Annals of Botany 78 : 255266, 1996

Exogenous Auxin Effects on Lateral Bud Outgrowth in Decapitated Shoots

M O R R I S G. C L I N E
The Ohio State Uniersity, Department of Plant Biology, Columbus, Ohio 43210, USA
Received : 5 July 1995

Accepted : 11 March 1996

Key words : Apical dominance, lateral bud outgrowth, axillary bud, auxin, IAA, decapitation, Vicia faba, Ipomoea nil,
Pisum satium, Phaseolus ulgaris, Lycopersion esculentum, dgt, Coleus blumei, Arabidopsis thaliana, Helianthus
annuus, ThimannSkoog.

INTRODUCTION
Apically derived auxin in shoots is generally thought to
control apical dominance either directly via entry into
lateral buds with subsequent repression of outgrowth or
indirectly via some other mechanism, i.e. activation of a
second inhibitor messenger, auxin-cytokinin ratio, secondary growth substances, nutrient diversion, etc. (Martin,
1987 ; Cline, 1991 ; Bangerth, 1994 ; Stafstrom, 1995 ; Tamas,
1995). As studies of control mechanisms of developmental
responses in plants have progressed, particularly with the
use of molecular and genetic approaches (Cline, 1994), there
has been increased interest directed towards the role of
auxin in apical dominance, and with the degree of branching
serving as a possible indicator of auxin activity (Tepfer,
1984 ; Lincoln, Britton and Estelle, 1990 ; Shen et al., 1990 ;
Estruch et al., 1991 ; Klee and Romano, 1994).
Because of this increased attention concerning auxin in
apical dominance and inasmuch as the precise mechanism
of auxin action in apical dominance has yet to be elucidated,
a closer scrutiny of the evidence supporting auxin involvement in the control of lateral bud outgrowth is needed.
The strongest evidence supporting either a direct or an
indirect role for apically-derived auxin in controlling apical
dominance comes from the classical ThimannSkoog experiment with Vicia faba (Thimann and Skoog, 1933). They
demonstrated that auxin applied in agar blocks every 6 h to
the stump of a decapitated Vicia faba stem inhibited the
outgrowth of lateral buds situated lower on the stem.
Thimann (1937) initially proposed the direct inhibition
hypothesis which presumes that auxin enters the lateral bud
and directly inhibits its outgrowth. Subsequently, he
modified his views to include indirect auxin action via
involvement with cytokinins (Sachs and Thimann, 1967) or
0305-7364}96}08025512 $18.00}0

ethylene (Russell and Thimann, 1988) as well as effects of

light (Thimann, 1977).
While many investigators have substantiated and extended the results of the ThimannSkoog experiment (i.e.
exogenous auxin repression of lateral bud outgrowth in
decapitated shoots), others have reported anomalous effects
and there have been questions about the direct role of auxin
in apical dominance (Jacobs et al., 1959 ; Brown, McAlpine
and Kormanik, 1967 ; Ali and Fletcher, 1970 ; Shein and
Jackson, 1972 ; Hillman, 1984). Wareing and Phillips (1981)
stated it is now generally considered that auxin does not
exert its inhibitory effect on lateral buds in such a direct
manner as that originally proposed by Thimann . Some
workers (Gregory and Veale, 1957 ; McIntyre, 1977) have
proposed nutrition to be the most important controlling
factor in apical dominance. To date, no one has reported
evidence of auxin restoration of apical dominance in
Arabidopsis via the ThimannSkoog experiment.
The question has also been raised as to how the terminal
bud can continue to grow while simultaneously generating
inhibitory concentrations of auxin to the outgrowth of
lateral buds below. Klee et al. (1987) have found that
overproduction of auxin in petunia via transformation with
IAA biosynthetic genes of Agrobacterium eliminates branching. With a similar transformed system, Romano, Cooper
and Klee (1993) have reported an increase in apical
dominance in Arabidopsis as measured by a reduction in
fresh weight of secondary inflorescences. However, this
auxin overproduction occurs in every cell of the organism
rather than being localized in buds of adjacent nodes as
might occur in a natural system. In addition, Lincoln et al.
(1990) reported that total number of inflorescences arising
from the rosette does not differ greatly between axr1 [the
bushy auxin-resistant mutant] and wildtype even though
# 1996 Annals of Botany Company

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In 1933 Thimann and Skoog demonstrated exogenous auxin repression of lateral bud outgrowth in decapitated shoots
of Vicia faba. This evidence has given strong support for a role of auxin in apical dominance. Most, but not all,
investigators have confirmed Thimann and Skoogs results. In the present study, auxin treatments were carried out
on ten different species or plant types, many of which were treated with auxin in different forms, media and under
different light conditions. The ThimannSkoog experiment did work for most species (i.e. exogenous auxin did repress
bud outgrowth) including the dgt tomato mutant which is known to be insensitive to auxin in certain responses. Toxic
auxin symptoms were observed in some but not all species. The ThimannSkoog experiment did not work for
greenhouse-grown Coleus or for Arabidopsis. Light was shown to reduce apical dominance in Coleus and Ipomoea nil.
# 1996 Annals of Botany Company

