Annals of Botany 94: 913917, 2004

doi:10.1093/aob/mch213, available online at www.aob.oupjournals.org

TECHNICAL NOTE

Statistical Recognition of Random and Regular Phyllotactic Patterns

B E R N A R D J E U N E 1 and D E N I S B A R A B E 2,*
1
Laboratoire de Cytologie Experimentale et Morphogenese vegetale, Universite Pierre et Marie Curie,
Bat. N2, 4 place Jussieu, 75 252 Paris Cedex 05, France and 2Institut de recherche en biologie vegetale,
Jardin botanique de Montreal, 4101 Sherbrooke Est, Montreal, Canada H1X 2B2

Received: 7 June 2004 Returned for revision: 12 July 2004 Accepted: 20 August 2004 Published electronically: 11 October 2004

Aims A statistical method used in ecology is adapted to characterize the degree of order in phyllotactic systems.

Scope The test consists of subdividing a planar projection of the stem apical meristem into 16 sectors and counting
the number of primordia appearing in each. By dividing the sum of squared deviations by the mean number of
primordia per sector the chi-square (c2) is obtained. When there are a total number of 20 primordia, if the c2 is less
than 626, the phyllotaxis is spiral; if it is between 626 and 275 the phyllotaxis is random; and if it is greater than
275, the phyllotaxis is distichous or whorled (level of significance a = 5 %). It is also possible to remove one or more
sectors. If there are k sectors, the two critical values delimiting the random zone will be found in a c2 table for k 1
degrees of freedom.

Conclusions The method is applied to the analysis of sho mutants described by Itoh et al. in 2000 (Plant Cell 12:
21612174). The results obtained are in agreement with the theoretical analysis showing that a whorled or spiral
phyllotactic system may contain a certain number of randomly distributed elements without losing its regular
global structure. 2004 Annals of Botany Company

Key words: Shoot apex, phyllotaxis, models, mutant, chi-square.

INTRODUCTION degree of order in a phyllotactic system. With this formula,

a spiral system will appear as disorganized even though it is
The mechanisms involved in the regulation of phyllotactic
a well-ordered pattern.
patterns have been studied intensively from an experimental
With respect to the problem of entropy, Jean (1994, 1998)
and theoretical point of view (Jean, 1994; Jean and Barabe,
developed a model based on a concept of hierarchy to
1998; Reinhardt and Kuhlemeir, 2001). Phyllotactic sys-
represent the various spiral phyllotactic patterns (m, n),
tems are generally described by using parameters such as
where (m, n) represent the visible parastichy pairs. He
divergence angle, plastochrone ratio, and number of sets of
used an entropy-like function Eb based on the set of hier-
opposed visible spirals (parastichies). Theoretical models
archies to calculate their energetic cost. In this model (Jean,
are based on geometric regularities appearing at the level of
1994), parameter Eb represents the production cost of each
shoot apical meristem (SAM). However, the discovery of
type (m, n) of spiral pattern. The value of Eb increases as the
genes that promote random changes in phyllotactic patterns
number of parastichies increases, and there is no maximum
provides a new perspective as far as the analysis of phyllo-
value. However, in Jeans model it is not possible to deter-
taxis is concerned (Callos and Medford, 1994; Itoh et al.,
mine the entropy of the phyllotactic system without know-
2000; Reinhardt and Kuhlemeir, 2001). The main problem
ing the number of visible opposed parastichies (m, n).
with the analysis of phyllotactic mutants is the character-
Thus, to adequately classify phyllotactic patterns as ran-
ization of phyllotactic systems and their degree of order.
dom a statistical method allowing the degree of order in
Recently Itoh et al. (2000) analysed the pattern of leaf
a phyllotactic system to be characterized is needed. This
initiation in the shoot organization (sho) mutants in rice.
method should be able to quantify with precision the degree
Leaf primordia of sho mutants were initiated at nearly ran-
to which phyllotactic mutants are altered. To solve this pro-
dom positions on the shoot apex with a mean divergence
blem it is proposed to adapt a method, already used in ecol-
angle of 110 . To quantify the degree of order of phyllo-
ogy to study the dispersal of plants or animals, to the field of
tactic patterns appearing in these mutants it was proposed to
phyllotaxis. With this method it is possible to determine,
use partition entropy in the context of information theory.
within a confidence interval, to which phyllotactic pattern
However, it has been demonstrated recently that partition
a given phyllotactic organization belongs. After demonstrat-
entropy is not able to discriminate between an organized
ing how this statistical method can be adapted to study phyl-
spiral system, e.g. with a divergence angle of 1375 , and a
lotactic patterns, it is applied to the quantitative analysis of
random phyllotactic system (Barabe and Jeune, 2004).
sho mutants described by Itoh et al. (2000).
Therefore, partition entropy does not provide an adequate
means to quantify the alterations in phyllotactic patterns.
With the exception of distichous and whorled patterns, the
partition entropy cannot give an exact representation of the METHODS
The following method is derived from a statistical test
* For correspondence. E-mail denis.barabe@umontreal.ca frequently used in ecology to analyse spatial dispersal
Annals of Botany 94/6, Annals of Botany Company 2004; all rights reserved
914 Jeune and Barabe Statistical Recognition of Phyllotactic Patterns
Divergence : 13751
F I G . 1. Theoretical spiral phyllotactic system subdivided into 16 sectors.
Circles represent individual primordia and radiating lines represent sectors.
of plants or animals (Fowler et al., 1998; Lefranc et al.,
2001).
Let x be the number of taxa counted in a quadrat having a
negligible surface area in relation to the territory under
study, and k be the number of quadrats. If the dispersion
of taxa is random, the distribution of x will follow a Poisson
law. Therefore the normal approximation of this law implies
that the ratio
Pk
x 2
i1 xi 

