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INTRODUCTION
The monotypic genus Canacomyrica is an endan-
gered shrub or small tree, endemic to the ultramafic (ser-
pentine) soils of New Caledonia (Herbert, in press).
Since its description by Guillaumin (1941; see also
Guillaumin, 1939), who placed it in Myricaceae, there
has been confusion over its morphology and doubt about
its affinity. Prior to this study, little research had been
carried out into the biology of this species and basic
information, such as chromosome number, was un-
known.
Most modern taxonomists consider Myricaceae to
comprise three genera: Myrica L. (ditypic); Comptonia
L’Hérit. ex Aiton (monotypic); and Morella Lour. (c. 50
species) (Wilbur, 1994; Polhill & Verdcourt, 2000;
Herbert, 2005a). All three genera share features such as a
simple bifid style, no perianth and an orthotropous ovule.
In his original description, Guillaumin (1941) listed a
number of anomalous features that distinguish
Canacomyrica monticola from all other Myricaceae
species: androdioecious breeding system (male flowers
and hermaphrodite flowers on separate plants); lamellu-
lar, laciniate style; fleshy perianth; inferior ovary; and
ovule hanging from a long funicle with an inferior
micropyle. He later modified his description, stating that
the inflorescences were co-sexual and the ovule anat-
ropous (Guillaumin, 1948).
In a study of the floral morphology of Canaco-
myrica, Leroy (1949) showed that Guillaumin had incor-
rectly described several characters, stating that the flow-
ers were unisexual with the female flowers bearing ster-
ile anthers and the ovule was erect, sessile, and
orthotropous with a superior micropyle. On the basis of
this re-evaluation of morphology, Leroy (1949) accepted
the placement of Canacomyrica in Myricaceae. How-
ever, in recognition of the remaining unusual features he
placed it in the new subfamily Canacomyricoideae. The
work of Leroy (1949) appears to have been largely over-
looked, resulting in continued misinterpretation of the
morphology, particularly regarding the true nature of the
flowers (e.g., Macdonald, 1977, 1989; Cronquist, 1981;
but see Kubitzki, 1993b).
Several authors (e.g., Elias, 1971; Raven & Axelrod,
1974) have questioned the inclusion of Canacomyrica in
Myricaceae based on its distinctive morphology and dis-
tribution (Myricaceae as previously circumscribed does
not extend into Australasia). Thorne (1973) placed it in
349
Herbert & al. • Systematic position of Canacomyrica55 (2) • May 2006: 349–357
Nuclear and plastid DNA sequences confirm the placement of the enigmatic
Canacomyrica monticola in Myricaceae
Jane Herbert1, Mark W. Chase2, Michael Möller3& Richard J. Abbott4
1School of Biology, Sir Harold Mitchell Building, University of St. Andrews, KY16 9TH, U.K.
janeherbert2@yahoo.co.uk (author for correspondence).
2Molecular Systematics Section, Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9
3DS, U.K.
3Royal Botanic Garden, Edinburgh, 20A Inverleith Row, Edinburgh, EH3 5LR, U.K.
4School of Biology, Sir Harold Mitchell Building, University of St. Andrews, KY16 9TH, U.K.
Phylogenetic analyses of DNA sequences of the plastid rbcL gene were used to obtain a phylogenetic frame-
work for the New Caledonian endemic genus Canacomyrica (monotypic). A further analysis of selected gen-
era within Fagales combining DNA sequences of the rbcL gene, plastid trnL-F region and nuclear ITS region
was also performed. In all analyses Canacomyrica fell into a well-supported clade in which it occupied a posi-
tion as sister to the remaining genera of Myricaceae. A chromosome number of 2n = 16, consistent with
Myricaceae, is reported for Canacomyrica for the first time. On the basis of the phylogenetic data and numer-
ous shared morphological features, the original placement of Canacomyrica in Myricaceae is accepted.
Canacomyrica is distinguished from other members of the family by the presence of staminodes in the female
flower, a six-lobed perianth and a lamellular, laciniate style. The affinities of Myricaceae within Fagales are
re-evaluated in the light of the unusual morphological features of Canacomyrica. The significance of stamin-
odes in Canacomyrica is discussed with reference to pollination syndrome and incomplete suppression of male
function in Myricaceae. A lectotype is designated for the name Canacomyrica monticola Guillaumin.
