A PHYLOGENY OF ARCTOSTAPHYLOS (ERICACEAE) INFERRED FROM NUCLEAR RIBOSOMAL ITS SEQUENCES

2009·17 MB·English

by  Gregory A Wahlert

A PHYLOGENY
OF ARCTOSTAPHYLOS INFERRED
(ERICACEAE)
FROM
NUCLEAR
RIBOSOMAL SEQUENCES
ITS
Thomas
Gregory Wahlert
Parker
A.
V.
Ohio San
University Francisco State University
Ohio 4570 San USA.
Athens, U.S.A. Francisco, California 94132,
1,
[email protected]
[email protected]
CVasey
Michael
Department
of Environmental
Studies
of Santa Cruz
University California,
Santa
Cruz, California 95064,
U.S.A.
INTRODUCTION
Arctostaphylos Adans. (Ericaceae) composed of 108 taxa and the most species-rich genus of subfamily
is ca. is
Arbutoideae Nied. (Wells 2000). Except Spreng.—which has circumboreal
for Arctostaphylos uva-ursi (L.) a
—
distribution the remaining species are confined to western North America and Mexico. In California,
occur communities, montane
species of Arctostaphylos in a variety of plant including chaparral, scrub,
mixed- coniferous serpentine barrens, and oak savannas. At low
forests, latitudes, Arctostaphylos uva-ursi
montane
occurs in regions Rockies, Appalachians, Alps, Himalayas) and in coastal pine barrens and
(e.g.,
of Texas
Journal of the Botanical Research Institute 3(2)
674
New (Donn.Sm.) represents
P.V.Wells
dunes and Michigan, USA). A. uva-ursi subsp. cratericola
Jersey
(e.g.,
highlands Guatemala.
population in the volcanic of
disjunct
a
Arbutus
Arbutoideae with other genera: Arctous Nied., L.,
placed subfamily five
in
Arctostaphylos
is
&
Breckon Molecular
Zucc, Small, and Xylococcus Nutt. (Diggs 1981).
Ornithostaphylos
Comarostaphylis
between basalmost
Arbutoideae the
and morphological phylogenetic studies in the Ericaceae place the
Markos presented phylogenetic
(Kron (1998)
Monotropoideae and the rest of the family et 2002). et al.
al.
spacer (nrlTS)
from nuclear ribosomal internally transcribed
inferred
16 species of Arctostaphylos
trees for
was congruent
not
The two-clade topology recovered in their trees
and 28S ribosomal sequences.
nuclear
G.Don
monophyly species
with two subgenera of Wells (2000) nor did support the of the A. hooheri
the
it
Boykin presented nrlTS phylogeny of 38 taxa
(Markos (2005) a
complex Pungens 1998).
in section et et al.
al.
congruent (with respect
two-clade phylogenies are
recovered a two-clade topology. Further, these
that also
20
with phenogram constructed from flavonoid data for species of Arctostaphylos
composition)
species a
to
(Denford
1981).
CA
The centered about San Francisco, in the California Floris-
center of diversity tor Arctostaphylos is
from Mendocino County Santa Barbara
The and Coast Ranges of California, to
Province. coastal areas
tic
endemic
has high percentage of
(Markos al Arctostaphylos a
County, particularly rich in taxa et 1998).
are
considered
many of which are edaphic specialists (Wells 2000). Nearly half of all California taxa are
taxa,
may
modern
(CNPS The taxa be the result of a radiation
endangered 2009). diversity of
threatened, or
rare,
end Miocene Ma) (Edwards 2004). Factors contributing to the diversi-
since the of the (5.3
of Arctostaphylos
an frequency
increase in
genus include adaptation to California's diverse substrates, fire
of the local
fication
and contemporaneous expansion of
Mediterranean the
the end of the Pliocene, the onset of a climate,
at
&
Edwards
Axelrod Axelrod 1989; 2004).
(Raven 1978;
chaparral into forested areas
The
base
and two processes driving speciation in Arctostaphylos.
Hybridization polyploidization are
=
chromosome number x = with most species being either diploid (In 26) or tetraploid
in Arctostaphylos 13,
is
and evidence the allopolyploid
= provide morphological cytological for
Schierenbeck
(1992)
(2n et
52). al.
homoploid hybrid
mewukha Merriam. Many other taxa have been proposed as arising via specia-
origin of A.
&
Knight Keeley
Dobzhansky Schmid 1968; Kruckeberg 1977; 1985;
1968;
1953; Gottlieb et
tion al.
(e.g.,
&
Massihi 1994; Parker Vasey 2004). For a group of 46 edaphic specialists or endemic taxa, Wells (2000)
subsp.
ploidy observed in Arctostaphylos uva-ursi coactilis
proposed hybrid origin for 37. Differing levels
a
&
which
and
= 52 and suggests recent recurring polyploidization, is
(2n 78)
Fernald J.F.Macbr. 26, 39,
common Brochmann
2004).
plants (Rosatti 1981; et
in arctic al.
