EFFECT OF GROWTH SUBSTANCES ON THE

ABSCISS LAYER IN LEAVES OF COLEUS'

R. MAURICE MYERS
(WITH NINE FIGURES)

Introduction
LAIBACH (i i) reportedthat orchidpolliniainsertedin the splitend ofpetiolesof
debladedleaves ofcoleus and certainotherplants delayed abscissionofthepetioles
II-20 days longerthan that of untreatedpetioles. LA RuE (I2) found that in-
doleaceticacid (heteroauxin),pollen,and urinedelayed petiole fall fora fewdays
whentheywereapplied to debladed petiolesof coleus leaves. A studyof the effect
ofgrowthsubstancesin relationto otherfactorsknownto affectabscissionseemed
desirable.
The earlierliteratureon abscission is extensiveand will not be discussed here.
It is reviewedby KENDALL (9), LLOYD (3, I4, I5), and SAMPSON (i8). It is gen-
erallyagreed that abscissionis usually precededby the developmentof an absciss
layerfollowedby the disintegrationof certainlamellae in the walls of cells in this
layer. SAMPSON found that an absciss layer started to formin the thirdpair of
leaves below the terminalbud of coleus plants bearing eightpairs of leaves, and
that the layers were not mature until these leaves were the sixthpair below the
terminalbud. He foundthat at the timeof abscissionthe lamellae of the primary
wall adjacent to themiddlelamellae disintegratedbeforethe middlelamellae. The
cellulosepropertiesof these lamellae disappeared and a test forpectic compounds
became more intense. These facts led him to describeabscission as a change of
cellulose to pectose and later to pectic acid which resultedin an excess of pectic
acid over the amountof calcium necessaryto maintainthe solidityof the middle
lamellae of the cell wall in the absciss layer. His inferencesas to the natureof the
changes in cellulose,however,were made in the absence of adequate chemical
data. Even today the changesthat may occur in these compoundsin cells under
the action of enzymesare not well enough known to admit a more detailed de-
scription.
Material and methods
Several thousand plants of Coleus blumeiBenth (var. Golden Bedder) were
propagated fromcuttingsin a greenhouseduring the winterand spring. Since
coleus has oppositeleaves, certaintreatmentscan be performedon one leaf while
I A fulleraccountof thisresearchis available,see I7.
323] [BotanicalGazette,vol. 102

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324 BOTANICAL GAZETTE [DECEMBER

the otherof the same age is leftas a control. At any particulartime the plants
usually have eight pairs of leaves, not counting those folded togetherin the
terminalbud. Eight to tenplantswereused foreach experiment,and each experi-
mentwas repeatedseveral times.
A numberof different treatmentswereused and these are describedseparately.
The growthsubstancesused were carefullyweighedand mixed with the quantity
of anhydrouslanolinnecessaryto make the desiredconcentrations.The mixtures
wereheateduntilthelanolinmeltedand the substanceswerecompletelydispersed.
Fresh mixtureswere made at frequentintervals,althoughit was foundthat the
growthsubstancesremainedactive in thelanolinforseveralyears. The amountof
paste applied was not measured,but about the same amount was applied to each
part of the plant. Controlswere treatedsimilarlywith anhydrouslanolin,and all
cut surfaceswere coated withlanolin to preventdesiccation.
Data forthepre-abscissionperiodwerecollecteddaily. The pots containingthe
plants were tapped gentlyat that time,as petioles in which abscissionwas prac-
ticallycompleteoftenremainedattached to the stem forseveral days by a small
piece of dried tissue.
The internalchangeswerefollowedby theuse offreehandand rotarymicrotome
sections. Freehand sectionswerefrequentlyemployedbecause the changestaking
place during abscission could be better observed. Longitudinal sections cut
throughthe stem and petioles at each node gave comparableviews of the absciss
layersof the pairs of opposed leaves.
Investigation
GROWTH OF LEAVES
In order to discover any possible relationshipbetween development of the
leaves, growthof the absciss layer,and subsequentabscission,the relativeratesof
developmentof the different parts of a leaf were measured.
All leaves on several plants were markedoffinto 4-mm.squares with a special
rubberstamp. As a check,leaves of otherplants were markedoffin 4-mm. seg-
mentswith a small brush and India ink. Microscopicexaminationof the cells of
the petioles,especially those close to the absciss layer,were also made.
The growthof a coleus leaf (fig.I) is similarto that of tobacco as describedby
AVERY (i), but it is more basipetal. There is an increasinggradientof expansion
fromthe tip of the leaf to the base of the blade. A similargradientof expansion
occursin themidriband petiole,almostto its base. By the timea leafhas attained
one-fifth its finalsize the tip has practicallyceased expanding. The greatestexpan-
sion is in the second and thirdpairs ofleaves below the terminalbud; considerably
less in the fourthpair; and in the fifth,expansionof the blade has ceased but the
petioles of these leaves and the sixth and seventh pairs of leaves continue to

