氰化物是目前世界范围内最常使用的提取黄金和白银等贵重金属的沥滤剂,其对自然生态环境的污染和破坏以及对人畜和其它生物的毒性作用是众所周知的.本试验用一自行设计的生物反应器来观察黄豆(Glycine max(L)Merr.)和玉米(Zea mays L.)对氰化物污染土壤的原位修复的可能性.室温条件下(23.0～26.0℃),低浓度的氰化物污染液对(≤45.5 CN mg·L-1)二种测试植物的生长没有产生任何毒性作用;而在高浓度的氰化物试验组(≥91.0 CNmg·L-1),二种测试植物的生长都出现了明显的滞长现象(生长率下降大于10%),但没有观察到其它毒性反应.同时二种测试植物的叶片细胞用来测定植物细胞线粒体中的氰丙氨酸合成酶(β-cyanoalanine synthase)转化氰化物的潜力.实验是在一封闭的玻璃器皿(100mL)中进行的(100mL的氰化钾溶液中加入1.5g(鲜重)植物的叶片,氰化钾溶液的浓度大约1.0 CNmg·L-1).在为期28 h的时间内,水溶液中超过90%的氰化物被植物的叶片去除;黄豆和玉米的的叶片细胞对氰化物去除率分别测定为4.43mg CN·kg-1(鲜重)·h-1和3.42mg CN·kg-1(鲜重)·h-1.本实验结果表明,植物对氰化物污染的土壤原位修复方法是一种可行的和有效的选择.
Cyanide is a high-volume production chemical and the most commonly used leaching reagant for gold and silver extraction. Its environmental behavior and fate is of significant concern because of its high toxicity. This paper presents an investigation of the feasibility of phytoremediation as an in- situ technique to treat cyanide-contaminated soil. Corn(Zea mays L.)and soybean(Glycine max(L.)Merr.)were used. Results from this tests indicated that low dose of cyanide(≤45.5 CN detectable effects under room temperature(23.0～26.0℃). Significant reduction in the plant growth rates(≥10%)was found at high cyanide concentration in the irrigation water(≥91.0 CN mg·L-1)used. The potential of detached leaves to remove cyanide was also investigated. Excised plant leaves(1.5 g fresh weight)were kept in a glass vessel with with 100 mL spiked aqueous solution(deionized oxygen-saturated water)at 23.5℃ for 28 h. The disappearance of cyanide from the aqueous solution was tracked photometrically. More than 90% cyanide in the aqueous solution was removed by plant materials over a 28-h exposure period. The cyanide removal capacity was determined to be 4.43 and 3.42 mg CN·kg fresh weight-1·hour-1 for soybean and corn, respectively. Resutls from this investigation indicated that plants and plant materials were all found to be able to efficiently remove cyanide from the contaminated medium through their internal enzyme systems. In conclusion, cyanide elimination with plants seems to be a feasible option for cleaning soils contaminated with cyanide or gold and silver mine waste and wastewater.
全 文 ： 万方数据
exceptionknownsofar131．Asparagine，anon．toxic
essentialamionacid，wastheonlymetabolitedetectedin
experimentsi volving14C．1abeledCN’ forboth
cyanogemcandnon。cyanogenlcplants‘””i．
Thecurrentapproachwastofeedtoxiccyanide
wasteothecandidateplants．Indeed，phytoremediation
hasbecomea cost—effectiveoptionforcleaningupthe
environmentalco minatedsites．Itisconsideredtobea
“GreenRevolution”．Thefirstfull．scaleapplicationwas
carriedoutinDenmarkona formergasworkssite
contaminatedwithcyanide．Evenmorepromisingisthe
applicationfor emediatinggoldmines16J．Successful
phytoremediationwillrequiretheselectionof
appropriatepl nts．Thebeststrategyisthereforet test
thelocalvegetation．inparticularcy nogenlcplants
(thosethataretoxicthemselvesbecauseth yproduce
cyanide)andfast—growingspecies．Thebestspeciesmay
growinthewildoncyanidecontaminatedsites．Inthis
study,twocyanogenicplantswereusedtotesthe
feasibilityofplantstoremovecyanidefromthe
contaminatedsoil．
1 Materialsandmethods
1．1Toxicitytestsofcyanidewithplants