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ClineExogenous Auxin Effects in Decapitated Shoots

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F. 1. Lateral bud outgrowth approximately 1 week following decapitation. A, Vicia faba L. (Broad Windsor Bean). B, Helianthus annuus L.
(Mammoth Grey-striped Sunflower). C, Ipomoea nil L. Roth (Japanese Morning Glory). D, Pisum satium L. [Little Marvel Pea (dwarf )], and
E, Thomas Laxton Pea (normal). F, Phaseolus ulgaris L. (Blue Lake Bean). The stem stumps of all decapitated plants shown here were treated
with lanolin except for Japanese Morning Glory (C) which has a taped cotton swab for treatment with aqueous solution.

the number of lateral branches did greatly increase in the

former. Beveridge, Ross and Murfet (1994) reported that
grafting of non-branching wildtype stocks to the branching
rms-2 pea mutant scions did not normalize endogenous IAA
levels even though it did inhibit branching.

White (1976) stated that discrepancies in the results of

different workers using apparently similar methods of
application of IAA to decapitated plants of a single variety
of Phaseolus obviously require some explanation . He
carried out experiments, focusing on this single species with

ClineExogenous Auxin Effects in Decapitated Shoots

variations in plant age, concentrations and quantity of IAA,
type of lanolin, region of application and time of year. His
results showed that the effectiveness of IAA applied did
vary significantly with those conditions. He also found that
in many instances, IAA did completely replace the main
shoot with respect to correlative inhibition of lateral bud
growth.
The objective of the present study was similar to that of
Whites except that instead of analysing a single species, the
focus here was to compare responses to exogenous IAA in
repressing lateral bud outgrowth in ten different species or
plant types, including the mutant dgt tomato which is
known to be insensitive to auxin promotion of certain
responses, and Arabidopsis thaliana. In addition, the effects
of auxin in different concentrations (both aqueous and in
lanolin), forms (IAA, -NAA and -NAA) and under
different light conditions were studied in a number of these
species.

Seeds of Vicia faba L. (Broad Windsor Bean), Helianthus

annuus L. (Mammoth Grey-striped Sunflower), Ipomoea nil
L. Roth, strain violet (syn. Pharbitis nil ) (Japanese Morning
Glory), Pisum satium L. [Little Marvel (dwarf ) and Thomas
Laxton (normal) Pea], Phaseolus ulgaris L. (Blue Lake
Bean) and Lycopersicon esculentum, Mill., strain VNF8 and
the mutant diageotropica (dgt) tomato were germinated in
Pro-mix, a general purpose peat-vermiculite growing medium. The seeds of Ipomoea were scarified in concentrated
sulphuric acid for 40 min and soaked overnight in running
water and germinated in Petri dishes before planting. Coleus
blumei Benth. was propagated from stem cuttings. All the
plants except the normal pea were grown in a greenhouse
(1632 C) during the winter}spring of 199394 and the
spring}summer of 1995 with supplementary General Electric
400 watt mercury vapour lamps (total irradiance : up to
3300 mol m# s"). All the above plants, except Coleus,
were also grown in growth rooms (2729 C) under continuous light (General Electric, Power Groove cool white
fluorescent and incandescent sources 25450 mol m# s").
The ages of the plants at the times of decapitation and the
beginning of the auxin treatments varied with the growing
conditions (i.e. in greenhouse and growth rooms) and the
developmental status of the lateral buds. The ages (in d)
ranged as follows : Vicia faba, 1213 ; Helianthus annuus,
1927 ; Ipomoea nil, 1232 ; Pisum satium, dwarf, 1021,
normal, 1516 ; Phaseolus, 1018 ; Lycopersicon esculentum,
VNF8, 2754 ; dgt, 3968 ; Coleus (from time of stem
cutting), 2137 ; Arabidopsis thaliana, 3552. Intact plants
with elongating lateral buds were excluded. Shoots were
decapitated with a razor blade about 1 cm above the base of
the lateral bud at the node indicated for each species in the
Results section (Fig. 1). Excluding the cotyledons, nodes
were counted upward from the base of the plant. There was
some variation in this distance depending upon the species
and the relevant internode lengths. It was necessary that
there be sufficient distance between the lateral bud and the
decapitated stem stump above so that auxin could be

