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.

Two new ceramides from the stems of Piper betle L.

Two new ceramides,(2S,3S,4R)-2-N-[(2 R)-2 -hydroxypentacosanoylamino]-nonacosane-1,3,4-triol(1) and(2S,3S,4R,8E)-2- N-[(2 R)-2 -hydroxytetracosanoylamino]-8-eicosylene-1,3,4-triol(2) have been isolated from the stems of Piper betle L.collected from Baoshan city of Yunnan Province in China.Their structures were determined by spectroscopic and chemical methods.

An Investigation of the Vegetative Anatomy of <i>Piper sarmentosum</i>, and a Comparison with the Anatomy of <i>Piper betle</i>(Piperaceae)

This paper describes the detailed anatomy and micromorphology of the vegetative parts of Piper sarmentosum Roxb. (synonym, P. lolot C.DC.), which is a southeast Asian medicinal plant valued for its medicinal and culinary uses. The study also compares the anatomy of the leaf, petiole, stem and root of P. sarmentosum with those of the well-characterized P. betle L. The leaves of both the species are morphologically comparable and have similar culinary uses. The present study showed that the leaves of both the species possess more or less comparable anatomical features with a few differences;whereas, the microscopic features of the petiole, stem, and root of the two species are distinctive.

Rooting of Stem Cuttings with Different Indole 3 Butyric Acid (IBA) Treatments and Development of Micropropagation Protocol for <i>Piper betle</i>L. Node Culture

The present study, conducted during 2016 and 2017 seasons, aimed to investigate the effect of IBA on rooting of Piper betle L. stem cuttings (softwood and semi-hardwood). The experiment was undertaken in misting house field 2 UPM using the sand media to determine the adventitious roots initiation and development using the histological method. The cuttings were treated with different IBA concentrations (0, 500, 1000, 1500 and 2000 mg/L). The nodes explants were used in the development of a protocol for in vitro propagation of P. betle L., with different concentrations of Clorox with different times of immersion (20% Clorox 10 minutes, 30% Clorox 10 minutes, 20% Clorox 20 minutes, and 30% 20 minutes). In multiplication of the plantlets, Murashige and Skoog (MS) medium with different concentrations of BAP (0, 0.5, 1.0, 2.0 mg/L) were used to investigate the rooting of the explants. The results indicated that the types of the cuttings were different in the rooting capacity and the length of the roots. Moreover, it was found that in comparison with the control treatment, by a rise in the concentrations of the IBA, there was a significant upsurge in the rooting percentage, the root diameter, and the number of the roots. The results indicated that the types of cutting with 1000, 1500 and 2000 mg/L IBA perform better in the root percentage (100%) in the semi hardwood cuttings. The best results, however, were 2000 mg/L IBA in the semi hardwood cuttings, with the number of the roots to be 35.05, and the fresh weight of the roots to be 3.94 g, the dry weight of the roots to be 0.33 g, the length of the roots to be 391.88 cm, the roots diameter to be 1.21 mm, the surface area of the roots to be 121.83 cm2, and the root volume to be 2.99 cm3. Nonetheless, the optimal concentration of Clorox with the time immersion was 20% with the 20-minute immersion time, which produced a shoot induction percentage of 30% dead explants and a mean number of 70.00 shoots per explant and the optimal concentration of benzylaminopurine (BAP) at 1.0 mg/L. It is of note that a shoot induction percentage of 22.29% and a mean number of 4.1% number of auxiliary bud per treatment. P. betle shoots in MS medium without PGR MS (0.0) yielded a good rooting.

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.

Hydroxychavicol,a polyphenol from Piper betle leaf extract,induces cell cycle arrest and apoptosis in TP53-resistant HT-29 colon cancer cells

