Phycologia (1997) Volume 36 (3), 195-207

Hierarchical analysis of reproductive potential in

Mazzaella laminarioides (Gigartinaceae, Rhodophyta)
BERNABE SANTELICES AND ENRIQUE MARTINEZ A.

Departamento de Ecologfa, Facultad de Ciencias Biol6gicas, Pontificia Universidad Cat6lica de Chile,

Casilla 114-D, Santiago, Chile

I
BERNABE SANTELICES AND ENR QUE A. MARTiNEZ. 1997. Hierarchical analysis of reproductive potential in Mazzaella lami­
36:
narioides (Gigartinaceae, Rhodophyta). Phycologia 195-207.

Estimations of reproductive effort in seaweeds are important for evaluation of the effects of ecological parameters on fitness
as well as for prediction of colonization capacity and competitive potential. Such estimations must consider several levels
of resolution. At the coarsest scale of analysis, fertile blade abundance must be measured. At a finer level of analysis,
cystocarps and tetrasporangial sori must be quantified within blade populations. At the finest level of analysis, the condition
of the spores, including their size, stage of maturation, and germination potential should be considered. This study tests
whether the integration of these various scales of analysis into a single algorithm might improve the estimations of repro­
ductive potential. Application of this analysis to populations of Mazzaella laminarioides (Bory) Fredericq revealed new
patterns of distribution at the three hierarchical levels analyzed and new trends across hierarchical levels. The pattern of
distribution of fertile blades in the field is characterized by high spatial and seasonal heterogeneity. The size structure of
fertile blades revealed a minimum size necessary to achieve reproductive maturity, and the density per unit area of cystocarpic
and tetrasporangial sori was higher in larger blades than in smaller ones. No clear pattern of maturation of reproductive
structures was found within blades when sampled along axes from apex to base and from side to side. At the finest level
of analysis, it was found that variation in color of reproductive bodies correlated with different spore morphologies and
germination rates, thus suggesting a maturation trend. The resulting algorithm used to estimate reproductive potential inte­
grated infonnation from all three levels of analysis. Reduction of variability by making assumptions of homogeneous
germination potential, homogeneous blade sizes, or homogeneous spatio-temporal distribution significantly influenced esti­
mates of the number of spores produced per unit area of habitat.

INTRODUCTION At a finer level, the abundance of cystocarps and tetraspor­

Reproductive effort in seaweeds is often difficult to measure
(reviewed in DeWreede & Klinger 1988; Santelices 1990).
Reproductive bodies in many species are single celled and can
be distinguished from other kinds of cells only after micro­
scopic observation. In addition, reproductive structures may
have short temporal persistence, making them difficult to de­
tect and measure in the field. In spite of these difficulties,
measurement of reproductive potential is important for eval­
uating the effects of ecological parameters on fitness, coloni­
zation capacity, and competitive potential of a given species.
Within the Rhodophyta, the family Gigartinaceae contains
species with easily distinguishable reproductive structures.
Cystocarps and tetrasporangial sori are visible with the naked
eye, and each sorus can be dissected and its spores released
and counted. Probably due to such methodological advantag­
es, the reproductive biology of this group has received more
attention than has that of other red algae. Studies have been
performed on haploid-diploid ratios, colonization processes,
and potential for spore production (e.g. Mathieson
1975; Hansen & Doyle 1976; Hansen 1977; Bhattacharya
1985; Dyck et at. 1985; Hannach & &
Burns
angial sori must be quantified within individual blades. The
heterogeneity of blade size and potential variation in stages
of maturation must be considered, especially in gigartinacean
species, which frequently show significant size variation
(Martinez & Santelices 1992). At the finest level of analysis,
spore size, maturation state, and potential for germination
should all be considered. Many of these aspects have been
neglected in previous studies, and no one has included all
three levels of resolution in the same study. In this work we
test whether the integration of coarse and finer scales of anal­
ysis into a single algorithm will improve our capacity to es­
timate reproductive potential. We considered the following: 1)
distribution, abundance and size structure of fertile blades in
the field; 2) distribution, abundance, and maturity of repro­
ductive structures (cystocarps and tetrasporangial sori) across
a mature blade; and 3) morphology, density, maturity, and
germination rates of individual propagules (carpospores and
tetraspores).
In doing analyses at different hierarchical levels, more sta­

Santelices 1985; May

1986; Westermeier et at. 1987; McLachlan et al. 1988; Luxoro
& Santelices 1989).
Examination of the variability patterns in the above studies
suggested that surveys on reproductive potential in macroal­
gae must consider several levels of resolution. At the coarsest
scale of analysis, fertile blade abundance must be measured.
tistical predictability and generalization is obtained as one
moves up the hierarchy to larger and larger aggregations,
while 'sweeping under the rug' details of variation within the
aggregate (Levin 1992). The best known cases that show the
effects of changing scales of analysis on resulting patterns are
in the field of spatial structure of patches in foraging inter­
actions (Kotliar & Wiens 1990) and landscape studies (La­
vorel et at. 1993; Wiens 1993); fractal geometry and spectral
analysis are also good examples. For community ecology, the