1995)—
hybrid Rieseberg par-
Morphological intermediacy not always a reliable indicator of origin (e.g.,
is
and polyploidization in Arctostaphylos has lead to
introgression
that
in Arctostaphylos. likely
ticularly It is
based on morphological characters.
and taxonomic that are
misleading phylogenetic inferences classifications
been
nrlTS region has
and A.Gray, the bi-parentally inherited,
In perennial species of Paeonia L. Sidalcea
In
taxa (Sang 1995; Whittall et 2000).
used corroborate parentage of allopolyploid et al. al.
successfully to
may
evidence
provide additional
sequencing maternally inherited chloroplast
angiosperms, of the
direct
Wahlert
(2005)
by maternal parent (Koontz 2004). In Arctostaphylos,
hybrid origin identifying the et al.
of
trnS-trnG
and spacers atpB-rbcL, trnL-trnF,
intron the intergenic
sequenced chloroplast regions (trnL
five
The combined
and resulting
geographically distant species.
and 12 morphologically divergent
trnH-psbA)
for
Thus,
and had parsimony informative positions. direct
alignment was 1,836 base pairs (bp) long just three
confirm maternal
could not be used to infer relationships or to
sequencing of the chloroplast in Arctostaphylos
parentage in hybrids species.
appear influence the rate of speciation. In California's
Different history strategies in Arctostaphylos to
life
and Ceanothus (Rhamnaceae) are the only genera of
chaparral communities, Arctostaphylos L.
fire-prone
mode
reproduction (Wells
have evolved an obligate seeding of
shrubs containing species that
sclerophyllous
and
populations
response
In these genera, in about two thirds of the species, adults are killed in to fire
1969).
but
by from seed banks. Most chaparral shrubs re-sprout after a stand-replacing fire,
recruitment
recover
Wahlert phylogeny and
et ITS Arctostaphylos 675
al.,
in species of Arctostaphylos and Ceanothus, seeds are stored in a persistent seed bank from which they
soil
germinate only after fire (Parker and Kelly 1990). Therefore, each post-fire generation of obligate seeders a
is
product of sexual reproduction, whereas facultative sprouters regenerate primarily from vegetative re-sprouts
&
A
from
(but also to a lesser degree seed) (Keeley Keeley 1981). higher rate of morphological
diversifica-
and endemism 70%
supported by
tion local for seeders is the fact that ca. of Arctostaphylos taxa are obligate
seeders (Wells 1969). Generation-time theory predicts that taxa with shorter generation times have
will
DNA
higher molecular compared
rates of evolution in neutral regions taxa with longer generation times
to
&
Andreasen
(e.g., Baldwin 2001). For that reason, it is hypothesized that in Arctostaphylos, there will be a
among
higher ITS sequence compared
rate of interspecific variability obligate seeders to facultative sprouters.
The primary was
goal of this study to estimate the phylogeny of Arctostaphylos with extensive taxon
sampling using nrlTS sequences. Increased taxon sampling could refine species composition of each
clade,
might reveal additional subclades, and could allow for more rigorous testing of the monophyly of certain
we
infrageneric taxa. In addition, sought to corroborate hybrid origin in putative hybrid species and
to a
test
hypothesis higher compared
of rates of nucleotide substitution in obligate seeders
to facultative sprouters.
AND METHODS
MATERIALS
Taxon
Sampling
The
ingroup included a total of 98 accessions representing 83 unique taxa. The remaining 15 accessions
were
multiple samples of morphologically diverse and/or widely distributed species: Arctostaphylos
uva-ursi,
HBK,
A. patula Greene, A. pungens A. manzanita Parry, A. glandulosa Eastw., and A. tomentosa (Pursh) Lindl.
The
outgroup contained
six species representing the other five genera in the Arbutoideae. All sequences
generated for this study were new, except for two accessions of Arctostaphylos and outgroup which
five taxa,
were downloaded from GenBank (Appendix Taxonomy
Wells with names,
follows (2000) geographic
1).
GenBank
voucher specimen and numbers Appendix
origin, data, accession given in All accessions
se-
1.
quenced by
for this study are represented voucher specimens deposited at the Harry D. Thiers Herbarium
San MO.
