LaboratoryManual

BIOL160

Title:___________________________________________________________________
________________________________________________________________________
AuthorsName:___________________________________________________
BiologyDepartment,HendrixCollege,Conway,AR72032

Theintegrationandcoordinationofprocessesinvolvedinthegrowthanddevelopmentof
awholeplantareregulatedbyavarietyofsubstances.Fivemajorkindsofplanthormonesare
generallyrecognized:auxins,gibberellins,cytokinins,ethylene,andgrowthinhibitors(abscisic
acidandperhapsothers).Thetermauxin,gibberellin,orcytokininisagenericname
referringtoagroupofsubstancesthatcausethesamephysiologicalresponsesandaresimilarin
chemicalstructure.Forexample,morethaneightydifferentsubstancesareknownas
gibberellins,basedonsimilaritiesintheirchemicalstructures.Manyofthesegibberellinsare
biologicallyactive,however,somegibberellinsappeartohavenobiologicalactivity(Ecklund,
1999).
Dwarfismisoccasionallyencounteredinplantspecies.Ifthesmallsizeoftheplantsis
duetoageneticmutation,itcanbeperpetuatedinastrainorvarietyofaspeciesbyselective
breeding,anditusuallyistheresultofreducedelongationofinternodes.Inmanydwarfplant
varieties,butnotall,theapplicationofagibberellincausestheplantstodevelopasnormaltall
plants,suggestingthatdwarfismisduetoablockingibberellinproductioninthedwarves
(Ecklund,1999).
Apicaldominanceisaphenomenoninwhichthetipoftheshootofanherbaceous
angiospermisabletosuppressthegrowthofaxillarybuds.Thissuppressionmaintainsasingle
stemwithlittleornobranching.Ithasbeenshownthatanauxinproducedbytheapicalbud
movesdownthestemandinhibitsthegrowthoftheaxillary(lateral)buds.
Thegoalsofthisexperimentaremany.Wewilltestthehypothesisthattheapplicationof
gibberellinstodwarfpeaplants(PisumsativumL.,var.littlemarvel)willcausethemtodevelop
asnormaltallplants.Wewillalsoinvestigatetheresponseofthepeaplantstoincreasing
concentrationsofgibberellins.Wehypothesizethat___________________________________
_____________________________________________________________________________.
Finally,wewilltesttheresponseofthedwarfpeaplantstoauxinandobservethephenomenon
ofapicaldominance.
MATERIALSANDMETHODS
Gibberellinexperiment
Totesttheeffectsofgibberellinsonthegrowthofdwarfpeaplants(PisumsativumL.,
var.littlemarvel),Iselectedonecontainer(14cmindiameter)of___________________

LaboratoryManual

BIOL160

(number)sixweekoldplants.Ialsoobtainedadropperbottlecontainingoneofthefollowing
concentrationsofgibberellin:0.001,0.01,0.1,0(control),1.0,10,or100gGA3perdropor
oneoftheunknowns:extract,extractdiluted1/10,orextractdiluted1/100.Ithenlabeledthe
containerofplantswithmyname,mylabday,andmychosentreatment.
Beforetreatingtheplantswithmygibberellinsolution,Ilabeledalltheplantsinthe
containerusingflagsofmaskingtapeattachedtotoothpicks.Eachplantwasgivenanumber
suchthatitcouldbeindividuallyidentified,anditsassociatedflagwaspushedintothedirtnext
totheplant.Then,thetotalnumbersofvisiblenodesoneachplantaswellaseachoftheir
heights(initialstemlength)wererecorded.Theheightwasmeasuredbetweenthetopofthe
cotyledonarynodeandthetopofthehighestvisiblenode.
Ithencarefullyseparatedtheappressedstipulesofthehighestvisiblenodeofeachplant
andappliedonedropofmygibberellinsolution.Imadesurethatthedropwasheldbetweenthe
twoimmatureleaves,butifitfelloff,Iappliedanotherdrop.Theplantswereplacedona
mobilegrowthcartandcheckeddailyforwaterneeds.
Thefollowingweek,Iobtainedmycontainerofplantsandagainrecordedthenumberof
visiblenodesandtheheight(finalstemlength)ofeachplant.Ithencalculatedthepercent
changeinstemlengthforalloftheplantsbysubtractingtheinitialstemlengthfromthefinal
stemlength,dividingthisnumberbytheinitialstemlength,andmultiplyingby100%[(final
stemlengthinitialstemlength)/initialstemlengthx100%].Ameanandstandarddeviationof
percentchangeinstemlengthwascalculated.Ialsocalculatedthenumberofnodesaddedfor
eachplantandameanandstandarddeviationforthesedata.
Otherinvestigatorsrepeatedtheaboveproceduresusingdifferentgibberellin
concentrations(0.001,0.01,0.1,0,1.0,10.0,and100.0gGA3perdroporextract,extract1/10,
andextract1/100).Igatheredthemeanpercent(+/1S.D.)changesinheightandthemean
number(+/1S.D.)ofnodesaddedfromtheseinvestigatorsandcombinedthemwithmyown.I
thengraphedthepercentchangeinheightvs.gibberellin(GA3)concentrationonsemiloggraph
paper,andusingthegraph,IestimatedtheconcentrationsofGA3ineachoftheextracts.
Auxinexperiment
Toobservethephenomenonofapicaldominance,Iobtainedtwocontainers(14cmin
diameter),eachwith___________________(number)ofsixweekolddwarfpeaplants(Pisum
sativumL.,var.littlemarvel).Ilabeledeachcontainerofplantswithmyname,mylabday,and
eitherauxinorcontrol,andlabeledeachplantineachpotwithanumber.Todoso,Iplaceda
maskingtapetoothpickflagnexttoeachplant.
Icuta1.5x1.5cmsquareofaluminumfoilforeachplantinbothtubs,exceptfortwo
andcreatedcapsfortheplantsbycenteringeachpieceofaluminumfoiloverthetopofa
woodendowelandfoldingdowntheedges.Leavingoneplantineachcontainerintact,Icutthe
topsofftheotherplants,justunderthefourthnodeabovethecotyledons.Usingthebroadendof
atoothpick,Iplacedagloboflanolinlargeenoughtocoverthecutsurfaceoneachofthecut
plantsinthecontrolcontainer.Similarly,Iplacedaglobofauxin(IAA)inlanolinoneachof
thecutplantsintheauxincontainer.
Eachcutplantinbothcontainerswasthencappedwithapieceofaluminumfoil.Lastly,
thenumberofaxillarybudsthatweregreaterthanthreemillimetersinlengthoneachplantwas
recorded.Theplantswerethenplacedonagrowthcartandcheckeddailyforwaterneeds.

LaboratoryManual

BIOL160

Thefollowingweek,Iobtainedthetwocontainersofplantsfortheauxinexperiment.I
removedthealuminumfoilcapsandrecordedthenumberofaxillarybudsgreaterthanthree
millimetersinlengthoneachplant.Icalculatedthetotalnumberofaxillarybudsgreaterthan
threemillimetersinlengthforthecontrolandtheauxintubsandcomparedtheseresultstothose
ofotherinvestigators.
RESULTS
Thissectionistobecompletedandsubmittedatthenextlabmeeting.
DISCUSSION
Thissectionistobecompletedandsubmittedatthenextlabmeeting.
LITERATURECITED
ECKLUND,PR.1999.Chapter17Angiospermgrowthanddevelopment.InJ.C.Glaseand
P.R.Ecklund[eds.],Investigativebiology:alaboratorytext,311334.CornellBiology
Department,Ithaca,NewYork,USA.