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I 940] MYERS-COLEUS 325

elongate. The base of the petiolesdistal to the absciss layerhas the longestperiod
of expansionof the entireleaf, althoughit does not elongate as much as the seg-
mentsof the petioleimmediatelyabove it. The base of the blade has the greatest
amountof expansion,whichis attainedbeforeelongationof the petiole has ceased.
Growthof the abscisslayer
begins in the partially en- /
larged and vacuolated cells
in the base of the petiole of
the thirdpair of leaves below
the terminalbud and contin- |
ues until these leaves have a
become the sixth pair. The i
blade is 85-90 per cent ex- -? ---/ --1 - \-
panded before the absciss t j
layer begins to develop. yI X
There is considerablecell en- /
largementin thepetioledistal
to the absciss layer. The
cellsbecomelargerthanthose
of the stemcortexat the base
of thepetiole. The epidermal
cellsdistalto the absciss layer
bcoell grealtly
thelongatedlandr FIG. i.-Place of expansion in growth of typical coleus leaf
become greatly elongated and
do not increase much in width. The base of the petiole proximalto the absciss
layer and a part of the stem cortexcontributeslittle to the increasein lengthof
the petiole.
DEVELOPMENT OF ABSCISS LAYER AND ABSCISSION

Under ordinaryconditionsthe loss of leaves of coleus probably is always the

resultof two processes: growthof an absciss layer,followedby dissolutionof cer-
tain lamellae in thecell walls in thislayer. Under certainconditions,eitherprocess
may occur withoutthe other.
The absciss layer is formedat the base of the petiole in a plane normalto the
opposed leaves. It usually begins developmentin the third pair of leaves and
maturesin the sixth and seventhpairs. Its initiationis firstevident by the ap-
pearance of a few transversedivided cells in the epidermisand peripheryof the
cortex. In a shorttimeit appears as a ringof dividingcells around the base of the
petiole. These cells continueto divide,and as morecells become involvedthe ring
gradually widens and thickens,finallyextending across all the tissues of the
petiole except the xylem. Ten to fifteenlayers of cells may be involved in the

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326 BOTANICAL GAZETTE [DECEMBER

outerpart,but onlysix to eightlayersin the centralpart of the petiole.The

is aboutthesamethickness
abscisslayer,however, sincethecellsare
throughout,
ofdifferentsizes.
Whensectionsarecutat rightanglesto theplaneoftheabscisslayerit has the
appearanceofa meristem. The cell contentsstaindarkerthantheothercellsin
thecortex, havemoreprominent nuclei,and lacklargevacuoles.As it nearsma-
turitythecellcontentsbecomegranularand the cell wallsless distinct.At ma-
turityit becomesexternallyevidentby the slightdepressionin thepetiole(the

.~ ~ ~ ~~~~~q 4

AA

FIGS. 2, 3.-Fig. 2 (left),longisection

throughbase ofpetioleofsixthpairofleavesbelowterminal
bud,showing matureabscisslayers.Fig.3 (right),sameofpetioleoffirst
pairofleavesbelowterminal
bud;leafdebladed3 months previously.

abscissionzone),sincethecellson eithersideofit continueenlarging whilethose

in theabscisslayerdividewithbut littleenlargement.
In theyounger leavesthecellsin theregionwheretheabscisslayerwilllater
formare indistinguishablefromtheothercorticalcellsof thepetiole.Beginning
withthethirdpairofleavesbelowtheterminal bud,however, a distinctdifference
becomesevident.The cellsofthepetioledistalto theabscisslayerbecomemuch
largerand moreelongatedthanthoseoftheabscissionzoneand stemcortex.An
abscisslayerthatis matureis shownin figure2.
The finalstepin theseparation
oftheleaffromthestemis thedissolution ofa
portionofthewallsofsomeofthecellsin theabscisslayer.Shortly beforeabscis-
sion takesplace, thepetiole-and eventuallythe entireleaf-beginsto lose its
greencolor,and at thetimeofabscissionitis usuallypaleyellow.Whena breakin

