全 文 ：天然产物研究与开发 NatProdResDev2009, 21:744-747
文章编号:1001-6880(2009)05-0744-04
　
　
　ReceivedApril14, 2008;AcceptedMay15, 2008
　FoundationItem:ThisworkwasfinancialysupportedbytheNational
KeyTechnologyR＆DProgramofChina(No:2008BADA5B03 and
2007BAD57B02)andBingtuanDoctoralFoundationofChina(No:
2006JC09).
＊CorrespondingauthorTel:86-10-62731199;E-mail:lgzhou@cau.edu.
cn
六种藜科植物提取物对植物病原菌的抑制活性
杨红兵 1, 2 ,周亚明2 ,刘　浩 2 ,杜　华 2 ,马占鸿 2 ,李　春 1 ,周立刚 2＊
1石河子大学化学化工学院 , 石河子 832003;2中国农业大学农学与生物技术学院 ,北京 100193
摘　要:研究了角果藜(Ceratocarpusarenarius)、盐穗木(Halostachyscaspica)、里海盐爪爪(Kalidiumcaspicum)、叉
毛蓬(Petrosimoniasibirica)、盐角草(Salicorniaeuropaea)和小叶碱蓬(Suaedamicrophyla)等六种新疆藜科植物提
取物及其不同极性萃取部分对根癌土壤杆菌(Agrobacteriumtumefaciens)、黄瓜角斑病菌(Pseudomonaslachry-
mans)、番茄疮痂病菌(Xanthomonasvesicatoria)等植物病原细菌以及杨树溃疡病菌(Botryosphaeriadothidea)、棉
花枯萎病菌(Fusariumoxysporumf.sp.vasinfectum)、稻纹枯病菌(Rhizoctoniasolani)等病原真菌的抑制活性。结
果显示角果藜 、叉毛蓬和盐角草乙醇粗提物表现出一定抗细菌活性 , 其中以叉毛蓬和盐角草提取物对黄瓜角斑
病菌的抑制活性最强。多数植物提取物及其不同极性萃取部分对杨树溃疡病菌表现出强的抑制活性。抗真菌
活性成分主要存在于供试植物的石油醚 、氯仿和正丁醇萃取部分中 , 提示活性成分为极性中等的化合物。角果
藜和盐角草乙醇粗提物及其不同极性萃取部分对供试真菌有较好的抑制活性。
关键词:角果藜;盐穗木;里海盐爪爪;叉毛蓬;盐角草;小叶碱蓬;提取物;植物病原菌;抑制活性
中图分类号:Q946.887;R285;S482.2 文献标识码:A
InhibitoryActivityofExtractsandFractionsfrom
SixChenopodiaceousPlantsonPlantPathogens
YANGHong-bing1, 2 , ZHOUYa-ming2 , LIUHao2 , DUHua2 , MAZhan-hong2 , LIChun1 , ZHOULi-gang2＊
1SchoolofChemistryandChemicalEngineering, ShiheziUniversity, Shihezi832003 , China;
2ColegeofAgronomyandBiotechnology, ChinaAgriculturalUniversity, Beijing100193 , China
Abstract:Crudeethanolextractsanddifferentpolarfractionsofsixchenopodiaceousplantspecies(i.e.Ceratocarpus
arenarius, Halostachyscaspica, Kalidiumcapsicum, Petrosimoniasibirica, SalicorniaeuropaeaandSuaedamicrophyla)in
XinjiangofChinawerestudiedfortheirinhibitoryactivityagainstplantpathogenicbacteria(i.e.Agrobacteriumtumefa-
ciens, Pseudomonaslachrymans, Xanthomonasvesicatoria)andpathogenicfungi(i.e.Botryosphaeriadothidea, Fusarium
oxysporumf.sp.vasinfectum, Rhizoctoiansolani).TheextractsofC.arenarius, P.sibiricaandS.europaeashowedthean-
tibacterialactivity.P.sibiricaandS.europaeaextractsweredemonstratedthestrongestinhibitoryactivityonbacterium
P.lachrymans.MostoftheextractsandfractionsshowedstrongactivityonB.dothidea.Antifungalcompoundsexistedin
theeitherpetroleumetherfractionorchloroformone.TheethanolcrudeextractsandfractionsofC.arenariusandS.euro-
paeashowedthestrongestantifungalactivityagainstaltestfungi.
