Botanical Journal of the Linnean Society, 2011, 167, 281–300. With 4 figures
Molecular phylogenetics of the juno irises, Iris
subgenus Scorpiris (Iridaceae), based on six
plastid markers
NURSEL IKINCI1*†, TONY HALL2†, M. DOLORES LLEDÓ2, JAMES J. CLARKSON2,
NICO TILLIE2,3, ARNIS SEISUMS4, TAKESHI SAITO5, MADELINE HARLEY2 and
MARK W. CHASE2
1
Abant Izzet Baysal Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, 14280-Bolu, Turkey
Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3DS, UK
3
Witmolen 12, 2645 GK, Delftgauw, The Netherlands
4
The National Botanic Garden, 1 Miera Str., Salaspils, LV 2169, Latvia
5
Department of Environmental Science and Technology, Meijo University, Nagoya 468-8502, Japan
2
Received 22 February 2011; revised 20 July 2011; accepted for publication 22 July 2011
Relationships among the roughly 55 species of Iris subgenus Scorpiris have been studied. A matrix of six plastid
DNA regions (matK, rpl14-rps8 spacer, infA-rpl36 spacer, trnE-trnT spacer, trnL intron and trnL-F spacer) was
produced from 57 accessions (52 taxa) and analysed with both parsimony and Bayesian methods. Five major
clades are identified, of which four have strong geographical correlations, whereas the fifth corresponds to Iris
section Physocaulon. In our results, several species are placed with species not previously considered to be
related, although, in some cases, there are morphological characters that suggest that these newly indicated
relationships are reasonable. For some of the other oddly grouped species, we can only assume that remarkable
parallelisms in morphology have occurred or hybridization is involved. Presently, with plastid DNA as our only
comprehensive data resource, we are not able to evaluate more thoroughly these more puzzling associations
of species. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167,
281–300.
ADDITIONAL KEYWORDS: Afghanistan – Eurasia – Juno – karyotype – Kopet Dag – morphological
parallelisms – pollen morphology.
INTRODUCTION
Iris L. is the largest genus in Iridaceae, comprising
approximately 270 species distributed in Eurasia,
North Africa and North America. Subgeneric classifications of Iris have been based mainly on form of
subterranean storage organs and occurrence of a
beard on the outer perianth segments (or ‘falls’).
Mathew (1989), basing his groupings on Lawrence
(1953) and Rodionenko (1961), divided Iris into six
*Corresponding author. E-mail: [email protected]
†These authors contributed equally.
subgenera, in part according to their underground
storage organs: subgenus Iris, subgenus Limniris
(Tausch) Spach, subgenus Nepalensis (Dykes)
G.H.M.Lawr., subgenus Xiphium (Mill.) Spach, subgenus Scorpiris Spach and subgenus Hermodactyloides Spach. Those having bulbs with fleshy roots
during the dormant stage were placed under subgenus Scorpiris. Subgenus Scorpiris (synonym genus
Juno Tratt.), known colloquially as ‘the junos’, has
roughly 55 species and is the largest (Mathew, 2001).
Juno irises have bulbs with papery tunics and usually
fleshy, sometimes thick, storage roots when dormant
(Fig. 4). The channelled bifacial (two-sided) leaves are
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
281
282
N. IKINCI ET AL.
A
B
C
D
E
F
G
H
I
Figure 1. Floral traits of Iris subgenus Scorpiris. A, I. cycloglossa. B, I. rosenbachiana. C, I. magnifica. D, I. persica. E,
I. hippolyti. F, I. warleyensis. G, I. orchioides. H, I. parvula. I, I. narbutii.
arranged distichously (Mathew, 1997). In subgenus
Scorpiris, the three inner perianth segments (or ‘standards’) are generally reduced, and occasionally they
are almost whisker-like; they are usually held at or
below the horizontal and often strongly reflexed. Relative to the other species of this subgenus, Iris cycloglossa Wendelbo has unusually large, rounded
laminae (or ‘blades’) to standards that are held above
the horizontal, as well as exceptionally large, rounded
blades to the falls (Fig. 1).
Rodionenko (1961) studied 24 species of subgenus
Scorpiris, placing them in genus Juno. He found that
the most common pollen type is spheroidal in shape
and clypeate, with the reticulate exine divided into a
number of discrete polygonal plates, generally from
four- to six-sided but most often pentagonal (footballlike; Fig. 2A). However, a few species have pollen that
is zona–aperturate, with the reticulate exine divided
into two hemispherical halves (hamburger-like;
Fig. 2B). A third type, found in only two Mediterra-
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
283
Figure 2. The three pollen types observed in Iris subgenus Scorpiris. A, football-like, I. stenophylla. B, hamburger-like,
I. aitchisonii. C, spiny, I. planifolia; scale bar, 50 mm.
nean species, has spheroidal, clavate pollen (spiny;
Fig. 2C), the incomplete exine comprising club-like
protuberances, with a spiral or winding aperture.
Rodionenko (1961), within his treatment of Juno,
proposed its subdivision into several sections and
series based primarily on storage organ, pollen and
seed morphology. Of these groupings, the two major
divisions [section Juno (Tratt.) Benth. and section
Physocaulon (Rodion.) B.Mathew & Wendelbo] were
accepted by Wendelbo & Mathew (1975). Rodionenko
created section Acanthospora Rodion. for the two
species with clavate pollen: Iris planifolia (Mill.) Fiori
& Paol. and I. palaestina (Baker) Boiss. Later he
(Rodionenko, 1994) isolated I. cycloglossa in its own
section: section Wendelboa Rodion. The recognition of
these last two sections and Rodionenko’s other infrasectional divisions remains debatable.
Chromosome variation is complex in Iris, and
neopolyploidy is an important process for the genus
(Goldblatt & Takei, 1997). Subgenus Scorpiris is also
cytologically diverse, with counts ranging from
2n = 14 (Gustafsson & Wendelbo, 1975) to 2n = 50
(Simonet, 1952), especially relative to the species of,
for example Iris section Oncocyclus (Siemss.) Baker,
which are exclusively 2n = 20 (Avishai & Zohary,
1977).
Only one species, Iris planifolia, is found in Mediterranean Europe and North Africa, whereas all other
members of the subgenus are distributed from central
Turkey and Transcaucasus, south through southern
Israel and Jordan as far as Saudi Arabia (I. postii
Mouterde), east through western Asia to northern
Pakistan and as far as Kashmir (I. aitchisonii (Baker)
Boiss.) and north through Central Asia, primarily the
Pamir-Alai and Tien-Shan ranges (Mathew, 1997).