applied to the stump without contact with the bud. Auxin

was applied to the stem stump twice daily beginning
immediately after decapitation in approximately 150 l
doses as IAA, - or -NAA at concentrations ranging from
10' to 10$ in 005 % Tween 20 aqueous solution in a
taped cotton swab (Fig. 1 C) or as IAA in lanolin, 01, 05,
1 %. Treatments normally extended from 4 to 10 d with
daily measurements of the lateral bud situated at the highest
node below the point of decapitation. Two exceptions were
Vicia faba and Pisum satium (normal) where the largest
rather than highest lateral bud was used to determine bud
outgrowth. Each treatment was usually carried out with
four to nine plants. At least two experiments with each of
the plant types were carried out in both the greenhouse and
the growth room with the exceptions indicated above.
For certain light experiments (as indicated in the Results
section), Ipomoea seedlings were moved from growth rooms
to out-of-doors (irradiance under shade screens, up to
210 mol m# s" and in the open, up to 6900 mol m# s")
during the summers of 1992 and 1994 and Coleus plants
propagated from stem cuttings were grown in growth rooms
with greatly reduced irradiance from General Electric cool
white fluorescent lamps (2530 mol m# s"). There were
usually four to ten Coleus and Ipomoea plants in each of the
auxin treatments. In the outside shade screen experiments,
there were eight to 11 Ipomoea plants per treatment. Seeds
of Arabidopsis thaliana (strain CS 1072 Chi-O from the Ohio
State University Arabidopsis Biological Resource Center)
were germinated in water-saturated Magik-moss potting
soil containing perlite and vermiculite in greenhouse
following a 2-d treatment in the refrigerator (03 C). After
several weeks, some seedlings were moved to the growth
room (2729 C) under continuous light while the remaining
were kept in the greenhouse.

RESULTS
Auxin significantly inhibited lateral bud outgrowth following decapitation when applied to the stem stump of most
of the species tested [Ipomoea nil, Helianthus annuus,
Lycopersicon esculentum (VNF8), Pisum satium (Little
Marvel and Thomas Laxton), and Vicia faba] at concentrations of 10& and}or 10% in aqueous solution or 01
and}or 1 % in lanolin (Figs 2, 4 ; Table 1). Inhibition was
also found in the tomato mutant dgt (Fig. 4) which is
insensitive to auxin with regard hypocotyl elongation and
ethylene production (Kelly and Bradford, 1986). In Ipomoea,
Helianthus and dgt tomato there were no obvious toxic
effects of exogenous auxin applications observed in any
experiments. In Vicia faba and Pisum, occasional auxininduced aberrations in stem and leaf growth were observed
whereas in the VNF8 tomato such aberrations were more
common.
Auxin had no effect on restoring apical dominance in
decapitated greenhouse-grown Coleus (Fig. 5) or Arabidopsis
(Fig. 3, Tables 2 and 3) and only a partial effect on bean
(Fig. 2 F). -NAA was generally more potent than IAA in
inhibiting lateral bud outgrowth. -NAA, the inactive auxin

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MATERIALS AND METHODS

257

258

ClineExogenous Auxin Effects in Decapitated Shoots

Bud length (mm)

30

Decap
control

Decap
control

20
15

20

IAA
4
10 M

10
0.1%
IAA

10

5
-NAA 10 M

1%
IAA
1

20

Decap
control

IAA

12
8

10

IAA

10

IAA

10

IAA

25

12

Intact

Decap
control

Bud length (mm)

10

16

4
M
M

0.1% IAA
Intact
control
1

Decap
control

30
Decap
control

20
20
15

1%
IAA

10

10

0.1% IAA

1% IAA
Intact
1

Day

Intact control
1

Day

F. 2. Auxin repression of lateral bud outgrowth over 3 or 4 d following decapitation. A, Vicia faba (growth room, n 4). B, Helianthus annuus
(growth room, n 5). C, Ipomoea nil (growth room, n 34). D, Pisum satium (dwarf ) (growth room, n 56). E, Pisum satium (normal)
(growth room, n 7). F, Phaseolus ulgaris (greenhouse, n 4). Decap, Decapitation. Vertical lines represents.d.

analogue, which was tested on seven of the ten plant types,

usually had little or no effect on apical dominance in most
experiments. With some species [Vicia faba ; Pisum (Little
Marvel), Helianthus annuus], the lack of effect of auxin in
aqueous solution may have been due to a penetration or
metabolism problem. Auxin in lanolin was always effective.
In the following section are descriptions of apical
dominance in the ten plant types, and their responses to
decapitation and to auxin treatments under growth room
and greenhouse conditions. The term strong apical dominance as used here signifies little or no lateral bud outgrowth
in intact plants. Medium signifies some bud growth and
weak indicates substantial and continuing lateral bud
growth in intact plants.

Vicia faba (Broad Windsor Bean)

The intact plant had weak to medium apical dominance.
Lateral buds on nearly half of the greenhouse plants and on
a quarter of the growth room plants had grown out before
the start of the auxin treatment. These plants had to be
excluded. Decapitation above the third node resulted, most
often, in the outgrowth of the lateral buds at the basal node
but sometimes at the upper nodes (Fig. 1 A). All three lateral
buds were repressed by the auxin treatment with the top
(third) bud (closest to the auxin application) inhibited the
most. Consistent repressive effects on bud growth were
found with 1 % IAA in lanolin and partially at 01 % (Fig.
2 A). There was some swelling and abnormal curving of

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Bud length (mm)

ClineExogenous Auxin Effects in Decapitated Shoots

T 1. Outgrowth of lateral buds (mms.d.) oer 3 or 4 d
following decapitation and treatment of shoot stump with
IAA, -NAA, -NAA in aqueous solution or IAA in lanolin.
Decap., decapitation
Days after decapitation
0