195
196 Phycologia, Vol. 36 (3), 1997
analysis has focused on the scale at which biotic interactions
are replaced by climatic constraints on large parameters such
as productivity, especially in marine environments (Menge
Olson 1990). In our study we apply a hierarchical approach
to studying the reproductive biology of Mazzaella lamina­
& blade area. Densities of cystocarps and tetrasporangial sori
rioides (Bory) Fredericq, examining it at the level of spores,
blades, and populations. Thus, for one population of this gi­
gartinacean alga, we intend to evaluate the effects of adding
and removing variability of different levels, using a simple
algorithm that translates across scales.
METHODS
Study site and sampling protocols
All samples were collected at Matanza (33°56'S, 71°53'W ) on
the coastline of central Chile. Nine areas of c. 25 m2 each
were chosen as permanent study sites. These areas were dis­
tributed along a homogeneous 1-km long stretch of rocky
shore. In the study sites, Mazzaella laminarioides exhibits
100% cover during most of the year (Santelices
buena 1987).
& Noram­
Within each of the nine study sites, 10 areas, each 900 cm2,
were scraped to remove all blades of Mazzaella laminarioides.
Sampling was done once per season for a year (January, May,
August, and November 1993). The samples were kept cool,
and in the laboratory the blades were sorted into three cate­
gories: cystocarpic, tetrasporic blades, and 'other'. The 'other'
category included all males, infertile females, and females that
were reproductive (e.g. with trichogynes) but had not yet
formed cystocarps. All fertile blades collected were used to
measure reproductive potential.
Distribution of fertile blades
The mean abundances of fertile blades of each karyological
phase from each of the nine sampling areas during each sea­
son were compared using a Kruskal-Wallis test (Siegel
Castellan 1988).
& fertile season (autumn) were measured using light microscopy.
Sizes of fertile blades
Blade length and width were measured for at least half of the
blades collected in each quadrat. Surface area of blades was
estimated using the linear regression between blade area and
length
(Martinez
X&
width dimension, as described in a previous study
Santelices 1992). Intra- and inter-seasonal com­
parisons of size distributions of vegetative and fertile blades
were done using two-sample Kolmogorov-Smimov tests (Sie­
gel &Castellan 1988).
For each season, the mean sizes of fertile blades collected
from sampling areas having more than four fertile blades were
compared using a Kruskal-Wallis test and the a posteriori
Tukey test among sample pairs.
Distribution of reproductive bodies
Each season, about 40% of the surface area of fertile blades
of all sizes and from both karyological phases (n =
22-197)
were examined under a stereomicroscope to score the density
of reproductive bodies. Mean density of cystocarps and te­
trasporangial sori was calculated by counting sori along five
1-cm wide linear transects extending from the base to the tip
of the blade, and then extrapolating these densities to the total
were compared among seasons using a Kruskal-Wallis test.
For all seasons, mean density of reproductive bodies and its
variability (coefficient of variation) was correlated with blade
size, using a linear regression model.
Trends in differential distribution of reproductive bodies
within blades of different sizes were tested using 20 blades of
two extreme size classes: small cystocarpic blades (mean area
= 6.2 cm2, standard error (SE) = 2.3), small tetrasporic blades
(mean area
(mean area
== =
7.1 cm2, SE = 2.2), large cystocarpic blades
blades (mean area
44.2 cm2, SE = 2.3), and large tetrasporic
33.6 cm2, SE = 2.2). For each blade, the
density of reproductive bodies was sampled at four extreme
positions: blade apex (apical portion), blade center (medial
portion), blade base just above the short stipe (basal portion),
and to one side of the medial sample, next to the blade border
(lateral portion). After checking that the assumptions for this
parametric test were met by the data, ANOVA was used to
compare densities at these four positions.
Maturation and fertility of reproductive bodies
A representative number of fertile blades (20-35) from each
season were examined. All reproductive bodies were classified
according to color type as viewed under a stereomicroscope.
The color types distinguished were red, pale red, yellowish,
and colorless. The number of sori of each color found along
five longitudinal sections running from the base to the tip of
each blade was used to calculate total number of sori of a
given color in the blade and in the population of blades at a
given season. To relate color type to the size of the blade, the
abundance of reproductive bodies of each color was surveyed
in the above-described subsamples of large and small fertile
blades.
The diameters of 60 spores of each reproductive phase and
color type from large fertile blades obtained during the most
Spores per reproductive body were counted for 20 cystocarps
and 20 tetrasporangial sori of each color. Cystocarps and sori
were homogenized and the spores were left for 30 min in 0.25
ml of 0.25% NaC!. Then they were centrifuged at 10,000 rpm
for 15 min and resuspended in 0.05 ml of 0.25% methylene
blue. Spores were counted using a Neubauer chamber under
a light microscope.
Germination rates of spores obtained from reproductive
bodies of each phase and color type from blades collected
during autumn (1995) were evaluated after incubation of
spores in 30 Petri dishes filled with sterile and filtered culture
medium for 48 h (Correa & McLachlan 1991). The incubation
conditions for spore germination were 50 j.Lmol m-2 S-1 of
photon irradiance, 13°C, and 12 h of daily light.
Algorithms
Using the information gathered as described above, a single
algorithm was used to obtain estimations of the number of
spores (carpospores and tetraspores per m2) produced season­
ally by the population of M. laminarioides at Matanza. The
values for the two phases and for the four seasons were added
to give an annual reproductive potential (RP) value. The fol-
 Santelices & MartInez: Reproductive potential in Mazzaella laminarioides 197

lowing algorithm was used to incorporate the variability ob­

served at different levels of analysis:
----

0�W 20
--
25 � $
I---G---tJ-i 9/<;>$ 2. 0
= 2: 2:4 RP
16

i=i j=1
sjrijgj!i'
()z 1.6 6 ----

= = rij = = j =
where Sj spores per reproductive body for each of the four
color types (j 1-4), reproductive bodies per blade, for
�
Z::>CD 15 0 --