Junos have adapted to these predominantly mountainous regions, with their hot, dry summers and cold
winters, where precipitation is restricted to autumn/
spring rains and spring snowmelt (Hall, 1998).
Previous molecular phylogenetic studies that
assessed the position of Iris have included those of
Reeves et al. (2001) and Goldblatt et al. (2008). Both
used multiple plastid DNA regions to assess phylogenetic relationships and biogeography and divergence
times of Iridaceae. Phylogenetics of Iris has been
broadly assessed by Tillie, Chase & Hall (2001) and
Wilson (2004, 2009, 2011) using various plastid DNA
regions; these studies showed that subgenus Scorpiris
is deeply embedded within Iris and therefore should
not be considered a distinct genus, Juno. More narrowly focused studies have examined Iris series Californicae (Diels) G.H.M.Lawr. [Wilson, 2003; the only
study thus far to use the nuclear ribosomal internal
transcribed spacer (ITS) DNA region] and Siberian
Iris spp. (Makarevitch et al., 2003; also using plastid
DNA).
As summarized above, there is no comprehensive
molecular phylogenetic study of subgenus Scorpiris.
The study of Wilson (2004) sampled only six species
from subgenus Scorpiris, whereas Tillie et al. (2001)
included 11 species. In a later paper by Wilson (2011),
in which nine juno species were sampled, subgenus
Scorpiris is shown to be monophyletic only if Iris
falcifolia Bunge is included, but that Central Asian
rhizomatous species is morphologically a member of
subgenus Iris (Hall, 2011), and the exact identity of
Wilson’s material of I. falcifolia is questionable. The
present study is the first detailed work on molecular
phylogenetics and systematics of subgenus Scorpiris.
It should be noted that approximately 10 juno taxa,
primarily from Afghanistan and adjacent Pakistan,
are not in cultivation (see under ‘Species not included
in this study’ below) and thus could not be sampled.
The main objective of this study is to unravel evolutionary relationships among the species of subgenus
Scorpiris. We used DNA sequences from the following
regions: matK, rpl14-rps8 spacer, infA-rpl36 spacer,
trnE-trnT spacer, trnL intron and trnL-F spacer.
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
284
N. IKINCI ET AL.
Nuclear ribosomal spacer (ITS nrDNA) sequences
have not been used because in Iris and many other
Iridaceae these spacers exist as multiple copies, and
no single copy type seems to be found in all species,
thus making ITS rDNA an unsuitable marker in Iris.
(AB Gene, Epsom, UK). Amplified products were purified using NucleoSpin Extract II PCR purification kit
(Machery-Nagel GmbH & Co. KG, Duren, Germany)
following the manufacturer’s protocols.
MATERIAL AND METHODS
DNA EXTRACTION
Cycle sequencing reactions were performed using the
BigDye Terminator Kit ver. 3.1 (Applied Biosystems
Inc.). Cycle sequencing products were cleaned using
the ethanol–EDTA precipitation method (protocol provided by Applied Biosystems Inc.). Cleaned samples
were analysed on an ABI 3 capillary DNA sequencer,
using the manufacturer’s protocols.
The raw sequences were edited and contigs
assembled using Sequencher version 4.1 (Gene Codes
Inc., Ann Arbor, MI, USA). They were aligned by eye
in the matrix following the guidelines provided by
Kelchner (2000). All sequences have been submitted
to GenBank (accession numbers in Table 1).
Maximum parsimony (MP) was used to analyse the
combined plastid data in PAUP ver. 4.0b10 (Swofford,
2002). All changes were assessed as unordered and
equally weighted (Fitch parsimony; Fitch, 1971).
Indels were coded as missing data and thus not
included in the analyses. Most parsimonious trees
(MP) were obtained using 1000 replicates of random
taxon addition sequence and tree bisection–
reconnection (TBR) branch swapping saving multiple
trees (MULTREES), with a limit of ten trees per
replicate. Bootstrap analyses (Felsenstein, 1985) were
carried out using 1000 replicates with TBR swapping,
and a limit of ten trees retained for each replicate.
Bayesian analysis was carried out using MrBayes
version 3.1.2 (Huelsenbeck & Ronquist, 2001). An
HKY85 model was specified in which all transitions
and transversions have potentially different rates.
More complex models were also tested, but these
provided the same tree with similar posterior probabilities (PP; results not shown). Analyses were performed with 500 000 generations of Monte Carlo
Markov chains with equal rates and a sampling frequency of ten. Three separate runs were performed to
ensure that the analysis was producing consistent
results. Microsoft Excel was used to plot generation
number against lnL to find the ‘burn-in’. Trees of low
PP were deleted, and all remaining trees were
imported into PAUP ver. 4.0b. A majority rule consensus tree was produced showing the frequencies (i.e.
posterior probabilities) of all observed bi-partitions.
SEQUENCE
DNA was extracted from living specimens cultivated
in the Alpine Unit at the Royal Botanic Gardens, Kew
(Table 1) or from seeds. Vouchers are photographs
taken of flowering specimens and mounted on herbarium sheets, stored in the Herbarium of the Royal
Botanic Gardens, Kew (K). For extractions, 1 g of
fresh tissue was used. DNA was extracted using a
modified Doyle & Doyle (1987) 2 ¥ cetyl trimethylammonium bromide (CTAB) method. DNA was precipitated in chilled ethanol (-20 °C) for at least 24 h and
then re-suspended in 1.55 g mL–1 caesium chloride/
ethidium bromide. Samples were then purified using
a density-dependent gradient, followed by removal
of the ethidium bromide with butanol and caesium
chloride by dialysis; all DNA is stored in 10 mM
Tris-EDTA (TE) (pH 8) -80 °C in the DNA Bank at
the Royal Botanic Gardens, Kew; these samples are
available to other researchers upon request (http://
data.kew.org/dnabank/homepage.html).
MARKER
SELECTION/AMPLIFICATION
Twelve plastid markers selected from those studied in
Shaw et al. (2007) were evaluated, and the most variable four of these were amplified for the whole data
set of 60 Iris taxa: 52 taxa from subgenus Scorpiris
and eight outgroup taxa from other subgenera of
Iris (subgenus Iris, subgenus Nepalensis (Dykes)
G.H.M.Lawr. and subgenus Limniris (Tausch) Spach).
The rpl14 and rpl36 regions were amplified with
primers rpl14-rps8 and infA-rpl36 (Shaw et al., 2007).