KEYWORDS:
Canacomyrica, dioecy, Fagales, Myricaceae, New Caledonia, rbcL sequences, staminodes.
his list of Taxa Incertae Sedis, later including it in Myr-
icaceae without comment (Thorne, 1992a, b, 2000).
Others have accepted the original placement of Canaco-
myrica (Cronquist, 1981; Kubitzki, 1993b; Takhtajan,
1997). The recent placement of Canacomyrica in a sepa-
rate family (Canacomyricaceae; Doweld, 2000) added
little insight into the problem of the systematic position
of this taxon.
To date Canacomyrica has not been included in a
phylogenetic study, but it has been suggested that its
anomalous features may be primitive in Myricaceae
(Macdonald, 1989; Carlquist, 2002). If this is the case,
then a better understanding of the relationship of
Canacomyrica to Myricaceae may shed light on the
affinities of the family. Based on molecular data,
Myricaceae are placed in Fagales (sensu APG, 1998,
2003) in the nitrogen-fixing clade, which are nested
within the eurosid I clade (fabids) of the eudicots (Chase
& al., 1993; Soltis & al., 1995, 2000; Swensen, 1996 ;
APG, 1998, 2003). The expanded concept of Fagales
comprises Nothofagaceae (sister to the rest of the order),
Fagaceae, Ticodendraceae, Betulaceae, Casuarinaceae,
Juglandaceae (including Rhoipteleaceae) and Myrica-
ceae. With one exception (Maggia & Bousquet, 1994),
all recent molecular studies of Fagales have reported
Fagaceae (s.s.) as sister to the remaining five families
(the core “higher” hamamelids of Manos & Steele,
1997). Manos & Steele (1997) showed Myricaceae to be
sister to Betulaceae, Ticodendraceae and Casuarinaceae.
Li & al. (2002) placed it sister to all other core “higher”
hamamelids. In an analysis that sampled widely from
throughout Fagales, Li & al. (2004) placed Myricaceae
as sister to only Juglandaceae.
Many studies have employed phylogenetic analysis
of DNA sequences of the plastid rbcL gene (ribulose bis-
phosphate carboxylase/oxygenase, large subunit) to as-
sess the affinities of taxonomically enigmatic angio-
sperm taxa (e.g., Fay & al., 1997; Chase & al., 2002;
Längström & Chase, 2002; Sosa & Chase, 2003; Whit-
lock & al., 2003). A wide range of rbcL sequences are
now available (Savolainen & al., 2000), permitting
analyses across a diverse array of flowering plants.
In this study we used rbcL sequences to investigate
the phylogenetic relationships of Canacomyrica. We
chose to combine rbcL sequences with two more variable
regions: the plastid trnL-F region (consisting of the trnL
intron and the trnL-trnF intergenic spacer) and the nucle-
ar ITS region (consisting of the internal transcribed spac-
er region of the 18S-5.8S-26S nuclear ribosomal cistron).
Many studies have shown the trnL-F region to be useful
in resolving relationships above family level (e.g.,
Richardson & al., 2000; Li & al., 2002). Although ITS
has been widely used for resolving sub-familial level
relationships (Alvarez & al., 2003), it is seldom used at
higher taxonomic levels due to alignment difficulties.
The availability of ITS sequence data for Fagales species
motivated its use in this study as an additional source of
potentially informative characters from a different
genome.
MATERIALS AND METHODS
Plant material. —
Silica gel-dried leaf material
(Chase & Hills, 1991) was obtained for C. monticola,
Myrica hartwegii Watson, Morella cordifolia (L.)
Killick, and Comptonia peregrina (L.) L’Hérit. Voucher
information and GenBank accession numbers for plant
material used in this study are listed in the Appendix.
DNA was extracted using a 2X CTAB method adapted
from (Doyle & Doyle, 1990), modified to include a wash
with ammonium acetate (7.5 M NH
4
AC; Weising & al.,
1995) to remove impurities co-precipitated with the
DNA.