Keywords:Ceratocarpusarenarius;Halostachyscaspica;Kalidiumcapsicum;Petrosimoniasibirica;Salicorniaeuropaea;
Suaedamicrophyla;plantpathogen;inhibitoryactivity
Introduction
Chenopodiaceousplantsarewidelydistributedinarid
andsalinaarea.Thereare184chenopodiaceousspecies
belongingto38 generainChina.Mostofthem are
foundinXinjiang, northwestofChina[ 1, 2] .Themajor
constituentsisolatedsofarfromchenopodiaceousplants
arealkaloids, terpenoidsandflavonoids, whichshowed
avarietyofbiologicalactivitiesandecologicalfunctions
[ 3] .Fewplantsofthisfamilyhavebeenscreenedto
showantimicrobialpotential[ 4-6] .Sixchenopodiaceous
plantspecies, i.e.CeratocarpusarenariusL., Ha-
lostachyscaspica Bieb., Kalidium capsicum Ung.-
Sternb., PetrosimoniasibiricaBunge, Salicorniaeuro-
paeaL.andSuaedamicrophylaPal., arerichbiore-
sourcesmainlydistributedinXinjiangofChina[ 1] , and
notstudiedwithrespecttotheirantimicrobialactivity
againstplantpathogens.Weatemptedinthisstudyto
screenantimicrobialchenopodiaceousplantextracts
thathavepotentialforagriculturalapplicationtomini-
mizecroplossescausedbypathogensofagricultural
importance.
MaterialsandMethods
Plantmaterials
Theaerialpartsofsixchenopodiaceousplantspecies
werecolectedinAugust2007 atMushroomLakeRes-
ervoir, 20kmawayfromNorthShihezi, XinjiangofChi-
na.ThesampleswereauthenticatedasCeratocarpus
arenariusL.(C.ar), HalostachyscaspicaBieb.(H.
ca), KalidiumcaspicumUng.-Sternb.(K.ca), Petrosi-
moniasibiricaL.(P.si), SalicorniaeuropaeaL.(S.
eu)andSuaedamicrophylaPal.(S.mi)byProf.
PingYanattheColegeofLifeSciencesofShiheziU-
niversityinChina.Thevoucherspecimensweredeposi-
tedintheColegeofAgronomyandBiotechnologyof
ChinaAgriculturalUniversity.
Fungalandbacterialstrains
Plantpathogenicbacterialstrains, Agrobateriumtumefa-
ciens, PseudomonaslachrymansandXanthomonasvesi-
catoria, andplantpathogenicfungalstrains, Botryo-
sphaeriadothidea, Fusariumoxysporiumf.sp.vasinfec-
tumandRhizoctoniasolani, werefromtheDepartment
ofPlantPathologyofChinaAgriculturalUniversity.Al
bacteriawereactivatedonaLuria-Bertani(LB)agar
medium(sodiumchloride5, peptone10, yeastextract
5, agar20g/Lindistiledwater)plate, andthensub-
culturedonLBagarmediumat28℃ inthedarkatev-
erytwo-day sintervalspriortoantibacterialtest.Al
fungiwereactivatedonapotatodextroseagar(PDA)
medium(potato200, glucose20, agar20 g/Lindis-
tiledwater), andthensubculturedonPDAmediumat
25 ℃inthedarkateveryfourtoseven-day sintervals
priortoantifungaltest.
Preparationofplantcrudeextractsandfractions
Theplantmaterialsweredriedinshadeatroomtem-
perature.Firstofal, each200 gpoweredsamplewas
extractedfirstusing95%(v/v)ethanol(3×1000mL)
at85 ℃ for4 h.Themaceratedplantsuspensionwas
thenfiltered.Thecombinedfiltratewasconcentrated
underreducedpressureat50 ℃ toafordaresidueas
anethanolcrudeextract(abbreviatedasEt.extr.).A
majorityoftheethanolextractwasdissolvedinhotdis-
tiledwater(300 mL)togetasuspensionsolution,
whichwasfractionatedbypetroleumether(3 ×200
mL), chloroform(3×200 mL)andn-butanol(3×200
mL)toafordpetroleumetherfraction(Pe.fract.),
chloroform fraction(Chl.fract.)n-butanolfraction
(But.fract.)andaqueousfraction(Aq.fract.).The
yieldsofplantextractsandfractionsweresummarized
inTable1.
