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用于植物组织染色的一些本地草本植物染料的筛选

2020-09-30 来源:乌哈旅游
Journal ofForestry Research(2010)21(1):81—84 D0I l0.1007/Sl 1676.O10.0014—2 Screening of some indigenous herbal dyes for use in plant histological staining A.J.Akinloye・H.C.Illoh・A.0.Olagoke Received:2009—07—3 1; Accepted:2009—09—05 ◎Northeast Forestry University and Springer—Verlag Berlin Heidelberg 2010 Abstract:The efficacies of some indigenous herbal dyes for use in staining plant materials were examined to obtain non—toxic,eco—fiendlry and cheap stains for use in plant histology.Dye extracts from Bixa orellana,Curcuma domestica,Lonchocarpus cyanescens and Pterocar— pus osun were used to stain wood sections using the existing standard staining procedures with little modiicatfion.All the extracts had affinity for the fibre and vessel elements except the extract from L.cyanescens. (Banderanayake 1 999).These products are also relied upon by a great number of people living in the cities.Extracts from plants are used in various fields in Nigeria and other developing coun— tries.They are used as folkloric medicine re.g.bush medicine used in Herbal remedy),pesticides and insecticides,dyes for textile materials,as well as stains for biological specimens.Plant dyes are utilized locally to dye cloths,domestic fowls,house The extracts from C domestica and B orellana had higher selectivity than those ofP osun for ibrfe.From the results ofthe absorbance curves. each of the dye extracts from all speciese had minimum of two peaks, indicating that they had two or more colour imparting chromophores except dye extract from C domestica.All the dye extracts were acidic with pH range of 3.77 to 6.77.Therefore,this study shows that dye ex— tracts from B.orellana.C domestica and P.OSUH could be solitarily or in loor and wallf and household utensils.They are also used to pre— serve animal skin,decorating human body for differentiating anima1. Plants yield many colours for dyeing a variety of articles. Some trees and many herbs produce dyestuffs,and hitherto the bulk ofworld’s dyes can be produced from plant sources.In facts, plants have provided an important source of pigments and tannin combination with artificial dyes for plant histological staining. Keywords:herbal dyes;indigenous herbs;ecofriendly stains;histologi— caI staining;wood sections orf the world(Lillie 1 969).Plant dyas are found in the root,root bark,1eaves,flowers,stem,stem bark,fruit skins and nutshells. Some plants may have more than one colour or sources of colour among the various parts and/or at various stages of their growth or development.These dyes are capable of improving the con— trast of plant sections,so that distinction can be made between lntroduetioll Forest dwellers depend on plants for tannins,dyes,food as raw materials for a wide variety of traditional products and artifacts one cel1 and the other or between cel1 and its constituents. However,the applications of synthetic dyes are presently lim— ited because of their hazardous effects on human and animal health.Bhuyan and Saikia(2004)asserted that the prevalent withdrawal from the use of some of these synthetic dyes is very connected with the detection of their hazardous nature.Also, The online version is available at htto:/ A.J.Akinloye Plant Anatomy Research Laboratory,Department of Botany and Mi— most developing