BiologyIB DesignLaboratory

Studentsfullname 02/03/10

InvestigatingtherelationshipbetweensoilsalinityandgrowthintwoPhilippinerice(Oryza sativaL.)varieties:DinoradoandIR841.

ResearchQuestion:Whatistherelationshipbetweenthesalinityofthesoilandthechangeinthedry massofrice( OryzasativaL.)seedlingsinthetwodifferentvarieties,DinoradoandIR841,whengrown tomaturegrainstage? Backgroundinformation:Soilsalinityhasasignificanteffectonriceyieldsbyseveralmechanismsand consequentlyposesamajorobstacletoriceproductionworldwide1.Althoughothercropshave exhibitedinhibitionthatisspecifictotheproportionofsoilions,osmoticpressurehasbeenshowntobe themostsignificantfactorregardingsalinitytoaffectthedrymassofriceatthematuregrainstage2. Althoughriceexhibitsanionspecificinhibitionbychlorineatthefloweringstage2,thematuregrain stageultimatelydeterminesyields,andthereforewillbethestageofgrowthinvestigated.Allyield relatedcharacteristics,suchasfertility,tilllength,paniclenumberandpaniclelengthareaffectedbysoil salinity3.Thisstudywillinvestigatetheeffectofsalinityonthefinalyieldofoneplantcyclebymeasuring thedrymassofthematuregrains. Otherfactorsaffectingphotosyntheticrateandplantgrowthmustalsobeconsidered.Theintensityand colorcompositionofthelightreceivedbythegrowingrice,forinstance.Differentplantvarietieshave differentactionspectraandthereforewillperformbetteratdifferentwavelengths4.External temperatureandroottemperaturealsoaffectricegrowth.Externalairtemperaturecouldaffectthe rateofenzymaticreactionsintheplant,suchasthefixationofCO2byRuBPcarboxylaseintheCalvin cycle5,whereasroottemperatureaffectsricegrowthbyinfluencingrootporosityandthereforenutrient intake5.Soilcomposition,includingtheavailabilityandproportionNPKcompounds,humusdeposits,the microbiologicalcompositionofthesoil6,moisturetensions,oxygen7andcarbondioxide5levelsallaffect ricegrowthbyaffectingphysiologyandlevelsoflimitingfactorsinkeymetabolicreactions. Hypotheses:AsthesalinityofthesolutionsusedtoirrigateOryzasativaL.increases,theplantsfrom bothvarietieswilldecreasebecauseoftheeffectofthesolutionsdecreasedosmoticpressure.A decreasedosmoticpressurelimitofsolutionssurroundingplantrootssuppressestheplants transpirationstreamandthereforelimitsitsabilitytotransportmineralsandNPKcompoundsfromthe rootstotherestoftheplant.Whenirrigationsolutionsbecomehypotonic,thedrymassoftheplants maynolongerbeabletogrowpastacertainstageofgerminationbecausethetranspirationstreamwill besosignificantlyinterrupted.
1

Zheng,Linghe,andMichaelC.Shannon."SalinityEffectsonSeedlingGrowthandYieldComponentsofRice."Crop Science40(2000):9961003. CropScienceJournal.org.CropScienceSocietyofAmerica.Web.Feb.2010. <http://crop.scijournals.org/cgi/content/full/40/4/996>. 2 Ehrler,William."SomeEffectsofSalinityonRice."BotanicalGazzette122.2(1960):10204.JSTOR.Web.Feb.2010. <http://www.jstor.org/pss/2473354>. 3 Shereen,Aisha,S.Mumataz,S.Raza,M.A.Khan,andS.Solangi."SalinityEffectonSeedlingGrowthandYield ComponentsofDifferentInbredRiceLines."PakistaniBotanicalJournal37.1(2005):13139.Print. 4 Allott,Andrew.IBDiplomaProgrammeBiologyCourseCompanion.NewYork:OxfordUP,USA,2007.Print. 5 Setter,T.L.,andH.Greenway."GrowthReductionsofRiceatLowRootTemperature:DecreasesinNutrientUptakeand DevelopmentofChlorosis."OxfordJournalofExperimentalBotany39(1998):81129.OxfordUniversityPress.Web.Feb. 2010.<http://jxb.oxfordjournals.org/cgi/content/abstract/39/6/811?ck=nck>. 6 Studiesonnutrientuptakeofriceandcharacteristicsofsoilmicroorganismsinalongtermfertilizationexperimentsfor irrigatedrice."JournalofZhejiangUniversityScience6.2(2005):14754.PubMed.Web. <http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1389631/>. 7 Patrick,W.,andW.Fontenot."GrowthandMineralCompositionofRiceatVariousSoilMoistureTensionsandOxygen Levels."AgronomyJournal68(1976):32529.Web.Feb.2010.<http://agron.scijournals.org/cgi/content/ abstract/68/2/325>.