0
214
2713
1611

0
354
4314
2314

0
94
0
0

0
207
12
0

255
86
42
31
83
82
31
52

41
136
113
31
72

41
4511
238
115
259

solution (10& to 10$ , Fig. 2 C) and IAA in lanolin (01

and 1 %, Table 1) significantly inhibited lateral bud
outgrowth following decapitation with no toxic auxin
symptoms observed except at 10$ IAA. As Fig. 2 C
indicates, although there was no effect at 10' IAA, there
was increasing repression of axillary bud outgrowth from
10& to 10$ .
Pisum sativum [Little Marel (dwarf ) and Thomas
Laxton (normal) Pea]
The intact plants had moderate apical dominance.
Following decapitation above one of the higher nodes
(usually the fifth), many of the buds at the lower nodes
would simultaneously grow out to considerable lengths
(Fig. 1 D, E). In the dwarf pea, the bud at the highest node
usually grew out most rapidly followed by the buds at the
second and third highest nodes (Fig. 1 D). In the normal
pea, it was the bud at the second or third node from the base
which grew out more rapidly followed by those of the fourth
and fifth nodes, respectively (Fig. 1 E).
Auxin did significantly restore apical dominance in both
the dwarf and the normal pea plants following decapitation.
Significant inhibition of bud outgrowth was obtained in
dwarf pea with 10% -NAA but not with 10% IAA in
aqueous solution (Table 1). IAA in lanolin at 01 % was
effective in both dwarf (Fig. 2 D) and normal (Fig. 2 E) types
although some swelling in the stem stump and abnormal
stem curvature were observed in the normal peas and in one
experiment with the dwarf peas.
Phaseolus vulgaris (Blue Lake Bean)

stems which accompanied the auxin treatment. Neither IAA

nor -NAA in aqueous solution were very effective in
repressing axillary bud outgrowth (Table 1).

The intact plant had very strong apical dominance with

absolutely no sprouting of lateral buds. Outgrowth of buds
in the greenhouse was not observed until nearly a week
following decapitation above the first node, whereas it was
observed in the growth room within 1 or 2 d (Fig. 1 B).
Release from apical dominance could be inhibited by 10&
-NAA (Fig. 2 B) or by 01 % IAA in lanolin (Table 1). IAA
in aqueous solution was ineffective at 10% (Fig. 2 B). No
toxic auxin effects were observed.

The intact plant had weak to medium apical dominance.

Its apical dominance has been described as incomplete
(Tamas, 1987). The axillary buds subtended by the primary
leaves (the first node, above which we decapitated) were
more inhibited than the buds at the second node subtended
by the first trifoliate leaf. The bean plant grows rapidly, the
internodes are large and the plant is easy to work with.
Decapitation definitely accelerated lateral bud outgrowth
(Fig. 1 F) but the inhibitory effect of IAA (1 % in lanolin,
Fig. 2 F and 10% in solution, Table 1) applied to the stem
stump was only partial. -NAA at 10% had a stronger
effect (Table 1). No toxic effects of auxin were observed
except for a little swelling and bleaching of the stem stumps
upon which the auxin was directly applied.

Ipomoea nil (Japanese Morning Glory)

Coleus

The intact plant had medium to strong apical dominance.

There was no axillary bud outgrowth in the growth room or
in winter-grown greenhouse plants. There was slight
outgrowth of cotyledonary buds of greenhouse plants in the
spring (data not shown). Decapitation above the second
node nearly always resulted in the rapid outgrowth of the
bud at the node (Fig. 1 C). The bud at the first node also
often exhibited a small amount of temporary growth.
Both IAA and -NAA (data not shown) in aqueous

The intact Coleus plant had weak apical dominance. It

had short internodes, grew slowly and was bushy. The basal
branches were of greater length than the higher branches
because of their greater age. These plants were propagated
from cuttings. Many intact plants had to be excluded from
the study because of extensive axillary bud development
beginning at the basal nodes. Only plants with very small
lateral buds (! 34 mm) were selected. At the time of
decapitation, the lateral buds of the lowest of the top three

Helianthus annuus (Mammoth Grey-striped Sunflower)

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Vicia faba, growth room, n 4

Intact
0
0
0
Decap. control
0
12
83
IAA 10%
0
55 1410
-NAA 10%
0
12
56
Helianthus annuus, growth room, n 45
Intact
0
0
0
Decap. control
0
11
32
IAA 01 % lanolin
0
0
0
IAA 05 % lanolin
0
0
0
Ipomoea nil, growth room, n 10
Decap. control
10 31
81
IAA 01 % lanolin
20 21
32
IAA 1 % lanolin
10 20
21
Pisum satium (dwarf ), greenhouse, n 4
Intact
31 31
31
Decap. Control
21 31
51
31 41
52
IAA 10%
-NAA 10%
21 21
21
-NAA 10%
21 31
42
Phaseolus ulgaris, growth room, n 4
Intact
30 41
40
Decap. control
40 72 113
IAA 10%
31 52
82
-NAA 10%
30 41
62
-NAA 10%
31 52
92