countries can no longer afford the ever increas— ing cost of synthetic dyes(Avwioro et a1.2005a).These identi・ led problfems associated with the use of synthetic dyes now prompted the search for low—cost dyes,especially of biological origin,that will be effective,easy to use,biodegradable and safe crobiology,University of Ibadan,Oyo State 20000 1,Nigeria H.C.Illoh to both the human and environment health. Department of Botany,Obafcmi Awolowo University,Ile—Ife,Osun State 220005,Nigeria Natural vegetable dyes now gain prominence all over the world,especially in the developing countries due to their eco- A.O.Olagoke f Department of Foresty and Wood Technolrogy,Federal University of Technology,Akure,Ondo State 340001,Nigeria Email:akinlove iohnson@vahoo COUI friendly nature and hence,their promising usage was used as histopathological stains.This is typified by the most used dye haematoxylin obtained from the Mexican tree,Haematoxylon campechianum rBaker et a1.1 976).There are the numerous that have examined the potential of natural dyes for use in histopa— Resoonsible editor:Zhu Hon thology(Avwioro et a1.2005a&b:Eom et a1.200 1:Padhy et a1. ^、 Springer 82 1990;and Garg et a1.19911.Gaur et a1.f1998)also stressed that herbal stains from Saffron safifower and henna could be suc— cessfully utilized for differentiating inactive living and dead nematodes during bioassay and other investigations.Therefore, in the present study,we investigated the efifcacies of four herbal dyes in a bid to obtain non—toxic.eco—friendly and cheap stains ofr use in plant histological studies. Materials&methods Collection and preparation of materials The materials used for extraction of dyes in this work include: matured seeds of B#ca orellaHa,collected at the Polytechnic, Ibadan,Nigeria;dried heartwood of Pterocarpus OSHH,collected in Awosun Village,Ife North Local Government Area.Osun State Nigeria;rhizome of o rcuma domestica collected in the Plant Nursery Botany and Microbiology Department University of lbadan,Nigeria;and the young leaves of Lonchocarpus cya— neseens collected along Road 1,University oflbadan,Nigeria. Ripe,dried fruits of B.orellana were harvested.The seeds were shelled from the pods and winnowed to remove the husk, dust and other foreign particles and then kept in polyethylene bag Matured dried heartwood of P OSRl'1 was milled into powder using milling machine after thorough cleaning.The powder wood was kept in polyethylene bag. Fresh matured rhizomes of C domestica was mashed using pestle and mortar after thorough cleaning.The mashed rhizomes were sun—dried to reduce the moisture content before it was kept in polyethylene bag.The young leaves of L.cyanescens were collected,rinsed with distilled water to remove dirt.This was air— dried in the Laboratory.The dried leaves were ground using mortar and pestle.The mashed leaves were sun—dried to reduce the moisture content after the sample was kept in polyethylene bag.A1l the Plant materials used were identified at the Forest Herbarium Ibadan(FHI1 Extraction ofthe dyes Dyes were extracted from 1 kg each of matured heartwood ofB. orel ̄na seeds C domestica rhizomes.P.0s“ and young leaves of L.cyanescens using sohxlet extractor with ethanol as solvent. The dye yield was 205 g forB.orellana,202 g forP osHn,252 g ofre domestica and 105 gforL.cyanescens. Staining procedure and slide preparation