Primers trnE (UUC) and trnT (GGU) (Ebert &
Peakall, 2009) were used to amplify the trnE-T
region. The matK region was amplified using the
primers -19F (Kores et al., 2000) and 2R (Johnson &
Soltis, 1995). The last regions, trnL intron and trnL-F
intergenic spacer, were amplified as one piece with
the c and f primers of Taberlet et al. (1991). With the
exception of matK, these regions were sequenced with
the same primers used for amplification; sequencing
of this region was carried out using the PCR primers
plus two internal primers, 458F (Molvray, Kores &
Chase, 2000) and 1326R (Sun, McLewin & Fay, 2001).
Target regions were amplified in a Gene AmpTM
9700 PCR system (Applied Biosystems Inc., Warrington, UK) using ReddyMix PCR Mastermix at
2.5 mM magnesium chloride (MgCl2) concentration
PRODUCTION/ANALYSES
RESULTS
The six regions were amplified for all 52 taxa (57
accessions) from subgenus Scorpiris and eight outgroup taxa from other subgenera of Iris (Table 1). The
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
Chase I-87; Kew 1982–3679. Pakistan: North-West
Frontier Prov., Sirikot Hills
Chase 11470; Kew 1998–4341. Uzbekistan: Pskem Range,
Pskem Valley, Sjemessas, c. 2500 m
No voucher: Kew 2008–1995. Armenia: Syunik District,
near Schvanidzor
Hall, T. image 83–3441: Kew 1983–3441. Iran: Bakhtaran
(Kermanshah), Kuh-e Bozab, 2300 m
Hall, T. image 77–2757; Kew 1977–2757. Afghanistan:
Kataghan Prov., 11 km S of Farkhar, 2100–2400 m
Hall, T. image 02–150: Kew 2002–150. Tadzhikistan: near
Duschanbe
Chase 11220: Kew 1999–3340. Tadzhikistan: Sanglok
Range, c. 1600 m
Hall, T. image 96–2662; Kew 1996–2662. Uzbekistan: W
Tien-Shan, Baschkhizilsai Valley
Hall, T. image 82–3698; Kew 1982–3698. Armenia: near
Sevan Lake, 2100 m
Chase 11472; Kew 1985–2299. Turkey: B9 Van Prov.,
108 km west of Van towards Bitlis, 1750 m
Chase I-122 ; Kew 1977–2765. Afghanistan: Herat Prov.,
Kotal-e Mir Ali, 1680 m
Chase I-25: Kew 1988–2789. Jordan: Desert Highway 53,
45 km south of Maan
Chase 10921; Kew 1982–3876. Turkmenistan: Kopet Dag,
near Dushak, 1900–2000 m
Chase I-46: Kew 1983–3440. Iran: Gorgan, Golestan Forest
near Almeh, 1600 m
Chase I-216: Kew 1985–3374. Turkey: B6 Sivas Prov.,
50 km east of Sivas
Chase I-217; Kew 1990–2446. Turkey: B5 Nevşehir Prov.,
between Zelve and Avanos, 1000 m
Hall, T. image 97–6377; Kew 1997–6377. Kirgizstan:
Fergana Range, Kugart Valley, c. 2200 m
Hall, T. image 00–4213; Kew 2000–4213. Uzbekistan: S
Kyzyl Kum Desert, near village of Koktscha, 400 m
Chase 10922; Kew 1998–4342. Iran: 10k m south-east of
Servestan, 1700 m
Iris aitchisonii
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
I. hymenospatha subsp.
hymenospatha
I. hippolyti
I. graeberiana
I. galatica*
I. galatica
I. fosteriana*
I. fosteriana
I. edomensis
I. caucasica ssp.
caucasica
I. caucasica subsp.
turcica
I. cycloglossa
I. capnoides
I. aff. bucharica
I. bucharica
I. cf. baldshuanica
I. aucheri
I. atropatana
I. albomarginata
Voucher and provenance data (from RBG Kew database)
Taxa
FR870390
I-122
10922
37878
11222
I-217
I-216
I-46
10921
FR870436
FR870419
FR870432
FR870381
FR870392
FR870416
FR870429
11472
I-25
FR870377
FR870418
FR870439
FR870389
FR870400
FR870384
FR870431
FR870413
FR870402
matK
13033
11221
11220
13116
13227
11218
36518
11470
I-87
DNA bank
accession
FR870713
FR870696
FR870709
FR870658
FR870669
FR870693
FR870667
FR870706
FR870654
FR870695
FR870716
FR870666
FR870677
FR870661
FR870708
FR870690
FR870679
rpl14-rps8 IGS,
rps8 gene &
rps8-rpL36 IGS
Table 1. List of taxa used in this study, their RBG Kew DNA Bank accession numbers and GenBank accession numbers
FR870581
FR870564
FR870577
FR870526
FR870537
FR870561
FR870535
FR870574
FR870522
FR870563
FR870584
FR870534
FR870545
FR870529
FR870576
FR870558
FR870547
trnE-trnT
spacer
FR870647
FR870630
FR870643
FR870592
FR870603
FR870627
FR870601
FR870640
FR870588
FR870629
FR870650
FR870600
FR870611
FR870595
FR870642
FR870624
FR870613
trnL intron
and trnL-F
spacer
MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
285
Hall, T. image 98–4343; Kew 1998–4343. Iran: near Arak,
Mt Kuh-e Burfhane
Hall, T. image 00–4218; Kew 2000–4218. Uzbekistan:
Tschatkal Range, Schavzasai Valley, above village of Tut,
c. 1000 m
Chase 10924: Kew 1978–3606. Turkmenistan: Kopet Dag,
20 km west of Ashkabad
Hall, T. image 93–3295; Kew 1993–3295. Kazakhstan: Mt
Kyndiktas, east end of the Kurdai Pass, c. 90 km north
of Bishkek, 900 m
Hall, T. image 91–1475; Kew 1991–1475. Tadzhikistan:
South Pamir-Alai, Tabakchi Mts near Kurgan Tube,
c. 700 m
Chase I-93; Kew1989-1787. Uzbekistan: Alay Range, south
of Fergana
Chase I-77; Kew 1984–1505. Uzbekistan: Samarkand Mts,
Takhta-Karacha Pass, by the Kashkadarya Spring,
1650 m
Hall, T. image 96–2664; Kew 1996–2664. Uzbekistan: near
Dzhizak, Timurlan Gate
Chase I-96 K; Kew 1982–4802. Afghanistan: Hindu-Kush,
Salang Pass, 2000 m
Chase I-28; Kew 1984–8346. Uzbekistan: Samarkand Mts,
Samarkand Road about 10 km from Aman Kutan Valley,
800 m
Chase I-29; Kew 1989–2503. Uzbekistan: southern part of
country, Baisuntau Range
Hall, T. image 00–4227; Kew 2000–4227. Uzbekistan:
Fergana Valley, mountains north-east of Aravan, 1100 m
Hall, T. image AG-13991; T. Hall: private collection.