R=

x
is distributed as a c2 having k  1 degrees of freedom (see
Appendix). A confidence interval bounds the zone inside
which the null hypothesis is not rejected (hypothesis of
random dispersal of taxa). Outside the limits of this interval,
if the value of
X
k
SSx = xi 
x2
i1
is very low, the dispersion will be regular. If the value of SSX
is very high, the dispersion will be aggregated (SSX = sum of
squared deviations).
This test can easily be adapted to analyse phyllotactic
patterns. A planar projection of the shoot apical meristem
(SAM) is subdivided into k equal sectors (k = 2, 4, 8, 16,
. . . ), each one containing x foliar primordia (Fig. 1). If all
primordia (n) in SAM are dispersed randomly, x will follow
a binomial distribution:


1
X = B n,
k
Additionally, if n and k are high enough, this distribution
converges to the Poisson law of parameter n/k. When the
T A B L E 1. Comparison of the probability values between a
binomial distribution and a Poisson distribution when n = 20
primordia and k = 16 sectors (X = number of primordia in
a sector)
0 1 2 3 4
P (Binomial) 0.2751 0.3667 0.2323 0.0929 0.0291
P (Poisson) 0.2865 0.3581 0.2238 0.0933 0.0291
value of n is near 20, a value that can easily be obtained
experimentally, and k = 16, the approximation is valid
because the probability values do not differ by more than
1 % between the two types of distribution (Table 1).
Since we know the total number of primordia (n) and the
number of sectors (k), it is of course possible to use the
normal approximation of the exact binomial distribution to
calculate the expectation. However, the degree of precision
obtained by using the binomial law does not compensate for
the increase in calculation necessary to obtain this precision.
As it has been shown above, the Poisson law, estimated by
using the mean number of primordia appearing in each
sector, is completely satisfactory for our purposes.
RESULTS
Table 2 comprises angles of divergence corresponding to
distichous (180 ) or whorled patterns (60 , 120 ), spiral
patterns (7796 , 995 , 13751 , 15114 ), belonging to
normal and anomalous phyllotactic series in the sense of
Jean (1994) and hypothetical spiral patterns (100 , 130 ).
When a SAM containing more than 20 primordia is divided
into 16 sectors (d.f. = 15) (Fig. 1), the different phyllotactic
patterns are readily distinguishable (Table 2). For example,
when d.f. = 15, the random phyllotactic pattern is the only
one appearing in the zone of random dispersion (Fig. 2). The
dots located below the confidence interval correspond to a
uniform dispersion (spiral phyllotaxis), and those located
above the confidence interval to an aggregated dispersion
(distichous or whorled phyllotaxis).
However, when the SAM is divided in four or eight
sectors, aggregated or uniform patterns are observed inside
the limits of the random phyllotactic zone. To obtain a
statistically significant delimitation between different
types of patterns, the number of sectors must be 16 with
20 or more primordia (Table 2). However, when there are 16
sectors and more than 50 primordia it becomes difficult to
statistically separate uniform phyllotactic patterns charater-
ized by even numbers such as 100 from random patterns.
This is not the case for spiral patterns characterized by
angles of divergence of 7796 , 995 , 13751 or 15114 .
The preceding results show it is possible to statistically
differentiate between distichous or whorled patterns (aggre-
gated dispersion), spiral patterns (uniform dispersion) and
random patterns (random dispersion). However, is it still
possible to separate these three types of phyllotactic patterns
 Jeune and Barabe Statistical Recognition of Phyllotactic Patterns 915
T A B L E 2. c2 values for different angles of divergence (top row), different number of sectors ({16}, {8}, {4}) and different number
of primordia (left column) (a = 5 %)