PCR amplification and sequencing. —
Ampli-
fication of the rbcL gene was carried out in two overlap-
ping fragments using the forward primers 1F and 636F
and the reverse primers 724R and 1460R (Fay & al.,
1997). Amplification of the ITS1-5.8S-ITS2 region was
carried out using the forward primer ITS5 and the
reverse primer ITS4 (White & al., 1990). Amplification
of the trnL-F region was carried out using the forward
primer c and the reverse primer f (Taberlet & al., 1991).
PCR reactions of 50 !l contained: 2 !l DNA tem-
plate, 0.2 mM of each dNTP, 0.3 !M of each primer, 2
units Taq polymerase (Bioline, London, U.K.), 2 mM
MgCl
2
, and 5 !l reaction buffer (160 mM (NH
4
)
2
SO
4
,
670 mM Tris HCl, 0.1% Tween 20, pH 8.8). The follow-
ing PCR profile was used for both rbcL and trnL-F: 1
cycle at 94ºC for 4 min; 30 cycles at 94ºC for 45 s, 55ºC
for 45 s and 72ºC for 3 min; 1 cycle at 72ºC for 10 min.
The PCR profile used for the ITS region was as follows:
1 cycle at 94ºC for 3 min; 30 cycles at 94ºC for 1 min,
55ºC for 1 min and 72ºC for 1 min 30 s; 1 cycle at 72ºC
for 5 min. The resulting PCR products were purified
using QIAquick purification kits (QIAGEN Ltd., Craw-
ley, U.K.) according to the manufacturer’s instructions.
Sequencing reactions of 20 !l contained the CEQ
™
DTCS Quick Start Kit (Beckman Coulter Ltd, High
Wycombe, U.K.) and the following primers: the same
primers as in the PCR for the rbcL gene; primers ITS2,
ITS3, ITS4 and ITS5 (White & al., 1990) for the ITS
region; and primers c, d, e and f (Taberlet & al., 1991) for
the trnL-F region. Reactions were performed using the
following PCR profile: 25 cycles at 96ºC for 10 s, 50ºC
for 5 s, 60ºC for 4 min. The sequence reaction products
were cleaned according to manufacturer’s instructions
before being run on a CEQ
™
8000 Genetic Analysis
Herbert & al. • Systematic position of Canacomyrica 55 (2) • May 2006: 349–357
350
System (Beckman Coulter Ltd, High Wycombe, U.K.).
Forward and reverse sequences were manually assem-
bled in Chromas version 2.12 (Technelysium Pty. Ltd.,
Helensvale, Australia).
Alignment and analyses. —
A preliminary study
of over 500 eudicot taxa using the Savolainen & al.
(2000) matrix was first performed to establish the broad-
scale relationships of Canacomyrica (results not shown).
Based on these results an rbcL dataset of 35 taxa (the
“narrow rbcL” dataset) was assembled from the fabid
(eurosid I) clade. Fabales taxa were chosen as outgroup.
New sequences included in the narrow rbcL dataset are
listed in Appendix; the remainder were previously pub-
lished (Savolainen & al., 2000).
A combined dataset of rbcL, ITS and trnL-F se-
quences focusing on 11 Fagales taxa (the “combined
Fagales” dataset) was assembled using new sequences
produced in this study and previously published
sequences (Appendix). Following the approach of
authors such as Wiens (1998) and Reeves & al. (2001) it
was considered appropriate to combine these datasets
after separate analyses showed there to be no strongly
supported (>85%) incongruent clades among the individ-
ual datasets (results not shown). Fagaceae taxa were used
as outgroup in this dataset. Alignment of all sequences
was carried out by eye following the guidelines in
Kelchner (2000). Alignment of rbcL sequences required
no gaps. Alignment of the trnL-F sequences required the
insertion of gaps. Alignment of the ITS sequences
required the insertion of gaps and the exclusion of some
regions where alignment was ambiguous. An aligned
matrix may be obtained on request from MWC (m.chase-
@kew.org.)
Phylogenetic analyses were performed using PAUP*
Herbert & al. • Systematic position of Canacomyrica55 (2) • May 2006: 349–357
351
Table 1. Comparison of morphological characters of Fagales (excluding Nothofagus) (Goldberg, 1986; Wilson &
Johnson, 1989; Kubitzki, 1993a, b; Zheng-yi & al., 1999).