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I9401 MYERS-COLEUS 327

the lowerpart ofthe base of thepetioleis evidentthe abscissionprocessesare well

advanced, and in a few days separationis completed.
Usually disintegration of cell walls is evidentin two or threelayersin the distal
part of the absciss layer. The process apparentlybegins in the cells in the lower
side of the petiole. As the middlelamellae of these cells dissolve,the longitudinal
shrinkageof the cortexpulls themslightlyapart and theirexposed surfacesround
up as a resultof turgor.Once the firstbreak is made the dissolutioncontinues
ratherrapidly. Dissolutioncontinuesthroughthe absciss layer and graduallyex-
tends throughthe epidermalcells and throughthe cortex,until all that supports
the petiole is the upper part of the cortexand xylem. Finally these corticalcells
separate,the xylemruptures,and the leaf falls.
EFFECT OF REMOVING THE BLADES
Investigators(3, 4, 7, I9, I5, i8, 20, 21, 22, 23) have shown that abscissionis
acceleratedby high and low light intensities,different lengthsof day, high and
low water supply, high and freezingtemperatures,low concentrationsof anes-
thetics,toxic concentrationsof acids and salts, ethylenegas, woundingof and
completeremovalof the blade.
To followthe internalchangesthat occur afterdebladingand to comparethese
changeswiththoseoccurringin an intactleaf,all the blades of one groupofplants
wereremovedand in a second groupone blade of each pair ofleaves was removed,
the oppositeleaf remainingintact. Sections cut throughthe absciss layersof the
youngerleaves usually indicated that cell divisionof debladed petioles had been
accelerated. In the fifthand lowerpairs of leaves accelerationof cell divisionwas
not so evident. In these older leaves the absciss layerswere matureor almost so,
and debladingdid not cause additional cell division. The deblading,however,al-
ways acceleratedabscission of the petioles.
There were several variations. Occasionally abscission occurred without de-
velopmentofthe absciss layer. Abscissionofdebladed petiolesofthe third,fourth,
and fifthpairs of leaves oftenoccurredwithoutfurtherdevelopmentof the im-
matureabsciss layers,beyond that of the opposed intact leaves. The cell walls in
the debladed petioles were less distinct,the cell contentsmoregranularand more
readilystained-the usual conditionsjust priorto abscission. In the firstpetiole
below the terminalbud, in which no absciss layer was visible at the time of de-
blading,abscission oftentook place withoutdevelopmentof an absciss layer. It
was frequentlyobserved that theseyoung petiolesdid not increasein size and did
not abscise afterdeblading,or abscised onlyaftermorethan 3 months,whenthey
werein thesame positionas the sixthpair ofleaves. In such petiolesa well-defined
absciss layer had developed (fig.3), but dissolutionof the cell wall lamellae was
not takingplace. The factorscausing this long delay have not been discovered.

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328 BOTANICAL GAZETTE [DECEMBER

Afterdeblading,the youngestpetioles did not abscise as soon as the oldest

petioles,and therewas a gradientin time betweenthem (fig.5). At the time of
debladingthe absciss layerswere in various stages of development,fromnone in
the firsttwo pairs of leaves to one that was maturein the sixthpair. Apparently
the lengthof timebeforeabscissionunderthese conditionsis relatedto the degree
of maturityof the absciss layer or to the age of the leaf. Since debladingusually
acceleratesdevelopmentof the absciss layer, it matures more rapidly in older
leaves whereit is already well advanced. DOUBT (3) found that treatingcoleus
withethylenegas caused the oldest leaves to abscise first.

EFFECT OF GROWTH SUBSTANCES ON DEVELOPMENT

OF ABSCISS LAYER AND ON ABSCISSION
OF DEBLADED PETIOLES

In one groupof plants a blade was removedfromone of two oppositeleaves at

each node of the stemand a i per cent mixtureof heteroauxinapplied to the ends
of the debladed petioles (fig.4). By cuttingfreehandsections it was possible to
compare the absciss layers and abscission phenomena in a treated debladed
petiolewith those in the petiole of the opposite intact leaf.
Developmentof the absciss layers in the treateddebladed petioles was similar
to that in the petioles of intact leaves. The time elapsing beforeabscission oc-
curredwas nearlythe same (fig.4), exceptin the oldestleaves whichdropped l-io
days soonerthan the treated debladed petioles. As will be shown later, this fact
may be correlatedwith the smalleramountof growthsubstance comingfromthe
blades of the older leaves, indicatingthat heteroauxindelays the finalstages of
abscission.
In anotherexperimentall the blades wereremovedfromthe plants and a i per
centheteroauxinmixturewas applied to one petioleat each node, and the opposite
petiole smeared with plain lanolin. Treatment of the debladed petioles usually
delayed developmentof the immatureabsciss layers,and abscission. It retarded
initiationof the absciss layer and its growthat any stage of development. The
increasein the pre-abscissionperiod depended upon the stage of developmentof
the absciss layers and the timeofyear (fig.6). The morematurethe absciss layer
the shorterthe time beforeabscission. During May the abscission time was less
than that for the leaves debladed in January. In the sixth,seventh,and eighth
pairs of leaves the absciss layers were so well developed that therewas littledif-
ferencein theirappearance in the treated and untreatedpetioles. Usually there
was decided delay in the growthof the absciss layers in the young petioles,but
occasionally no differencecould be seen. These facts indicate that either the
heteroauxindid not always delay developmentof the absciss layer,or the develop-
ment of the absciss layer was not always accelerated by removal of the blade.

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I940] MYERS-COLEUS 329

Deblading a leaf stopped elongationof the petiole. Diameter measurements

werenot made, but it was apparentthat therewas littleor no increasein diameter.