Francisco which
at State University (SFSU), except for Arctous erythrocarpa, deposited
at
is
DNA
PCR
and
Extraction, Amplification, Sequencing
DNA
Genomic was from herbarium DNeasy™ (QIAGEN
extracted material using Plant Mini Kits Corp.,
Alameda, CA, The was
USA). nrlTS region amplified using Arctostaphylos-speciRc primers designed
for this
GGAAGTAAAAGTCGTAACAAGG
CTGGGGTCGCAATCAAGG.
study ITS 4-M = and ITS 5-M =
(5' to
3'):
PCR
amplifications for the ITS region were carried out under the following conditions: an denatur-
initial
ation step (92°C, 2 min) followed by 35 cycles of denaturation (92°C, 30 annealing (58°C, 30 and
sec),
sec),
PCR
elongation (72°C, 2 min), with a final extension step (72°C, 5 min). amplifications were performed in
DNA
PCR
25 pi reactions containing 10-100 ng genomic and 0.25 Expand Taq High System™
pi Fidelity
Mg
PCR
2+
with buffer and (Roche Diagnostics, Indianapolis, IN, USA). products were visualized on an
MO
PCR (MO
agarose gel and purified using BIO UltraClean™ Clean-up Kits BIO Laboratories, Carlsbad,
CA, USA). Cycle-sequencing reactions used the same primers, annealing temperature, and
additive as
for
PCR
and employed BigDye®
fluorescent dye-labeled chemistry (BigDye Terminator Cycle Sequencing
Kit,
Applied Biosystems, Foster City, CA, USA). Cycle-sequenced products were separated and visualized on an
DNA
When
ABI 377 sequencer (Applied Biosystems, Foster CA, USA). polymorphic
City, a nucleotide posi-
DNA
was was
tion detected, the taxon sequenced a second time to confirm the polymorphism. sequencing
was
performed San
the Francisco State University Conservation Genetics Laboratory.
at
Phylogenetic Analyses
Due
low and
to rates of nucleotide substitution few insertions/deletions, ITS sequences were aligned
easily
Ann Maximum
automatically using Sequencher version (Gene Codes, Arbor, MI, USA). parsimony
4.1
(MP) was conducted on two 98 and
analysis data sets: the total data set for Arctostaphylos sequences,
1) all
reduced with removed had polymorphic
2) a data set 17 taxa that nucleotide positions. Forty-four ingroup
Texas
Journal of the Botanical Research Institute of 3(2)
76
6
"Gray Leaf" haplotype),
which shared ITS sequences, were treated as a single "taxon" (the
identical
taxa,
MP
MP
used
was and analyses
number both analyses 55 38, respectively.
taxa in
therefore the actual of
space multiple islands of
Olmstead and Palmer (1994) search the tree for
search strategy of to
the heuristic
was
PAUP* carried out
implemented 4.0bl0 (Swofford 2002),
in
shortest This search strategy, as ver.
trees.
used in subsequent searches: 1,000
from each starting trees
with saved step as 1)
in four steps, the trees
MulTrees tree-bisection
neighbor interchange (NNI), Off,
random sequence nearest 2)
addition replicates,
A
On. consensus
On, and TBR, MulTrees tree
MulTrees NNI, MulTrees strict
reconnection (TBR), Off, 4)
3)
branch support phylogenetic trees
from each Internal of
from saved search.
was calculated the shortest trees
TBR
with
search
using heuristic
1985) a
was estimated with 100 bootstrap (BS) replicates (Felsenstein full
from BS
random Results the analysis
MulTrees and 10 stepwise addition replicates.
branch swapping, on,
were weighted and unordered,
equally
summarized consensus Nucleotide characters
were in tree.
a strict
and were missing
indels treated as data.
98 and 6 outgroup taxa was 600 bp in length (0%
The sequence matrix the ingroup accessions
aligned for
A
bp
one
deletion
and 62 parsimony informative characters. at
missing with 127 variable positions
data),
&
Merriam,
(W.Knight Gankin) A. nissenana
P.V.Wells,
403 was malloryi
detected Arctostaphylos
position for
sequence—
"Gray
an nrlTS the
Forty-four taxa shared identical
and three subspecies of A. viscida Parry.
used throughout
Appendix phrase "Gray Leaf haplotype" this
Leaf" haplotype— and are listed in 2 (the is
among
morphology sharing
uniform taxa this
shared sequence and does not imply a
study
to refer to this
from
ranged
ingroup nrlTS sequences to
The uncorrected pairwise distances for the
sequence).
identical
maximum
parsimoni-
100,000 equally
reached of
Parsimony complete ITS data a
2.4%. analysis of the set
with consistency index (CI) of 0.837 (0.753
ending generation, a
ous of length 178 before initial tree
trees
A
shown)
consensus
(not
uninformative and retention index (RI) of 0.881. strict tree
excluding characters)
American
North
A. uva-
accessions of
polytomy and small clade of six
three clades: a all
revealed a large 1)
and containing
and (59% clade
(58% clade containing A. pumila Nutt. A. tomentosa BS), 3) a
ursi BS), 2) a (1)
&
(90%
mewukka and M.C.Vasey V.T.Parker BS).
ohloneana
A.
A.
from superimposed
sequence electropherograms
Polymorphic nucleotide positions were detected in the
A.
(SNAPs) and were confirmed in 17 taxa (18% of ingroup): A. otayensis,
patterns of nucleotide additivity
&
A.
mewukka, Wiesl.), A. viscida ssp. viscida,
drupacea Parry (P.VWells), A. A. pilosula (Jeps.
pringlei ssp.
manzanita
(Gankin) P.VWells, A.