each blade size interval and color type (i

== ==
gj
1-16, 1-4),
germination rates (0-1) for spores of each color type (j
1-4), and J: blades of each size interval per 1 m2 of �W 10 1.2�
substratum (i 1-16).
In order to incorporate size variation and patch structure of >5 0. 8 �
M. laminarioides in blade density estimations, the mean abun­
dance of blades of all size intervals at each of the nine sam­ �w�
J SU AU WI SP 10.4
=
pled sites (area 0.01 m2) were added. This density value
was then extrapolated to one square meter. After all estima­
tions were done, the final result was expressed as the number
of spores with a capacity for germination that were produced
per square meter. SEASON
In order to evaluate the effect of partial elimination of the
observed variability on the estimations of reproductive poten­
tial, four types of modifications were used to simplify the
Fig. 1. Relative abundance of cystocarpic and tetrasporic phases as
percentage of the number of blades in the population of Mazzaella
laminarioides sampled at Matanza. Total number of sampled blades
was 6800 in summer (SU), 2900 in autumn (AU), 3 100 in winter
described algorithm: 1) elimination of variability in spore den­
sity associated with color type (i.e. assuming that the four
color types of reproductive bodies have similar spore densi­
err
(WI), and 5500 in spring (SP). The ratio is the relative abundance
of fertile cystocarpic and tetrasporic blades.

ties); 2) elimination of variability in density of reproductive

bodies associated with blade size (i.e. assuming that the 16
blade size intervals have a density of reproductive bodies
equal to the average of the three largest blades); 3) steps 1
and 2 combined; and 4) elimination of variability in spore
viability associated with color types (i.e. assuming 100% ger­
mination rate for all spore types). The effect of each simpli­
fication was expressed as deviations (in %) from the result
obtained when all the variability found was included in the
calculation.
RESULTS
Distribution of fertile blades
remained, though in low numbers, only in those areas with
highest autumn representation (areas 8 and 9). By contrast,
fertile cystocarpic blades persisted in low quantities in most
sampling areas (Fig. 2).
Size of fertile blades
The surface area of juvenile blades of M. laminarioides
ranged from 1-2 cm2 to usually less than 100 cm2 (Fig. 3),
whereas the surface area of reproductive blades ranged from
10 cm2 to more than 160 cm2. The size structure of infertile
and fertile blades (Fig. 3) is noticeably skewed to the three
smallest classes. However, there were significantly more larger
mature blades than vegetative ones, irrespective of phase and

The relative abundance of fertile blades of Mazzaella lami­

narioides varied seasonally (Fig. 1); abundance was greatest
(2l.8%) in autumn and lowest (2.6%) in summer. Fertile cys­
tocarpic blades were more abundant than tetrasporic blades in
all seasons and especially in autumn (Fig. 1). The cystocarpic
to tetrasporic ratio (Crr) for the whole population during the
four seasons was significantly different from 1 : 1 (Chi
= = P=
squared test, X2 9.529, df 3, 0.023).
During the seasons of greatest fertility (i.e. autumn and win­
ter), fertile blades of both phases occurred in all the sampling
areas, but they differed in their relative abundance (Fig. 2).
Cystocarpic blades in sampling areas 3 and 5 were almost
twice as abundant during autumn as those from areas 2 and 6
season (Kolmogorov-Smimov tests for two samples, p
0.001). During summer, the population structure became less
skewed as a result of increased representation of the inter­
<
mediate size classes, especially among cystocarpic blades.
Seasonal comparisons of size structure of cystocarpic blades
indicated that the summer population was significantly differ­
ent from populations collected during the other three seasons
(Kolmogorov-Smimov test for two samples, Dmax
=P= =
0.003 for the summer-autumn comparison; Dmax
0.002 for the summer-winter comparison; and Dmax
== = PP
0.309,
0.374,
0.317,
0.023 for the summer-spring comparison). The size dis­
tribution of tetrasporic blades in summer also differed from
other seasons, but only the summer-winter comparison was
statistically significant (Dmax = 0.318, P=
0.034). There were

test, p< = = P=
(Kruskal-Wallis KW 8.795, p 0.032; Tukey a posteriori
0.05). Tetrasporic blades were more abundant during
autumn in sampling areas 8 and 9 than in sampling areas 3
and 5 (Kruskal-Wallis KW = 9.086, 0.028; Tukey a
no significant differences in the size distributions of tetra­
sporic versus cystocarpic blades at any season.
Mean fertile blade area showed high spatial heterogeneity
(Fig. 4). Summer cystocarpic blades of area 8 were signifi­
<
posteriori test, p 0.05).
During the season of least fertility (summer), fertile tetra­
sporangial blades disappeared from most of the beds; they
cantly larger than those of all other areas (Tukey a posteriori
test, p <
0.05), whereas tetrasporic blades did not show sig­
nificant differences among sites (Fig. 4). In autumn, cystocar-
198 Phycoiogia, Vol. 36 (3), 1997

2.5 CYSTOCARPI C BLADES

summer 2. 5 TETRASPORI C BLADES
summer
2.0 2.0
1.5 1.5
2.0 2.0
. 05 0.o.5o �======�����
-?ft.-... 2.0 2. 5 autumn
2.0
U>-Z 2.1.50 1.5
W=> 0.5 2.0
oW0:: 0.2.05 00..50
���� 2. 5 =: ::=
winter 2.0 -------w-in-t-e'r
,---
- - - :=: :: �: ��
u. 2.0
w> 1.5 1.5 I
�<....J 2.0 2.0
W0:: 0.O.o5 LJ 0.5
2.5 spr i n g 0.
2. 0
5 ,--- spr i n g ----�
2.0 2.0
1.5 1.5
2.0 2.0
0.0.05
""' "0-" -
0.o.05 -I-----=-.�-=-,JLULL,.-
1 2 3 4 56 789
+-- "-.-= """'-"' SAMPLING AREAS 1 2 3 4 56 789

