声波-盐析辅助蒸馏提取黑胡椒精油方法优化及活性分析

采用超声波-盐析辅助水蒸气蒸馏法提取黑胡椒(Piper nigrum L.)精油,利用响应面法确定了最佳的液料比、超声时间、NaCl浓度等提取参数。结果表明,超声波辅助蒸馏法提取黑胡椒精油的最佳条件参数为液料比17∶1、超声时间13 min、NaCl浓度2.4%。在此条件下黑胡椒精油的得率为1.79%。对使用此方法获得的黑胡椒精油进行了抗氧化和抑菌活性检测,结果表明提取物具有一定的抗氧化能力,且对ABTS的清除能力远强于对DPPH的清除能力。同时,提取物对金黄色葡萄球菌和青枯假单胞菌均具有较强的生长抑制功能。

超声波辅助亚临界水提取荜茇总黄酮工艺优化及其抗氧化活性评价

采用超声波协同亚临界水技术进行荜茇中黄酮类成分的提取条件研究,并对其不同部位的总黄酮提取物进行抗氧化性对比分析。结果表明,超声波协同亚临界水提取荜茇整株总黄酮最佳工艺条件为超声温度50℃,超声功率240 W,超声时间40 min,料液比1∶30(g/mL),亚临界水提取温度125℃,亚临界水提取时间为40 min,在此工艺条件下荜茇整株总黄酮最高提取量为(30±0.05)mg/g。不同部位总黄酮提取物对DPPH自由基和超氧阴离子自由基清除能力的大小顺序为果穗>叶>根>茎,DPPH自由基清除率和超氧阴离子自由基清除率的IC50值分别为荜茇果穗37.96、304.33μg/mL,荜茇叶44.55、384.36μg/mL,荜茇根47.68、461.54μg/mL,荜茇茎130.60、487.80μg/mL。采用超声波协同亚临界水提取荜茇整株总黄酮的工艺条件合理,提取得到的总黄酮具有良好的抗氧化性。

HPLC法测定蒙药甘露益肝胶囊中胡椒碱的含量

目的:建立蒙药甘露益肝胶囊中胡椒碱的含量测定方法。方法:采用高效液相色谱法。应用DiamonsilC18色谱柱(250 mm×4.6mm,5μm)色谱柱;以甲醇-水(77:23)为流动相;流速为0.8ml.min-1;检测波长为343nm;柱温为室温。结果:胡椒碱在0.083 2μg^0.499 2μg范围内,与峰面积具有良好的线性关系,r=0.9998;平均回收率为99.75%、RSD为2.54%(n=6)。结论:该方法精密度高,分离度好,可用于蒙药甘露益肝胶囊中胡椒碱的含量测定。

流式细胞术估测胡椒属植物基因组大小方法的建立及应用

为建立胡椒属资源基因组大小流式细胞术测定方法,本研究筛选了6种细胞核提取液和3种不同部位叶片,以已知基因组大小的黄瓜为标样,运用流式细胞术对胡椒属资源基因组大小进行了测定。结果表明:改良细胞核提取液和倒一叶是适合胡椒流式细胞术的提取液和取样部位;栽培胡椒品种古晋、Aman和EMAS胡椒的基因组大小为667.94~719.32 Mb,而热引1号、柬埔寨小叶种、班尼约尔1号和73F5的基因组大小为759.69~785.38 Mb,因此不同栽培品种之间基因组大小存在差异;推算基因组大于900 Mb的石楠藤和山蒟的染色体条数应该是大于52且为13的倍数,基因组大小在600~800 Mb范围内种质的染色体数可能是52条。杂交种不但具有与栽培胡椒和黄花胡椒相近的表型,其基因组大小也几乎等于栽培胡椒和黄花胡椒的平均值,推测杂交种可能是栽培胡椒和黄花胡椒的杂交后代。本研究可为胡椒属资源基因组大小测定提供方法,也将为胡椒属种质资源评价及远源杂交等研究提供技术基础。

