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MegParmelee
Mrs.Norris
APEnvironmentalScience
6February2015
Collaborators:AlphonsusBrown,DevlinSantos,CharlesBaker
LD50Lab
I:Introduction
InthefrigidwintersoneoftendoesnotseeinthemoderateclimateofNorthCarolina,wateronthe
surfaceoftheroadscanfreezeover.Thisisobviouslyahazardforanycarsthatdriveontheseslippery
streets.Incasessuchatthis,saltisappliedtotheroadsinordertopreventtheasphaltfromicingover.
Thisisbecausesaltlowersboththemeltingandfreezingpointofice(WhyDoTheyUseSaltToMelt
Ice?).Whilethisincreasesthesafetyofpeoplewhochoosetodriveindangerousweather,theuseof
saltonhighwayscouldhavepotentiallyadverseeffectsonthevegetationthatgrowsneartheseroads.In
theLD50(doseresponse)lab,differentconcentrationsofsaltwereintroducedtoradishseedsinorder
todetermineiftheirgrowthwasaffected.LD50isthemedianlethaldosethathastheabilitytokillhalf
oftheorganismsinatestpopulation,measuredinmgofpesticideperkgofbodyweightaccordingto
theEnvironmentalProtectionAgency.TheLD50ofasubstancecanbegraphedinwhatsknownasa
doseresponsecurve,whichisaplotofdatathatshowstheeffectsdifferentdosesofatoxicagentona
groupoftestorganisms.Inordertodeterminehowdifferentconcentrationsofsaltaffectedtheseeds
growth,serialdilutionswereperformedtocreate5uniqueconcentrationsofsaltinwater.Aserial
dilutionisdefinedasanydilutionwheretheconcentrationdecreasesbythesamequantityineach
successivestep(Kotrla,Tony).

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II:Hypothesis
Ifradishseedsareaddedtowaterwithdifferentconcentrationsofsalt,theseedsinthelowest
concentrationofsaltwillbemorelikelytogerminateandexperiencethehighestamountofgrowthin
radiclelength.Thesaltconcentrationof24mgNaCl/LH
OwillbetheLD50ofradishseeds.
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III:Variables
Inthisexperiment,theindependentvariablewastheconcentrationofsaltinthewater.The
dependentvariableswerethenumberofseedsthatgerminatedandthelengthoftheradicleafter45
days.Thecontrolwastheseedsinthepetridishwiththedistilledwaterandtheexperimentalgroupwas
the50seedsthatwereinthedisheswiththevaryingsaltconcentrations.
IV:Materials&Methods
50mldistilledwater
20mlofwaterwith24gNaCl/LH
O
2
60radishseeds(inthiscaseFrenchBreakfastwasused)
6petridishes
1papertowelroll
1. Using20mloftheoriginalconcentrationofsalt,performaserialdilutiontocreatefive
differentsaltconcentrations,plusasixthwithplainwater
2. Countout60seeds,10foreachwatersample
3. Put10seedsintoafoldedpapertowel,spacetheseedsoutevenlyonthepapertowelsothat
theydonottoucheachother

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4. Wetthepapertowel/seedsandsealinapetridish,besuretolabelthedisheswiththesalt
concentrationforthatsample
5. After45days,examinetheseeds,countingthenumberofeachthatgerminated
6. Forthegerminatedseeds,measurethelengthoftheradicle
V:Data&DataAnalysis
Afterthelabwascompleted,thepercentofseedsgerminatedandthelengthoftheradicleswas
collectedandorganizedintoachart.
Concentration
sofsalt
solution
(mg/L)

%of
seeds
germinate
d

Replicatesradiclelength
(mm)

12345678910

T=mean
radiclelength

Differencein
radiclelength:
CT

C=control

100%

60,25,50,40,25,75,15,30,
55,30

40.5

1.5

60%

15,30,25,10,

10.5

30

10%

1,

0.1

40.4

0%

40.5

12

0%

40.5

24

0%

40.5

Theseresultswerethenput
intotwographs.Inthe
EffectsofSaltonRadish
Seedsgraph,itisapparent
thatthehigherthe
concentrationofsalt,the