Turkey: north-eastern part of country, A8 Artvin Prov.,
Yusufeli, around Öğdem, 1635 m
Hall, T. image 98–4353; Kew 1998–4353. Uzbekistan:
south-west Pamir-Alai, Chulbair Mts, near village of
Sina
Hall, T. image 02–2744; Kew 2002–2744. Turkey: C4
Mersin (Içel) Prov., just south of Gökbelen, on main road
from Gülnar to Silifke, 1250 m (but data suspect – more
likely collected in Syria)
I. hymenospatha subsp.
leptoneura
I. inconspicua
I. nusairiensis
I. nicolai
I. nezahatiae
I. narynensis
I. narbutii*
I. narbutii
I. microglossa
I. maracandica
I. magnifica
I. linifolia
I. leptorrhiza
I. kuschakewiczii
I. kopetdagensis
Voucher and provenance data (from RBG Kew database)
Taxa
Table 1. Continued
15843
11235
36519
13032
FR870376
FR870379
FR870382
FR870380
FR870388
I-28
I-29
FR870404
I-96
FR870420
FR870406
I-77
37876
FR870378
FR870408
FR870430
I-93
I-74
11226
FR870433
FR870412
37877
10924
FR870385
matK
11224
DNA bank
accession
FR870653
FR870656
FR870659
FR870657
FR870665
FR870681
FR870697
FR870683
FR870655
FR870685
FR870707
FR870710
FR870689
FR870662
rpl14-rps8 IGS,
rps8 gene &
rps8-rpL36 IGS
FR870521
FR870524
FR870527
FR870525
FR870533
FR870549
FR870565
FR870551
FR870523
FR870553
FR870575
FR870578
FR870557
FR870530
trnE-trnT
spacer
FR870587
FR870590
FR870593
FR870591
FR870599
FR870615
FR870631
FR870617
FR870589
FR870619
FR870641
FR870644
FR870623
FR870596
trnL intron
and trnL-F
spacer
286
N. IKINCI ET AL.
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
I. subdecolorata
I. stenophylla subsp.
stenophylla
I. stocksii
I. stenophylla subsp.
allisonii
I. rosenbachiana
I. regis-uzziae
I. pseudocaucasica s.s.
I. pseudocapnoides
I. postii*
I. postii
I. popovii
I. planifolia
I. persica
I. aff. parvula
I. parvula s.s.
I. palaestina
I. orchioides
I. aff. nusairiensis
Hall, T. image 03–2998; Kew 2003–2998. Turkey: B7
Malatya Prov., between Malatya and Pütürge, Kubbe
geçidi,1900 m
Hall, T. image 96–2663; Kew 1996–2663. Uzbekistan:
western Tien-Shan, Baschkhizilsai Valley
Hall, T. image 02–183; Kew 2002–183. Israel: coastal
plain, 50 km north towards Tel Aviv, Biniamina (Sharon)
Hall, T. image 02–210; Kew 2002–210. Uzbekistan:
southern part, Kugi-Tang, above the village of
Hodjaankan
Chase I-92; Kew 1991–1330. Tadzhikistan: Hissar Range,
near village of Takob
Hall, T. image 90–2096; Kew 1990–2096. Turkey: B7
Elaziğ Prov., 6 km east from Pertek, 1260 m
Chase I-20; Kew 1992–40. Italy: Sicily, Prov. Palermo,
8 km along road from Piana degli Albanesi to Ficuzza,
800 m
Chase I-38 K; Kew 1991–1478. Tadzhikistan: southern
Pamir-Alai, near village of Tovil-Dara, 2100 m
No DNA Bank voucher; Kew 1988–2783. Jordan: 6–8 km
north-east of Qasr Kharana (no voucher, but the only
juno iris recorded for this area)
Chase I-73; Kew 1983–2159. Saudi Arabia: Turayf Camp,
north-east of Turayf, 884 m
Hall, T. image 84–1511; Kew 1984–1511. Uzbekistan:
Aktash Mtn near Tashkent, c. 1300 m
Hall, T. image 98–4358; Kew 1998–4358. Iran: Tochal Mts,
north of Teheran
Chase I-24; Kew 1984–4829. Jordan: southern part, Ras
an-Nagb
Chase I-30; Kew 1992–1999. Tadzhikistan: Sanglok Range,
c. 1700 m
Hall, T. image 00–4231; Kew 2000–4231. Turkey:
southwestern part, C3 Antalya Prov., south of Akseki,
900 m
Hall, T. image 00–3729; Kew 2000–3729. Turkey: C4
Konya Prov., 30 km from Konya to Seydişehir, 1600 m
Hall, T. image 10–1073; Kew 2010–1073. Afghanistan:
Helmand Prov., Kajaki, near reservoir
Chase I-239; Kew 1990–2580. Kazakhstan: Chu-Ili Mts,
west of Kurdai Pass, c. 900 m
I-239
11245
13112
I-30
I-24
11238
11236
I-73
13049
FR870691
FR870651
FR870374
FR870414
FR870678
FR870675
FR870688
FR870686
FR870671
FR870668
FR870652
FR870692
FR870687
FR870660
FR870704
FR870676
FR870664
FR870694
FR870699
FR870401
FR870398
FR870411
FR870409
FR870394
FR870391
FR870375
FR870415
FR870410
I-20
I-38
FR870383
FR870427
FR870399
FR870387
FR870417
FR870422
13045
I-92
13115
13118
11237
21829
FR870559
FR870519
FR870546
FR870543
FR870556
FR870554
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MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
287
31373
I-131
I-133
I-132
I-147
I-136
I-82
I-90
I-137
I-88
I-75
I-45
11257
11253
11252
11247
I-98
11248
11246
DNA bank
accession
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matK
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rpl14-rps8 IGS,
rps8 gene &
rps8-rpL36 IGS
*Second samples of indicated species used to verify the position of the species (not included in Figure 3 – see text).