Humboldt CA,
from A. manzanita ssp.
manzanita manzanita Co., roofii
ssp.
CA, x
from San Diego A.
Munz, manzanita bowermanii, A. pungens Co., pacifica
(Eastw.) A. ssp.
laevigata
ssp.
CA,
from San Diego A. myrti-
Co.,
glandulosa
A. glandulosa
edmundsii J.T.Howell, ssp.
Roof, A. nissenana, A.
within
The number polymorphic nucleotide positions
and Howell. of
nummularia A. hispidula
A.
(1)
folia,
Of
4.3% these 17 taxa, 12 are tetraploids
an ranged from 6 (0.7% of variable positions).
individual to
1 to
A. manzanita
drupacea,
hybrid-derived or introgressed taxa (A. pringlei ssp.
and/or hybrids. In four putative
showed
and sequence electropherograms near-perfect
manzanita A. hispidula)
A. ssp. laevigata,
ssp.
roofii,
and
hybrids (Table
between synapomorphic nucleotides of suspected parents
1).
patterns of additivity
rem
MP
A with polymorphic nucleotide positions
second was conducted with the 17 taxa
analysis
with 56 parsimony informative chara
The sequence matrix had 126 variable positions
aligned
resulting
consensus 350 equally
MP of
two-clade topology in the
reduced ITS data recovered a strict
analysis of the set
= CI = CI excluding uninformative characters = 0.677,
parsimonious length 169, 0.817,
trees (Fig. (tree
1)
(77%
and Clade was moderately well supported
= was weakly supported (55% BS) 2
Clade
RI
0.858).
1
uva-u
containing the six accessions of A.
Within Clade two subclades were recovered: a clade
1)
BS).
1,
and
nummularia A.Gray,
(59% and containing three subspecies of A. stanfordiana Parry, A.
BS) a clade
2)
44 with
P.VWells (76% In Clade the Gray Leaf "taxon" representing the taxa identic
mendocinoensis BS).
2,
were pungens from Arizona, A. austral
sequences out in a polytomy while three clades resolved: 1) A.
fell
Wahlert et ITS phylogeny and Arctostaphylos
al.,
G A G
G/A A/G G/T
G T
154
and
A. pringlei subsp. pringlei (71% BS), A. pumila and A. tomentosa subsp. tomentosa (65% and A.
2) BS),
3)
and
(59%
malloryi A. subsp. mariposa
visrida BS).
Of 44
an Gray
the species sharing identical Leaf haplotype, five are facultative sprouters (A. glandulosa
and
ssp. cushingiana, A. glandulosa ssp. glaucomollis, A. tomentosa ssp. daciticola, A. tomentosa ssp.
insulicola,
—
A. tomentosa ssp. rosea) and the rest are obligate seeders. The low variability of the nrlTS region and the
both and an sequence—
fact that obligate seeders several sprouters shared identical precluded a of higher
test
compared
nucleotide substitution rates in seeders sprouters.
to
The phylogeny inferred from the nrlTS data reveals resolution than had been expected mor-
sets less for this
and The
phologically diverse widely distributed genus. species composition of Clades and 2 recovered in
1
MP
the analysis of the reduced nrlTS data set largely congruent with the two-clade molecular phylogenies
is
Markos and Boykin and phenogram
Denford
of et al. (1998) et al. (2005), the flavonoid of (1981).
MP
None
of the subclades recovered in analyses have a discrete pattern of geographical distribution
mapped
that can be onto the phylogeny. Although the circumarctic distribution of Arctostaphylos uva-ursi
is
unique in the genus, this species also distributed along the coastal areas and mountains of western North
is
America where
co-occurs with other taxa A. columbiana Piper, A. imbricata Eastw., A. Neither
patula).
it (e.g.,
and mapped
are there discrete readily interpretable morphological or cy tological synapomorphies that can be
onto this phylogeny to support the present taxonomic groupings of Wells (2000) or the subclades that were
Boykin
recovered in the phylogenetic trees. et (2005) found that the character states of inflorescence bracts,
al.
and monophyly
bark, distribution of stomata, ploidy level were not consistent with of certain infrageneric
and some more
groups that states have arisen than once in the genus. Because Wells' (2000)
classification
on
of Arctostaphylos based potentially homoplasious morphological characters, discordance between his
is
and
the nrlTS phylogeny not unexpected.
classification
is
The
occurrence polymorphic showing between
of nucleotide positions patterns of additivity synapo-
morphic and
nucleotides of putative hybrids suspected parents suggestive of hybrid origin in four taxa
is
many
more
However, polymorphic showing between and
(Table patterns of additivity parents hybrid
sites
1).
would
needed
offspring to confirm hybrid origin for the taxa in Table Sang 1995). Such
i 1 et
(e.g., al.
of Texas
Journal of the Botanical Research Institute 3(2)
Michigan
uva-ursi
New
Jersey
uva-ursi
Washington
uva-ursi
Colorado
uva-ursi
Wyoming
uva-ursi
mendocinoensis
nummularia
decumbens
subsp.
stanfordiana
stanfordiana subsp. raichei
stanfordiana
subsp.
stanfordiana
ohioneana
Nevada
patula
hookeri
hookeri subsp.