Character Fagaceae Myricaceae Canacomyrica Juglandaceae Betulaceae Casuarinaceae
N-fixing root Absent Present Unknown Absent Present Present
nodules
Stipules Present Absent or Absent Absent (present Present Absent
foliaceous in Rhoiptelea)
Style Linear, short Linear, Lamellular, Plumose or Linear, Linear,
elongate laciniate lamellular elongate elongate
Inflorescence Usually lax Erect, spicate Erect, spicate Lax Lax Erect,
structure spicate
Ovule Anatropous Orthotropous Orthotropous Orthotropous Anatropous Orthotropous
(hemitropous
in Rhoiptelea)
Fruit size and Nut (>1 cm Drupe (to 3 cm Drupe (to 5 mm Winged nutlet (to 9 Winged nutlet (to Samara (2–12
structure long) borne diam.) usually diam.) with a mm diam.) or large 5 mm diam.) or mm long) en-
in cupule covered with smooth fleshy nut (to 6 cm diam.) nut (to 2 cm closed in woody
fleshy papillae pericarp with much lobed diam.) enclosed bracteoles
cotyledons in enlarged bracts
Leaf form Simple Simple or Simple Pinnately or impari- Simple Simple, reduced
pinnatifid pinnately compound
Tepals/perianth 6 tepals Absent 6 lobed perianth 0–4 tepals (0–)1–4(–6) tepals Absent
Integuments 2 (1) 1 2 1 (2) 1, 2 2
Chromosome nr. x = 12, 11, 22 x = 8 x = 8 x = 16 x = 8, 14 x = 8, 14
Fig. 1. One of the 12 most parsimonious trees produced
from analysis of the narrow rbcL dataset. Branch lengths
are shown above the branches (DELTRAN optimization),
bootstrap percentages "50 are shown in bold below the
branches. Asterisks indicate groups not present in the
strict consensus tree.
*
*
11
96
89
69
97
74
53
65
73
69
81
94
64
57
99
50
58
64
97
96
95
92
73
68
24
72
14
3
7
7
8
7
721
28
21
28
35
48
27
11
11 20
34
721
44
19
8
42
7
7
8
10
11 3
5
2
7
1
81
7
5
5
7
27
4
11
11
5
7
6
8
10
13
15
20
9
20 310
9
7
3
Morella
Myrica
Comptonia
Canacomyrica
Juglans
Carya
Rhoiptelea
Alnus
Carpinus
Corylus
Betula
Casuarina
Allocasuarina
Ceuthostoma
Gymnostoma
Ticodendron
Fagus
Quercus
Trigonobalanus
Chrysolepis
Nothofagus
Celtis
Humulus
Cecropia
Ficus
Ulmus
Elaeagnus
Rhamnus
Dryas
Rosa
Spiraea
Cucurbita
Datisca
Pisum
Albizia
Myricaceae
Juglandaceae
Betulaceae
Casuarinaceae
Ticodendraceae
Fagaceae
Nothofagaceae
Celtidaceae
Cannabaceae
Urticaceae
Moraceae
Ulmaceae
Elaeagnaceae
Rhamnaceae
Rosaceae
Cucurbitaceae
Datiscaceae
Fabaceae (outgroup)
Fagales
Rosales
Cucurbitales
100
100
100
Version 4.0b10 (Swofford, 1998). Maximum parsimony
analysis was carried out with tree-bisection-reconnection
(TBR) and saving multiple trees (MulTrees). Maximum
parsimony trees were obtained using the heuristic search
option for the narrow rbcL dataset (Fitch parsimony).
The small size of the combined Fagales dataset made it
possible to carry out a branch and bound search. Branch
lengths were calculated using the delayed transformation
(DELTRAN) optimization.
Support for the clades obtained was assessed using
bootstrap analysis (Felsenstein, 1985) performed using
the heuristic search option and 1000 replicates for the
narrow rbcL dataset and using the branch and bound
search option and 100 replicates for the combined
Fagales dataset. The following categories were used to
describe bootstrap percentage (BP) results: 50–74, weak
support; 75–84, moderate support; 85–100, strong sup-
port.