Table1　Yieldsofplantcrudeextractsandfractions
Plant
Et.
extr
.(%)
Pe.
fract.
(%)
Chl.
fract.
(%)
But.
fract.
(%)
Aq.
fract.
(%)
C.ar 5.07 0.95 1.33 1.42 0.91
H.ca 27.27 5.45 1.10 6.75 9.11
K.ca 21.65 5.38 1.38 5.73 7.31
P.si 7.37 1.86 0.37 1.65 1.95
S.eu 15.13 5.19 0.53 4.13 5.06
S.mi 26.19 8.15 1.77 5.24 9.70
Antibacterialasay
Anagarweldifusionmethod[ 7] wasusedtoevaluate
antibacterialactivityoftheplantextracts.Briefly, 150
μLofthetestbacterialdilution(109 CFU/mL)was
addedinto15mLofLBagarat45℃, thenmixedthor-
oughlyandpouredintoa9cmdiameterPetridishcon-
taining10mLofsolidified3% wateragar.Aftersolidi-
fication, sevencircularequidistantwels(5 mmindi-
ameter)weremadeintheLBagarlayerusingsterile
corkborers.Theextractsweredissolvedindimethyl
sulfoxide(DMSO)toafinalconcentrationof100 mg/
mL.Then50μLofeachextractsolutionwasaddedinto
thewels.DMSOwasusedasanegativecontroland
streptomycinsulfateasapositivecontrol.Theplates
wereincubatedfor24 hat37 ℃, andtheinhibition
zonesweremeasured.Altestswereperformedintripli-
cateandrepeatedtwice.
745Vol.21　　　　　　YANGHong-bing, etal:InhibitoryActivityofExtractsandFractionsfromSixChenopodiaceousPlantsonPlantPathogens　
Antifungalassay
Toevaluatetheantifungalactivityofplantextractsand
fractions, amycelialradialgrowthinhibitiontestwas
used[ 8] .Eachdriedextractwasdissolvedinsomeor-
ganicsolvent(i.e.ethanolassolventforB.dothidea
andR.solanitest;DMSOassolventforF.oxysporumf.
sp.vasinfectum test).Each dissolvedextractwas
pouredintotheassayflaskcontainingahotsterilized
PDAmediumat50-60 ℃.Afterthoroughlymixing, a-
bout15mLoftreatedmediumwaspouredintoaPetri
dish(90mmdiameter).Theconcentrationoftheorgan-
icsolventinmediumwas1%(byvolume)withafinal
concentrationoftheextractsinmediumas1.00 mg/
mLrespectively.Adisc(5 mmdiameter)offourto
seven-day-oldpathogenmycelialculturewasasepticaly
transferredtothecenterofthesolidifiedPDAmedium
inplates.After3d(R.solani), 5 d(R.dothidea)and
10 d(F.oxysporumf.sp.vasinfectum)ofinoculation
respectivelyat25℃ inthedark, mycelialgrowthofthe
pathogenwasmeasured, andanaveragewastakenof
twomeasurementscrosswisemadeoneachplate.Nega-
tivecontrolplatesweretreatedwiththesamequantity
oforganicsolventmixed withPDA.Carbendazim
(Aldrich, USA)with0.01 mg/mLinmediumwas
usedasareferencefungicide.MycelialgrowthinPDA
mediumcontainingplantextractsanddiferentpolar
fractionswascomparedwiththegrowthofthepathogen
inplatescontainingorganicsolvent(control).Eachtest
wasperformedinfivereplicatesandrepeatedtwice.