Seedsofcorn(ZeamaysL．)andsoybeans(Glycine
max(L．)Merr．)weresewninplasticcontainersfilied
withsandandsoilbeforesawingthexperimentalplant
seeds．Thethicknessofthesandandthetopsoillayer
were3cmand6cm，respectively．A11soilsweretaken
fromtheHAUcampusinChangsha．Thesoilshadbeen
previouslyanalyzed，butnode ectablemountsof
cyanidehadbeenfound．Thesecontainersw eall
placedinanincubatorwithaconstanttemperatureof
23．5"12untileavesandshootsappeared．Seedsof
soybeanswerelocallyavailablespecieswhilethecom
wasobtainedfromtheInstituteofCropGermplasm
ResourcesoftheChineseAcademyofAgricultural
SciencesinBeijing
Thetoxicitytestswerecarriedoutinthelaboratory
atroomtemperatureof23．0～26．0。Cundernatural
sunlight．Eachplasticontainerhadoneplantandsix
replicateswereconductedin eachtestinggroup．
Potassiumcyanidesolutionwith9．1．18．2，45．5and91．0
CNmg·L-‘werepreparedinthelaboratoryusing0．9066
CNg·L．1stocksolutionofKCN．Incontrol，tapwater
wasused．ThepHofthespikedsolutionwasadjustedto
6．9withsodiumhydroxide．Noattemptwasmadeto
selecthomogeneousplantmaterialsfora11testinggroups
(whichisdifficult)．butineachtestinggroupthereisa
smalldifferenceamongplantmaterials．Eachcontainer
waswateredwith10b15mLoftherespectivesolution
everydaywithinthentiretestperiod，dependingonthe
transpirationofplants．Notethateachtestinggroup
addedthesamevolumeofspikedaqueoussolutionin
ordertokeepthesamecyanidedose．Theheightof
plantswasallmeasuredatthebeginningandthendof
thetests．Thedurationofthistestwas10days．
1．2Removaltestsofcyanidewithdetachedleaves
Sealedglassvesselswereusedcontainingcyanide
solutionandplantleaves．Plantmaterialswerewashed
bytapwaterbeforeth xcision．Excisedleaveswerecut
intopieces，preciselywe ghed(1．5gfreshweight)and
placedinaflaskof100mL．then100mLofspiked
aqueoussolution(deion&edoxygen—saturatedwater)
wereadded．ThepHofspikedsolutionwasadjustedto
6．9withsodiumhydroxide．Theflasksw reclosedwith
aglasstopperandallplacedinanincubatorwitha
constanttemperatureof23．5℃．Threereplicateswere
conductedineachtestinggroup．Oneco trolwaswith
cyanide．butwithoutplantleavesandtheotherwaswith
plantleaves，butwithoutcyanide，toquantifytheffects
of photolysis，volatilization，hydrolysisand／or
degradationbymicroorganisms，andtochecktherelease
Table1Averagegrowthofcornplantsexposedtocyanideintheirrigationwater．Valuesrem ansofplantheightfromsix
replicates(inparenthesis：standarddeviation)．
CyanideconcentrationInitialaverageheight Finalverageheight Averagegrowth Reductionofgrowth
(CNmg·L．1) (cm) (cm) (cm) (％)
0 6．43(0．i033) 14．05(0．3209) 7．62(O．3189)0
9．1 6．55(0．15l7) 14．00(0．4290) 7．45(O．4637) 2．23
18．2 6．47(O．2160) 13．97(0．3983) 7．50(O．6033) 1．57
45．5 6．43(0．2066) 13．75(0．．2881) 7．32(O．4070) 3．94
11：Q 垒：i!lQ：兰丝12 1兰：兰Z(Q：≥垒ZQ2 主：12(Q：兰2112 兰!：兰2
Table2Averagegrowthofsoybeanplantsexposedtocyanideintheirrigationwater．Valuesrem ansofplantheightfromsix
replicates(inparenthesis：standarddevi tion)．
CyanideconcentrationInitialaverageheight Finalverageheight Averagegrowth Reductionofgrowth
(CNmg·L．1)(era) (cm) (cm) (％1
9．1 8．55(0．1517) 12．35(0．8191) 3．80(0．9487) I．30
18．2 8．60(0．1673) 12．25(0．8758) 3．65(0．8313) 5．19
45．5 8．58(O．2137) 12．17(0．7118) 3．59(0．6047) 6．75
21：Q 墨：墨Z lQ：兰Q堑2 11：三il垒：垒丝兰2 兰：2垒lQ：Z!熊 兰墨：：!