The extracted dyes were tested on wood sections of Cola gi2日n— tea which was collected from the Botanical Garden.University of Ibadan.Nigeria.The plant sections were stained in the ex. tracted dyes for 3 minutes.For simple staining,the sections were washed/rinsed in water before treatment in series of ethano】to remove water molecules(dehydration process)and to remove excess stain(differentiation process in acidiifed ethano1).The 垒Springer Journal ofForestry Research(2010)2l(1):8l-84 dehvdrated and differentiated sections were transferred into ab— solute xylene in two series to remove last rtaces ofwater,to clear the section(making it more transparent)and to remove last rtaces of ethanol and since xylene is the solvent of the mountant rDPX) used.It prevents cloudiness of the slide and it makes slide to dry fast since xylene is a volatile solvent.The section is mounted on glass slide in DPX mountant. For the double staining.each section was stained first in dye- stuffs extracted.The sections were rinsed in water and then counter stained in Alcian blue.The process of dehydration,dif- ferentiation and clearing was done as for the simple staining as explained above and the section mounted in DPX mountant on the slide. Microscopy Miscroscopical observation of each slide was made and recorded. Photomicrographs of the slides were made using Olympus photomicroscope with analog camera. Spectrophotometric analysis Each of the extracted dves was crystallized to obtain the solid dye.The 0.0002 g of each sample was dissolved in 1 0 mL of Absolute ethano1 and 1 0 mL of Absolute ethanol was served as blank.Each of the dye samples was run on UV/visible spectro— photometers,which automatically subtracted the effect of the blank and plotted the graph of Absorbance against wavelength. pH determination The pH of each dye was determined with the use of digital BMF/ pH meter. Results Performance of dye extracts on wood sections The dye extract from B.orellana was crystallized easily and imparted its orange colour discriminately on the fibre and vessel members.Though the dye extract from C domestica did not crystallize easily because of the high oil content.it is much more discriminatory in imparting its brilliant yellow colour on fibre and vessel members.The crystal of the dye extract from L.cya. nescens absorbed moisture readily from the atmosphere and it is ipso facto suspected to be rich in sugar content.It did impart no visible colour on any cel1.Also。the dye extract from P.OSUn imparted its red colour on all cells but fibre and vessel members took the stain more deeply Additionally dye extracts from B. orellana.Cdomestica and OSbln arehighly specificwhenused in double staining wim Alcian blue.The Alcian blue stained parenchyma and thin wal1 cells while dye extracts from B.orel- lana,C domestica and P.osun stained fibre and vessel members (Fig.1—3 . Journal ofForestry Research(2010)21(1):81-84 The inability of L.cyanescens to impart any visible colour shows that L.cyanescens cannot be relied upon as a plant histo. 