Chromosome count. —
Root tip preparations
were made from living seedlings of Canacomyrica
housed in the research collection at the Royal Botanic
Garden, Edinburgh (RBGE). Root tips were treated, fol-
lowing the protocol of Jong & Möller (2000), in
colchicine (0.05%) for 4 hours at room temperature, or in
saturated aqueous 1-bromonapthalene for 2 to 8 hours at
room temperature, or in 0.002 M 8-hydroxyquinoline for
4 to 8 hours at 12ºC. They were variously stained in
Feulgen’s reagent (Fox, 1969), lacto-propionic orcein or
aceto carmine, but no method gave satisfactorily intense
staining. After the root tips had been softened in an en-
zyme solution of 5% pectinase and 5% cellulase at 35ºC
for 20 min and mounted on slides, they were viewed
using phase-contrast (Zeiss, Axiophot) at 40#1.6 or 100#
magnification. Images were captured digitally.
RESULTS
Sequencing. —
The narrow rbcL matrix contained
35 taxa and 1343 characters, of which 206 were poten-
tially parsimony informative. Maximum parsimony
analysis of the narrow rbcL dataset produced 12 most
parsimonious trees (tree length 869 steps, CI = 0.51, RI
= 0.58). One of the 12 trees is shown in Fig. 1 with
branch lengths above the branches (DELTRAN opti-
mization) and bootstrap percentages (BP) equal to or
greater than 50 shown below the branches.
In all trees Canacomyrica is sister to the other
Myricaceae taxa in a strongly supported clade (92 BP).
In Fig. 1 (asterisks indicate nodes not present in all trees)
and in the strict consensus tree (not shown), Myricaceae
form a polytomy with two other clades, the first com-
prising Juglandaceae (including Rhoiptelea) and the sec-
ond Betulaceae, Casuarinaceae and Ticodendraceae.
The combined Fagales matrix contained 11 taxa and
2728 characters, of which 210 were potentially parsimo-
ny informative. The rbcL data contributed 51 potentially
parsimony informative characters (total 1286 charac-
ters), the trnL-F data contributed 75 potentially parsimo-
ny informative characters (total 1014 characters) and the
ITS data contributed 84 potentially parsimony informa-
tive characters (total 428 characters). In our combined
analysis 40% of informative characters were from the
ITS region. Of a total length of 664 aligned characters
(including gaps) sequenced for the region, we were
unable to use 236 characters (36%; 183 at the start of
ITS1, 53 at the start of ITS2) due to alignment difficul-
ties. Although the ITS region is seldom used above the
genus level (but see Loockerman & al., 2003; Muellner
& al., 2003) our results, in accordance with the findings
of Coleman (2003), suggest that this region has utility at
higher taxonomic levels within Fagales. Maximum parsi-
mony analysis of the combined Fagales dataset produced
a single most parsimonious tree (tree length 754, CI =
0.82, RI = 0.67). Figure 2 shows the single most parsi-
monious tree with branch lengths (DELTRAN optimiza-
tion) above the branches, and bootstrap percentages
equal to or greater than 50 shown below the branches.
Herbert & al. • Systematic position of Canacomyrica 55 (2) • May 2006: 349–357
352
Fig. 2. The single most parsimonious tree produced from
analysis of the combined Fagales dataset. Branch
lengths are shown above the branches (DELTRAN opti-
mization), bootstrap percentages "50 are shown in bold
below the branches. Asterisks indicate groups with boot-
strap percentage of less than 50.
Myricaceae
Juglandaceae
Betulaceae
Casuarinaceae
Fagaceae (outgroup)
Fagus
Quercus
Casuarina
Carpinus
Alnus
Rhoiptelea
Juglans
Canacomyrica
Comptonia
Myrica
Morella
2
91
15
12
8
29
100
41
48
100
13
33
47
31
100
41
56
33
18
*157
36
97
71
71
10
99
*
In the single most-parsimonious tree, Canacomyrica
is sister to a strongly supported clade (100 BP) compris-
ing the rest of Myricaceae. Monophyly of Myricaceae s.l.
(including Canacomyrica) is also strongly supported
(100 BP). Myricaceae s.l. are sister to Juglandaceae, but
this relationship received bootstrap support of less than
50%.
Chromosome count. —
The root material avail-
able for this study was of inferior quality due to the dif-
ficulty of cultivation for this rare material, which subse-
quently died. Under phase-contrast, chromosomes were
clearly discernible in a small number of metaphase or
pro-metaphase cells. Figure 3 shows a total of 16 chro-
mosomes (2n= 16) counted for Canacomyrica montico-
la.