1% heteroauxin

40 C

35 O^ ^ Deblode& leaf treated Pladr lanolin Plain lanolin

3wth 1% heteroc-in 20

Intact leaf 0
012 5r 1
20 ' 3

alo

5 ~~~~~6
4 51 ~~~~~~5
PIlI
al di L.-j -% heteroaaooin 150

151
F Debladed leaf trsated
wth 1I/oheter~o.0auin3IS
ps Untreated debladed leaif a ot
6 I

and35tio0 S5

l'i~~~~~~~~~~~~~~~~~3

/r\

'01,
(~~~.~~~i~~I)
teopstinatefLe

lavs tredi auryadi ay

/
i.7
nc
Lanolin
in

amont
dic Lan ainnli
Lanolin

ogrwhsusacdifinfrmlve
heteraoauin
1%~~~~~~~~~~~~~
a.
K1. ~~~~~~~~~~~~~~~~a. ~~~~~~~~~~~~~~~~~b

8
(1?) t

II ~~~~~~.15 "Lanolin~Lanli
las~~~~~~~~~~Dcs
tube
01C~~~~~~~~~~~~~~~
I
tube

heteraoauin

A treated debladed petiole, however, usually

ahe nolrt o elo Lanongaeasmcha
FIGS. 4-9.-Fig. 4, pre-abscissionperiod of pairs of untreateddebladed petioles; numbersreferto
positionof leaves below terminalbud. Started March 2. Fig. 5, same oftreateddebladed petiolesand
opposed intactleaves. StartedMay 2. Fig. 6, same of treateddebladed petiolesand opposeduntreated
debladedleaves. StartedinJanuaryand in May. Fig. 7,amountofgrowthsubstancediffusing fromleaves
and petiolestreatedwith i.o per cent heteroauxin;testscarriedout in March. Averageof io tests. (In
gammas ofheteroauxinequivalents;controlswithagar containingioo gammasofheteroauxinper liter.)
Fig. 8, methodoftreatingleaves (dottedlinesindicateportionremoved). Fig. 9, methodoftreatingleaves
and petioles.

A treateddebladed petiole,however,usually elongatedas much as the petiolesof

the opposite intact leaf. Likewise the increasein diameterwas similar. MAI (i6)

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330 BOTANICAL GAZETTE [DECEMBER

had similarresultswhenhe treateddebladed petioleswithpollen. The amountof

enlargementdependedon the age of the leaf. Many of the youngerpetioles con-
tinuedto elongateand reachedor exceeded the dimensionsof the petioles of the
oppositeintactleaves. The petiolesof the firstleaves below the terminalbud did
not enlargeas much as the petioles of the second and thirdleaves. There was
littleelongationof the olderpetioles,but therewas usually morethan that in the
oppositeintactleaves.
These experimentsindicatethat the heteroauxinhas an effecton elongationof
the petioles,developmentof the absciss layers,and on abscission of the petioles
similarin kind to that of the blade and nearlyequal to it in magnitude.
In otherexperimentsconcentrationsof I, 0.5, and o. i per cent heteroauxinwere
used. The i and 0.5 per cent concentrationswereabout equally effective in delay-
ing abscissionbut about 25 per cent more effectivethan the o.i per cent concen-
tration.
Othergrowthsubstances,indolepropionicand indolebutyricacids, wereapplied
to debladed petioles in i per cent concentrations.The indolebutyricacid was
about as effectiveas heteroauxinin delaying abscission,but the indolepropionic
acid was about 25 per cent less effective.

MEASUREMENTS OF GROWTH SUBSTANCES DIFFUSING FROM LEAVES

Measurementsof growthsubstancesdiffusing fromcoleus leaves weremade by
the double decapitationAvena technique (24). The leaves were severedfromthe
plants,the cut ends wiped withmoistfilterpaper and attached to a sheet of agar
witha drop of0.5 per cent gelatin. At the end of 2 hoursthe leaves wereremoved,
the agar allowed to stand for40 minutesand then cut into twelve equal blocks
and applied to decapitated Avena coleoptiles. Tests were made forthe amount of
phytohormones diffusingfrompetiolestreatedwith i per cent heteroauxinon the
distal ends.
The greatestamount of phytohormones diffusedfromthe upper leaves (fig.7).
It is in these leaves that expansionoccurs most rapidly. There was less diffusing
fromthe olderleaves whereexpansionhad almost ceased. Usually therewas none
diffusingfromthe eighthpair of leaves. AVERY (2), workingwith tobacco, and
GOODWIN (5) with two species of Solidago, had similarresults. The quantity of
substancesdiffusing fromthetreatedpetiolesexceededthatcomingfromuntreated
leaves but was not much greaterthan that comingfromthe second pair of leaves
duringthe spring.
Leaves tested duringthe winterand early springgave very weak or negative
tests forgrowthsubstances; duringthe late springthe amountincreased consider-
ably. GUSTAFSON (6) had similarresultswith certainfruits. It is interestingto
note that abscissiontook place moreslowlyduringthe winterwhen the amountof

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I940] MYERS-COLEUS 33I

phytohormones was low than duringthe late springwhenit was higher,and that a
i per cent concentration was moreeffectivein inhibitingabscissionin the summer
than in the winter. Treated petioles remainedon the plants much longerin the
winterthan in the summer,but the effectiveness of the treatmentis based upon
the behaviorof the controlsforeach season.
During the winter(Januaryto March) the second untreateddebladed petioles
abscised in an average of 2 Idays, and the treateddebladed petiolesabscised in an
average of 57 days (fig.6). In the spring(May to June)similarpetiolesabscised in
i6 and 32 days, respectively(fig.6) and in the summer(Juneto August) in 4 days
and 30 days, respectively.The abscissionof intactleaves was considerablyslower
during the winterthan in the spring and summer. During the wintermonths
growthof the entireplant was also much slowerthan duringthe springand sum-
mer. It is obvious that internalconditionsotherthan the growthsubstances are
also primaryfactorsin abscission.