CA
Calaveras
patula Co.,
pungens
California
hearstiorum
hookeri subsp.
sensitiva
densiflora
CA
Plumas
patuia Co.,
Guatemala
uva-ursi
pungens
Arizona
n-|=::
australis
A
subsp.
pringlei pringlei
I
tomentosa
tomentosa subsp.
mailoryi
mariposa
subsp.
viscida
Gray Leaf haplotype
rainbowensis
glandulosa
nevadensis
tomentosa subsp. Crustacea
(1)
tomentosa subsp. Crustacea
(2)
manzanitaXA.
stanfordiana
tomentosa subsp. subcordata
klamathensis
subsp. pulchella
A. viscida
-
Alaska
Arctous rubra
-
Arctous erythrocarpa Siberia
-
Comarostaphyiis
diversifolia
-
Ornithostaphylos oppositifoiia
-
Arbutus menziesii
-
Xylococcus
bicolor
-
3
Wahlert phylogeny and
et ITS Arctostaphylos 679
al.,
polymorphisms
hybrid where both
suggest: 1) recent origin parental alleles are maintained in the hybrid
progeny, between genomes
different existing parental
2) alleles in tetraploids, recently substituted nucle-
3)
have been homogenized
otides that not fully via concerted evolution, or intra-individual nrlTS paralogs
4)
(Wendel Koontz
et 1995; 2004; Bailey 2003).
et et
al. al. al.
The 44
taxa sharing the Gray Leaf haplotype morphologically and composed both
are diverse are of
and
obligate seeders facultative sprouters (Appendix Sprouting species sharing the Gray Leaf haplotype
2).
by
represented
are five tetraploid taxa of the Arctostaphylos glandulosa-tomentosa complex, whereas most of
As
the other taxa are diploid obligate seeders. a result, a hypothesis of greater nrlTS nucleotide substitu-
tion rates in short generation-time seeders long generation-time sprouters could not be These
vs.
tested.
common
results suggest ancestry of "Gray Leaf" taxa, and by extension, the remaining taxa in Clade
all
2.
common
phylogenetic reconstruction based on nrlTS an
If of Arctostaphylos is accurate inference of ancestry,
much
then there has been morphology, and
diversification in the cytology, ecological life-histories of taxa
in Clade 2 without corresponding nrlTS sequence
divergence.
Resolving phylogenies and
species-level difficult in perennial, reticulating, putatively recently
is di-
genera such and
versified as Arctostaphylos other genera containing species that are widely distributed and
morphologically diverse Ceanothus (Rhamnaceae), Hardig 2000, Leucaena
(Fabaceae), Bailey
(e.g., et al. et
al.
Low
2004, Inga (Fabaceae), Richardson et 2001). interspecific sequence variability in the nrlTS region in
al.
may
Arctostaphylos be due several factors, including recent diversification in California since the end of the
Miocene, long generation time between sexual reproduction genome homogenization
events, via introgres-
and
sion, rapid concerted evolution of nuclear ribosomal ITS arrays in both diploids and
tetraploids.
The show
from and
results this study others that direct sequencing of the nrlTS region and the chloroplast
DNA
provide insufficiently variable sequences needed to resolve phylogenetic relationships in
Arctostaphylos.
Other PCR-based techniques have been used in Arctostaphylos without success. For example, the presence
RAPDs
endophytic
of fungi in the sclerophyllous leaves of Arctostaphylos prevented the use of (randomly
DNAs) comm,
amplified polymorphic and sequence
inter-simple repeats (ISSRs) Reading, Wahlert
(C. pers.
unpubl. Another approach RFLPs
using fragment polymorphisms)
data). (restriction length of the chloroplast
may
genome
prove mewukka
useful in corroborating the hybrid origin of A. Schierenbeck, comm.).
(K. pers.
Bi-parentally inherited nuclear genes with adequate sequence needed phylogeny
variability are to estimate in
Sequences from copy from anonymous
Arctostaphylos. single- or low- nuclear genes or nuclear regions using
may
sequence characterized amplified regions (SCARs) provide means phylogeny and
a resolve the
to to
many
tease apart patterns of reticulation in Arctostaphylos. Until then, fascinating questions of polyploidiza-
tion, hybrid and ecological speciation, biogeography, and ecology go unanswered remarkable
will in
this
APPENDIX
1
dGenBankac
oucherspeci numbers
vithv Tien information an cession
for nrlTS
y D.Thiers He rbarium at San Fra Cisco State U niversity (SFSU), except for
Outgroup. &
Arbutus menziesii Pursh*, AF086828; Arctous rubra (Rehder E.H.Wilson) Nakai*, AF091 944; Arctous erythrocarpa
MO
number
Small., Russian Siberia, accession 5196245, GQ281006; Comarostaphylis Greene*, AF091947;
diversifolia (Parry)
Ornithostaphylos AF297794 and AF091962 and
oppositifolia (Parry) Small*, (ITS 5.8S), (5.8S ITS Xyiococcus bicolor Nutt*,
1 2);
AF091948.