Fig. 2.Seasonal variation in the spatial distribution of the relative abundance of fertile blades of Mazzaella laminarioides (relative to all fertile
and infertile sampled blades) in the nine sampling areas at Matanza.

pic blades from areas 2, 4, and 5 were significantly smaller
than blades from areas 6, 7, and 9, whereas tetrasporic blades
from area 6 were significantly larger than blades from all other
sampling areas (Fig. 4). In winter, the size of cystocarpic
blades did not differ significantly throughout the nine sam­
pling areas, but tetrasporic blades did show some significant
differences in extreme sizes. Thus, blades from area 4 were
the smallest and those from area 6 were the largest (Fig. 4).
Significant differences were found in spring: cystocarpic
blades from area 9 were the smallest, those from areas 1 and
4 were the largest, and tetrasporic blades from areas 5 and 9
were smaller than those from area 6 (Fig. 4). Thus, the size
of fertile blades showed seasonal and spatial differences. Fur­
thermore, the data show that the largest blades do not neces-
 &
Santelices Mart{nez: Reproductive potential in Mazzaella laminarioides 199

100 VEGETATIV E BLADES

,--- summer 45 FERTI ----, ,- LE BLADES summer
80 5<=5t8
n=2997 30 J<=35t25
n=44
60 LD
Q7 J< =
n=2724t16
40 15
20Oj�I ��n�� ������ ���

wo 80 100 �--- autumn 45 -� -- �- autumn

z 60
5<=7t10
n=2234 30 ()5<=21t18
�LD5<=21t19 n=197
4:o 40 Q7 n=179
Z 20 15
::J(l)
4:
100 �--- wi n ter ---� winter
80 J<n=2593
=4t5 ()J<=16t14
�LD5<=18t17 n=60
15 Q7 n=80
20
100 ,--- spri n g 45 ----� ,- spring
80 n 5<n=389=8t11 30 ()� 5<=22t16 n=46
60 I LD
Q7 J<=20t12
n=44
40 15
20O
0 30 50S I Z70��n�n�
1BLADE 0.,.-'-"' ,1Jil.3.l...<2- 0 L1'Lrl. 5����---I�-"Y-S0 Inil-"'-,-l-Z7n0El-,l.Ln�(cm2)90���--__I
90E (cm2)1 0 >120 0 +----1BLADE 1 0 >120
Fig. 3. Relative abundance of vegetative and fertile blades of different size classes of Mazzaella laminarioides. Percentages and mean values
refer to sampled blades; ::': value refers to standard deviation. Open bars. cystocarpic blades; hatched bars. tetrasporic blades. Only upper values
are indicated for each size interval on the x-axis.
200 Phycoiogia, Vol. 36 (3), 1997

sarily occur in the areas with highest abundance of fertile

blades of the same phase (compare Figs 2 and 4).
10080 ,-
SUMMER
C: KW
=1 2.3*
Distribution of reproductive bodies
T: KW
=O.3
The density of reproductive bodies varied seasonally (Fig. 5).
Cystocarps were significantly denser in summer and autumn
60
than in winter and spring (Kruskal-Wallis KW
1 X = = 40.9, P <
10-8; a posteriori Tukey test, p < 0.05). Tetrasporangial
40
sori were significantly denser in autumn than in winter or
spring (KW 2 l .8, P < 1 X
10-6; a posteriori Tukey test, p
< 0.05), but there were no significant differences between
summer and autumn, nor between summer and winter or 1 2 3 4 5 6 7 89
spring (a posteriori Tukey test, p >
0.05).
Large blades have a greater density of cystocarpic and te­
trasporangial sori than do small blades (Fig. 6). In addition,
100
-NE 80 ,-
AUTUMN
C: KW
=59.1 *
larger blades have a more even distribution of reproductive
bodies as the coefficient of variation of their abundance cor­
relates negatively with blade size (Fig. 7), at least during au­
-CJ 60 T: KW
=24.0*

tumn, the most fertile season (Y

for cystocarpic blades and Y ==
96.0 - 0.174X, P
76.3 - 0.41X, P == 0.015
0.023 for
tetrasporic blades). No significant differences in density of
<W 40
reproductive bodies were observed among apical, medial, ba­
sal, and lateral parts of small tetrasporic or cystocarpic blades a= O20
(Table 1). Large and small cystocarpic blades show lines of
two to 15 cystocarps at different stages of maturation, sug­ < 100 1 2 3
�+_L¥L
,-4 5 6 7 89

gesting localized waves of maturation across the blade (Fig.