MOVEMENT OF THE INHIBITING EFFECT

Experimentswere designed to determinewhetherthe inhibitingeffectof the
blade moved across, up, or down the stem. In one group of plants one blade of
each pair ofleaves was removed,the oppositeleaf remainingintact,and in another
groupof plants both blades were removed. There was no significantdifference in
the time of abscissionor in the developmentof the absciss layers in the debladed
petiolesof the firstgroupand in the second group. This indicatesthat thereis no
movementof the inhibitingeffectof the blade across the stem to the absciss layer
of the oppositeleaf, at least in significantamounts.
In anotherexperimentall blades were removedfromtwo groupsof plants. All
petiolesof one groupremaineduntreatedexcept fora coating of plain lanolin. In
anothergroupone petioleofeach pair was treatedwith i per centheteroauxin,the
opposite petiole being smeared with plain lanolin. There was little differencein
the developmentof the absciss layersor in the timeof abscissionof the untreated
petioles of these two groups.
One per cent heteroauxinplaced on the sides of the stem of plants fromwhich
all the blades had been removed did not cause significantinhibitionof the de-
bladed petioles, eitherabove or below the treated part. It has been shown by
SNOW (ig) and others that under natural conditionsthe movementof growth
substances is polar, that is, down the stem. AVERY (2) showed that phyto-
hormonesmoved down the leaf throughthe veins and midrib. HITCHCOCK and
ZIMMERMAN (8), however,showed that the application of certain syntheticsub-
stances in water to the soil of potted plants caused root formationon the stem,
epinastyof leaves, and othereffects,indicatinga movementup into the stem and
leaves. They wereunable to securethese effects,except near the point of applica-

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332 BOTANICAL GAZETTE [DECEMBER

tion,whentheyused the mixturein lanolin. When cuttingswereplaced in a strong

solutionof heteroauxin(I: 30,000) for4 hours and the blades removedtherewas
delay of a fewdays in the abscissionof the petiolesas comparedwith comparable
cuttingsin distilledwater.
When approach graftsweremade by splicingone of a pair of debladed petioles
to an intact leaf of anotherplant, abscissionof the graftedpetioles was delayed
not more than 2 days longer than the controls,although both petioles usually
abscised in about the same time. This may indicate some movementof plant
hormoneacross the graft.No graftsformeda union.
These data indicate that there is usually no significantmovementof the in-
hibitingeffectof the blade or of applied substanceacross or up or down the stem.
When cuttingsare placed in a strongheteroauxinsolution there is apparently
some movementup the stem and into the petioles.
Later evidence will show that the inhibitingeffectof applied hormone,and
presumablyalso of the blade, is correlatedwith the amount of substance trans-
located. Furtherevidence will be given indicatingthat under certain conditions
thereis no significantmovementof the inhibitingeffectfroman intact leaf to the
oppositeintact leaf.
EFFECT OF PLACING PLANTS IN CONTINUOUS DARKNESS

ZIMMERMAN and HITCHCOCK (25) foundthat tomatoplants kept in the dark for
5 days did not show geotropiceffects,but curved down when heteroauxinwas
applied to the stem. They mentionedthat if the assumption of KEEBLE et al.
(io)-that the upward growthof plants placed in a horizontalpositionis due to a
redistribution of substance is correct,plants that no longergrowupward should
not contain growthsubstances.
AVERY (2) reportedthat phytohormones disappeared fromtobacco plants after
5 days in the dark. ZIMMERMAN and HITCHCOCK(25), however, reportedthat
tobacco plants i8 inchestall stillelongatedafter3o days in the dark. They found
to account forthiscontinuedgrowthiftherecan be "no growthwithout
it difficult
the presenceof hormones." Using plants withstorageorgans (potatoes), theyob-
tained geotropiceffectsand stem elongationaftermore than 6o days in the dark.
Starch and sugars were detected as long as growthcontinued.
Rapidly growingcoleus plants wereplaced in a dark box in June. At the end of
about io days the leaves began to abscise, theyoungestand oldestleaves abscising
first.No absciss layers were foundin the upper two pairs of leaves at the timeof
abscission. In the older leaves therewas apparentlysome accelerationof these
layers. Abscissionof intact leaves was delayed by the application of heteroauxin
i-iO days longer than the opposite intact untreatedleaves. There was usually
more elongationof the petioles and more increase in area of the blade of the