A
Ingroup. Arctostaphylos andersonii San Mateo CA, USA, & M.CVasey GQ280910;
A.Gray, Co., VT.Parker 764, auricutata Eastw.,
Contra Costa Co., CA, USA, M.CVasey 840, GQ280911; A. australis Eastw., Baja Norte, Mexico, M.CVasey GQ280912;
799, A.
Sonoma
bakeri Eastw. subsp. bakeri Eastw., Co., CA, USA, M.CVasey GQ28091 Sonoma
71, evis P.V.Wells,
A
Co., CA, USA, VT.Parker M.CVasey 548, GQ28091 canescens subsp. canescens Marin CA, USA, & M.CVasey
Eastw., Co., VT.Parker
4;
GQ28091
443, A. canescens subsp. sonomensis Lake CA, USA, M.CVasey GQ28091
(Eastw.) P.V.Wells, Co., 282, A. catalinae
5;
6;
&
Santa Barbara CA, USA, M.CVasey GQ280917; columbiana Humboldt
P.V.Wells, Co., VT.Parker 155, A. Piper, Co., CA, USA,
A
A
M.CVasey GQ280918; &
1030, confertifiora Eastw., Santa Barbara Co., CA, USA, VT.Parker M.CVasey 900, GQ280919; cruzen-
Journal of the Botanical Research Institute of Texas 3(2)
680
A
Sonoma M.CVasey
CA, USA,
GQ280920; densiflora M.S.Baker, Co.,
San Obispo CA, USA, VJParker et
Roof, Luis Co., al. 12,
sis
A
A & M.CVasey,
M.CVasey GQ280922; gabilanensisVT. Parker
Monterey CA, USA, 81
GQ280921 edmundsii J.T.Howe!!, Co., 9,
69,
;
Adams McMinn,
y ex
GQ280923; giandulosa subsp. cushingiana (Eastw.)
San Benito CA, US 1086, A.
Co.,
A
San Diego CA, USA,
CA USA MCVasey GQ280924; giandulosa subsp. giandulosa Eastw. Co.,
Co
1002, (1),
Santa Barbara
A & M.CVasey GQ280926;
San Benito CA, USA, VJ.Parker 456,
GQ280925; giandulosa subsp. giandulosa Co.,
M.CVasey
724, (2),
A
Angeles
GQ280927; g/auca Los
M.CVasey WOO, LindL,
Angeles CA, USA,
A os Co.,
giandulosa subsp.
c
A
A MCVtaey GQ280929;
/j/sp/^fa
Santa Cruz CA, USA, 407,
M.CVasey809, GQ280928; g/uf/nosa B.Schreib., Co.,
USA,
CA,
Co.,
A
Munz,
cultivated material;
MCVasey GQ280930; hookeri subsp. franciscana (Eastw.)
USA,
Norte CA, 360,
Howeil, Del Co.,
A
(Hoover & Roof)
hearstiorum
USA, GQ280931; hookeri subsp.
Arboretum, Golden Gate San Francisco, CA,
Strybing Park,
&
A
Cruz CA, USA, VJ.Parker
G.Don, Santa
Obispo USA, AF106817; hookeri subsp. hookeri Co.,
CA,
San
RV.Wells* Luis Co.,
A
GQ280933;
montana Marin CA, USA, Markos613, Zioote-
M.CVasey GQ280932; hookeri subsp. (Eastw.) P.V.Wells, Co., 5.
A.
55,
A
M.CVasey
Monterey CA, USA, 1050,
San Francisco CA, USA, AF1 0681 ftcxjver/ RV.Wells, Co.,
subsp. raven/7 P.V.Wells*, Co., 9;
ri
A
CA,
A M.CVasey GQ280935; /mu/ar/s Greene, Santa Barbara Co.,
Mateo USA,
San CA,
GQ280934; imbricata Eastw., Co., 18,
A & CA, USA, M.CVasey 756,
S.W.Edwards, Keeler-Wolf W.Knight, Siskiyou Co.,
Anonymous GQ280936; klamathensis
USA,
s.n.,
A
A (W.Knight & Gankin)
GQ280938;
M.CVasey malloryi
San Obispo CA, USA, 1046,
GQ280937; luciana RV.Wells, Luis Co.,
A
bowermanii Contra Costa
& GQ280939; manzanita subsp. Roof, Co.,
M.CVasey
PV CA, USA, VJ.Parker 834,
Shasta
Wells, Co.,
GQ280941
A M.CVasey
Glenn CA, USA, 21
M.CVasey GQ280940; manzanita subsp. elegans (Jeps.) RV.Wells, Co., 6,
CA, USA, 169, ;
A
& MCVtoey GQ280942; manzanita subsp.