WINTER
8c). Large tetrasporangial blades exhibit significantly more
sori in apical parts of blades than at the base (Table 1), but w 80 C:
T:
KW
=34.6*
KW
=7.4
no linear arrangement of sori was found (Fig. 8d).
c
Maturation and fertility of reproductive bodies

Cystocarps and tetrasporangial sori were sorted into four color

<.....I
+),
types: red (R pale red (R - ), yellowish (Y) and colorless
(C) (Fig. 8a-b). Generally, in small fertile blades « 10 cm2), IX)
yellowish and colorless reproductive bodies predominated
1 2 3 4 5 6 7 89
= ±
orless cystocarps
sori = ±50%
=
== ±± ±
(cystocarpic blades: yellowish cystocarps
21%
61% 4.3, col­
2.7; tetrasporic blades: yellowish
4.1, colorless sori 30% 3.9). In larger
100 ,-
j
blades, red reproductive bodies were the most abundant (cys­ 80 SPRING
C: KW
=1 4.7*
tocarpic blades: red cystocarps 40% 3.2; tetrasporic
T KW
=1 2.S-

error.
= ±
blades: red sori 31%± 3.1). All percentages are standard

The relative abundance of the various kinds of reproductive

60
bodies varied seasonally but only some of the differences were
+
significant (Fig. 9). The R type cystocarps increased signif­
40
icantly from summer to autumn, whereas the opposite trend
+
was shown for the C type cystocarps. The R and C types of
tetrasporangial sori also showed seasonal changes, with higher
1 2 3 4 5 6 7 89
abundance in winter and spring, respectively (Fig. 9).
The various kinds of carpospores and tetraspores (R +, Fig. 4.
Y, and C) exhibited significant differences in diameter (Fig.
R-, SAMPLING AREAS
Seasonal fluctuations in mean blade size of cystocarpic (open
10). Mean carpospore diameter increased from 1 l .6 !Lm (type bars) and tetrasporic blades (hatched bars) of Mazzaella laminarioides
C) to 20 !Lm (type R +);tetraspore diameters ranged from 15
+
!Lm (type Y) to 28.4 !Lm (types R and C). The population
in the nine sampled areas. Significant differences p (*, <
0.01) refer
to Kruskal-Wallis tests applied within cystocarpic (C) and tetrasporic
(T) blades. Vertical lines represent two standard errors.
of C type tetraspores was found to be bimodal, with modes
at 14 ± 1 !Lm and at 27 ±
1 !Lm. These were examined under
the microscope to be certain that the largest colorless spores
did not correspond to undivided tetrasporangia. diameter and density increments (Fig. 11). Carpospore density
Data on spore numbers per cystocarp followed a pattern
similar to diameter, suggesting that maturation implies both
ranged from 442
in the R + ± 450 in C type cystocarps to 3177 2492
type cystocarp. Mean tetraspore density ranged
±
 Santelices & Mart(nez: Reproductive potential in Mazzaella laminarioides �01

30 T 4000 -,-
1:(.)� wc:s .3000
(Dex:wa..
CDena. 20
-

u� 10 1
a..a::« ma.cx:<. 2000 o§b
enu>- -
0I-m 1000 060> . . 0
'
.Ci"

>-0 p'
'

4000 �
o SUMMERI AUTUIMN WINTIER SPRIING 60 (6c0m2) 100
40 AREA
o 20 BLADE
1:(.)� 30 wc:s 3000
CDa::a. COcx:wa..
oen...J 20 <(5Z 2000
(!)«z �a.o. . , . .
�o 10
a.. �I­
1000
�I­WI- WI- o
o SUMMER AUTUMN WINTER SPRING Fig. 6. o 20 40 60 60
BLADE AREA (cm2) 100
Relationship between the per-blade mt;an density of reproduc­

Fig. S. SEASON
Seasonal fluctuation of mean density of cystocarps and tetra­
tive bodies and blade area. Solid lines are curves fitted for actual
values. Segmented lines assume that the mean density of reproductive
sporangial sori of Mazzaella laminarioides. Vertical bars represent two bodies for blades smaller than 10 cm2 is also maintained for larger
standard errors. blades. Data include fertile blades collected in autumn.

from 885
of sori.
±
388 in Y type of sori to 2359±+ 988 in R type + four seasons, 68.2% ±
9.0 of the tetraspores and 63.8%
1104 of the carpospores were produced by blades of a size of
35 cm2 or less. The four modifications of the algorithm with
±
Germination was restricted mainly to R carpospores and
reduced variability at different levels of analysis resulted in
tetraspores and to R - carpospores (Fig 12). The Y type car­
overestimations of the reproductive potential of M. lamina­
pospores and the R - type tetraspores exhibited germination
rioides. These overestimations were slightly lower for the pre­
rates of less than 10%; they do not differ statistically from C
dicted number of tetraspores, ranging from 4%, when the
type spores, which did not germinate.
spore density per reproductive body was homogenized, to
148%, when germination was assumed to be 100% (Table 2).
Integration of information in algorithms
For carpospores, the overestimation ranged from 28%, after
All the observed variability, from spore quality to abundance reducing the variability observed for spore density per repro­
per blade size interval, was considered when the algorithm for ductive body, to 162%, when reduced variability was used for
estimating reproductive potential was used without modifica­ both density of spores and density of reproductive bodies.
tions. Such estimation gave a minimum number of spores pro­ For the algorithm used, the key factor for these estimations
duced annually at the sampled site of 2.9
m-2 ycl and 2.3 X X
108 carpospores
108 tetraspores m-2 yr-I• Averaging the
of reproductive potential was the variability observed in spore
germination rates among the four color types. Thus, maximum
202 Phycoiogia, Vol. 36 (3), 1997

Ta(I) ble 1.
Mean densities of reproductive bodies (per 0.5 cm2) of Maz­

::�
zaella laminarioides in apical (a), basal (b), medial (m), and lateral

o CYSTOCARPS (> parts of blades. Data include density (D) and standard error (SE)
of cystocarps and tetrasporangial sori of small « 10 cm2) and large
20 cm2) reproductive blades. Results of ANOYA (F values and
associated probability p) for the comparison among the four blade
I
portions are included. n = 10 for all values.
-
?fl-Z «oB 0 0
-
200
r=-0.174 D:!:: SE