This study aims to elucidate the antiproliferative mechanism of hydroxychavicol(HC).Its effects on cell cycle,apoptosis,and the expression of c-Jun N-terminal kinase(JNK)and P38 mitogen-activated protein kinase(MAPK)in HT-29 colon cancer cells were investigated.HC was isolated from Piper betle leaf(PBL)and verified by high-performance liquid chromatography(HPLC),nuclear magnetic resonance(NMR),and gas chromatography-mass spectrometry(GC-MS).The cytotoxic effects of the standard drug 5-fluorouracil(5-FU),PBL water extract,and HC on HT-29 cells were measured after 24,48,and 72 h of treatment.Cell cycle and apoptosis modulation by 5-FU and HC treatments were investigated up to 30 h.Changes in phosphorylated JNK(pJNK)and P38(pP38)MAPK expression were observed up to 18 h.The half maximal inhibitory concentration(IC_(50))values of HC(30μg/mL)and PBL water extract(380μg/mL)were achieved at 24 h,whereas the IC_(50)of 5-FU(50μmol/L)was obtained at 72 h.Cell cycle arrest at the G0/G1 phase in HC-treated cells was observed from12 h onwards.Higher apoptotic cell death in HC-treated cells compared to 5-FU-treated cells(P<0.05)was observed.High expression of pJNK and pP38 MAPK was observed at 12 h in HC-treated cells,but not in 5-FU-treated HT-29 cells(P<0.05).It is concluded that HC induces cell cycle arrest and apoptosis of HT-29 cells,with these actions possibly mediated by JNK and P38 MAPK.

Influence of phytochemicals in piper betle linn leaf extract on wound healing

Background:Wound healing has being extensively investigated over the world.Healing impairment is caused by many reasons including increasing of free-radicals-mediated damage,delaying in granulation tissue formation,reducing in angiogenesis and decreasing in collagen reorganization.These facts consequently lead to chronic wound healing.Piper betle Linn(Betle)leaves have been folklore used as an ingredient of drugs for cutaneous wound treatment.However,the effect of betle leaf on wound healing is not yet well elucidated.In this study,we aimed to investigate the healing efficacy of methanol leaf extract of Piper betle Linn on proliferation of fibroblast NIH3T3 cells as well as full-thickness burn and excision wounds in swiss mice.Methods:Scratch wound healing assays were conducted to examine the effects of betle leaf extract on healing activity of fibroblast cells.Burn and excision wounds on swiss mouse skins were created for investigating the wound healing progress caused by the betle leaf extract.Malondialdehyde(MDA)was also evaluated to examine the products of lipid hydroperoxide(LPO)under conditions of with or without betle leaf extract treatment.Results:The results of this study showed that Piper betle Linn leaf extract in methanol increased proliferation of NIH3T3 cells and promoted wound healing in vitro and in vivo with both burn wound and excision wound models.In addition,this extract significant decreased level of malondialdehyde(MDA)in liver of treated-mice compared with that in non-treated mice.Conclusions:Our results suggest that Piper betle Linn can be used as an ingredient in developing natural origin drugs for treatment of cutaneous wounds.

4种方法提取蒌叶鲜叶挥发性成分的比较分析

蒌叶鲜叶具有独特的辛香味,在我国南方地区常用作嚼槟榔青果的植物配料。为了探究蒌叶鲜叶的挥发性成分,采用气相色谱-串联质谱仪对不同方法(水蒸气蒸馏法、固相微萃取法、超声辅助正己烷法和超声辅助二氯甲烷法)提取的挥发性成分进行分析。结果显示,4种方法提取和鉴定出的挥发性成分共92种,相对含量较高为酚类、烯类、酯类和醇类,主要成分为蒌叶酚、乙酸丁香酚酯、香叶烯D、4-烯丙基邻苯二酚二乙酸酯、β-石竹烯、γ-衣兰油烯等;水蒸气蒸馏法提取的挥发性成分最多,共鉴定60种;固相微萃取法、超声辅助二氯甲烷和超声辅助正己烷法提取鉴定的挥发性成分分别为53种、42种和25种;4种提取方法共有组分17种,而固相微萃取法特有成分20种,水蒸气蒸馏法获得特有成分18种。不同提取方法获得挥发性成分的种类和含量差异较大,以挥发性活性成分为目的的提取方法,水蒸气蒸馏提取法较为合适,固相微萃取法结合水蒸气蒸馏法提取蒌叶鲜叶,能够获得更加全面的挥发性成分,可为蒌叶香气物质基础的开发与利用提供参考。

蒌叶茎化学成分研究

目的:研究蒌叶茎的化学成分。方法:应用各种色谱技术进行分离纯化,用理化及波谱分析方法确定化合物结构。结果:从蒌叶茎70%丙酮提取物的石油醚部分和乙酸乙酯部分分离得到9个化合物,分别鉴定为:6β-羟基-豆甾-4-烯-3-酮(1)、β-谷甾醇(2)、豆甾醇(3)、齐墩果酸(4)、23-hydroxyursan-12-en-28-oic acid(5)、β-谷甾醇-3-O-β-D-葡萄糖-6′-棕榈酸酯(6)、β-胡萝卜苷(7)、(2S)-4′-羟基-7-O-β-D-葡萄糖-2,3-二氢黄酮苷(8)和α-乙基葡萄糖(9)。结论:除化合物2之外,其余化合物均为首次从该植物中分离得到。