I. falcifolia
I. collettii var. acaulis
I. aff. decora
I. decora
I. staintonii
I. tectorum
I. japonica
I. zaprjagajevii
Outgroups
I. atropurpurea
I. milesii
I. willmottiana
I. warleyensis*
I. warleyensis
I. aff. vicaria
I. vicaria
I. aff. tubergeniana
Chase I-90; Kew 1989–1103 (no data)
Chase I-137; Kew 1981–8091. India: Himachal Pradesh,
near Taralla Saraj, 2743 m
Chase I-136; Kew 1990–374. Bhutan (no other data)
Chase I-82; Kew 1979–5129. Japan: Honshu, Niigata
Prefecture, Itoigawa City, Nechi, 60 m
Chase I-147; Kew 1992–2305. Nepal: Ganesh Himal,
Biram, Abuthum Lekh, 3750 m
Chase I-132; Kew 1974–4108. Nepal: Bagang Khola,
3050 m
Chase I-133; Kew 1981–4071. China: Sichuan, Shih-Mein,
1000 m
Chase I-131; Kew 1990–3105. China: Yunnan, Cang Shan
above Dali, near Zhong He Temple, 2591 m
Hall, T. image 06–1130; Kew 2006–1130. Uzbekistan:
Bokhara – Qarshi road
Hall, T. image 89–2504; Kew 1989–2504. Uzbekistan:
southern part, southern Kugi-Tang
Chase I-98; Kew 1986–3837 (no data)
Hall, T. image 96–2667; Kew 1996–2667. Uzbekistan:
western Tien-Shan, Baschkhizilsai Valley
Hall, T. image 93–3342; Kew 1993–3342. Tadzhikistan? (no
data)
Hall, T. image 99–3352; Kew 1999–3352. Uzbekistan:
southern part, Chulbair Mts, near village of Sina (locus
classicus), c. 2200 m
Hall, T. image 91–1479; Kew 1991–1479. Tadzhikistan:
Hissar Range, Maihura Valley, 2400 m
Hall, T. image 89–2507; Kew 1989–2507. Uzbekistan:
western Pamir-Alai, Susi-Stau Range
Chase I-45; Kew 1984–1510. Uzbekistan: Samarkand Mts,
summit of Takhta-Karacha Pass, 1700–1800 m
Chase I-75; Kew 1990–2579. Kazakhstan: Karatau, Sajasu
Valley, 1300 m
Chase I-88; Kew 1990–2312. Tadzhikistan: Pamir, 3205 m
I. svetlanae
I. tadshikorum
I. tubergeniana
Voucher and provenance data (from RBG Kew database)
Taxa
Table 1. Continued
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and trnL-F
spacer
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N. IKINCI ET AL.
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MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
aligned data set comprised 4599 characters, of which
580 were variable and 270 were potentially parsimony
informative. Analysis produced > 15 000 most parsimonious trees (the limit set before analysis) with a
length of 780 steps, consistency index (CI) = 0.82 and
retention index (RI) = 0.90. One of the most parsimonious trees (BP) is illustrated in Figure 3 with
bootstrap percentages (BP) and Bayesian posterior
probabilities (PP) provided below the (BP/PP) branches and branch lengths above; nodes not found in the
strict consensus tree are indicated by an arrowhead; in
the Bayesian tree, some of the unresolved taxa in the
parsimony analysis are resolved, but they receive PP
much less than 0.50 and are not indicated in Figure 3.
The analysis indicated that subgenus Scorpiris is
monophyletic (BP 100, PP 0.97; Fig. 3). The first
major clade (clade A) comprises species from section
Physocaulon and is sister to all other sampled
members of subgenus Scorpiris (BP 100, PP 0.97).
This section occurs in Afghanistan (Hindu-Kush) and
adjacent regions (including southern Pamir-Alai and
Kopet Dag). A smaller clade (clade B; BP 100,
PP 0.98) formed by accessions of I. bucharica Foster
and I. vicaria (Vved.) T.Hall & Seisums (see Appendix), from northeastern Afghanistan/Tadzhikistan
and Tadzhikistan/southern Uzbekistan, respectively,
is then sister to the remaining taxa, except perhaps
for I. microglossa Wendelbo (see below).
Iris microglossa, a morphologically isolated species
from northeastern Afghanistan, is part of a polytomy
with the other two other strongly supported major
clades (clades C and D with BP 91 and 96, respectively, both PP 0.97). Iris stocksii (Baker) Boiss.
(southeastern Afghanistan and western Pakistan),
I. aitchisonii (eastern Afghanistan, western Pakistan
and Kashmir) and I. cycloglossa (western Afghanistan) form a subclade (C1; BP 80, PP 0.86) that is
part of a basal trichotomy in the second major clade
(clade C), which is further divided into two subclades.
One of the two subclades (C2; BP 98, PP 0.94) consists entirely of junos with a western distribution
and contains two Turkish endemics, I. stenophylla
Hausskn. ex Baker and I. galatica Siehe, with the two
species that comprise the Acanthospora group:
I. planifolia, primarily from the western Mediterranean and North Africa and I. palaestina, from the
eastern Mediterranean. This subclade also includes
I. aucheri (Baker) Sealy and its close relatives from
the Near East, I. nusairiensis Mouterde and I. regisuzziae Feinbrun. The third subclade (C3; BP 93,
PP 0.93) consists of all the other western Asian junos,
including I. caucasica Hoffm. and I. nezahatiae Güner
& H.Duman (similar species except in flower colour),
I. persica L. (Fig. 1), I. atropatana Grossh. and I. hymenospatha B.Mathew & Wendelbo. Iris edomensis
Sealy and I. postii, from the Middle East, also fall in
289
this subclade. Iris fosteriana Aitch. & Baker, a highly
divergent species from Paropamissus in northwestern Afghanistan and the Kopet Dag of Iran and
Turkmenistan, is also placed within this subclade as
sister (BP 94, PP 0.98) to I. pseudocaucasica Grossh.,
from the Elburs range of Iran and mountainous
regions of Transcaucasus, the pair then sister to the
rest of subclade C3 (BP 100, PP 0.96), the species of
which are otherwise genetically similar and highly
unresolved.