DISCUSSION
Systematic implications. —
In all our analyses
Canacomyrica formed a strongly supported monophylet-
ic group with Myricaceae s.s. The combined use of DNA
sequences from three different regions gave greater boot-
strap support for this group than the analysis of rbcL
alone (100 BP versus 92 BP). There was insufficient
variability within the DNA markers used to resolve the
sister group of Myricaceae. In the single most parsimo-
nious tree produced by the combined analysis,
Juglandaceae were sister to Myricaceae s.l. (including
Canacomyrica), although this relationship received boot-
strap support of less than 50%. Li & al. (2004) recovered
a sister group relationship between Myricaceae (exclud-
ing Canacomyrica) and Juglandaceae with weak boot-
strap support (66 BP) and high Bayesian posterior prob-
ability (0.95 PP). A morphological phylogenetic study by
Hufford (1992) also supported a sister group relationship
between Myricaceae and Juglandaceae (but not
Rhoiptelea), and a close relationship between these fam-
ilies has been recognized previously by several authors
(e.g., Hjelmqvist, 1948; Leroy, 1949; Melchior, 1964;
Cronquist, 1981; Macdonald, 1989; Thorne, 1992a, b,
2000; Kubitzki, 1993b).
The principal differences between Myricaceae s.l.
and Juglandaceae are leaf form, inflorescence structure,
fruit morphology and size (Table 1). The two families
share the synapomorphies of chains of cuboidal crystal-
containing cells in the wood (Carlquist, 2002) and aro-
matic resin glands. The lamellular, laciniate style and
fleshy perianth that differentiate Canacomyrica from the
rest of Myricaceae are characters that can be found in
Juglandaceae (Table 1). The ovule in Canacomyrica is
bitegmic (Herbert, 2005b; A. Doweld, pers. comm.), a
feature that is shared with Rhoiptelea and may be sym-
plesiomorphic in Fagales.
The sister relationship of Canacomyrica to the other
Myricaceae genera, indicated in this study, is consistent
with the original proposal to include it in the family
(Guillaumin, 1939, 1941). Myricaceae are generally de-
fined by simple, entire (pinnatifid in Comptonia) leaves
with resinous (usually aromatic) “balloon” glands
(Chourey, 1974), short, erect inflorescences, solitary
ovules and small drupaceous fruits with small (typically
<10 mm diameter) seeds. Canacomyrica shares all these
features with other genera of Myricaceae. Furthermore,
studies have shown that Canacomyrica has pollen with a
Myrica-type aperture (Sundberg, 1985) and myricaceous
wood anatomy (Carlquist, 2002). The chromosome count
reported here for Canacomyrica (2n = 16) is consistent
with counts a basic chromosome number of x= 8 report-
ed for other members of Myricaceae, e.g., Myrica 2n=
16, 48, 96 (Löve, 1980, 1982; Morawetz & Samuel,
1989; Al-Bermani & al., 1993), Comptonia 2n = 32
(Löve, 1982), and Morella 2n= 16 (Oginuma & Tanaka,
1987).
The sister relationship between Canacomyrica and
Myricaceae s.s. is also consistent with the opinion of
Leroy (1949), who argued for subfamily status for
Canacomyrica. The presence of a long terminal branch
for Canacomyrica (Fig. 2) indicates significant diver-
gence between the two lineages as might be expected
Herbert & al. • Systematic position of Canacomyrica55 (2) • May 2006: 349–357
353
Fig. 3. Chromosomes of
Canacomyrica
monticola. Photo-
graphs A, B, and C are of the same cell taken at different
depths of focus to capture all the chromosomes present:
(A) clearly shows two chromosomes, these can be seen
less clearly in (B) overlaid by a third chromosome; (B)
clearly shows three chromosomes; (C) clearly shows ele-
ven chromosomes and less clearly the position of the
cluster of three shown in (B). Total = 16 chromosomes.
(D) Drawing of the metaphase spread illustrated in A to C,
showing the relative position of the 16 chromosomes.
given its unique morphological features. On the basis of
the strongly supported monophyly of Myricaceae s.l. in
our analyses and the numerous morphological features
shared between Canacomyrica and Myricaceae,
described above, we accept the original placement of
Canacomyrica in Myricaceae.