蒌叶抗氧化活性成分的提取及活性测定体系的优化

目的:研究蒌叶乙醇提取物的抗氧化活性。方法:采用微波辅助乙醇提取法提取蒌叶抗氧化活性成分,以鲁米诺化学发光体系测定蒌叶乙醇提取物清除羟自由基的效果,并对测定体系进行优化。结果:当料液比为13∶25(mg/mL),乙醇浓度为50%,提取温度为55℃,提取时间为25min,微波提取功率为600W时,蒌叶乙醇提取物对羟基自由基的清除效果最佳,清除率可达98.2%,清除率测定体系中,当pH10.18,Luminol浓度为0.8×10-4mol/L,H2O2浓度为0.3%时,体系测量精密高、稳定性和重复性好。结论:蒌叶乙醇提取物对·OH有很好的清除作用。

蒌叶茎中甘油酯和神经酰胺类化合物的研究

目的:研究蒌叶茎的化学成分.方法:应用各种色谱技术进行分离纯化,用理化及波谱分析方法鉴定化合物结构.结果:从蒌叶茎70%丙酮提取物中分离得到3个化合物,分别鉴定为:1-O-十六碳酰基甘油酯(1),1-O-β-D-半乳糖-(6→1)-α-D-半乳糖-2,3-O-十六烷酸甘油二酯(2),poke-weed cerebroside(3).结论:3个化合物均为首次从该植物中分离得到.

HS-SPME-GC-MS分析蒟酱叶挥发性成分

首次采用顶空固相微萃取和气质联用技术(HS-SPME-GCMS),结合保留指数法,用峰面积归一化法测定蒟酱叶中挥发性成分及相对百分含量。结果表明,从蒟酱叶中鉴别出27个化学成分,占峰面积的97.53%,化合物结构类型包括醛、酸、酯、酚、烯烃、芳香烃和(环)烷烃。2-甲氧基-5-甲基苯甲醛(42.89%)、胡椒酚醋酸酯(12.49%)、异丁香酚(13.42%)、4-烯丙基-1,2-二乙酰氧基苯(9.47%)、胡椒酚(2.89%)、γ-毕橙茄烯(2.74%)、丁香酚(2.66%)和乙酸异丁香酚酯(2.06%)是蒟酱叶的主要挥发性成分。

蒌叶乙醇提取物抗氧化及抗疲劳作用研究

以75%乙醇提取蒌叶提取物,运用鲁米诺化学发光体系测定提取物清除羟自由基的能力,并以小白鼠为研究对象,从运动耐力学和生化指标两方面,考察蒌叶乙醇提取物的抗疲劳活性。结果表明:蒌叶乙醇提取物对羟基自由基抑制的IC50为63.65μg·mL-1,其提取物能显著延长小鼠负重游泳时间,明显降低运动后血清尿素氮水平,具有降低小鼠运动后血乳酸曲线下面积的作用。蒌叶乙醇提取物具有抗氧化和缓解体力疲劳的作用,其提取物中抗氧化活性成分有较高的热和光稳定性。

Designer probiotics: Development and applications in gastrointestinal health

Given the increasing commercial and clinical relevance of probiotics, improving their stress tolerance profile and ability to overcome the physiochemical defences of the host is an important biological goal. Herein, I review the current state of the art in the design of engineered probiotic cultures, with a specific focus on their utility as therapeutics for the developing world; from the treatment of chronic and acute enteric infections, andtheir associated diarrhoeal complexes, to targeting HIV and application as novel mucosal vaccine delivery vehicles.