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shortertheradiclelength.Withcompletelydistilledwater,themeanradiclelengthwas40.5mm.When
thesaltconcentrationincreasedbyjust1.5mg/L,theaverageradiclelengthdecreasedby75%,and
whenthisconcentration
wasdoubledthelength
droppedtoatenthofa
millimeter.Ata
concentrationof6
mg/L,noneoftheseeds
grewaradicleandthis
trendcontinueduntil24
mg/L.Additionally,the
percentofseedsthatgerminatedwasgraphed.Theshapeofthegraphisnearlyidenticaltotheone
abovethereisnodatapast6mg/L.Thecontrolhadallofitsseedsgerminatewhiletheconcentrationof
1.5mg/Lhad60%ofitsgerminateandthe3mg/Lmerely10%.
VI:Conclusion
Afterananalysisoftheeffectsofsaltonseedgerminationandradiclelength,itisapparentthatthe
originalhypothesisisonlypartiallycorrect.Whileitwastruethattheseedsinthelowestconcentration
ofsaltexperiencedthemostgrowth,theLD50ofradishseedsismuchlowerthanthepredicted24mg
NaCl/LH
O.At6mg/L,noneoftheseedsgerminated,whichmeansthattheLD50hadtooccurprior
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tothispoint.Whenlookingatthedatacollectedinthistrial,itwouldappearthattheLD50ofradish
seedsisaround1.5mg/L.Thisisbecauseonlyalittleoverhalfoftheseedsinthedishwiththis
concentrationofsaltgerminatedandtheLD50ofanorganismisthedoserequiredtokill50percentof

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atestpopulation(LD50Values).However,theclassaverageforthatconcentrationwas80%,which
leavesonetobelievethat3mg/L,whichhadanaverageof43.75%,isthetrueLD50ofradishseeds.In
thedatashownabove,therewasonlya10%chanceoftheseedsshowinganysignsofgrowthatthis
point.Apotentialreasonforthisdiscrepancywouldbethefactsthatthegroupsuseddifferenttypeof
radishseeds.TheseedsportrayedinthisreportwereFrenchBreakfastradishestheycouldpossiblybe
moresensitivetosaltandhavealowerLD50thanotherkindsofradishes.Nonetheless,theeffectsthat
salthadonradishgrowthwereverysignificant.Thecontrolfeaturedradisheswhichgrewanaverageof
over40mmandassoonassaltwasaddedtothewater,theaveragelengthoftheradicledroppedto10.5
mm.Fromthis,theconclusioncanbedrawnthatsalthasadverseeffectsonthegrowthofradishseeds,
stuntingitandthenpreventingitalltogether.Thisobviouslyhassomedangerousimplicationsforthe
environment,andshowevidenceastowhysalinizationissodetrimentaltoplantgrowth.Thisis
somethingthatmustbebroughttothepublic.Approximatelymorethan22milliontonsofsaltare
scatteredontheroadsoftheU.S.annually(Stromberg,Joseph).Whenitrains,wheredoesallthissalt
go?Therunofffromhighwaysenterstheecosystemsthatsurroundit,slowingplantgrowthandeven
preventingit.However,duringthewintermonthssaltisessentialtomaintainsafetyontheroadsand
saltusecannotbecompletelystopped,thoughitcanandshouldbelimited.

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WorksCited
Kotrla,Tony."SerialDilutions."
SerialDilutions
.AustinCommunityCollege,n.d.Web.01Feb.2015.
"LethalDosage(LD50)Values."
EPA.gov
.EnvironmentalProtectionAgency,27June2012.Web.01
Feb.2015.
Stromberg,Joseph."WhatHappenstoAlltheSaltWeDumpOntheRoads?"
Smithsonian.com
.
SmithsonianInstitution,6Jan.2014.Web.06Feb.2015.
"WhyDoTheyUseSalttoMeltIceontheRoadintheWinter?"
HowStuffWorks
.HowStuffWorks.com,
n.d.Web.01Feb.2015.