万方数据
2期 于晓章，等：氰化物污染的植物修复可行性研究 99
Theconcentrationofspikedsolutionwasofcyanide
fromtheplantmaterials．
approximately1．0CNmg·L．1．Thepr senceof
cyanideintheaqueoussolutionwasmeasuredbefore
planttissuesw readded，afterwhicht ereductionof
cyanideconcentrationinsolutionwasquantifiedwithan
intervalof4h．Thedurationofthistestwas28h．
1．2Chemicalan ysis
ThestocksolutionofpotassiumcyanidefO．9066
CNg·L．1)waspreparedinthelaboratory．The
concentrationofcya ideintheaqueoussolutionw．a．、．s
determinedphotometricallybya standardmethodiZl．
ResultswereexpressedasCNmg·L-1．Notethatl mg
KCNequals0．4mgCN．Betweenl and5 mLwater
samplesweretakenandtransferredintoa 25mL
colorimetriccyl nder,andl％sodiumhydroxidewas
addedtothemarkof10mL．5．0mLofbuffersolution
containingpotassiumd hydrogenphosphate(17g·L‘)
andsodiumphosphate(17．75g·L-1)wereadd d．Then
0．2mLofl％(V／M1chloramine．Tsolutionwereadded．
Theflaskwasealedwithstopperandleftfor3 to5
minutesb foreadding5．0mLofthecolourreagent(a
mixtureof isonicotinica id and
3-methyl—l—phenyl一5一pyrazolonewiththeconce trations
of15g·L-‘and12．5g·L．1，respectively)．Theflaskwas
filledwithwatertothe25mLmarkandthoroughly
mixed．Finally．thecolorimetriccylindersw eallkeptin
a waterbathat32℃for40min．Theblanksolution
(cyanide-freewat r)waspreparedinthesameway．The
absorptionof lightat 638nmwasmeasured
(Photometer-721． ShanghaiAnalyticalInstrument
Company)ina cellofopticalp thlengthof10mm．
Standardsolutionsofpotassiumcyanidewereprepared
intherangeof0．007—0．181CNmg·L-1andusedforthe
calibration．Thedetectionl mitsofthismethodwas
determinedtob between0．004and0．25CNmg·L一．
dependingonthevolumeofthesample．
2 Resultsandiscussion
2．1 Toxicitytestsofcyanidetoplants
Toxicitytestsofcyanidetocoin(Zeam ysL．)and
soybeans(Glycinemax(L．)Merr．)wereinvestigated．
Tablesland2givethedetailsofaveragegrowthofcorn
andsoybeanplantsexposedtocyanidenthewaterfor
maintainingwatermoisturetoareasonablelevel．
Afterthe10daysexposureperiod，neitherde ectable
effects(>10％reductioningrowthrate)，norobserved
effects(chorosisofleaves)couldbefoundinbeththe
controlsandthexposedgroupsinthelowcyanidedose
(95．5CNmg·L‘)fors ybcanandcorn．Plantsll
100kedh althyandhadnoindicationofbeingpoisoned．
Whenexposedtohighdoseofcyanide仑91．0 CN
mg·L-1)，cornandsoybeanplanthadseveredetectable
toxicsymptoms，eg．，r ductionofgrowthrates，butno
obs rvedeffects(chorosisofleaves)wasfoundduring
thentiretestperiod．
2．2Removaltestsofcyanidewithdetachedleaves
Theresultsofthecyanideremovaltestwithleaves
areshowninTable3．Inthecontrolwithplantleaves
(centrel1)，nocyanidewasdetectedoverthentiretest
period．Noobviouschangeofthecyanideconcentration
inaqueoussolutionnthesecondcontrolwithoutplant
materials(centrel2、wasobservedovera28．hourperiod
ofexposure．Theseresultsindicatethattheremovalof
cyanidefromaqueoussolutionnalltestinggroupscan
beaccountedtotheuptakeandmetabolismbyleaves．
Cyanideinsealedvesselswithleavesofsoybcan
(Glycinemax(L．)Merr．)wasreducedfrom1．01l to
0．004CNmg·L-1duringthe28．hourperiod．while
leavesofCOITI(ZeamaysL．)degradedcyanidefrom
1．021to0．097CNmg·L11．Theamountofcyanidein
sealedvesselswasignificantlyreducedduringthe
presenceofplantleavesinthetestflasks．withthe
removalr tesof99．63％and91．41％ofthetotaldose
forsoybeanandcornovera28-hourperiod，respectively．
Itmaybequestionedwh therd tachedleavescan
keepuptheirnormalfunctioninginasealedvessel．Plant
leavesusedinthistestalldidbestundertheseconditions
andseemedhealthyforthentiretestperiod．On-going
gasexchangecouldbeobservedintheformofbubbles．
whensunlightfellontheflasks．Thismethodwasalso
successfullyusedin ourpreviousexperimentof
persistence．．IofMI BEagainstmetabolismbyDanish
vegetation。”．
2．3 Determinationof hemaximumcyanideremoval
capacity
Ifanenzymeconvertsonechemical(thesubstrate)
intoanother(theproducO，thevelocityof henzymatic
reactioncanbedescribedbyMichaelis．Mentenkin tics．
v=(vⅢ。宰回／(KM+回
Table3Measuredcyanideconcentrationin queoussolutionwithplantleaves．Valuesarem ansofCNconcentrationfromthI屯e
replicates(inparenthesis：standarddeviation)．
=。——_·——————霎翌L翌曼二星：}_——≤S盥』篁∑』三∑———磐塑翌曼：生121竺丛里墨：．垦：12 19堕!坚：兰：12 1竺堕婴曼：生!!