The final major clade (clade D; BP 96, PP 0.97)
includes all the remaining Central Asian species
(Pamir-Alai and Tien-Shan). This clade is also divided
into two subclades. The first subclade (D1; BP 80,
PP 0.92) has Iris capnoides (Vved.) T.Hall & Seisums
(see Appendix) as sister (BP 98, PP 0.98) to all other
species in this subclade; I. orchioides Carrière (Fig. 1)
and I. aff. tubergeniana Foster form a pair (with
BP 100, PP 0.99) that is sister (BP 99, PP 0.99) to a
subclade that is internally genetically depauperate
and highly unresolved; this subclade is composed of:
I. graeberiana Sealy (which is sister to the others
in Bayesian analysis, PP 0.95), I. albomarginata
R.C.Foster, I. inconspicua (Vved.) T.Hall & Seisums
(see Appendix), I. willmottiana Foster, I. tubergeniana
s.s., I. pseudocapnoides Ruksans, I. subdecolorata
Vved. and I. kuschakewiczii B.Fedtsch. The second
subclade (D2; BP 100, PP 0.97) has a basal polytomy
with I. parvula (Vved.) T.Hall & Seisums s.s. (Fig. 1;
see also Appendix), I. magnifica Vved., I. narynensis
O.Fedtsch. plus (BP 60) I. aff. parvula, I. tadshikorum
Vved., I. linifolia (Regel) O.Fedtsch. and a further
subclade (BP 80, PP 0.96) that contains three closely
related species, I. maracandica (Vved.) Wendelbo,
I. svetlanae (Vved.) T.Hall & Seisums (see Appendix),
I. hippolyti (Vved.) Kamelin (Fig. 1) and a final pair
(BP 62, PP 0.97) formed by I. narbutii O.Fedtsch. (a
morphologically isolated species from the Pamir-Alai
and western Tien-Shan; Fig. 1) and I. warleyensis
Foster (Fig. 1) from the western Pamir-Alai. The
Bayesian analysis positions I. narynensis with I. aff.
parvula (BP 62, PP 0.97) and I. linifolia with I. parvula s.s. and I. tadshikorum (PP 0.96).
For the Bayesian analysis, the ‘burn-in’, 5000 trees
of low likelihood were omitted from the consensus
tree. As indicated by the posterior probabilities provided above, the Bayesian results are almost identical
with those from the parsimony analysis.
DISCUSSION
The precise position of subgenus Scorpiris within Iris,
and within Iridaceae as a whole, has been contentious. In this study, our arrangement of the tree
(Fig. 3) positions subgenus Scorpiris as sister to the
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
290
N. IKINCI ET AL.
2
5
0
4
3
1
0.96
1
62/0.97
16
100/
0.97
7
62/0.97 1
1
60/
0.93
2
80/0.96
2
2
2
0
0
1
0
2
4
96/0.97
1
1
80/0.92
1
0.79
4
93/
0.93
9
100/
0.96
6
98/
0.98
5
99/
0.99
8
2
75/0.98
8
4
98/0.94
4
80/0.86
16
13
100/0.98
16
100/0.97
17
100/0.97
13
3
8
8
3
68/0.93
0.35
4
91/
0.97
2
0.57
19
12
8
3
1
3
94/0.98
81/0.93
3
78/0.93
1
2
1
1
3
1
0.58
1
9
100/
0.97
1
0.98
4
10
100/0.99
20
7
1
0.12
2
75/
0.79
0.95
3
3
4
3
11
1
5
2
82/0.98
3
5
3
97/0.97
6
4
97/0.97
8
2
11
59/0.93
17
3
3
1
2
65/0.94 1
6
3
15
2
2
100/0.97
1
1
61/0.97 12
7
9
100/0.99
4
49
1
100/0.96 61
1
15
3
8
0.44
6
0.93 7
1
17
97/0.96
0.74
100/0.95 26
1
19
0.46 37
linifolia
parvula
tadshikorum
narynensis
aff. parvula
magnifica
narbutii
warleyensis
svetlanae
hippolyti
maracandica
pseudocapnoides
tubergeniana
willmottiana
inconspicua
albomarginata
subdecolorata
kuschakewiczii
graeberiana
orchioides
aff. tubergeniana
capnoides
postii
caucasica
nezahatiae
caucasica subsp. turcica
hymenospatha subsp. leptoneura
persica
edomensis
atropatana
hymenospatha
fosteriana
pseudocaucasica
nusairiensis
aucheri
aff. nusairiensis
regis-uzziae
galatica
palaestina
planifolia
stenophylla subsp. allisonii
stenophylla
stocksii
cycloglossa
aitchisonii
microglossa
bucharica
vicaria
aff. bucharica
aff. vicaria
nicolai
cf. baldshuanica
zaprjagajevii
rosenbachiana
popovii
leptorrhiza
kopetdagensis
atropurpurea
falcifolia
staintonii
aff. decora
collettii var. acaulis
decora
milesii
tectorum
japonica
D2
D1
C3
C2
C1
B
A
outgroups
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
291
Figure 3. One of the most parsimonious trees for Iris subgenus Scorpiris. The numbers above branches are Fitch branch
lengths (DELTRAN optimization). Bootstrap percentages > 50 and Bayesian posterior probabilities are given below
branches for supported clades (BP, PP); if no bootstrap percentage is provided, then that node received < 50%; some groups
resolved in the Bayesian tree (with much less than PP 0.50) are shown here as polytomies. (Table 1).
䉳
rest of Iris, but the arrangement of the outgroups was
carried out for convenience and does not reflect the
actual position of the subgenus within Iris. In addition to their unusual flower and root morphology,
junos also have a unique leaf anatomy (Rudall &
Mathew, 1993), being the only members of Iridaceae
with entirely bifacial leaves (upper and lower surfaces
with different underlying anatomy), even at the seedling stage. Other studies (Tillie et al., 2001; Wilson,
2004, 2009, 2011) have shown that this leaf condition
is a reversal from the unifacial leaves (both surfaces
with the same anatomy) typical for Iridaceae and
that subgenus Scorpiris is clearly deeply embedded
within Iris, thus making recognition of genus Juno
inappropriate.
This subgenus exhibits an enormous diversity of
characters: overall dimensions (some species are only
a few centimetres tall and virtually stemless, whereas
Iris magnifica may reach a metre in height), bulb and
storage root shape, foliage form and dimensions,
flower colour (there can be bewildering variation,
even within a single species), flower size and shape
and crest development (Fig. 1).
Generally, the molecular results indicate an isolated position for section Physocaulon (primarily
Afghanistan, but extending into adjacent regions that
include parts of Iran, Turkmenistan, western Pakistan and Tadjikistan) and support the separation of
northeastern species of section Juno (almost exclusively Pamir-Alai and Tien-Shan in Central Asia)
from western species of section Juno (western Asia
and the Mediterranean), whereas a small disparate
group of southeastern (primarily Hindu-Kush) species
of section Juno appears closer to the western species.
Because of the tuber-like stem base, Wendelbo &
Mathew (1975) considered section Physocaulon to be
the most morphologically advanced group of junos, a
hypothesis that does not involve knowing their position in a phylogenetic tree.