Antifungalactivitywasrecordedintermsofpercentage
colonyinhibitionandcalculatedaccordingtoourprevi-
ousreport[ 5, 6] .Percentage(%)ofmycelialgrowthin-
hibitionwasdeterminedas[ (Dc -Dt)/Dc] ×100,
whereDcistheaveragediameterincreaseoffungalcol-
onywithcontrol, andDtistheaveragediameterin-
creaseofafungalcolonywithtreatment.
ResultsandAnalysis
Inhibitoryactivityoftheextractsonplantpathogenic
bacteriaAsshowninTable1, theethanolcrudeextracts
ofC.arenarius, P.capsicumandS.eusopaeashowedin-
hibitoryactivityagainstalthreetestbacteria.Ofthem,
theextractsofP.sibiricaandS.europaeapossessedthe
strongestinhibitoryactivityonP.lachrymans.Otherex-
tracts(i.e.H.caspica, K.capsicumandS.microphyla
extracts)wereinactiveontestbacteria.
Table2　Antibacterialactivityoftheplantextracts
Treatment PlantpathogenicbacteriumA.tumefaciens P.lachrymans X.vesicatoria
C.arEt.extr. + + +
H.caEt.extr. - - -
K.caEt.extr. - - -
P.siEt.extr. + ++ +
S.euEt.extr. + ++ +
S.miEt.extr. - - -
Streptomycinsulfate +++ +++ +++
　　Note:Thequantityoftheextractorstreptomycinsulfateineachwel
was5mg, DMSOassolventwasfoundtoshownoinhibitionontestedbac-
teria.– indicatednoinhibitionwasobserved, +indicatedthediameterof
inhibitoryzonetobe5-10mm, ++indicatedthediameterofinhibitory
zonetobe10-15 mm, and +++indicatedthediameterofinhibitory
zonetobemorethan15mm.
Inhibitoryactivityoftheplantextractsandfrac-
tionsonplantpathogenicfungi
AsshowninFigs.1 to3, mostoftheplantextractsand
fractionsshowedantifungalactivityonB.dothidea, the
poplarstemblistercankerpathogeninthreetestfungi.
Antifungalcompoundsmainlyexistedinthepetroleum
etherfraction, chloroformfractionandn-butanolfrac-
tion.Ofalextractsandfractions, theethanolcrudeex-
tractsanddiferentpolarfractionsofC.arenariusand
S.europaeashowedthestrongestantifungalactivityon
testfungi.
Discussion
Syntheticfungicidesorbactericideshavebeenproven
efectivelyincontrolingplantdiseases.However, their
usagehasbeenincreasinglyrestrictedduetoenviron-
mentalandhealthconcerns[ 9] .Higherplantscanpro-
duceagreatdiversityofsecondarymetabolites, manyof
whichhaveantimicrobialefects[ 10, 11] .Itremainsevi-
dentthatplantnaturalproductsstilhaveanenormous
potentialinmodernagrochemicalresearch.Overthe
pastdecadetherehasbeenanelevatedinterestin
searchingforantimicrobialagentsofplantorigin, as
welasinisolatingandidentifyingantimicrobialcom-
poundswithpossibleuseinintegratedplantprotection
anddiseasemanagementprograms[ 12] .
746 NatProdResDev　　　　　　　　　　　　　　　　　　　　 　Vol.21
Sixchenopodiaceousplantspecieswerescreenedfor
theantimicrobialactivityagainstplantpathogensinthis
work.Therewerenopublishedreportsonthechemistry
andbiologicalactivityonthesesixplantsexceptK.
capsicumstudiedforitsphenolics[ 13] andS.europaea
studiedforitsflavonoids[ 14] .Themostoftheextracts
andfractionsofsixchenopodiaceousplantspeciesfrom
XinjiangofChinashowedantimicrobialactivityon
plantpathogens.BycombiningtheresultsshowninTa-
ble2 andFigs.1 to3, muchmoreantimicrobialcom-
poundswithdiferentpolarityexistinC.arenariusand
S.europaea, suggestingthattheymaybeusedasanti-
microbialagentsfortheplantdiseasescontrol.Further
investigationofantimicrobialcompoundsintheseplants
isofinteresting.
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747Vol.21　　　　　　YANGHong-bing, etal:InhibitoryActivityofExtractsandFractionsfromSixChenopodiaceousPlantsonPlantPathogens