蒌叶细菌性疫病的症状和病原菌鉴定

本文描述了海南省某蒌叶种植基地发生细菌性疫病的症状,根据对该病原菌的形态、染色反应、培养性状、生理生化特性、寄主范围等测定结果,将该病原菌鉴定为甘蓝黄单胞菌萎叶致病变种[Xanthom onas campestrispv.betlicola(Patel,Ku lkarn i etDhande1951)Dye 1978.]。

胡椒属野生近缘种‘芦子’鉴定评价及创新利用

以大叶种胡椒(Type Lampong)为对照,对胡椒属野生近缘种质资源‘芦子’(Piper betle L.)的植物学性状、农艺性状及抗逆性状(抗寒、抗病)等主要性状进行鉴定评价。结果表明:野生近缘种质‘芦子’长势强,冠幅大,果穗长,座果率高,抗寒、抗病能力较强,农艺性状及抗性明显优异大叶种胡椒;通过对‘芦子’种质资源的无性繁殖品种改良技术进行探究,说明‘芦子’野生近缘种质资源是胡椒品种改良较好的中间材料,将为胡椒种质资源创新利用及胡椒产业发展奠定良好的基因基础。

Antibacterial activities of ethanol extracts of Philippine medicinal plants against multidrug-resistant bacteria

Objective: To investigate the antibacterial activities of crude ethanol extracts of 12 Philippine medicinal plants.Methods: Crude ethanol extracts from 12 Philippine medicinal plants were evaluated for their antibacterial activity against methicillin-resistant Staphylococcus aureus, vancomycinresistant Enterococcus, extended spectrum β-lactamase-producing, carbapenem-resistant Enterobacteriaceae and metallo-β-lactamase-producing Pseudomonas aeruginosa and Acinetobacter baumannii. Results: The leaf extracts of Psidium guajava, Phyllanthus niruri, Ehretia microphylla and Piper betle(P. betle) showed antibacterial activity against the Gram-positive methicillinresistant Staphylococcus aureus and vancomycin-resistant Enterococcus. P. betle showed the highest antibacterial activity for these bacteria in the disk diffusion(16-33 mm inhibition diameter), minimum inhibitory concentration(19-156 μg/m L) and minimum bactericidal concentration(312 μg/m L) assays. P. betle leaf extracts only showed remarkable antibacterial activity for all the Gram-negative multidrug-resistant bacteria(extended spectrum β-lactamaseproducing, carbapenem-resistant Enterobacteriaceae and metallo-β-lactamase-producing) in the disk diffusion(17-21 mm inhibition diameter), minimum inhibitory concentration(312-625 μg/m L) and minimum bactericidal concentration(312-625 μg/m L) assays. Conclusions: P. betle had the greatest potential value against both Gram-negative and Grampositive multidrug-resistant bacteria. Favorable antagonistic activities were also exhibited by the ethanol extracts of Psidium guajava, Phyllanthus niruri and Ehretia microphylla.

蒌叶挥发油的提取及抗氧化和抑菌活性研究

采用水蒸气提取法对蒌叶挥发油进行提取,在单因素实验的基础上,以正交实验筛选最佳的提取工艺条件;同时考察蒌叶挥发油抗氧化和抑菌活性。结果表明,蒌叶挥发油提取的最佳工艺条件为:料液比m_(干蒌叶粉)∶V_水=50∶1000(g/m L),超声时间为30 min,超声温度70℃,超声功率192 W,加热回流提取4 h,蒌叶挥发油的得率为2.96%。蒌叶挥发油清除DPPH·和·OH的IC_(50)分别为33.19、71.11μg/m L;挥发油对金黄色葡萄球菌、枯草杆菌、大肠杆菌、蜡状芽孢杆菌、四联球菌、藤黄八叠球菌、白色葡萄球菌、黑曲霉、毛霉、青霉均有明显的抗菌活性,其MIC分别为0.625、1.25、0.625、0.625、0.625、2.50、1.25、0.313、0.625、0.313 mg/m L,故蒌叶挥发油具有较强的抗氧化和抑菌活性。

蒌叶的化学成分及生物活性研究进展

蒌叶作为"药食同源"植物,其味辛、微甘、性温,可以治疗风寒咳嗽、胃寒痛、湿疹等症,其叶具有强烈而辛辣的芳香气味,在亚洲广泛用于咀嚼剂。化学分析表明,蒌叶中含有蛋白质、淀粉、脂肪等营养成分及萜类、生物碱、多酚、黄酮类等代谢成分;现代药理学研究表明,蒌叶具有抑菌、抗氧化、抗癌、抗虫、抗炎、抗疟疾、抗糖基化等作用。本文综述了蒌叶的物质基础和生物活性,旨在为蒌叶资源的进一步研究、合理开发利用提供参考依据。