259

260

ClineExogenous Auxin Effects in Decapitated Shoots

nodes were removed. Decapitation above the third node of

these greenhouse plants had only a small effect on
accelerating bud outgrowth. Auxin (01 % and 1 % in
lanolin) application to the stem stumps had no repressing
effect on lateral bud outgrowth of greenhouse plants (Fig.
5).
Lycopersicon esculentum [VNF8 (normal ) and dgt
(mutant) tomato]
The intact plants of both dgt and VNF8 had medium
apical dominance with little or no lateral branching. Neither
the dgt nor the VNF8 in our growth room or greenhouse
conditions were categorized as having reduced apical
dominance or as being bushy. In a few experiments, some

lateral bud development was noticed in intact plants. The

VNF8 plants were larger than the dgt plants. The shoot of
the young dgt seedling initially grows more or less vertically.
After several months, the shoot gradually assumes a
horizontal orientation. The seedling shoots of dgt, although
not yet at horizontal growth stage, were floppy and hence
were staked in an upright position for convenience in auxin
treatment. Since horizontal positioning tends to weaken
apical dominance (Prasad and Cline, 1985 b), such staking,
if anything, would have a countering effect to anomalous
bud outgrowth.
Following decapitation, which was usually carried out
above the fifth node, several of the lateral buds on any given
dgt or VNF8 plant vigorously sprouted more or less
simultaneously (Fig. 3). This most often occurred at the

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F. 3. Upper left and right, VNF8 (wildtype) and dgt (mutant) tomatoes about a week following decapitation showing lateral bud outgrowth.
The stem stumps have been treated with lanolin. Lower left, Arabidopsis, 1 week following decapitation ; A, decapitation above the first node ; B,
intact control ; C, decapitation above first node with 1 % IAA on stump of main stem ; Lower right, Coleus showing the branch-promoting effect
of greenhouse high light (B) as compared with indoor low light (A).

261

ClineExogenous Auxin Effects in Decapitated Shoots

B

Bud length (mm)

-NAA
105M

16

Decap
control

12

15

IAA

Decap
control

10

-NAANA
A105M
Intact

10
5

-NAA 105M IAA 104M

-NAA
104M
3 Intact
2

C
20

Decap
control

25

Decap
control

15
IAA 1%

20

0.1% IAA
10

IAA 0.1%
10

0.5% IAA
1% IAA

Intact
2

4
Day

4
Day

F. 4. Auxin repression of tomato [VNF8 (A, C) and dgt (B, D)] lateral bud outgrowth in growth room following decapitation. Auxin in aqueous
solution, A, B (n 45, 4). Auxin in lanolin, C, D (n 34, 4). Decap, decapitation. Vertical lines represents.d.

Bud length (mm)

10

1%
IAA
Decap
control

Intact
control

4
2

4
Day

F. 5. Lateral bud growth in Coleus blumei over 6 d following

decapitation and stem stump treatment with 1 % IAA (greenhouse,
n 910). Vertical lines represent s.d.

upper-most nodes but sometimes at one of the lower nodes.

The points on the curves in the graphs represent the mean
length of the highest lateral bud on each plant. The young
axillary buds of tomato were difficult to observe and to
measure because of their small size and narrow shape.
The treatment with exogenous auxin [10& -NAA
and}or 10% IAA in aqueous solution (Fig. 4 A, B) and
01 % IAA in lanolin, Fig. 4 C, D] did, for the most part,
significantly inhibit axillary bud outgrowth in both plant
types. IAA in aqueous solution had variable effects on
VNF8 but -NAA, down to 10& , had consistent
repressive effects on bud outgrowth (Fig. 4 A). -NAA had

no inhibitory effect. In several instances it appeared even to

promote bud growth. IAA at 10% and 10& -NAA
generally restored apical dominance in dgt (Fig. 4 B and
data not shown). IAA in lanolin (both at 01 and 1 %) was
quite effective in repressing bud outgrowth in both VNF8
and dgt (Fig. 4 C, D). Hence, no essential difference in
sensitivity could be positively detected between dgt and
VNF8 in the response to auxin in repressing lateral bud
outgrowth except for the toxic auxin effect mentioned
previously for VNF8.
Arabidopsis thaliana
Arabidopsis is a facultative long-day plant with weak
apical dominance consisting of a rosette from which arises
a single branching floral shoot or inflorescence followed by
the generation of several surrounding axillary or secondary
inflorescences also arising from the rosette. The measurement of changes in apical dominance status is complex
because of the two types of branching, lateral shoots
developing from the main shoot and axillary inflorescences
developing from the rosette and the rapidity with which
they occur. The lateral buds (shoots) of the main shoot
begin emerging almost as soon as the main shoot begins to
bolt. In the many mutants and strains of Arabidopsis now
available, there exists a wide range of branching habits.
The CS 1072 Chi-O strain used in the present study has
stronger apical dominance than most Arabidopsis types
inasmuch as it generally lacks axillary inflorescences.
Decapitation of the main floral shoot resulted in increased

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Bud length (mm)

IAA 105M

20

262

ClineExogenous Auxin Effects in Decapitated Shoots

T 2. Effect of 1 % IAA on lanolin on Arabidopsis. Outgrowth of first lateral bud (mms.d.) and axillary inflorescence
1 week following decapitation of the main shoot and treatment of the shoot stump with 1 % IAA in lanolin. Decapitation was
either 35 mm aboe the first node or 35 mm aboe the base of the main shoot (below the first node) as indicated. Axillary
inflorescence deelopment is gien as a percentage of the plants sprouting one or more axillary inflorescences. *Main shoot
length
Decap above first node Decap below first node
Intact
plant