{16} 60 77.96 99.5 100 120 130 137.51 151.14 180 Ran
10 18.8 6 6 9.2 44.4 6 6 6 70 15.6
20 34.4 4 4 4 87.2 5.6 4 2.4 140 13.6
50 83.76 1.84 1.84 5.04 216.88 3.12 1.84 0.56 350 10.8
100 166.88 1.12 0.8 9.12 433.44 4.96 1.12 0.48 700 19.4
{8} 60 77.96 99.5 100 120 130 137.51 151.14 180 Ran
10 4.4 1.2 2.8 2.8 17.2 2.8 1.2 1.2 30 9.2
20 7.2 1.6 0.8 1.6 33.6 3.2 0.8 1.6 60 3.2
50 16.88 0.56 0.24 2.48 83.44 1.2 0.56 0.24 150 5.0
100 33.44 0.32 0.32 4.48 166.72 1.92 0.16 0.32 300 4.8
{4} 60 77.96 99.5 100 120 130 137.51 151.14 180 Ran
10 1.2 0.4 0.4 0.4 3.6 0.4 0.4 0.4 10 2
20 2 0.4 0 0.4 6.8 0 0.4 0.4 20 0
50 5.2 0.08 0.08 0.72 16.72 0.08 0.08 0.08 50 0.72
100 10.32 0.08 0.08 1.44 33.36 0 0.08 0.08 100 3.6

{16} c2 < 626 = spiral; 626 275 = random; c2 > 275 = distichous or whorled.
{8} c2 < 169 = spiral; 169 160 = random; c2 > 160 = distichous or whorled.
{4} c2 < 022 = spiral; 022 935 = random; c2 > 935 = distichous or whorled.
Bold indicates random phyllotaxis; italic indicates distichous or whorled phyllotaxis; normal type indicates spiral phyllotaxis.

40 T A B L E 3. c2 values for 50 primordia of which 10, 20 or 30

60
120 primordia are distributed randomly
35 130
15114 137.51 77.96 99.5 100 130 151.14 60 120 180
Random 10 8.88 4.40 5.68 5.68 2.48 6.18 62 152.24 249.52
30 7796
995 20 8.24 6.18 5.04 6.96 5.68 8.24 38.96 83.76 134.96
100 30 10.16 10.16 6.96 6.96 8.24 6.96 20.40 33.84 57.52
25 180
13751 Values for which the distribution of primordia is not significantly different
2

20 from a random distribution are in bold type (k = 16 sectors; a = 5 %).