Small blades
F P
0I- a
m
Cystocarps
27.9 :!::
29.0 :!::!::: 6.8
6.3
0.887 0.457

<0::: b 2 1.6
36.6
7. 1
5.9
<> Tetrasporangial
sori

U.o 0 40 80 120 160

400
a
m
b
56.7:!::
59.0:!::
41.0 :!:: 10.6
8.0
7.1
1.027 0.392

I-ZW TETRASPORIC SORI I Cystocarps

60.3 9.3
Large blades

300
t)U. • • r=-0.168
I
a
m
b
36.3
26.5
36.9
:!::!::: 4.3
1.5
6. 1
2. 154 0. 110

U.W0 •
200 Tetrasporangial
sori
26.6 2.2

() a
m
b
63.2 :!::
47.3
36.4
:!::!::: 4.3
4.4
4.1
5.447 0.003

I 55.2 6.4

The distribution pattern of fertile blades indicates that most

40 AREA80 (cm2)120 160
o BLADE areas are dominated by one phase or the other. Seasonal
changes in fertility essentially correspond to changes in the

Fig. 7. Coefficient of variation for the density of reproductive struc­

tures of Mazzaella laminarioides as a function of blade size during
autumn.
overestimation was obtained for the total number of spores
(carpospores plus tetraspores) when germination was assumed
to be 100%. In this case, the overestimation was 156%, re­
sulting in a predicted number of spores that was one and one­
half times the result obtained when all variability was consid­
ered (Table 2).
DISCUSSION
The results gathered in this study provide new information at
all three hierarchical levels analyzed. Unlike the previous re­
port on the same species in the same sampling area (Santelices
& Norambuena 1987), reproductive blades of both karyolog­
ical phases were present throughout the year and did not show
seasonal disappearance. The difference probably resulted from
the much greater intensity of sampling effort applied in the
present study and suggests that studies measuring reproductive
potential of red algae should include intensive sampling dur­
ing the less fertile seasons to ensure adequate representation
of reproductive stages all year round.
number of fertile blades of a given phase within a given area.
No evidence of displacement of one phase by the other was
observed in this study. Our results suggest a pattern of distri­
bution essentially determined by the pattern of establishment
of the population and maintained by the perennial nature of
the holdfast and basal parts of the thalli (May 1986; Santelices
& Norambuena 1987; G6mez &
Westermeier 1991).
At another level of analysis, we found that blades could
become fertile at a size of about 10 cm2• However, the density
per unit area of cystocarps and tetrasporangial sori was higher
and more uniform in larger blades than in smaller ones. There­
fore, larger blades have more reproductive bodies not only
because they are large, but also because the reproductive bod­
ies are more densely packed. Tetrasporangial sori are signifi­
cantly more abundant at the blade tips. In this species, blade
elongation is by the activity of a meristematic region located
at the base of the blade (Santelices &
Norambuena 1987).
Therefore, tetrasporangia maturation is related to age of the
blade parts, but no clear pattern of maturation from tip to
bottom or from side to side was found in the cystocarpic
blades. The occurrence of lines of cystocarps may suggest
highly localized waves of carpogonial differentiation within
the blade. Alternatively, it may suggest an inhibitive effect of
the maturing cystocarp on neighboring carpogonia such that
a pattern of evenly spaced rows of mature cystocarps is pro­
duced. Even though cystocarpic development has been de-
 Santelices & Martinez: Reproductive potential in Mazzaella laminarioides 203

FigFisg8sa8-ad,. b.
= C.
Reproductive bodies of Mazzaella laminarioides, light microscopy.
Cross sections of reproductive bodies showing the different color types: red = R +, pale red = R -, yellowish = Y, and colorless

FiFigFiFisggg8...c888-cbad....
Cystocarps. Scale bar = I mm. 0.1
Tetrasporangial sori. Scale bar =
Surface view of reproductive bodies.
mm.

Cystocarps are typically arranged in straight or curved rows (arrows). Scale bar = I mm.
Fig. 8d. Tetrasporangial sori are not arranged in lines. Scale bar = Imm.
204 Phycoiogia, Vol. 36 (3), 1997

CYSTOCARPS,------, TETRASPORANGI
SORI RED A L
80 80

60
RED 60

40 40

20

0
SU AU WI SP SU AU WI SP

80 .----R=-IA,::-OL-=E::-cR=-E=O=-- 80
PALE RED
60 60

--� 40
-�-
40

UZw UZw
20

C!SZ::J p U AU WI SP C!S
-,------.Y"'E=L.-L"'O,.....W"""'IS"'H..- Z::J<
0
SU AU WI SP

< 80 80,- -
--------
�� E
�L
I �LO�WW.S
I �H

>w
-
60

40
60

>w I
40
nIiI II
� � I
Wa::....J 20
....Ja::W
20
II
o
SU AU WI SP SU AU WI SP

80 -,----7C70�LO�R�L�E=S�S� 80
COLORLESS
60 60

40 T 40

20

SU AU WI SP
0
a b ab
SU AU WI SP

Fig. 9. SEASON SEASON

Seasonal variation of the relative abundance of the four color types of reproductive bodies of Mazzaella laminarioides. Different letters
indicate significant differences among seasons (p < 0.05,
Tukey a posteriori test). Vertical bars represent 2 standard errors.
 Santelices & Martinez: Reproductive potential in Mazzaella laminarioides 205