Control

1 % IAA

Control

1 % IAA

Decap above first node Decap below first node

Intact
plant

Control

Growth room
n
First Lateral Bud
Axial Inflorescences

1 % IAA

Control

1 % IAA

Greenhouse

23521*
0

14524
71 %

13229
33 %

100 %

100 %

12457*
0

8729
60 %

7852
60 %

100 %

100 %

Growth room
Intact control
Intact IAA 10%
Decap. control
Decap. IAA 10%
Greenhouse
Intact control
Intact IAA 10%
Decap. control
Decap. IAA 10%

Main
shoot

Axillary
inflorescence
length (mms.d.)

Axillary
inflorescence
percentage

7
7
7
8

23521
21420

3823
7143
8141

0
38
100
100

5
9
5
10

12457
10946

0
2531
2124

0
0
100
60

outgrowth of lateral (buds) shoots from the main shoot as

well as the subsequent generation of axillary inflorescences
from the base of the plant (Fig. 3). If the point of
decapitation of the main shoot was above the lowest lateral
emerging shoot (bud), then that shoot grew out to a much
greater extent than in the intact plant and took over as the
main shoot. If the decapitation of the main shoot was below
the lowest lateral emerging shoot (bud), then several axillary
inflorescences grew out from the base of the plant. The
greenhouse plants exhibited somewhat stronger apical
dominance with respect to axillary inflorescence outgrowth
than did the growth room plants.
In essentially no plant did auxin, either as 1 % IAA in
lanolin applied to the decapitated stump of main shoot or as
10% IAA spray applied every other day to the whole
plant, have any effect in restoring apical dominance, i.e., in
inhibiting lateral branching of the main shoot or the
outgrowth of the axillary inflorescences (Fig. 3, Tables 2
and 3). In the case of decapitation above the first node, there
appeared to be some auxin repression in the percentage of
axillary inflorescence development in the growth room
(Table 2) but this was not confirmed in the greenhouse nor
in the more direct test when IAA was applied following
decapitation below the first node.

Attempts to restore apical dominance with auxin spray

applied to the small and extremely bushy trp 1-1 tryptophanrequiring auxotrophic mutant (suggestive of an auxin defect,
Last et al., 1992) were also unsuccessful (data not shown).
Influence of light
Past observation (Hosokawa et al., 1990) indicated that
when Ipomoea nil was propagated in growth rooms under a
mixture of cool white fluorescent and incandescent sources
(irradiance : 25450 mol m# s"), a 30-d-old plant was
characteristically tall (4070 cm) without branching. However, when the plant was grown outdoors (irradiance : up to
6900 mol m# s"), the plant was short, often with such a
proliferation of branching that the study of auxin effects on
bud outgrowth was impossible to carry out since most buds
had substantially elongated at a very early developmental
stage. Therefore, Ipomoea plants were grown indoors until
the age of 14 d, at which time they were moved outdoors
(still in pots), decapitated and stem stumps treated with
auxin in lanolin for 4 d. There appeared to be little or no
difference in sensitivity in auxin repression of axillary bud
outgrowth between indoor plants at low irradiance and
indoor plants moved outdoors at high irradiance at the time

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T 3. Effect of 10% M IAA spray on Arabidopsis. Outgrowth of axillary inflorescences 1 week following decapitation of
the main shoot 35 mm aboe the base of the plant which was sprayed eery other day beginning 15 d before decapitation with
10% M IAA. Axillary inflorescence deelopment is gien as a percentage of plants sprouting one or more axillary inflorescences

263

ClineExogenous Auxin Effects in Decapitated Shoots

T 4. Outgrowth of lateral buds (mms.d.) of Ipomoea
nil plants oer 3 or 4 d following decapitation and treatment
of shoot stump with IAA in lanolin either in growth room
(25450 mol m# s") or outdoors (up to 6900 mol m# s")

32.1
100

200

37.6

10
20
10

31
21
20

81
32
21

255
86
42

21
30
31

31
31
81

61
31
31

123
61
31

236
113
52

75
Weight (g)

Growth room, n 10
Decap. control
IAA 01 %
IAA 1 %
Outdoors, n 48
Decap. control
IAA 01 %
IAA 1 %

50

100

3.7

50

Weight (g)

Height (cm)

F. 7. Bar graphs comparing total fresh weights and heights of

Ipomoea nil plants grown outside in heavy shade (+) and in the open
(*). n 811.

2.6

22.6

16.7

that they were hardly recognizable as Ipomoea nil plants.

Both the lateral bud length and the total fresh weights of the
shaded plants were very much reduced as compared to those
of unshaded plants (Figs 6 and 7). At the fifth, sixth and

20.5
17.9

30

25.5

27.3

Bud length (cm)

40

0.3
0.1

0.2
0

13

0.4
0.06

2.1
0.2

2.6

9 10 11 12
Node number

0.9
0.06

0.7
0.7

0.9
0.4

0.2

3.5
1.3

24.2

1.4
0.2

0.4

0.3

15.7

0.5

4.9

0.1

10

2.5

18.2

9.6

1.8

20

14

15

16

17

18

19

20

F. 6. Bar graphs comparing lateral bud length (cms.d.) of Ipomoea nil plants grown outside in heavy shade (+) and in the open (*) at different
nodes numbering up from the base of the shoot. n 811.