Section Physocaulon (clade A) is further divided
into two well-supported subclades, equivalent to Rodionenko’s (1961) Juno series Physocaulon (synonym
series Drepanophyllae Rodion.) and Juno series
Rosenbachianae Rodion., although we do not accept
his more recent (Rodionenko, 1994) grouping of
species within that section. Features of the section
include a persistent, tuber-like base of the flower
stem (giving rise to the name of the section; Fig. 4),
which, along with the bulb, becomes an important
part of the storage organ, seeds with a prominent
white aril, radish-like roots that are markedly
swollen above and tapering abruptly below, and falls
usually with more-or-less parallel-sided claws (‘hafts’)
without wings (lateral extensions), the margins of
which may be either downturned or upturned and
gutter-like.
The Physocaulon subgroup, with narrow hafts to the
falls and standards that are greatly reduced and
almost whisker-like, contains Iris kopetdagensis
(Vved.) B.Mathew & Wendelbo, one of only five species
with zona-aperturate pollen (T. Saito, M. Harley &
T. Hall, unpubl. data). Rodionenko (1961, 1994) considered that only two species he placed in section
Physocaulon exhibited pollen grains of this type
(I. drepanophylla Aitch. & Baker, not sampled here,
and I. kopetdagensis), segregating them in the above
subgroup, but he was undoubtedly studying material
of I. kopetdagensis wrongly identified as I. drepanophylla; confidently identified material of the latter
species, in fact, has clypeate pollen (T. Saito, M. Harley
& T. Hall, unpubl. data). Although distinct in pollen
morphology, they are otherwise close species (Gustafsson & Wendelbo, 1975; Wendelbo & Mathew, 1975),
and there is little doubt that they both belong in the
Physocaulon subgroup (see also under ‘Species not
included in this study’ below). The affinities of I. leptorrhiza Vved. have been uncertain, mainly because of
its poorly known morphology. Rodionenko (1961)
studied I. leptorrhiza but he omitted to place it in any
of his Juno groups, although later it was placed in
section Juno (Rodionenko, 1994). In contrast, the
molecular study by Tillie et al. (2001) suggested an
affinity with I. nicolai Vved., the only other sampled
species of section Physocaulon. These new results not
only support the later report, but also show that
I. leptorrhiza is sister (BP 100, PP 0.99) to I. kopetdagensis in this subclade. Observations on recently
introduced material reveal that I. leptorrhiza has a
physocaulon during dormancy. It has been distinguished, also, as the only juno lacking storage roots
(e.g. Vvedensky, 1971); however, both T. Hall and A.
Seisums have observed the occasional tiny, radish-like
root during its dormant phase, as well as roots that are
soft-textured and absent during aestivation. Although
information is lacking on the absence/presence of an
aril to its seeds, this placement of I. leptorrhiza also
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
292
N. IKINCI ET AL.
Figure 4. Underground storage organ differences in Iris subgenus Scorpiris: left (I. aucheri) , fairly typical for section
Juno; right (I. nicolai), that found in the species of sect. Physocaulon.
has support from its floral and foliar morphology. The
species has clypeate pollen (T. Saito, M. Harley & T.
Hall, unpubl. data). The above two sister species,
placed by this study in the Physocaulon subgroup, as
well as those not sampled but highly likely to belong
here, have distinct internodes at the fruiting stage,
bracts that remain green throughout the vegetative
period and similarities in their flower morphology;
these would appear to be more reliable characters to
delimit this subgroup.
The Rosenbachianae subgroup consists of those
species with generally larger, more paddle-shaped
standards and falls with slightly widened hafts that
are invariably downturned at the margins; Iris cf.
baldshuanica O.Fedtsch., I. rosenbachiana Regel
(Fig. 1), I. popovii Vved., I. nicolai and I. zaprjagajevii
(N.V.Abramov) T.Hall & Seisums (see Appendix) form
a well-supported clade (BP 100, PP 0.97). These
results are supported by their generally similar morphology (compressed foliage internodes, membranous
bracts and flowering considerably in advance of leaf
development) and confirm that this taxonomic category is a natural subgroup. The species studied here
all have clypeate pollen (T. Saito, M. Harley & T. Hall,
unpubl. data).
One of the more unexpected results of this study is
the isolated position of a well-supported clade (B;
BP 100, PP 0.98) comprising just two species, Iris
bucharica and I. vicaria, each represented by two
samples. Both species are members of section Juno,
which contains all taxa with a flower stem dying off to
the base (so without a physocaulon), apart from the
two species of the Acanthospora group [also without a
physocaulon but separated by Rodionenko (1961) on
account of their distinctive pollen-surface architecture]. Iris bucharica and I. vicaria both have a welldeveloped stem and wingless flowers, but most
authors/taxonomists would have expected them to be
placed with other species of the eastern alliance.
However, support for their precise position within the
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
major polytomy formed by clade C (western species)
and clade D (eastern species) is lacking, and they
could still end up associated with other eastern taxa,
perhaps as sister to clade D. We note in this study
that even with the limited samples used, the groupings obtained in clade B do not correspond to the
widely accepted separation of I. vicaria and I. bucharica based merely on their flower colour differences
(white to blue–violet stained or pale yellow to rich
yellow, respectively). Chromosome data (M. Johnson
& A. Seisums, unpubl. data) show variation in
numbers and even ploidy that also do not correlate
with this view. Both species require further study.
The highly isolated position of Iris microglossa
(with winged falls), potentially as sister to all other
taxa of section Juno (a section that contains a huge
range of characters; Wendelbo & Mathew, 1975) and
the Acanthospora group, confirms its distinctive
nature. It also belongs to section Juno but has little in
common with either of those other highly divergent,
primarily Afghan/Hindu-Kush species I. cycloglossa
and I. aitchisonii which, although grouped in a sister
clade to the whole of the western alliance of species
(with I. stocksii, which is paired with I. cycloglossa),
are also morphologically distinct from one another,
especially in floral morphology; both have karyotypes
distinct from other junos but with some previously
unappreciated similarities to each other (Gustafsson
& Wendelbo, 1975). These two relic species (I. cycloglossa and I. aitchisonii) sometimes have branching
stems, a character found in no other juno species,
although this character is often absent from both
cultivated and wild specimens. They also have zona–
aperturate (hamburger-like) pollen (T. Saito, M.