A Sonoma CA, USA, Itftotef 508,
manzanita subsp. glaucescens RV.Wells, Co.,
Men-
manzanita
GQ280943; manzanita subsp. Parry
M.CVasey
Munz, Contra Costa CA, USA, 894, A. (1
faewgaro (Eastw.) Co., ),
A
Humboldt CA, USA, M.CVasey
manzanita
GQ280944; manzanita subsp. Co.,
VJParker&M.CVasey369,
docino CA, USA, (2),
Co.,
A
manzanita
A Amador M.CVasey GQ280946; subsp.
manzanita CA, USA, 767,
GQ280945; manzanita subsp. Co.,
851, (3),
A
Glenn CA,
MCViray GQ280947; manzanita subsp. (Gankin) RV.Wells, Co.,
Sonoma
CA, USA, 424, roo/7/
manzanita
Co.,
(4)
A
x
A A medfo
GQ280949;
Sonoma M.CVasey
x CA, USA, 425,
GQ280948; manzon/to stanfbrdlana, Co.,
M.CVasey883,
USA,
A
Mendocino CA, USA, G.Wahlert
M.CVasey GQ280950; mendocinoensis RV.Wells, Co.,
Mendocino USA,
CA, 253,
Greene,
Co.,
A
Mateo
A San
GQ280952; montaraensis Roof, Co.,
GQ280951 mewukka Merriam, Tuolumne Co., CA, USA, M.CVasey 575,
s.n.,
;
A
A GQ280954; morroensis
M.CVasey
GQ280953; montereyensis Hoover, Monterey Co., CA, USA, 656,
M.CVasey
USA, 743,
CA,'
A
Amador M.CVasey
USA,
CA,
& Obispo USA, Bode GQ280955; myrtifolia Parry, Co.,
San CA, 823,
Wiesl. Schreib., Luis Co., K.
B.
A
CA,
A M.CVasey GQ280957; n/ssenana Merriam, Placer Co.,
757,
GQ280956; nevadensis A.Gra<
733, \,
'
A
A
& M.CVasey 5 GQ280959;
ob/s-
Mendocino USA,
nummutaria CA, VJ.Parker
GQ280958; A.Gray, Co., 16,
M.CVasey
USA,
892,
A
M.CVasey & Santa Cruz CA, USA,
ohloneana Co.,
M.CVasey GQ280960; V.T.Parker,
Monterey CA, USA, 1052,
Co.,
A
Bode GQ280962; otayens/s
GQ28096 Obispo CA, USA, 780,
& M.CVasey *an Luis Co., K.
VJParker
1 1 1,
A
M.CVasey
M.CVasey GQ280963; x pacifica Roof, San Mateo Co., CA, USA, 20,
& San Diego CA, USA, 748,
Wiesl. Schreib., Co.,
A
Alameda
& M.CVasey GQ280965; pallida Eastw.,
A Monterey CA, USA, VJ.Parker 459,
GQ280964- pajaroensis J.E.Adams, Co.,
A
A
M.CVasey GQ280967; patu-
Nevada, USA,
Greene White 626,
M.CVasey GQ280966; patu/a Pine, Co.,
CA, USA, 436,
Co., (1 ),
& M.CVasey GQ280969;
USA,
Wood GQ2S0968; patula Calaveras Co., CA, VJ.Parker 770,
Plumas CA, USA, M. 316, A.
(3),
Co.,
1a
(2),
A
A drupacea Riverside
& San Obispo CA, USA, M.CVasey 233, GQ280970; pringlei subsp. Parry,
Luis Co.,
Jeps. Wiesl.,
p/'/osu/a
A
A
M.CVasey GQ280972;
Pima Arizona, USA, 232,
M.CVasey GQ280971 subsp. pringlei Parry, Co.,
USA, pringlei
CA, 993,
Co.,
;
& GQ280973; A.pungens HBK San Bernardino Co., CA, USA,
M.CVasey
terey CA, USA, VJParker 677, (1),
Co.,
GQ280975; A.pungens Pima Arizona,
M.CVasey Co.,
GQ28097 A.pungens San Diego CA, USA, 696,
M.CVasey Co., (3),
194, (2),
'4;
A
A
GQ280977; rainbowensis
CA, USA, VJParker
Santa Barbara etai.
GQ280976; pur/ss/ma RV.Wells, Co., 2,
M.CVasey228,
USA,
M.CVasey
San Mateo CA, USA,
Bode GQ280978; regismontana Eastw., Co.,
&Massihi,San Diego CA, USA,/C 775, A.