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35.3

22.8

25

of decapitation and auxin treatment (Table 4). Hence, there

was no immediate effect of high irradiance on exogenous
auxin restoration of apical dominance in decapitated plants.
When 16}17-d-old Ipomoea plants in pots were transplanted to outdoor plots under heavy shading (irradiance :
up to 210 mol m# s"), they appeared, after 1934 d, very
similar in overall form and height (i.e. tall without branching)
to indoor plants of approximately the same age (no data
given). Whereas those plants transplanted to outdoor plots
under full sunlight had, in time, such an extensive
proliferation of lower branches together with reduced height

50

150

Height (cm)

Days following decapitation

264

ClineExogenous Auxin Effects in Decapitated Shoots

Length (mm)

Decap
control
12
0.1%
IAA
8
1% IAA
4
Intact
2

6
Day

10

F. 8.Partial auxin repression of lateral bud outgrowth in Coleus at

reduced irradiance (2530 mol m# s") following decapitation over
10 d. Vertical lines represents.d. n 10.

DISCUSSION
The results of this study corroborated the Thimann-Skoog
(1933) experiment, that exogenously applied auxin to the
stem stump of a decapitated plant does restore apical
dominance for many plants under most conditions. This is
a classic example wherein the results one obtains depend
upon the particular plant system used and upon the
conditions employed. There were exceptions where the
Thimann-Skoog experiment did not work. In those plants
with weak apical dominance such as Coleus or Arabidopsis,
auxin had little or no effect on repressing bud outgrowth in
decapitated plants. Hence, it is understandable why Jacobs
et al. (1959) were not able to detect repressive auxin effects
on the release apical dominance with greenhouse-grown
Coleus plants. In plants with incomplete apical dominance
such as Phaseolus, auxin had only a partially inhibiting
effect on axillary bud growth. Hence, it is understandable
why Hillman (1984) questioned the role of auxin in apical
dominance in this species. Certain environmental conditions
can proliferate branching to an extent which makes it
impossible to carry out the Thimann-Skoog experiment.

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seventh nodes of the shade plants (Fig. 6), there was some
anomalous branching due to the fact that the shoot tips had
impinged against the overhanging shade screen, had bent
over and had released the buds at those nodes. Floral buds
were found to be much more prevalent in the sun plants
than in the shade plants. Since the spectral quality of
sunlight in both groups of plants presumably was the same
(i.e., the plastic shade screens would not be expected to
cause any such changes), the pronounced weakening of
apical dominance as exhibited by the proliferation of
branching in the sun plants could be attributed to increases
in irradiance (fluence) and not to changes in light quality.
When Coleus plants were grown indoors at greatly reduced
light levels (irradiance : 2530 mol m# s"), branching in
intact plants was much decreased compared to that of
greenhouse plants (Fig. 3) but increased lateral bud
outgrowth did occur at the highest node following decapitation. Definite but partial inhibitory effects of 01 and
1 % IAA in lanolin were detected in one out of five
experiments carried out at the reduced light level (Fig. 8).

Increased irradiance levels can greatly weaken apical

dominance (Gregory and Veal, 1957 ; Anderson, 1976).
High wind velocity (via thigmomorphic effects) or alterations in the direction of gravity can also release apical
dominance (Prasad and Cline, 1985 a, b).
Although Thimann, Sachs and Mathur (1971) reported
auxin in aqueous solution to be more effective than in
lanolin with Coleus and Pisum, in this study it was found
otherwise. This might have been due to different methods
used for applying the aqueous solution. Thimann employed
a fine piece of tubing over the end of the stem for continuous
flow whereas two doses of approximately 150 l were
applied daily by pipette to a cotton swab taped to the end
of the stem in the present study (Fig. 1 C). The auxin
concentration (14001700 units) which Thimann and Skoog
(1933) used to completely inhibit bud outgrowth in Vicia
faba is presumed to be equivalent to approximately 1 % in
lanolin or to 610# in aqueous solution (Stafstrom,
1993).
In retrospect, Vicia faba, the plant system which Thimann
and Skoog (1933) used in their classic study, probably was
not the best for studying apical dominance. First, it has
weak apical dominance and many plants had to be excluded
in the present study because they were already branching
before the time designated for decapitation, particularly in
the greenhouse plants. Second, following decapitation, it
was usually the bud at the basal node instead of the highest
node which sprouted first. This complicated the situation
because of the greater distance between the site of auxin
application (the decapitated stem stump) and the potentially
active axillary buds than when the highest bud sprouted
first.
It was observed in some plants where several Vicia buds
sprouted simultaneously that the higher buds located closer
to the site of auxin application were repressed the most. This
suggested that the auxin concentration probably decreased
gradually during transport away from its original source.
However, V. faba was sensitive to auxin and its apical
dominance was restored by auxin application. Those
concentrations of exogenous auxin (011 % IAA) which
inhibited apical dominance release also exhibited toxic
auxin effects.
The Thimann-Skoog experiment is carried out with the
greatest facility in a fast growing plant with large internodes,
moderate to strong apical dominance, easily observed
axillary buds, and where only the highest lateral bud grows
out following decapitation. Of the ten plants tested, Ipomoea
nil conformed most closely to the expectations of the
Thimann-Skoog experiment, followed by Helianthus annuus
which, however, was somewhat hindered by its slow growth
in our greenhouse conditions. Pea (Thomas Laxton) worked
well in many ways except for the uncertainty in which lateral
bud would sprout following decapitation. It was usual for
several buds to sprout simultaneously, which complicated
both the execution of the experiment and the interpretation
of the data.
In Arabidopsis, the complete lack of effect of auxin on
restoring apical dominance following decapitation may be
due in part to the fact that the upright shoot of Arabidopsis
is a floral shoot. All experiments done with other species in