 IndependentVariable:Concentrationofsalinesolution( addedtosoilsampleeverytwo days.Fivelevelsoftreatmentwillbeused:0.00M,0.25M,0.50M,0.75Mand1.00M. DependentVariable:Thedrymassoftheeachseedlingperleveloftreatment( aftergrowing for12weeksirrigatedinsalinesolution. Table 1: Controlled variables and methods for controlling and monitoring them. Variable Methodtocontrol Methodtomonitor
LightIntensity andColor Composition Temperature ofrootsand externalair: TrayswillbeplacedinthesameareasoftheHighSchool courtyardandwillthereforereceivethesamefluctuationsof lightcompositionandintensity. Alltrayswillbekeptatgroundlevelinthesame2X2mspace. Therefore,variationsinexternalairtemperatureshouldbe negligiblebetweentrays.Consequently,roottemperatures shouldbenegligiblydifferentasthesoilisthesamevolumeand depthineachtrayandeachtrayismadeofthesamematerial andthereforewillhavethesameinsulatingproperties. Onedataloggingprobelightintensity sensortobeplacedbesideeachtray.

Soil composition:

Soilhumidity:

Seedling health:

Seedling distributions:

Onedataloggingtemperatureprobein eachtray;astheexternalairtemperature andtheroottemperatureareclosely linkedwhenthericeisplantedineach traynotemperatureprobeswillbeused tomeasureexternalairtemperatureover eachtrayduetotheavailabilityofthe equipmentatschool. Inordertocontrolforsoilcompositionandmoisture,32kgof TheconcentrationofNPKnutrientswill soilwillbeorderedfromthesamesupplier.Thesoilwillthenbe beevaluatedusingtheLaMotteNPKSoil o heated250 Cinanovenfor10minutesinbatchesof3.2kgin TestKit . ordertosterilizethesoil,neutralizingtheeffectofvaried densitiesofmicrobiologicalfaunaandmoisturecontent.Dry, powderedsoilisalsomucheasiertotumbleandfacilitatesthe mixingofsoilparticlestoimprovethehomogeneousnessofthe distributionofnitrogen,phosphorusandpotassium(NPK) compounds.Thesoilwillberemovedandanyclumpscrushed sothatitcanbemoreeffectivelyhomogenisedwhenturnedin atumblerfor5minutes. Thetraywillbefloodedto1cmabovethesoillevel. Everydaythewaterlevelwillbe Apilotstudywillbeconductedbyplacingatrayofthesame measuredat6.00pm(closetosunset) dimensionswith1cmofwateroutintheHighSchoolcourtyard witharuler,andthetraywillbeflooded from5.00am6.00pmandmeasuringthechangeinwater to1cmabovethesoillevel. level.Thereforeestimationsofthevolumeofdistilledwater andmassofsaltnecessaryfortheexperimentwilldependof theresultsofthisstudy. Inordertocontrolforgeneticvariationamongseedlings,or otherfactorspredisposingseedlingstounusuallyloworhigh levelsofgrowth,seedswillbegrownfor5days.Soilwillbe preparedasdescribedintheproceduretocontrolsoil compositionandirrigated,accordingtotheprocedureto controlsoilhumidity,withtapwaterfromasinglesource.Five seedlingsthatfallwithin 1cmofthemodalheightofall seedlingsgrownwillbeselectedforeachleveloftreatment. Anyseedlingswithunusuallylightordarkleafcolorationwillbe excludedfromthisselection. Seedswillbeevenlyspacedasillustratedinfigure1and seedlingsspacedasillustratedinfigure2inordertoensure maximalandevenlydistributedspacefortherootgrowthof eachseedlingandplant.

 Diagrams: Figure1:Arrangementofseedsinseedbed.Figure2:Arrangementofselectedseedlings 20 cm 20 cm 9.42 cm 9.42 cm Rice seed 9.42 cm 20 cm 20 cm 9.42 cm 9.42 cm 6.67 cm Figure3:BirdseyeviewoftrayarrangementincentreofHScourtyard. Key: =30cm =OryzasativaL.Dinorado =OryzasativaL.IR841 0.00M 0.25M 0.50M 0.75M 1.00M 0.25M =Concentrationofsalinesolutionused. =Opaque1X1mtablecoveringlaptop. 0.00M 0.25M 0.50M 0.75M 1.00M nsh =Temperatureprobeconnectedtolaptop viaHUB. Onemortarandpestle. andappropriateHUBhardwareto Materials: One5kilogramsoiltumblermodify connect10probestoonecomputer. 32kilogramsofsoil. araffletickettumblerifnecessary. 1.8kilogramsofNaCl. Twospades. 26litresofwater(incontainer) Onesoilsack. 45Dinoradoand45IR841seeds. Onedigitalweighingscale. 3 Apparatus: Ten100cm beakers. Ten20X20X8cmtrays. Onestirringrod. Five20X20X8cmsteeloventrays. Onespatula. OneLaMotteNPKSoilTestKit . Twoopaque1X1mtable. Oneovenwithgastankand Twolaptopswithdatalogging thermostat. software. 20dataloggingtemperatureprobes Procedures: 1) Measuretheweightofthesoilandsteeltraybeforebaking. 2) Ovenbakethe32kgofsoilin10batchesof3.2kgat250oCfor10minutestosterilizesoiland standardisemoisturecontentbyevaporatingwaterinsoil. 3) Measurethepercentageweightlostbytheevaporationofwater.Calculatetheresultingmassof drysoilthatshouldbeapportionedtoeachtray. 4) Aseachbatchisremovedcrushsoilclumpswithpestle(andmortarifnecessary)tohelpsoil reachapowderyconsistency(seetable1forreasons). 5) Tumblethesoilcontinuouslyfor5minutesin5kilogrambatches.Aftereachbatchleave approximatelyhalfofthesoilinthetumblerbeforeaddingasmuchnewlydriedsoiltothe tumblerasmixedsoiltakenoutthisistohelpensurethatthesoilishomogenizedacrossthe batches,notsimplywithineachbatch. 3