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I940] MYERS-COLEUS 333

youngerintact treated leaves than in the opposite intact leaves. The untreated
plants had lost most of theirleaves after 2i days, but the stem and remaining
petioles were still elongatingand grewupward when placed in a horizontalposi-
tion. Applicationof growthsubstancesalso caused bendingof the stem. Shortly
afterthisthe plantsbegan to rot and the experimentwas discarded. Decay usual-
ly tookplace at the tip ofthe stemand in leaves that had not abscised. It affected
entireleaves but ceased at a point wherethe absciss layer usually formed.It did
not involve any stem tissue untillater.
Coleus plants were placed in shaded parts of the greenhouse.One of each pair
of leaves at the fivelower nodes below the terminalbud was inclosed in a black
sateen bag,2the opposite leaves being inclosed in similarwhitebags. The leaves
in the black bags abscised I-4 weeks beforethe leaves in the white bags. Other
plants withtheleaves similarlyinclosedin whiteand black bags, and not in either
type of bag, wereplaced in a dark box. The leaves of these threegroupsof plants
abscised in about the same time. These data indicatethat the blade is most effec-
tive in delaying abscission when it is exposed to light. The effectis not trans-
located froma leaf in the lightto the opposite one inclosedin a black bag in suf-
ficientquantitiesto cause a significantdelay in abscission.

EFFECT OF REMOVING CERTAIN PORTIONS OF THE BLADE

LA RUE (I2) reportedthat as littleas one-fourth ofthe blade of a coleus leafleft
intactwas sufficient to inhibitabscissionof the petiole,but he did not indicatethe
part of the blade removedor the extentof the inhibition.In a series of experi-
ments to check the relative effectiveness of differentparts of the blade, various
parts and amountswere removed. Plants bearingseven pairs of leaves wereused
in each experiment:
I. At each of the fivelower nodes one leaf of each pair was debladed (fig.8a).

At each of the fivelowernodes,therefore, therewas a debladed petioleopposed by

a petioleplus the midrib. A narrowridgeof the blade leftattached to the midrib
of theyoungerleaves expanded considerably,so the two upperpairs ofleaves were
not used. The pre-abscissionperiodofpetiolesbearingthe midribswas usually I-2
days longerthan the debladed petioles.
2. All the blade except a small portionof the tip was strippedfromthe midrib

of one leaf at each of the fivelower nodes. All the blade of the opposed leaf was
removedexcept a portionat the base of the blade about equal in area to the tip of
the opposite leaf (fig.8b). At each node, therefore,a leaf with only the extreme
tip of the blade intact opposed a leaf witha small portionof the base of the blade
intact. The base of the blade delayed abscission I2-6 days longer than the tip
2
A Westonlightmeterplaced insidethebags gave no readingwhenheldat a distanceof io inchesfrom
a 6o-wattbulb.

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334 BOTANICAL GAZETTE [DECEMBER

of the blade. The areas of the opposed leaves were equal only at the beginningof
the experiment. The tips did not expand, but therewas some expansionof the
bases, especiallyof the youngerleaves.
3. Incisions were made throughthe blade on each side of the midribof a leaf
at each of the five lower nodes. These incisions extended fromthe base of the
blade almost to the tip, leaving an area at the end of the blade undisturbedand
with the lower part of the blade detached fromthe midribbut connectedto the
tip of the blade (fig.8c). The blades of the opposed leaves were removedfromthe
midribexceptforan area at the tips about equal to that ofthe undisturbedportion
of the oppositeleaf. The tip of the blade plus the restof the blade detachedfrom
the midribdelayed abscission longer than the tip of the blade plus the midrib.
The difference was less than i day forthe older leaves to 5 days forsome of the
upper leaves.
4. One-halfof the upper part of the blade of a leaf at each of the fivelower
nodes was removed. About seven-eighthsof the upper part of the blade of the
oppositeleaves was removed,so that at these nodes a leaf withone-halfthe blade
intactopposed a leaf withone-eighththe blade intact (fig.8d). The largerportion
of the blade delayed abscission longer than the smallerportions,the difference
rangingfrom6 to 8 days.
These data indicate that the base of the blade is more effectivethan the tip in
delayingabscission. The midribis not very effective.One-halfthe blade will de-
lay abscissionlongerthan one-eighthof the base of the blade.

EFFECT OF CERTAIN TREATMENTS OF THE PETIOLES

The followingexperimentswere set up to compare the movementof the effect

of the blade in delayingabscissionwiththe movementof the effectof heteroauxin
in delayingabscission:
i. Two deep incisionswere made in the upper and lowersides of the petiole of
a leafat each node ofthe fivelowernodes of coleus plants (fig.9a). These incisions
cut throughmost of the petiole's tissueand leftonlya narrowbridgeof tissuecon-
necting the two parts of the petioles and completelysevered the semicircular
vascular system. The blades were thereforeattached to the petioles by a bridge
of cells but withoutdirectvascular connections.Lanolin was rubbed into the in-
cisionsand the leaves were supportedin the usual positionwith a piece of tinfoil
wrapped around the petioles. Usually the movementof water fromthe stem and
leaves was greatly reduced by these incisionsand the leaves soon wilted. The
blade of the oppositeleaf was removed. The debladed petiolesabscised 2-IO days
soonerthan the leaves withthe incisedpetioles. Some of the youngerleaves par-