--E::1. >­t-
a::w
enZW
.­w
�«
oen
QW
wc:o::
a::o
enC­ ena.
PALE RED YELTYPE
RED COLOR LOWISH COLORLESS RED PALE
RED

COLOR TYPE
YELLOW COLORLESS

Fig. 10. Mean spore diameter of the four color types for carpospores
(open bars) and tetraspores (hatched bars) of Mazzaella laminarioides.
Different letters indicate significant differences among color types (p
Fig. 1 . Mean number of spores per reproductive body of Mazzaella
laminarioides for cystocarps (open bars) and tetrasporangial sori
(hatched bars). Different letters indicate significant differences (p <
< 0.05,
errors.
Tukey a posteriori test). Vertical bars represent 2 standard 0.05, Tukey a posteriori test). Vertical bars represent 2 standard errors.

scribed in several members of the Gigartinaceae [see Hom­ study should be easily applicable to such cases. Results sug­
mersand &Fredericq ( 1990) for review], attention has focused
on the internal development of the carposporophyte and the
cystocarp, with less attention to the physiological and mor­
gest that integrating information from fine to coarse levels of
analysis reduces error by better representation of the sources
of variability at each of the hierarchical levels. Reducing
phological relations of maturing cystocarps in close proximity. sources of the error allows a more realistic estimate of repro­
The different colors of reproductive bodies is an important ductive potential.
new component for analysis. With the exception of the col­
orless tetrasporangial spores, color types correlated with dif­
ferent morphologies and germination rates, all suggesting a
maturation trend. Yellowish and colorless spores are more
100
abundant in small blades and in seasons when fewer blades
are reproductive (summer and spring). When pale spores are
present, spore abundance is low and size is small, and these
80
generally lack the capacity for germination. By contrast, red
and pale-red spores, which are abundant on larger blades and
--
#. 60
during the most fertile seasons, are larger and their germina­
tion rates are higher. The bimodality in size shown by the
colorless tetrasporangial sori included small, immature spores
0z
t:c 40
as well as large, mature but bleached spores. To our knowl­ z
edge, these relationships between spore morphology, appear­
ance, and germination have not been described previously
�a::
within this or related groups of red algae. Ignoring these as­
pects of reproduction has a profound effect on the results de­
rived from the algorithm we used to estimate reproductive
(!)w 20
potential, leading to estimations considerably higher than
those obtained when color types are included. 0
Many kinds of red and brown seaweeds show aspects of
reproductive morphology and field dispersion patterns of fer­
tile individuals that are similar to those of M. laminarioides.
RED REDPALE YELLOW COLORLESS
In many species, propagules are significantly smaller than the
COLOR TYPE
sori, and the sori are scattered over larger blade surfaces, bran­
chlets, or other parts of the thallus. Simultaneous considera­
Fig. 12. Germination rates of spores of Mazzaella laminarioides, in­
cluding carpospores (open bars) and tetraspores (hatched bars) of the
four color types. Different letters indicate significant differences (p<
tion of the several resolution levels involved is required to
measure reproductive potential. The method outlined in this
0.05,
rors.
Tukey a posteriori test). Vertical lines represent 2 standard er­
206 Phycoiogia, Vol. 36 (3), 1997

Table 2. Reproductive potential of Mazzaella laminarioides for each phase and season estimated with the algorithm unmodified (first column)
and with modifications that reduced variability in spore density (I),in density of reproductive bodies (2), in both parameters (3), and in
germination rates (4). Percentages in parentheses indicate the overestimation error involved under each simplification of the algorithm using the
unmodified algorithm as the base figure for comparison. See Methods for further explanation.

Phase and
season Unmodified
Reproductive potential

2
(X 108 spores m-2)

3 4

Cystocarpic
Summer 0.3 0.4 0.3 0.5 0.9
Autumn 1.8 2.3 2. 1 2.7 4.6
Winter 0.5 0.6 0.9 1.2 1.3
Spring 0.3 0.4 0.5 0.6 0.8
Annual totals (%) 2.9 3.7 3.8 5.0 7.6
(28%) (31%) (72%) (162%)
Tetrasporic
Summer 0.2 0.2 0.3 0.3 0.5
Autumn 1.0 l.l l.l 1.1 2.7
Winter 0.8 0.8 0.7 0.7 1.8
Spring 0.3 0.3 0.4 0.4 0.7
Annual totals (%) 2.3 2.4 2.5 2.5 5.7
(4%) (8%) (8%) (148%)
Grand totals (%) 5.2 6. 1 6.3 7.5 13.3
(9%) (2 1%) (44%) ( 156%)

Integration of various scales of analysis into a single al­ REFERENCES

gorithm calls attention to the need for additional field exper­
imental studies to improve estimation of reproductive poten­
BHATTACHARYA D. 1985. The demography of fronds of Chondrus
tial. The present study assumes one wave of spore production crispus Stackhouse. Journal of Experimental Marine Biology and
per season. Thus, colorless and yellow sori are assumed to
remain immature during the season of collection and to dis­
appear from one season to another. These assumptions are
Ecology 91: 2 17-23 1.
CORREA J.A. & McLACHLAN J.L. 199 1. Endophytic algae of Chon­
drus crispus (Rhodophyta). III. Host specificity. Journal of Phycol­

