Plant Growth Regulation 19: l-5, 1996.

@ 1996 Kluwer Academic Publishers. Printed in the Netherlands.

Indole-3-acetic acid transport in apical dominance: a quantitative approach.

Influence of endogenousand exogenousIAA apical source on inhibitory
power of IAA transport
A.A. Kotov
Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, ul. Botanicheskaya 35, Moscow, 127276,
Russia

Received 10 June 1995; accepted 12 August 1995

Key words: apical and lateral buds, apical dominance, correlative signal, IAA transport, leaves of apical buds,
Pisum sativum

Abstract

The relationship between the amount of indole-3-acetic acid transported (IAA transport) through the second node
of 7-day-old pea seedlings and the degree of inhibition of axillary bud outgrowth at the same node was studied.
For both the endogenous apical IAA source (leaves of apical bud) and the exogenous one (lanolin paste containing
0.25-l .Omg mL- ’ IAA) the slope of linear dependence between inhibitionand IAA transport was similar. However,
the same IAA transport induced different inhibitions, which were higher for the endogenous source. Moreover, the
apical bud induced higher inhibition at the same level of IAA transport when the 4th leaf was present than when it
was absent. Apparently, the source of IAA also may regulate the inhibitory power of IAA transported from it. IAA
transport appears to consists of “active” and “slightly active” one moving along different pathways.

Abbreviations: a and b - coefficients of linear regression of the type y = a + bx; (Y- confidence level of t-test;
ELISA - enzyme linked immunosorbent assay; CR:‘: - growth rate of the lateral bud of experimental/decapitated
(control) pea plants at the first and second days after treatment or decapitation; I - degree of inhibition of lateral
bud outgrowth; IAA - indole-3-acetic acid; Li,2,3 - the lengths of lateral bud at 1, 2 or 3rd day after treatment or
decapitation of pea plants; n - data number; r - correlation coefficient; T - amount of IAA transported through
the second node of pea plant for 3 hours; TIBA - 2, 3, Striiodobenzoic acid; t-test - statistical test used here to
compare slopes of linear regressions (y = a + bx) calculated as t = bl - bz / J[(se b1)2 + (se b2)2].

1. Introduction 2. Materials and methods

It is most likely that apically produced IAA acts in
apical dominance as a correlative signal via its active
basipetal transport [2,7]. Apically produced auxin may
suppress lateral bud growth by inhibiting auxin export
from these buds without penetrating them [lo, 12,
131. Cellular and molecular events contributing to this
mechanism require further investigation, but at present
there is no direct evidence that IAA transport alone is
necessary and sufficient for apical dominance [5, 61.
A quantitative approach to clarify the significance of
IAA transport as a correlative signal from exogenous
as well as endogenous apical sources is described.
The experiments were carried out on 7-day-old pea
(Pisum sativum L.) seedlings cv. Ul’yanovskii 68
grown in a growth chamber at 15 “C with a 14-h photo-
period at a light intensity of 10,000 lux at plant level.
After a treatment of the apical bud the amount of IAA
transported through the second node (IAA transport)
and the inhibition of elongation of the lateral bud at the
same node (inhibition) were determined (Figure 1).
IAA transport was quantified by ELISA [ 151of IAA
in the diffusate from 9-12 shoots. For this purpose, on
the second day after treatment, a plant was cut just
above the 2nd node and its basal end immersed in dis-
 17-day-old pea seedling 1

Leaflets of Treatment 1
apical bud ( excision of treatment
some leaflets (decapita
from apical tion)
bud)
0 Days after treatment
m- decapitation
Lateral ry Site of decapitation
Site of cutting for
diffusate sampling

(ZEl-
Diffusate
sampling

Figure I. Diagramatic view of the treatments used to investigate the relationship between IAA transport and the inhibition of lateral bud growth
in 7-d old pea (P. sarivum) plants.

tilled water (100 PL per plant) for 3 hours in darkness
at 15 “C. In parallel experiments using 9-12 plants, the
growth rate (GR) of the lateral buds and its’ inhibition
(I) were determined during the day preceding (GRi,
Ii) and subsequent (GR2, 12) to diffusate sampling.
Measurements of the lateral bud length were made
with a horizontal microscope at the first (Li), second
(L2) and third (Ls) day after experimental treatment
or decapitation. GRt and GR;! for experimental (GRe)
as well as for control (decapitated) plants (GRd) were
definedasGRi = lnL2 - 1nLi andGR;!= lnL3 - It&.
Ii and 12 were calculated as Ii = [(GR: - GRt)/GR$
x 100% and 12 = [(GR: - GR;)/GR$ x 100%. We
consider I as the mean of Ii and 12 to be a reliable esti-
mate of lateral bud inhibition at the moment of IAA
diffusate sampling.
2.1 Treatment 1
tions. In additional experiments we used plants with
the 4th internode alone representing a fully reduced
apical bud. The third leaf was removed in all exper-
iments. The experiments for both inhibition and IAA
transport determinations were repeated 4 times.
2.2 Treatment 2
In the second series, the apical IAA source was IAA
applied at concentrations ranging from 0.1 to 1.O mg
mL-’ in lanolin paste to the cut surface of the decap-
itated plants. For IAA transport determination, the
experiments were repeated twice. The relationship
between inhibition and IAA transport was interpreted
from their dependence on the IAA concentration
applied (Figure 3). In addition experiments, 10 mg
mL-’ 2,3,5triiodobenzoicacid (TIBA) was combined
with IAA applied at 3 concentrations.

In the first series of experiments, a surgically modi-

fied apical bud was used as a source of apical IAA.
It represented either the 4th or 5th leaf or an upper
apical part (6th and younger leaves) or their combina-