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I940] MYERS-COLEUS 335

tially recoveredand occasionally six to eight adventitiousroots grew fromthe

petiole,just distal to the upper incisions.
2. In a similarexperiment all the blades wereremovedfroma plant. Two deep
incisionsas describedin (i) weremade in a petiole at each of the fivelowernodes.
The distal ends of these petioles were treatedwith heteroauxin(fig.9b); the op-
posite petioles left untreated. The debladed petioles abscised 3-I4 days sooner
than the opposed treatedpetioles.
3. The blades were severedfroma leaf at each of the fivelower nodes. These
blades were then replaced on the stumpof the petiole and held in place with a
piece of glass tubing filledwith lanolin or melted agar. The tube fittedtightly
over the two portionsof the petiole and held them close together(fig.9c). The
opposed petioles were debladed. There was no significantdifference in the pre-
abscissionperiod of the debladed petioles and the petioleswith the blade severed
and replaced.
4. An experimentsimilarto (3) was set up withheteroauxinsubstitutedforthe
blade. All the blades wereremovedfromeach of the fivelowernodes. One petiole
at each node was severed in the middle and replaced on the stump and held in
place witha piece of glass tubingfilledwithlanolinor agar. The distal end of the
petiole was coated withheteroauxin(fig.9d). There was no significantdifference
in the time of abscissionof the opposed petioles.
These data indicate that the movementof the effectiveagent of the blade in
delayingabscissionis similarto the movementof heteroauxinsubstitutedforthe
blade, because: (a) the effectof the blades moves throughthe cells of the petiole
when the vascular systemis severed and the leaves wilted; (b) the heteroauxin
delays abscissionwhen it is applied to the end of a debladed petiole that has the
vascular systemsevered; (c) the effectof the blade does not move across a cut
surfacein significantamounts,as a blade severedand replacedon thepetiolesdoes
not delay abscission;and (d) the heteroauxindoes not move across a cut surfacein
a debladed petiole that has been severedand replaced on the petiole,in sufficient
quantitiesto delay abscission.
Summary
i.Followingthe reportsof othersthat growthsubstances and materials con-
tainingthemdelay abscissionof debladed petioles,a studyof theireffectsin rela-
tion to several otherfactorsthat affectabscissionwas made.
2. Growthof the coleus leaf is basipetal.
3. The blade is 85-90 per cent expanded beforethe absciss layer is initiatedin
the thirdpair of leaves and is maturein the sixthpair of leaves of plants bearing
eightpairs.

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336 BOTANICAL GAZETTE [DECEMBER

4. Abscissionusually occurs in the absciss layer,but under certaintreatments

it may occur withoutdevelopmentof this layer.
5. Older leaves usually abscise before younger leaves, and factors that ac-
celerateabscission cause older leaves to abscise first.
6. Removal of the blade accelerates abscission of all petioles, except the first
pair of leaves. A well-definedabsciss layer develops in these leaves afterseveral
months.
7. Partiallydebladed petioles do not abscise as soon as petiolesfromwhichthe
entireblade has been removed. Activelyexpandingportionsofthe blade are more
effectivein checkingabscission than the matureparts.
8. The basal half of the blade delays abscission 7-Io days longer than the
lowerone-eighthof the blade, but not so long as the intact blade.
9. A greateramount of phytohormonesdiffusesfrompetioles of the upper
leaves than fromthe lowerleaves.
io. Tests for growthsubstances diffusingfromleaves were negative or very
low duringthe winter.
ii. The upper intact leaves and the opposite debladed petioles treatedwith i
per cent heteroauxinusually abscise in about the same time. The lower intact
leaves abscise I-7 days before the treated debladed petioles, indicating that
heteroauxindelays the finalstages of abscission in addition to delayingdevelop-
ment of the absciss layer.
I2. Abscissionis more rapid in the springand summerthan in the winter.
I3. Heteroauxinapplied to debladed petioles has an effectsimilarto the blade
in delayingabscissionand favoringcontinueddevelopmentof the petioles.
I4. Development of the absciss layer and abscission is accelerated in leaves of
plants placed in the dark. Heteroauxin inhibitsthe abscission of these leaves.
I5. Heteroauxinwill pass throughdebladed petioles which have been severed
except forfouror fivelayersof outer corticalcells in sufficient quantitiesto delay
abscissionforseveral days.
i6. The effectof the blades passes throughcells of petioles severedin a similar
mannerand delays abscission.
I7. The pre-abscissionperiod of debladed petioles of cuttingsis increasedby
placing them in a strongsolutionof heteroauxin.
i8. One and 0.5 per cent concentrationsof heteroauxinare about equally effec-
tive in delayingabscissionbut are about 25 per cent moreeffectivethan a o. i per
cent concentration.
I9. Indolepropionicand indolebutyricacids have a similar effectin delaying
abscission.