based on the highly seasonal occurrence of fertile blades of

this species in central Chile (Santelices &
Norambuena 1987)
and on the increasing probabilities of blade disruption during
maturation and emptying of an increasing number of sori in
a given blade. However, these assumptions should be tested
in longer term field measurements. Such studies should pro­
vide information on the fate of the spores regarded as im­
mature within a season, on the average longevity of the fertile
blade, and on the potential occurrence of more than one wave
of spores produced by the same sorus. If yellowish and col­
orless spores mature within a season, the value of the ger­
mination factor (g) should be increased accordingly. On the
other hand, if the fertile blades can persist from one season
to another, a correction should be made in the blade density
factor if) to avoid counting in different seasons the same
ogy 27: 448-459.
DEWREEDE R. & KLINGER T.1988. Reproductive strategies in algae.
In: Plant Reproductive Ecology: Patterns and Strategies (Ed. by J.
Lovett-Doust & L. Lovett-Doust), pp. 267-284. Oxford University
Press, Oxford.
DYCK L., DEWREEDE R.E. & GARBARY D. 1985. Life history phases
in Iridaea cordata (Gigartinaceae): relative abundance and distri­
bution from British Columbia to California. Japanese Journal of
Phycology 3: 225-232.
G6MEZ I.M. & WESTERMElER R.C. 199 1. Frond regrowth from basal
disc in Iridaea laminarioides (Rhodophyta, Gigartinales) at Mehufn,
southern Chile. Marine Ecology Progress Series 73: 83-9 1.
HANNACH G. & SANTELICES B. 1985. Ecological differences between
the isomorphic reproductive phases of two species of Iridaea (Rho­
dophyta, Gigartinales). Marine Ecology Progress Series
303.
2:
HANSEN J.E. 1977. Ecology and natural history of Iridaea cordata
29 1-

spores that differ only in stage of maturation (e.g. yellow in

one season and red in the next). Finally, if the longevity and
(Gigartinales, Rhodophyta) growth. Journal of Phycology
402.
HANSEN J.E. & DOYLE W.T. 1976. Ecology and natural history of
13: 395-

the reproduction physiology of the fertile blade allows more

Iridaea cordata (Rhodophyta: Gigartinaceae): population structure.
than one seasonal wave of spore production per sorus, a new
factor should be added to the algorithm.
Journal of Phycology 12: 273-278.
HOMMERSAND M. H. & FREDERICQ S. 1990. Sexual reproduction and
cystocarp development. In: Biology of the Red Algae (Ed. by K.M.
Cole & R.G. Sheath), pp. 305-345. Cambridge University Press,
ACKNOWLEDGMENTS Cambridge.
KOTLIAR N . B . & WIENS J.A. 1990. Multiple scales of patchiness and

This study was supported by a SAREC-CONICYT grant (No. patch structure: a hierarchical framework for the study of hetero­
90-7) to the first author. Our appreciation is expressed to J.
Correa, M. Bobadilla, PSanchez, and M. Venegas for help in
geneity. Gikos 59:253-260.
LAVOREL S., GARDNER R.H. & O'NEILL V. 67:
1993. Analysis of patterns
in hierarchically structured landscapes. Gikos 52 1-528.
the field and in the laboratory analysis, and to A. Munoz for
LEVIN S.A. 1992. The problem of pattern and scale in ecology. Ecol­
improving the manuscript's grammar. We thank Dr J. Jones
and two anonymous reviewers for comments and criticisms
that improved the text.
ogy 73: 1943-1967.
LUXORa C. & SANTELICES B. 1989. Additional evidence for ecological
differences among isomorphic reproductive phases of Iridaea lam-
 & Santelices Mart{nez: Reproductive potential in Mazzaella laminarioides 207

206-21 2.
MARTiNEZ 1 9
E.A. & SANTELICES B. 2. -325:
inarioides (Rhodophyta, Gigartinales) . Journal of Phycology

/ 2 5: 5129-057. .
S ize hierarchy and the
factors in regulation of community structure. Trends in Ecology and
Evolution
SANTELICES B . Patterns of reproduction, d ispersal and recruit­

"Power Law" relationship in a coalescent seaweed. Journal of Phy­ ment in seaweeds. Oceanography and Marine Biology Annual Re­

cology 28: V.259-264. 1 975.

MATHIESON A.C. & BURNS R.L.
28: 17 -276. 1987. 151 152:
Ecological studies of economic
view
SANTELICES B. & NORAMBUENA R. A harvesting strategy for
Iridaea laminarioides in Central Chile. Hydrobiologia
red algae.
3 2 9-3 3 .
Growth and reproduction of natural and harvested
populations of Chondrus crispus Stackhouse in New Hampshire.
1P.J., 98 . 400 1987.
MAY G . 1986. 22: 448-455. 17: 1 37-156.
Journal of Experimental Marine Biology and Ecology
Life history variations in a predominantly gameto­
phytic population of Iridaea cordata (Gigartinaceae, Rhodophyta).
SIEGEL S. & CASTELLAN N . J .

WESTERMEIER R . , RIVERA
Nonparametric Statistics for the
Behavioral Sciences. McGraw-Hill, New York.
CHACANA M .
pp.
& G6MEZ I.
logical bases for management of Iridaea laminarioides Bory in
Bio­

Journal of Phycology
McLACHLAN J.L., LEWIS 198 . 45: 385-397. 19 3. 151 152: 313-328. 3:
N.r. & LAZO M.L. B i ological consid­
erations of Chondrus crispus Stackhouse (Rhodophyta, G igartina­
ceae) in the southern Gulf of St. Lawrence, Canada. Gayana
southern Chile. Hydrobiologia
WIENS I.A. Spatial scaling in ecology. Functional Ecology

29-54.
MENGE 1 9 0.
B .A. & OLSON A.M. 20
Role of scale and environmental Accepted February 1997