15 randomly distributed primordia does not change the pattern

10
5 and whorled patterns of 120 are particularly stable because
0 alter the general type of phyllotaxis.
0 2 4 6 8 10 12 14 16 18
Degrees of freedom
F I G . 2. c2 values for degrees of freedom ranging from 2 to 18. The
confidence interval (a = 5 %) bounds the zone inside which the
hypothesis of a random phyllotaxis is not rejected. Symbols represent
phyllotactic patterns.
when they are mixed with randomly distributed primordia?
This question has been addressed by determining the num-
ber of randomly distributed primordia that could be incor-
porated in a uniform spiral phyllotaxy to statistically obtain
a random phyllotactic pattern. With a total number of pri-
mordia equal to 50, it appears that the number of randomly
distributed primordia needed to modify a particular phyllo-
tactic pattern varies depending on the angle of divergence
(Table 3). For example, when ten primordia are randomly
distributed in a phyllotactic pattern characterized by an
angle of divergence of 13751 , this pattern cannot be sta-
tistically distinguished from a random pattern (Table 3). In
other cases (e.g. 60 , 7796 , 995 , 130 ) the addition of 20
significantly. This indicates that a regular phyllotactic sys-
tem may tolerate a certain amount of disorganization with-
out loosing its global characteristics. The distichous (180 )
the presence of 30 randomly distributed primordia does not
20 This same question can be addressed by adding primordia to
defined sectors within an otherwise random pattern. Consider,
for example, the distichous pattern found in maize (Jackson and
Hake, 1999) and rice (Itoh et al., 2000). Let us assume a
random distribution of 50 primordia, calculated with the
Poisson law of parameter (l = 50/16), and calculate the c2
values when primordia are progressively added in opposed
sectors one and nine (distichous position), and concomitantly
deleted in other sectors at random. It is found that the c2 values
increase proportionally to the number of primordia added
in sectors one and nine. Starting with a completely random
pattern, <20% of primordia (X = 8) are required in a dis-
tichous position (sectors one and nine) to obtain a
regular phyllotactic system, because the value of the c2
(2808 > 2750) falls outside the boundaries of the random
zone (Fig. 3).
A case study: sho mutants
In the sho mutants studied by Itoh et al. (2000), leaf
primordia were initiated at nearly random positions on
916 Jeune and Barabe Statistical Recognition of Phyllotactic Patterns
45
40
35
30
2
25
20
15
10
0 2 4 6 8 10 12 14
Number of distichous leaves
F I G . 3. Increase of c2 values when opposed primordia are regularly added
into sectors one and nine from a random distribution, such that the total
number of primordia remains equal to 50. The line at c2 value of 275 shows
the boundary between random and whorled patterns.
the shoot apex with a mean divergence angle of 110 . To
quantify the degree of order of phyllotactic patterns appear-
ing in these mutants, the method described above was
applied to the data previously published by Itoh et al.
(2000, fig. 6). For the purposes of the present analysis,
each mutant is considered as a single apex. If data were
taken on several apices, an ANOVA test would be necessary
to determine their homogeneity. Of course, in the case
of heterogeneity the present method is not applicable.
Here, only an example of the application of this statistical
test is shown.
With the SAM divided into 16 sectors, the phyllotactic
patterns appearing in mutants correspond statistically either
to an aggregative dispersion (distichous or whorled pattern;
sho2) or a random dispersion (sho1-1 and sho3). The
aggregative pattern is not surprising considering that the
wild-type pattern is distichous, and is suggestive of a develop-
mental constraint exerted by the initial phyllotactic pattern
on the types of patterns that can appear in phyllotactic
mutants. To test this, sectors one and nine, which comprise
the greatest number of primordia, were removed from sho2
data in the analysis. The newly obtained pattern (sho2P)
corresponds to a random phyllotaxis (Fig. 4). It appears that
phyllotactic mutants experience a shift from a distichous to
a random phyllotactic organization. Therefore, in the same
phyllotactic system, one may find regularly or randomly
distributed elements. This is in agreement with the present
analysis which shows that a distichous, whorled or spiral
phyllotactic system may contain a certain number of
randomly distributed elements without losing its regular
global structure.
DISCUSSION
The present analysis presents only a few examples of diver-
gence angles. A more complete table, taking into account a
35
SHO1
30 SHO2
SHO3
25 SHO2P
20
2
15
10
5
0
0 2 4 6 8 10 12 14 16 18 20
Degrees of freedom
F I G . 4. c2 values calculated for sho mutants (sho1-1, sho2, sho3) described
by Itoh et al. (2002) as a function of the degrees of freedom. In sho2P, sectors
one and nine were removed from sho2 data. The confidence interval (see two
lines on graph) (a = 5 %) bounds the zone inside which the hypothesis of
a random phyllotaxis is not rejected.
greater range of angles of divergence could be produced.
However, the purpose of this study was not to determine the
confidence interval for each angle but only to show that
different patterns can be distinguished by using a very
straightforward statistical analysis. This statistical test is
simple and accurate enough to determine a confidence inter-
val for different phyllotactic patterns.
In the preceding section it has been demonstrated that it
is possible to statistically separate three general types of
phyllotactic patterns by using a probability distribution. Of
course, in many cases, it may be easy to qualitatively
distinguish a whorled phyllotactic pattern from a spiral
or random pattern, particularly when whorls do not include
many primordia. However, it is more difficult to distin-
guish between a random pattern and a spiral pattern by
analysing their degree of order. For example, as has been
demonstrated previously, it is not possible to distinguish a
spiral pattern from a random pattern by using the formula
of partition entropy (Barabe and Jeune, 2004). On the other
hand, the use of a simple probability test may distinguish
between a random pattern and a spiral pattern. A statistical
analysis can also bring out a pattern that is not visible
a priori on a planar representation of the SAM. For
example, in the case of sho mutants, the predominace
of a distichous phyllotactic pattern is not recognizable
by visual observation of the schematic alignment of
primordia.
The method described above should be viewed as com-
plementary to already existing phyllotactic models. By
using this method, it is possible to statistically determine
to which phyllotactic pattern (distichous, whorled, spiral or
random) a particular type of phyllotactic organization
belongs. Subsequently, one can use a given deterministic
theoreotical model to quantitatively analyse the relationship
between phyllotactic parameters such as angle of diver-
gence, plastochrone ratio and number of parastichies in
the case of regular systems.
 Jeune and Barabe Statistical Recognition of Phyllotactic Patterns
The method presented above is a statistical tool that can
be used to analyse empirical results. It does not constitute a
theoretical model. However, let us note that there are no
stochastic general models of phyllotaxis. All theoretical
models developed up to now are deterministic and based
on regularities appearing in phyllotactic patterns (Jean,
1994; Jean and Barabe, 1998). It is hoped that the statistical
tool presented here will open new avenues of research for
the development of a probabilistic model of phyllotaxis.
917
Lefranc A. Jeune B, Thomas-Orillard M, Danchin E. 2001. Non-
independence of individuals in a population of Drosophila melanoga-
ster: effects on spatial distribution and dispersal. Comptes Rendus de
lAcademie des Sciences, Paris, Sciences de la vie/Life Sciences 324:
219227.
Rahman NA. 1968. A course in theoretical statistics (reprinted in 1978).
London: Charles Griffin & Co.
Reinhardt D, Kuhlemeier C. 2001. Phyllotaxis in higher plants. In:
McManus MT, Veit BE, eds. Meristematic tissues in plant growth
and development. Boca Raton, FL: CRC Press, 172212.