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I940] MYERS-COLEUS 337

All experimentsindicate that growthsubstancesfromthe blades influence

20.
the developmentof the absciss layer and the process of abscission,but theirrela-
tion to abscissionphenomenadoes not seem to be a simpleone.

The writerwishesto thank Dr. H. C. SAMPSONforsuggestingthe problem,for

his criticaladvice, and forsupervisingthe organizationof the results.
NORTHWESTERN UNIVERSITY
EVANSTON, ILLINOIS

LITERATURE CITED
I. ofthetobaccoleaf. Amer.Jour.Bot. 20: 565-
AVERY,G. S., JR.,Structureand development
592. I933.
2. , Differential ofa phytohormone
distribution in thedevelopingleafofNicotianaand
its relationto polarizedgrowth.Bull. Torr.Bot. Club 62:3I3-330. I935.
3. DOUBT, SARAH L., The responseofplantsto illuminating gas. BOT. GAZ.63:209-224. IQI7.
4. GOODSPEED, T. H., McGEE, J.H., and HODGSON, R. W., Note on the effectsof illuminating
gas and its constituentsin causingabscissionof flowersof Nicotianaand Citrus. Univ.
Calif. Publ. Bot. 5:439-450. i9i8.
5. GOODWIN, F. G., The role of auxin in leaf developmentin Solidagospecies. Amer.Jour.
Bot. 24:43-5I. I937.
6. GUSTAFSON,F. G., Auxindistribution in fruitsand its significancein fruitdevelopment.
Amer.Jour.Bot. 26: i89-i94. I939.
7. HEINCKE,A. J., Concerningthe sheddingof flowersand fruitsand otherabscissionphe-
nomenain apples and pears. Proc. Amer.Soc. Hort. Sci. I9I9:76-84. I9I9.
8. HITcHcocK, A. E., and ZIMMERMAN,P. W., Adsorption and movement of syntheticgrowth
substancesfromsoilas indicatedby theresponseofaerialparts. Contrib.Boyce Thompson
Inst. 7:447-476. I935.
9. KENDALL,J. N., Abscissionof flowersand fruitin theSolanaceae withspecialreferenceto
Nicotiana. Univ. Calif. Publ. Bot. 5:347-428. i9i8.
I0. KEEBLE, F., NELSON, M. G., and SNOW, R., The integration of plant behavior. IV. Geo-
tropismand growthsubstances. Proc. Roy. Soc. London,B io8:537-545. I93I.
II. LAIBACH, F., Wuchstoffversuche mitlebendenOrchideenpollinen.Ber. Deutsch.Bot. Ges.
5I:336-340. I933.
I2. ofauxinon abscissionofpetioles. Proc. Nat. Acad. Sci. 22:255-
LA RUE, C. D., The effect
259. I936.
I3. LLOYD, F. E., Abscission.Ottawa Nat. 28:5I-I05. I9I4.
I4. The abscissionof flowerbuds and fruitsof Gossypium
, and its relationto environ-
mentalchanges.Trans.Roy. Soc. Canada IO: 55-6i. I9I6.
I5. I Abscissionin Mirabilisjalapa. BOT. GAZ.6i: 2I3-230. I9I6.
I6. MAI, G., Korrelationsuntersuchungen an entspreitetenBlattstielenmittels lebenden
Orchideenpollinenals Wuchstoffquelle. Jahrb.wiss. Bot. 79:68I-7I3. I934.
I7. MYERS,R. M., The effect of growthsubstanceson the absciss layer in leaves of Coleus.
Ph.D. thesis,Ohio State University.Columbus,Ohio. I939.
I8. SAMPSON, H. C., Chemicalchangesaccompanyingabscissionin Coleusblumei.BOT. GAZ.
66:32-53. I9I8.

This content downloaded from 130.133.008.114 on November 27, 2016 18:07:24 PM

All use subject to University of Chicago Press Terms and Conditions (http://www.journals.uchicago.edu/t-and-c).
338 BOTANICAL GAZETTE [DECEMBER

I9. SNOW, R., Growth regulators in plants. New Phytol. 3I:336-354. I932.
20. WEISNER, J., Untersuchungen uiberdie herbstlicheEntlaubungder Holzgewachse.Lits.
Akad. Wissensch.Wien 64:465-509. i87I.
2I. , Uber Sommerlaubfall.
Ber. Deutsch. Bot. Ges. 22:64-72. I904.
22. Trieblaubfall. Ber. Deutsch. Bot. Ges. 22:3i6-323. I904.
,

23. , Hitzelaubfall. Ber. Deutsch. Bot. Ges. 23:4g-60. I1904.

24. WENT, F. W., and THIMANN, K. V., Phytohormones. New York. I937.
25. ZIMMERMAN, R. W., and HITCHCOCK, A. E., Effectoflightand darkon responsesofplants
to growthsubstances.Contrib.Boyce ThompsonInst. 8: 2I7-23I. I936.

This content downloaded from 130.133.008.114 on November 27, 2016 18:07:24 PM

All use subject to University of Chicago Press Terms and Conditions (http://www.journals.uchicago.edu/t-and-c).