1ogical stain when it was used on wood sections. imparting quality than L.cyanescens but not as good in colour impartation as C domestica and P.osun. Dye extracts from B.orellana and e domestica are much more discriminatory in imparting their colours on fibre.vesse1 Conclusions This study has established the fact that herbal stain from B.orel- members.1ignified cells and thick wal1 cells even without corn— bining with other stain.Dye extract from e domestica proved highly selective because it has inherent amn for fibre and lana.C domestica and P osun could be Successfully utilized for plant histology.Dye extracts from B.orellana,C.domestica and other lignified cells.B.orellana also has a high a衔nitv f0r fibre and other lignified cells but not as C domestica.When these dye extracts were used in combination with Alcian blue in double staining they become highly discriminatory and speciifc. The histochemical behaviour of P.osRn dye extract and Sa— franin O is similar because both of thcm imparted their colours indiscriminately on all cells but fibre,vessel members.other ligniifed cells and thick wall cells takes stain more deeply.Again both stains become tissue.specific when they are counter-stained with Alcian blue or any other stains that have amnitv for thin wall cells.However.尸.OSUn extract had pH 4.2 1 being acidic and Safranin O had pH 7.61 which falls between neutral and alkalinity.Phloroglucinol is a known stain for lignin and is acidic like al1 the extracted dyes with pH 4.97.Alcian blue is known to have affiniyt for thin wall cells and is acidic with pH 3.25. Therefore acidiyt and alkalinity may not be the best parameter to determine the amnnv of these dye extracts for tissues/cells. The absorption spectra of the crude dye extract from e do— mestica,P.osul ̄,B.orellana and L.cyanescens revealed a range of wavelength of absorption from 327 nnl to 506 nm.This fal1s within the visible region of the electromagnetic spectrum and shows the presence of colour imparting chromophore in the dye extracts.This chromophore may be a polynuclear hydrocarbon compound or a simple aromatic with conjugated side chain.Peak depicts the predominant colour being transmitted by an organic pigment when the complementary colour contained in the light passing through it has been absorbed. The peak Of a colour is determined by the predominant wavc— length of absorption in it.Except for C domestica which has only one peak,indicating that Curcuma domestica has only one colour imparting chromophore,others have two or more colour imparting chromophores.Popoola et a1.(1 994)made similar observation of a single peak on Zingiber ofifcinale,a member of the same family Zingiberaceae.with C domestica. The qualiyt of the colour of the extract is a product of the sharpness of its absorption spectrum which in the case 0f e如一 mestica and P osun is represented by narrow range of the inten. sity profile.The observation of a narrow absorption band for C domestica and P.osun in the spectrum is agreed with the bright. hess of the dyes in them.This was also observed in Z officinale by Popoola et a1.r 1 994).These two dye extracts imparted their colours brilliantly on fibre and vessel members.L.cyanescens dye extract has poor colour imparting feature and this could be attributed to its broad peak and Iow absorbance.No visible col— our was seen on the specimen stained wim its dye extract.Dye extract from B.orellana shows Iess broad peak and higher ab— sorbance than that of L.cyanescens.B.orellana has better colour 垒Springer P.osun are good replacement for Safranin O,Phloroglucinol and any stain with abiliyt to impart its colour on fibre and vessel elements.The results also revealed further that a high