ACKNOWLEDGEMENTS APPENDIX

We would like to thank Professor Christian Lacroix, Pro- If x follows a Poisson law, where its parameter l is esti-
fessor David Morse and Mr Stuart Hay for their valuable mated by the sample mean
x, then
comments on the manuscript. This research was supported X 
x
by grants from the Natural Sciences and Engineering Z= p

x
Research Council of Canada to D.B.
is approximately a standardized normal variable. The sum
REFERENCES of squares of k independent standardized normal variables
Barabe D, Jeune B. 2004. The use of entropy to analyse phyllotactic follows a c2 law with k  1 and not k degrees of freedom
mutants: a theoretical analysis. Plant Cell 16: 804806. because of the linear dependence of zi:
Callos JD, Medford JI. 1994. Organ positions and pattern formation in the
shoot. Plant Journal 6: 17.
X
k Pk
Fowler J, Cohen L, Jarvis P. 1998. Practical statistics for field biology, i1 xi 
x
2nd edn. Chichester: John Wiley & Sons.
zi = p =0
i1
x
Itoh J-I, Kitano H, Matsuoka M, Nagato Y. 2000. SHOOT ORGANIZA-
TION genes regulate shoot apical meristem organization and the pat-
tern of leaf primordium initiation in Rice. Plant Cell 12: 21612174. This reduces the numbers of degrees of freedom by one.
Jackson D, Hake S. 1999. Control of phyllotaxy in maize by the abphyl1 Then
gene. Development 126: 315329.
Jean RV. 1994. Phyllotaxis: a systemic study in plant morphogenesis. Pk
Cambridge: Cambridge University Press. i1 xi 
x2
Jean RV. 1998. Elementary rules of growth in phyllotaxis. In: Jean RV, R=
Barabe D, eds. Symmetry in plants. Singapore: World Scientific,
x
601618.
Jean RV, Barabe D. eds. 1998. Symmetry in plants. Singapore: World follows approximately a c2 law with k  1 degrees of
Scientific. freedom (Rahman, 1968).