Dioecy in Canacomyrica. —
Many authors (e.g.,
Macdonald, 1977; 1989; Cronquist, 1981; Zomlefer,
1994) have continued to follow Guillaumin’s (1941)
description of Canacomyrica in which he stated that
there are both hermaphrodite and male flowers, despite
later studies showing that the flowers are unisexual with
female flowers bearing six sterile anthers or staminodes
(Kubitzki, 1993b; Leroy, 1949). Based on observations
in the field and examination of a total of 50 flowers from
16 individual plants (Herbert, 2005b), we confirm that
male flowers and functionally female flowers are found
on separate plants in Canacomyrica. The confirmation
that Canacomyrica is dioecious is consistent with its
placement in Myricaceae (at least 75% of species are
dioecious), but staminodes are unknown in the rest of the
family.
In Canacomyrica, the catkin-like inflorescence, the
lax attachment of the anthers and the laciniate stigma
indicate that the plant is wind-pollinated. It could be
argued, however, that the staminodes in Canacomyrica
function as attractants for generalist pollinators.
Although Myricaceae are generally considered to be an
anemophilous family, in Hawai’i it has been observed
that introduced honey bees (Apis mellifera) visit the
flowers of Morella faya (Aiton) Wilbur (Vitousek &
Walker, 1989). Elsewhere, among the predominantly
wind-pollinated Fagales, staminodes have been recorded
in Lithocarpus Blume, Triganobalanus Forman, Quercus
subg. Cyclobalanopsis (Oersted) C. K. Schneider and all
Castanoideae (Fagaceae; Kaul, 1985; Kubitzki, 1993a).
In Lithocarpus, the presence of staminodes is acknowl-
edged to be associated with entomophily (Kaul, 1985)
and in Juglandaceae, insect pollination has been
observed in Platycarya (Endress, 1986). This raises the
possibility that Canacomyrica may be insect pollinated
and thus indicates an entomophilous ancestry for
Myricaceae. Such a suggestion is consistent with the
notion that wind-pollination in Fagales may have multi-
ple origins from insect-pollinated ancestral lineages
(Manos & al., 2001).
Alternatively, in the case of species in which monoe-
cy or dioecy are common, the occurrence of staminodes
may be due to incomplete suppression of male function
(Walker-Larsen & Harder, 2000). Incomplete suppres-
sion of male function is known in other Myricaceae, for
example: within a single population of Myrica gale L.
both dioecy and monoecy can occur, and sex expression
within a single stem has been reported to be unstable
from year to year (Lloyd, 1981); androgynous or mixed
inflorescences have been observed in several species of
Myrica and Morella; and, perhaps most significantly of
all, occasional functional stamens are often observed on
the fruit wall of Morella species (Macdonald, 1989; J.
Herbert, pers. obs.). In Canacomyrica the presence of
staminodes could be viewed as an extreme example of
the incomplete suppression of male function found
throughout Myricaceae.
Lectotypification. —
In the original description
of this species, Guillaumin (1941) listed several speci-
mens but the type was not indicated. Therefore, a lecto-
type is designated below. In addition, a detailed descrip-
tion of the species is given.
Canacomyrica monticola Guillaumin, Bull. Soc. Bot.
France 87: 300. 1941. – Lectotype (designated here):
NEW CALEDONIA, Noumea, Balansa 564 (P!).
Dioecious shrub or small tree to 7 m in height.
Unscented resinous glands present on leaves and inflo-
rescences. Leaves evergreen, typically to 14 cm long,
coriaceous, oblanceolate, serrate at apex (margin deeply
serrate in leaves of young plants), dark green above,
whitish below, trichomes absent; stomata anomocytic
(confirmed for this species on the basis of unpublished
data — R.S. Hill and R. Paull, University of Adelaide,
pers. comm.); stipules absent. Inflorescence an erect
spike; flowers spirally arranged and widely spaced on
floral stem, trichomes present on floral stem. Male flow-
ers surrounded by 3 bracts; 6 stamens, filaments free,
anthers dorsifixed; vestigial style and vestigial perianth
present. Female flowers surrounded by 3 bracts; 6 sta-
minodes present; style pink-red, bifid, lamellular with
laciniate margin; 6-lobed fleshy perianth present; ovary
sessile, 1-locular; ovule 1, erect, orthotropous, bitegmic.