ClineExogenous Auxin Effects in Decapitated Shoots

this study were carried out with vegetative shoots, or at least
shoots without the presence of visible floral buds. For
reasons which are not entirely clear, aging and reproduction
often have a weakening effect on apical dominance (Tamas,
1987).
dgt mutant tomato

Light
When irradiance is high, as outdoors, the lateral buds of
many species (except for those with very strong apical
dominance such as sunflower) will begin to grow out and to
proliferate, thus excluding the execution of the ThimannSkoog experiment which requires lateral buds to be in a
repressed state before decapitation and auxin treatment.
This proliferation of branching in outdoor-grown plants
was observed in the present study of Ipomoea nil, a plant
with moderate to strong apical dominance. From previous
indoor tests, it was clear that once vigorous axillary bud
growth had commenced, it was very difficult to repress, even
with high concentrations of auxin (data not shown). It is
also possible that other factors in the outdoor environment
besides light may have a weakening effect on apical
dominance.
When the irradiance is low, the lateral buds of many
species (except for those with very weak apical dominance
such as Arabidopsis) will be in a repressed state, thus
allowing for the execution of the ThimannSkoog experiment with subsequent inhibition of lateral bud outgrowth. In Coleus, a plant with weak apical dominance

where exogenous auxin is known to have no repressive effect

on lateral bud outgrowth following decapitation at high
irradiance in greenhouse environment (Jacobs et al., 1959),
marginal evidence was found in the present study to support
Thimanns contention (Thimann et al., 1971 ; Thimann,
1977) that auxin did inhibit bud growth when the irradiance
level was reduced (Fig. 8).
The fact that outdoor light control of apical dominance in
Ipomoea could be manipulated wholly by the use of shade
screens suggested that the response under the present
conditions was dependent upon changes in irradiance and
not in spectral differences. This result, while not excluding
the interaction with indirect light quality effects or with
auxin, is also consistent with the nutritional hypothesis of
apical dominance via increased photosynthate availability
for bud growth at high irradiance. The possibility that the
plastic shade screens could affect light quality and, hence,
other physiological processes cannot be ruled out.
Light is one of many stimuli (e.g. gravity, CO , nutrients,
#
etc.) which often will promote axillary bud outgrowth
(Hillman, 1984). The mechanism by which light induces
such outgrowth is unknown. Field and Jackson (1975) point
out some of the complexities involved in interpreting light
effects on apical dominance. Gregory and Veale (1957)
reported that auxin repression of axillary bud outgrowth
increased with decreasing light in Linum particularly at low
nitrogen levels. Thimann et al. (1971) suggested that high
light may promote the synthesis of cytokinins which in turn
may reverse the auxin effect. Accumulating evidence in the
literature for an indirect role of auxin in apical dominance
might also indicate the involvement of certain phytochromemediated processes, interaction with other hormones and
second messengers or by decreasing sensitivity to auxin.
CONCLUSION
The results of this study confirm that the ThimannSkoog
experiment does work for most species and suggests a
controlling role for auxin in apical dominance. Most likely
the role is indirect, perhaps involving cytokinins (Bangerth,
1994 ; Sandberg et al., 1995). It is hoped that the present
accelerating research involving both traditional physiological and molecular approaches with mutants will soon be
able to resolve these problems and more fully elucidate this
classic developmental phenomenon.
A C K N O W L E D G E M E N TS
Appreciation is expressed to Daniel Repicz for his diligent
efforts with the statistical data and graphs, Annabelle Chern
and Liang Shi for their competent assistance with the
preliminary dgt tomato experiments, to the Livingston Seed
Co. of Columbus, Ohio for their gracious donation of Vicia
faba seeds and to the Ohio State University Arabidopsis
Biological Resource Center for their generous help in the
donation of seeds and propagation of seedlings.
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If one accepts the auxin inhibition hypothesis as the

explanation for the mechanism of action of apical dominance then the lack of outgrowth of the axillary buds is
presumed to be due to repression imposed by apicallyproduced}basipetally transported auxin in the shoot. A
mutant such as the dgt tomato which has been demonstrated
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ethylene production (Kelly and Bradford, 1986) might also
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