perform- ance of these dye extracts is obtainable with the use of ethanol as a solvent for their extraction.This discovery will go a long way in reducing over—dependence on toxic.expensive and non. available exotic stains.Further research should be conducted on the analysis of the active chemical substances in the dye extracts. There is also a need to investigate the potential of the dye ex. tracts in detecting presence or absence of eell iDelusions and ergastic substnaces, References Avwioro OG,Aloamaka PC,Ojianya NU,Oduola T,Ekt)o EO.2005.Ex— tracts ofPterocarpus osHn as a histological stain for collagen fibres. .j. Biotechnol,4(5):460-462. Awvioro OG,Awoyemi FA,Oduola T.2005.A novel natural collagen and muscle stain from Morinda lucida extracts. P f.Med. ,4(2):44--48. Avwioro OG.2002.Histochemistry and t&sue pathology.】st cd.Ibadan. Nigeria:Clavedanun Press.PP.134—2】3. Baker FJ,Silverton RE.1 976 The theory of staining.In:Introduction to Medical Laboratory Technology,6th ed.London:Butterworths and Co, London,pp.385-391. Banderanayake WM.1 999.Traditional and medical uses of Mangoves.Man— groves andSaltMarslec,2f3):133—148 Bhuyan R,Saikia CN 2004 Isolation of colour components from native dye・ bearing plants in Northeastern India.Am Patho1.,164(5):873—877. Eom S,Shin D,Yoon K.2001.Improving the dyeability ofnatural colourants on cotton by cationization.Ind.JFibreTextRe.,26f4):425—431. Garg AS.Shenda and K.C Gupta 1991 Effect of mordants on colour of natural dye extracted from tissue lfowers urea mongsperma).Colourage, 38(2):50-53. Gaur HS,Chandel ST 1998.Use of indigenous non-toxic herbal dyes in staining nematodes.Ann PI.Protec Sci,6(2):189—2l9 Lillie RD,Conn’s HJ l 969.Biological Stains:A handbook on me nanIre and uses of the dyes employed in the biO1Ogica1 lab0ratOry,8 Ed.Ba1timOre: The W川am and Wi1kins COmpan y_pp.86一I I2 Padhy RN,Rathi D.199O.E f ct 0fmordants on dyeing ofc0tton with vege— tab1e dves Z f DyPr r r,23(25):27—28 P0pO0la AV,IpinmOrOti KO,OgunmOroti TO.1994 ExtractiOn,spec扛0一 sc0pic and colouring p0tentia1 studies 0f the dye in Ginger RhizOme (z g 6P q c 日 )JD口后_,&f 尺 ,37(5):217—22O. Chinese Abstracts 3 不论是颜色要是力学性能都发生了明显的变化。铁红和铁黑着色的试样 在整个老化过程中表现较好,颜料添加量约2.28%LL较适宜。图1表4参 13。 关键词:木塑复合材;颜料:老化:颜色:力学性能 CLc number:¥78l-2 Document code:A Article ID:1007—662X r2010)01—0059-04 Dol:1O.1007/sl1676.0l0—0009.z 20l0—0l—l0 印楝木屑压块物理及燃烧特性=Physical and combustion prope ̄ies of briquettes from sawdust of Azadirachta indica[刊,英]/Sotannde,O.A.i, Oluyege,A.O. and Abah,G.B. (1.Department of Forestry and Wildlife, Faculty of Agriculture,Universiyt of Maiduguri,P.M.B.1069 Maiduguri, Borno State,Nigeria;2.Department of Forestry and Wood Technology, Federal University of Technology,P.M.B.704 Akure,Ondo State,Nigeria) //Journal ofForestry Research一2010(1).63 ̄67 研究调查木薯淀粉和阿拉伯树胶粘合印楝木屑制成压块的特性。压 块在千斤顶平均压力为1O.7 kg・cm 制成。木屑与粘合剂的重量比分别为: 100:l5,100:25,100:35和100:45。测试压块的物理和燃烧特性。压块的 物理和燃烧特性的变化随粘合剂的种类和粘结剂的水平而变化 < 0.05)。结果表明:淀粉为粘合剂压制的木屑压块有较好的性能、密度、 耐用性、热值、固碳百分率、低灰和低挥发物分别为O.546 g・cm一、95.93%、 33.09MJ・kg‘。、84.70%、3.35% ̄11 11.95%;而阿拉伯树胶为粘合剂压制的 木屑压块的密度、耐用性、热值、固碳百分率、低灰和低挥发物分别为 0.425 g cm~、94 85%、32.76 MJ・kg~、87.30%、4.45%和8.75%。研制压 块是为了得到良好的燃料,当木屑与淀粉和阿拉伯树胶为粘合剂的比例 分别为100:25和100:35,能得到最佳木屑压块。图2表3参l6。 关键词:印楝;压块;粘结剂的水平;耐用性;燃烧特性 CLC number:S 759.8 Document code:A Article ID:1007——662X(2010)01-00635-05 DoI:1O.1007/s1 1676.010.001O一6 20l0—0l一1 l Pi lodyn在桉树活立木材性质评估中的应用=Use ofthe pilodyn for assess- ing wood prope ̄ies in standing trees of Eucalyptus clones[f0,英】/吴世军 , 徐建民 ,李光友 ,RISTO Vuokko ,陆钊华 ,李宝琦 ,王伟’(1.