6) Assoilismixedandtakenoutofthetumbleraddthemassofsoilcalculatedinstep3toeach tray. 7) Take0.1gsamples,composedofsmallersamplesfromdifferent5placesonthetopsoil(eachof whichisatleast2cmapart),fromeachtrayandusetheLaMotteNPKSoilTestKitto qualitativelyevaluatetheconcentrationofNPKcompounds 8) Usearulertoensurethatthesoilisevenlydistributedacrossthebottomofthecontainer. 9) Afterthesoilisdistributedtoeachcontainer,evenoutthesoilsuchthattheheightofthesoilin eachcontainerfallswithin cm(acrossthesurfaceofthecontainer)ofthemeanheightof thesoilineachcontainer. 10) Distributetheseedlingsasindicatedinfigure1,placingeachseed0.5cmbelowthesurface, coveringitbackupwiththedisplacedsoil. 11) Afterstephasbeencarriedoutforeachtray,slowlyfloodeachtrayuntilthewaterlevellies1 cmabovethesoilline,attemptingnottoexcessivelydisturbthetopsoil. 12) Afterstep10hasbeencarriedoutforeachtray,assemblethetraysandconnectthedata loggingapparatusasindicatedinfigure3,awayfromtheshadeofthetrees. 13) Waterthetrayseveryday,for5days,asindicatedintheproceduretocontrolsoilhumidity (table1). 14) Duringthesefivedays,prepare1000cm3ofeachsalinesolution(0.00M,0.25M,0.50M,0.75M, and1.00M). 15) After5days,selectseedlingsbytheprocedureoutlinedtocontrolseedlinghealth(table1). 16) Replantthe5selectedseedlingsfromeachleveloftreatmentasindicatedbyfigure2. 17) Waterthetrays,withthesalinesolutionspreparedinstep14,asindicatedintheprocedureto controlsoilhumidity(table1).Prepareapproximately800cm3ofeachsalinesolutioneach succeedingweekaccordingtotheamountnecessarytowatereachtraybytheprotocolto controlsoilhumidity(table1). 18) Attheendof12weeks,removeeachofthe5riceplantsfromeachlevelofanalysisandbake themevenlyspacedinasteelcupcakerack,oneplantpercupcakedivision,at150oCfor20 minutes. 19) Tarea100cm3beakeronthedigitalweighingscaleandcarefullytransferthedriedmassofthe onericeplacethebeaker.Recordthedatainthetablesbelowandrepeatthisstepforthe remainingriceplants.

Table 2: The relationship between the concentration of saline solution added to soil ( 0.05M) and the dry mass of the Oryza sativa L. Dinorado plant after it is grown for 12 weeks.
Concentrationofsalinesolution usedtofloodcrop(+/0.05M) Plant number 1 2 3 4 5 1 2 3 4 5 5 2 3 4 5 1 2 3 4 5 1 2 3 4 5 DrymassofOryzasativaL.IR841 plant(+/0.01g) AveragedrymassofOryzasativaL. IR841plant(+/0.01g) Standard Deviation

0.00

0.25

0.50

0.75

1.00

Table 3: The relationship between the concentration of saline solution added to soil ( 0.05M) and the dry mass of the Oryza sativa L. IR-841 plant after it is grown for 12 weeks.
Concentrationofsalinesolution usedtofloodcrop(+/0.05M) Plant number 1 2 3 4 5 1 2 3 4 5 5 2 3 4 5 1 2 3 4 5 1 2 3 4 5 DrymassofOryzasativaL.IR841plant (+/0.01g) AveragedrymassofOryza Standard sativaL.IR841plant(+/0.01g) Deviation

0.00

0.25

0.50

0.75

1.00