胡椒扦插苗对涝渍胁迫的生理响应

探究了胡椒扦插苗对涝渍胁迫的生理响应机制,旨在为胡椒抗涝渍栽培提供科学依据。以维持田间持水量的70%±5%为对照处理,采用双套盆法对生产上常用的胡椒扦插苗进行涝渍胁迫处理,分别测定涝渍胁迫1、3、6、9 d的超氧阴离子产生速率、过氧化氢和丙二醛含量、抗氧化酶活性、渗透调节物质和光合色素含量以及根系活力的变化。结果表明,涝渍胁迫1 d,胡椒扦插苗的脯氨酸含量比对照处理增加了27.24%,与对照组相比差异不显著(P>0.05),而超氧阴离子产生速率、叶绿素b、过氧化氢和丙二醛含量则明显上升,并显著高于对照组(P<0.05),其他生理参数测定值不同程度地低于对照处理,此时胡椒根尖开始褐变并坏死,而地上的植株部分没有受害症状,叶片仍呈深绿色。涝渍胁迫3 d,胡椒扦插苗的可溶性蛋白、脯氨酸和总叶绿素含量有小幅增加,但与对照组相比差异不显著(P>0.05)。SOD、POD活性明显上升,并显著高于对照组,叶绿素b含量虽有所降低,但仍显著高于对照组(P<0.05)。超氧阴离子产生速率、过氧化氢、丙二醛含量明显下降,但仍高于对照组,而超氧阴离子产生速率和丙二醛含量与对照组相比差异不显著(P>0.05),其他生理参数测定值不同程度地低于对照处理。此时,胡椒扦插苗的少部分根系已腐烂,少量的叶片褪绿变黄,无明显的萎蔫现象。涝渍胁迫6 d,胡椒扦插苗的SOD、POD、CAT、APX活性、可溶性蛋白和可溶性糖含量、叶绿素a/b和类胡萝卜素/总叶绿素比值以及根系活力不同程度地下降,并显著低于对照,而超氧阴离子产生速率、脯氨酸含量、叶绿素a和b含量、总叶绿素和类胡萝卜素含量、过氧化氢和丙二醛含量均有不同程度地上升,并显著高于对照组(P<0.05)。此时,胡椒扦插苗的绝大部分或全部根系已腐烂,大部分叶片或几乎全部叶片出现枯萎、变黄、脱落,最终植株死亡。研究结果表明,胡椒扦插苗对涝渍胁迫敏感,易受涝渍危害。试验结果初步揭示了胡椒怕涝渍的生理响应机制。

荜茇的本草考证

荜茇是一味经典的中草药,味辛,性热,具有温中散寒、下气止痛的功效。通过查阅历代本草、现代文献,从荜茇出处、名称、植物形态、产地、性味归经、炮制、功效、主治用量及禁忌等多方面进行梳理考证,以便为现代临床医师和研究者提供理论依据和研究方向。

新型HF-LPME-HPLC对蒙药荜茇生物碱含量测定研究

研究建立一种新型中空纤维膜液相微萃取联合高效液相色谱(HF-LPME-HPLC)测定蒙药荜茇中生物碱含量,对影响HF-LPME萃取效率的各项参数进行优化分析。中空纤维膜材料为偏氟乙烯纤维膜(空隙率>50%),正辛醇为萃取剂,萃取温度为室温,振荡速度为200 r/min,萃取时间为128 min。建立了新型HF-LPME的萃取方法,对蒙药荜茇中胡椒碱的R>5,与传统萃取方法相比,富集倍数为40倍。线性范围分别为0.006~0.01 mg/mL,相关系数(r)为0.9999。相比于传统的萃取方法,该方法使用极少量的有机溶剂,可高效萃取并测定蒙药荜茇中生物碱的含量,打破了仅能在环境和食品中应用的限制,可以在各类化学基础实验中推广应用。

基于正交实验法优化从荜茇中提取胡椒碱的工艺

基于正交实验,优化胡椒碱提取工艺并建立HPLC测定胡椒碱含量的方法。以荜茇为原料,在单因素实验设计的基础上,采用正交实验进行优化乙醇浓度、乙醇用量、提取时间及提取次数进行考察。胡椒碱的最佳提取工艺是65%乙醇,提取时间2 h,提取次数为3次,溶剂用量为6倍。胡椒碱在0.70~27.50μg/mL的范围内具有良好线性关系、相对标准偏差小于0.52%(n=5)、加标回收率为103.23%。胡椒碱的提取工艺合理、可行、胡椒碱的提取率较高,为荜茇的进一步开发提供了理论依据。

中国胡椒研究综述

胡椒自1947年引入中国,主要种植在海南、广东、云南、广西等省(区),随着胡椒生产的发展,对胡椒的研究也在不断深入。笔者从种质资源、繁殖技术、生理特性、根际微生物特性、耕作栽培、病虫害防治、产品加工、产品的综合利用和产业战略等方面对中国胡椒自引种以来的研究概况进行了综述,提出中国胡椒今后研究的方向,为从事胡椒科研、生产工作者提供参考。