Fruit a drupe to 1 cm diam., pericarp fleshy, white or
pink becoming black at maturity, endocarp hard.
Conclusion. —
In conclusion, we accept the place-
ment of Canacomyrica in Myricaceae on the basis of the
molecular data presented here. This decision is support-
ed by the confirmation of a dioecious breeding system
and a chromosome count of 2n= 16. The data are
ambiguous with regards to the sister group of
Myricaceae, but the clarification of the placement of
Canacomyrica enhances our understanding of pollina-
tion syndrome within the family, and further highlights
the morphological characters shared by Myricaceae and
Juglandaceae.
ACKNOWLEDGEMENTS
The authors thank L. Ronse Decraene (RBG, Edinburgh) for
Herbert & al. • Systematic position of Canacomyrica 55 (2) • May 2006: 349–357
354
comments on an earlier draft of this paper; S. Schuster (RBG,
Kew) for additional sequencing; P. Hollingsworth, M. Gardner
and A. Ponge (RBG, Edinburgh) for their help during fieldwork;
Administration, Province Sud, New Caledonia for permission to
collect; J. Dahl Regional Parks Botanic Garden, Berkeley (U.S.
A.), for supplying plant material; Administration of the Western
Cape Nature Conservation Board and the Cape Peninsula National
Park, South Africa, for permission to collect; and T. Meagher
(University of St. Andrews, U.K.) for providing plant material.
The first author was funded by a Natural Environment Research
Council studentship (ref.: NER/S/A/2000/03638) with additional
funding for field work from the Merlin Trust (Kettering, U.K.), the
Russell Trust (University of St. Andrews, U.K.) and the Davis
Expedition Fund (University of Edinburgh, U.K.).
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Appendix. Accessions sampled. Voucher information and GenBank accession number are given for sequences report-
ed here for the first time. Taxa for which sequences were previously published are listed with the original reference and
GenBank accession number.
Taxon, Voucher/Source, Genbank Acc. No.: rbcL, trnL-F, ITS.
Alnus firma Siebold & Zucc. (Betulaceae), Kamiya, unpubl., AB060562; Kamiya & Harada, unpubl., AB063524 and AB 063548;
Navarro & al., 2003, AJ251684. Canacomyrica monticola Guillaumin (Myricaceae), Herbert 934 (E), DQ310504; Herbert 934 (E),
DQ310508; Herbert 934 (E), DQ310500. Carpinus laxiflora (Siebold & Zucc.) Blume (Betulaceae), Kamiya & Harada, unpubl.,
AB060585; Kamiya & Harada, unpubl., AB063571 and AB063541; Yoo & al., 2002, AF432038. Casuarina equisetifolia L.
(Casuarinaceae), Sogo & al., 2001, AY033859; Li & al., 2002, AY147090; Steane, unpubl., AY864057. Comptonia peregrina (L.) L’Hérit.
(Myricaceae), Meagher s.n. (E), DQ310505; Meagher s.n. (E), DQ310509; Meagher s.n. (E), DQ310501. Fagus crenata Blume
(Fagaceae), Martin & al., 1993, L13339; Fujii & al., 2002, AB046508; Denk & al., 2002, AF456969. Juglans nigra L. (Juglandaceae),
Soltis & al., 1999, AF206785; Manos & al., 2001, AF303783; Potter & al., 2002, AF338491. Morella cordifolia (L.) Killick (Myricaceae),
Herbert 1007 (E), DQ310502; Herbert 1007 (E), DQ310506; Herbert 1007 (E), DQ310498. Myrica hartwegii Watson (Myricaceae),
Edwards 93 (RPBG), DQ310503; Edwards 93 (RPBG), DQ310507; Edwards 93 (RPBG), DQ310499. Quercus rubra L. (Fagaceae),
Bousquet & al., 1992, M58391; Gielly & al., 1994, X75707; Manos & al., 1999, AF098418. Rhoiptelea chiliantha Diels & Hand.-Mazz.
(Juglandaceae), Chen & al., 1998, AF017687; Manos & al., 2001, AF303773; Manos & al., 2001, AF303800.