中国林业 科学研究院热带林业研究所,广东广州510520;2.广西斯道拉恩索林业 有限公司,广西南宁530022)//Journal of Forestry Research.//Journal of Forestry Research.一2010(1).68-72 测定了建立在广西22个4年生桉树无性系的Pilodyn值、木材基本 密度、边材木材密度、心材木材密度和弹性模量。结果表明:Pilodyn值 为9.44—15.41mm、木材密度为O.3514-0.4913g・cm- 、弹性模量在3.94 GPa 到7.53GPa之间;Pilodyn值在不同处理、不同方向及不同参试无性系间 均存在极显著差异。Pilodyn值与各材性指标间的相关系数在一0.433到 .O.755之间,呈显著到极显著负相关。研究表明,Pilodyn方法可以预测 桉树活立木的木材密度和弹性模量。表6参28。 关键词:相关关系:桉树;弹性模量:Pilodyp;木材密度 CLC number:¥781 Document code:A Article ID:1007 ̄662X(2010)01・・0068—05 DoI:l0.1007/sl1676-010一O0l1—5 2OlO—Ol—l2 基于叶重的桉树单叶面积估计模型=Single leaf area estimation models based on leafweight ofeucalyptus in southern China[刊,英1/刁军 ,雷相东 。,洪玲霞 ,戎建涛‘,石强 (1.中国林业科学研究院资源信息研究所, 北京100091:2.深圳职业技术学院,深圳518055)//Journal of Forestry Research.*2010(1).73 ̄76 叶面积是树木生长模型和树木的生理过程的一个重要参数。根据叶 重量估计叶面积。本文以我国南方主要树种桉树为对象,实测了从25株 桉树上摘取的455片桉树叶子的重量和面积,分别建立了基于叶鲜重的 嫩叶与成熟叶单叶面积预测模型。其中80%的数据用于建模,20%的数据 用于模型验证。基于模型拟合效果、误差和残差分布,确定了成熟叶和 嫩叶的叶面积模型分别为幂函数和线性方程。经检验,模型精度均达95 ̄,/0 以上。建立的模型为估计单叶面积提供了一个简单可靠的方法,并且在 桉树的结构一功能模型研究中有应用潜力。图4表5参l5。 关键词:桉树;叶面积;叶重量;异速模型 CLC number:S71 8.43 Document code:A Article ID:1007—662x f2010)01—0073—04 DoI:l0.1007/s11676.010—00l2-4 2Ol0・0l-l3 紫椴花多糖的超声提取最佳条件和含量测定=Optimal ultrasonic extrac— tion condition and determination of polysaccharides in Tilia amurensis flow. ers[J:Jl,英]/穆立蔷1,2,刘立平1,马大龙 (1.东北林业大学林学院,哈尔滨 150040;2.黑龙江省科学院自然与生态研究所1//Journal of Forestry Re. search.一2010(1).77-80 设置7个萃取温度(25,35,45,55,65,75,85。C)和萃取时间(30,40,50, 6O,7O,8O,90 min)来选择超声提取紫椴花中多糖的最佳提取温度和提取 时间。利用分光光度法对紫椴花中的多糖含量进行了分离和测定。结果 表明:最佳提取条件为提取温度为75"C,提取时间为52 min 经蒽酮—硫 酸显色,于580 nm处测定,其多糖的含量为9.74%,RSD=0.47%fn=31。 此测定方法简便,样品溶液在4 h内显色稳定,重现性较好,平均回收率 为99.48% ̄1.01%,RSD=0.1 12%fn=3)。图l表3参13。 关键词:最佳条件;多糖;紫椴花;超声提取; CLC number:¥792.36 Document code:A Article ID:1007—662X r2010)01—0013—04 D0I:】O】007/sl1676—0l0一O0】3—3 20l0—0l-14 用于植物组织染色的一些本地草本植物染料的筛选=Screening of some indigenous herbal dyes for use in plant histological staining[刊,英]/A.J. Akinloye 1.H.C.Illoh .A.O.Olagoke* r1 Plnat Anatomy Research L曲。一 ratory,Department of Botany and Microbiology,University of Ibadan,Oyo State 200001,Nigeria;2.Department of Botany,Obafemi Awolowo Univer— sity,lie—Ife,Osun State 220005,Nigeria;3.Department of Forestry nad Wood Technology,Federal Universiyt of Technology,Akure,Ondo State 340001, Nigeria1//Journal ofForestry Research.-2010(1).81~84 为了改良有毒的、昂贵的、人造的非本地的染剂,研究用于植物组 织染色的一些无毒和环保型的草本植物染剂的功效。采用常规的染色方 法,利用从红木(Bixa orellana)、姜黄(Curcuma domestica)、约鲁巴 靛蓝(Lonchocarpus cyanescens)和紫檀(Pterocarpus OSUn)中提取的染 色剂用于木本植物切片的着色。除了从约鲁巴靛蓝中提取的染色剂夕}, 所有从植物中提取的染色剂都对纤维和导管分子有亲和性。从姜黄和红 木中提取的染色剂比从紫檀中提取的染色剂有更高的选则性。吸光率曲 线结果表明,除了从姜黄中提取的染色剂外,从植物中提取的染色剂有 两个小峰值,因此他们有两个或更多的染色团。染色剂显酸性(pH,3.77 .77)。研究表明,从红木,姜黄和紫檀中提取的染色剂可以单独或与 人工染剂结合用于植物组织染色。图3表2参12。 关键词:草本植物染料;本地草本植物;环保型染剂;组织染色;木本植 物切片 CLC number:Q945.8 Document code:A Article ID:1007-662X(2010)01・・0081-04 DoI:10.1007/sl l676.010—0014.2 2Ol0・0l-l5 孟加拉国曼尼普尔部落对部族药用植物的利用=Etnho—medicinal plnats use by the Manipuri tribal communiyt in Bangladesh[f0,英】/Md.Parvez Raua .Md.Shawkat Islam Sohel ,Sayma Akhter ,Md.Jahirul Islamz(1. Department of Forestry and Environmental Science,Shahjalal University of Science and Technology,Sylhet一3 l 14,Bangladesh;2.National Graduate 

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