Co.,
J.E.Keeley
A
A & San
& M.CVasey GQ280980; rudis Jeps. Wiesl., Luis
x Marin CA, USA, VJParker 276,
GQ280979; repens J.T.Howell, Co.,
557,
A
A GQ280982;
& Marin CA, USA, VJ.Parker 306,
M.CVasey GQ280981; sensitiva Jeps., Co.,
Obispo USA, VJParker 151,
Co., CA,'
A
decumbens
& GQ280983; stanfordiana Parry subsp.
M.CVasey
& Cruz CA, USA, VJParker 865,
Santa
Jeps. Wiesl., Co.,
silvicola
Mendocino
W.Knight,
Sonoma M.CVasey GQ280984; stanfordiana Parry subsp. raichei
CA, USA, 422, A.
(RV.Wells) RV.Wells, Co.,
A M.CVasey GQ280986;
Napa USA,
CA, 392,
M.CVasey GQ280985; stanfordiana Parry subsp. stanfordiana, Co.,
CA, USA, 277,
Co.,
A
GQ280987; tomentosa subsp.
Monterey CA, USA, VJ.Parker 584,
tomentosa subsp. Crustacea (Eastw.) RV.Wells Co.,
A. (1),
A
Obispo
San
M.CVasey GQ280988; tomentosa subsp. daciticola RV.Wells, Luis Co.,
Santa Cruz CA, USA, 786,
Crustacea Co.,
(2),
A & M.CVasey
USA,
Santa Barbara CA, VJ.Parker
M.CVasey GQ280989; tomentosa subsp. /nsu//co/a RV.Wells, Co.,
CA, USA, 1008,
A
A GQ280991; tomentosa
M.CVasey
Monterey CA, USA, 822,
GQ280990; tomentosa subsp. rosei (Eastw.) RV.Wells, Co.,
903,
A
& M.CVasey GQ280992; tomentosa subsp.
subcordata RV.Wells, Santa Barbara Co., CA, USA, VJParker 901,
subsp. (Eastw.)
A New
Ocean USA, VJParker
Monterey CA, USA, M.CVasey 245, GQ280993; uva-urs/ (L) Spreng., Co., Jersey,
tomentosa
Pursh, Co.,
A
A McEaghan GQ280995; Yellows-
uva-urs/,
SA, 933,
GQ280994; uva
437, ]
A
M.CVasey
Gunnison Colorado, USA,
& M.CVasey GQ280996; Co.,
Wyoming, uva-urs/,
USA, VJ.Parker 942,
tone National
Park,
A
A M.CVasey
Washington, USA, 340,
GQ280998; Clallam
GQ280997; Alaska, USA, J fi/sbop 684, ura-urs/, Co.,
uva-ursi,
643,
92-
A Garden,
from Guatemala, U.C. Botanical
material
cultivated
GQ280999; uva-ursi subsp. cratericola (J.D.Smith) RV.Wells,
A
A
GQ281 mariposa (Dudley) RV.Wells,
USA, M.CVasey 001 viscida subsp.
GQ281 Eastw, Marin CA, 783,
0545, 000; virgata Co.,
;
M Sonoma M.CVasey
(Howe CA, USA,
Tuolumne Co CA USA C GQ281 002; A.viscida subsp. pulchella RV.Wells, Co.,
\/asey 499,
1)
1
.
and
Wahlert phylogeny Arctostaphylos
et ITS
al.,
A
GQ281004;
er&M.CVasey849,
with sequences the "Gray Leaf'haplotype).
Forty-four taxa identical nrlTS
(i.e.,
canescens canescens, canescens
Arctostaphylos andersonii, A. auricuiata, A. bakeri ssp. bakeri, A. baker! ssp. sublaevis, A. ssp. A. s:
glandutosa cushingiana, glandule
sonomensis, A. catalinae, A. columbiana, A. confertiflora, A. cruzensis, A. gabilanensis, A. ssp. A.
montana,
ssp. glaucomollis, A. glauca, A. glutincsa, A. hookeri ssp. franciscana, A. hookeri ssp. A. hookeri ssp. ravenii, A. hooveri,
manzanita manzanita manzanita
manzanita
imbricata, A. insularis, A. luciana, A. ssp. elegans, A. ssp. glaucescens, A. ssp. (1),
x
tomentosa
obispoensis, A. osoensis, A. pajaroensis, A. pallida, A. purissima, A. regismontana, A. repens, A. silvicola, A. ssp. dacitico
We
and Harvey (Ohio reviewing manuscri]
thank Robert Patterson (SFSU) Ballard, University) for this
Jr.
and an anonymous
and providing comments improved David Keil (Cal Poly State University)
that
r<
it.
viewer provided helpful commentary that enhanced the paper. Frank Cipriano (Conservation Genetic
also
SFSU) and provided with sequencing. Oriane Hidalgo (Ohio University
Lab, Juniper Scribner assistance
translated the abstract into Spanish.
Unequal between annual and perennial lineages of checker
and 2001 evolutionary rates
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