；sontrol1 u u 0 0 0 0
Centrel2 1．021 1．019 1．022 1．018 1．019 1．020
(0．0048) (0．0435) (0．0392) (0．0048) (0．0102) (0．0051)
Glycinemax(L．)1．011 0．804 0．471 0．20l 0．022 0．004
Merr． (0．0050) (o．0097) (0．0097) (0．0098) (0．0191) f0．0039)
ZeamaysL． 1．021 0．736 0．584 0．464 0．126 0．097
一一 !Q2 鲤：垡丝Z2 IQ：丝!i! lQ：Q三2兰2 (Q：Q丝墨! (Q：Q12Q2
万方数据
生态科学 23卷
Wherevisthedisappearancevelo ity(masspertime)
ofthesubstrateofconcentrationS，PmaxiSthemaximum
disappearancevelocity,andKuisthehalf-saturation
constant(thesubstrateconcentrationwherethe
disappearancevelocityishalfthemaximum)．Figl gives
thedisappearancekinetics．Itisclearlyshownthatthe
disappearancecurveshowsl“order,whichmeansthe
substrateemovalCHIVeiSexponential．Inthistest．the
velocityofenzymereaction，themaximumcyanide
removalc pacity(Vmn)，istheslopeofthelinerphase，
expressedasamountsofcyanidemassremovalbyplant
materialspertime(ragCN·kgfreshweight。·hourl)．
Theremovale ocityofc anidewasdeterminedtob
4．43mgCN·kgfreshweightl·hour"‘forsoybeanand
corn， respectively．
1-2
l
乙0．8
己0．6
童0．4
§0．2
O
—O．2
1’i_e(h)
×Soybean·Corn
Fig1 Exponentialdis ppearancekinetics(firstorder)with
twoexponentialtrendlines．
2．4Comparisontoothert eatmentthods
Thehydrogenperoxideoxidation，INCOprocess，
bioremediationthroughmicroorganismsdalkaline
chlorinationrethecommonusedmethodstotreat
cyanide．contaminatedsites．Thefirstmethodyields
cyanate(OCN。)ornitrate(NO一3)，bothlesstoxican
cyanide．butneverthelessunde iredcompoundsi a
watershedIlUl．The烈COprocessdoesnott他at
thiocyannatea dferrocyanides，andcyanate(OCN)is
formed⋯I．Despitethev d ncethatbioremediationof
cyanidespossible．howeverthebact ialbreakdownof
cyanideproducesammonium(Nil：)thatisasomewhat
toxiccompoundtofish，andmayinduceeutrophication
oflakesandponds¨“．Thealkalinechlorinationof
cyanidelcadstotheproductionofcyanate(oCN)，while
chlorinatedproductsandfreechlorinemaybeemitted■
Theplantationsorfieldsforthetreatmentofmining
wastemaybedesignedinawaythatallwastewateris
consumedandtranspiredbytheplants．Thisalso
means’’zeroemissions’’(exceptwiththeharvested
product)．
Resultsfromtoxicitytestsindicatedthatlowdoses
ofcyanide(95．5CNmg·L-1)didnotcausetoxicityto
cornorsoybean．Theywerabletosurvivalwithout
observedandetectableeffects．Significantredu tioni
theplantgrowthratewasfoundatthehighcyanide
treatmentgroup(>91．0CNmgL-‘)，butnoobserved
effectwaseen．Additionally,resultsfromcyanide
removaltestsindicatedthatplantmaterialswerefoundto
beabletoefficientlyremove(andth tmeansdegrade
and／ormetabolize)cyanidefromtheaqu ousmedium
throughtheirntemalenzymesystems．Thissuggeststhat
cyanideeliminationbyplantsmaybeafeasibleoption
forcleaningupsoilscontaminatedwithcyanideorgold
andsilverminewasteandwastewater．
AcknowledgementsThankstotheInstituteofCropGermplasm
Resources，theChin seAcademyofAgriculturalScienceforthe
seedofcorn。JiangD osongforspeciesd terminationandSheree
PoindextenMcClemanforassistingw ththelanguageproblems．
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铷
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万方数据
氰化物污染的植物修复可行性研究
作者： 于晓章， Trapp Stefan
作者单位： 于晓章(湖南农业大学环境科学系,长沙,410128)， Trapp Stefan(丹麦技术大学,环境与资
源研究中心,DK-2800 Kgs. Lyngby,Denmark)
刊名： 生态科学
英文刊名： ECOLOGIC SCIENCE
年，卷(期)： 2004,23(2)
被引用次数： 2次
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