Harley & T. Hall, unpubl. data), whereas all other
species in section Juno have clypeate (football-like)
pollen. Initially, Rodionenko was unable to study
either I. cycloglossa (described in 1959) or I. aitchisonii, and his 1961 circumscription of genus Juno
section Juno contained exclusively species with the
latter type of pollen. Later he was able to study
I. cycloglossa and, based upon its zona–aperturate
pollen and other morphological characters, segregated
it into its own section: Juno section Wendelboa
Rodion. (Rodionenko, 1994); at the same time he
placed I. aitchisonii with other species of section
Juno. Like another divergent species, I. fosteriana (in
subclade C3), I. cycloglossa and I. aitchisonii have
somewhat strap-shaped foliage (a rare feature in the
subgenus; the majority of species have tapering, lanceolate to ovate–lanceolate leaves). It would appear
that members of section Juno from Afghanistan and
adjacent areas, especially the Hindu-Kush (I. microglossa and all three species placed in C1), are related
more closely to the western alliance than to Central
Asian species. That I. stocksii (central and southern
293
Afghanistan and adjacent Pakistan) is also placed
here in C1 is not unexpected, although its floral
morphology and semi-desert habitat would appear to
have more in common with compact I. persica and its
close relatives in subclade C3. Often the stem of
I. stocksii is completely concealed by foliage at anthesis; however, in fruit it does elongate and internodes
are clearly visible, as they are in both I. cycloglossa
and I. aitchisonii during flowering.
The western alliance (C) has two large, wellsupported subclades (C2 and C3). The first (C2) is
further divided into two. The first strongly supported
subclade consists of Iris stenophylla s.l. and the Acanthospora group. Johnson & Güner (2002) did not
recognize the separation of I. stenophylla into two
subspecies, but this species complex is heterogeneous
and clearly in need of further study; the two taxa
used in the present study are probably better
assigned to I. tauri Siehe (placed in synonymy with
I. stenophylla by Mathew, 1984, 1989). The two
closely related species of the Acanthospora group
(I. planifolia and I. palaestina), with clavate (spiny)
pollen (Rodionenko, 1987), are similar morphologically to other western species, especially to I. aucheri
in overall flower shape (T. Hall & A. Seisums, unpubl.
data). Despite their distinctive pollen architecture,
Rodionenko’s treatment of the Acanthospora group as
a separate section was viewed by Gustafsson & Wendelbo (1975) as questionable, and this has been found
unacceptable in the light of the present results. The
Acanthospora group plus I. stenophylla consist of
plants with compressed foliage internodes and winged
outer perianth segments. The second subclade of C2,
weakly supported, consists of I. aucheri and its relatives, I. regis-uzziae and I. nusairiensis, species with a
pronounced stem, lower leaves that form a sheath and
upper leaves that are erect, pale, inflated at the base
and somewhat bract-like in appearance, but enveloping the true bracts (or spathe valves) (T. Hall & A.
Seisums, unpubl. data). These species also exhibit
winged falls, although in I. regis-uzziae the lateral
extensions are occasionally not well expressed. Iris
aff. nusairiensis, restricted to a small area in southern Turkey, is genetically more similar in these
results to I. aucheri (primarily southeastern Turkey,
extending into adjacent parts of Iran, Iraq, Syria
and Jordan), although in morphology and karyotype
(A. Seisums, unpubl. data) it has more in common
with the compact western Syrian I. nusairiensis; this
trio of species forms a supported minor subclade
in our results (BP 68, PP 0.93). A totally unanticipated member of this second subclade within C2 is
I. galatica. This endemic of central Anatolia has
always been considered, along with I. stenophylla,
most closely related to I. persica (Mathew, 1989),
which falls into the subclade sister to that in which
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
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N. IKINCI ET AL.
I. galatica is placed by the molecular analysis; morphologically, I. persica and I. galatica are extremely
close, except for their bract features and some floral
morphological details. Dykes (1913) placed I. stenophylla and I. galatica in synonymy with I. persica,
treating them both as varieties of the last species, but
Mathew (1984, 1989, 1997) considered all three
species distinct. It should be noted that both I. galatica and I. stenophylla have falls that are ‘pinched’
between haft and blade, as if by forefinger and thumb;
however, this character has never been observed in
I. persica (T. Hall & A. Seisums, unpubl. data).
Iris galatica appears to have so little in common
with I. aucheri and its close relatives, in either morphology or karyotype (A. Seisums, unpubl. data), that
its proximity, albeit distant, is puzzling; that I. galatica and I. persica are not related is well supported by
these results. Because of this anomaly, a second
sample of I. galatica was sequenced, but this confirmed the unexpected results (results not shown)
from analysis of the first sample. Perhaps we are
looking at a species of potential hybrid origin, but as
only plastid DNA (maternally inherited) was used in
this study, this hypothesis remains to be explored
further. Parallelism in morphological traits is the
other possible explanation for this surprising placement of I. galatica.
The second major subclade (C3) of the western
alliance is also further subdivided, and bootstrap
support for this second western subclade is strong.
A highly divergent pair (BP 94, PP 0.98), Iris
pseudocaucasica and I. fosteriana (the latter species
with no previously recognized close affinities) is sister
(BP 100, PP 0.96) to all remaining taxa in this subclade. Mathew (1985) considered I. pseudocaucasica
and I. caucasica closely related; the latter is placed in
this internally highly unresolved subclade, but clearly
not with I. pseudocaucasica. Tillie et al. (2001) also
placed I. fosteriana with the western alliance,
although fewer juno species were included in that
study. A second accession of this species from a different part of its range was added (results not shown),
but this material confirmed its placement as shown
(Fig. 3).
Iris fosteriana, with elongated bulbs, extremely
fragile, long wiry roots, a distinct stem with strap-like
leaves, and wingless flowers with relatively large,
colourful, strongly reflexed standards, occupies an
important geographical position within subgenus
Scorpiris. It occurs in the Kopet Dag (northeastern
Iran and adjacent Turkmenistan) and in northwestern Afghanistan. The Kopet Dag is a phytogeographical bottleneck in the east-to-west distribution
of many other geophytes as well as these irises (with
regions both to the north and south of this range too
arid even for junos). Generally, such bottleneck
species have close relatives either to the east or to the
west, but rarely does a group have relatives on both
sides (e.g. the other juno from the Kopet Dag, I. kopetdagensis, has relatives only to the east). However,
we have seed-raised a spontaneous hybrid (named
‘Tehran
Beauty’)
between
I. fosteriana
and
I. pseudocaucasica s.s., which occurred initially in the
Tehran Botanic Garden. Although this is in itself not
proof of a close relationship between the two parental
species, it has been observed that garden hybrids
between the eastern and western representatives of
section Juno are extremely rare (T. Hall & A.
Seisums, unpubl. data). Seisums has produced the
only other substantiated hybrid (between I. orchioides
and I. caucasica), which nevertheless bridges this
apparent breeding barrier.
Among the remaining taxa in the second western
subclade (C3), we find both subspecies of Iris caucasica and the morphologically similar I. nezahatiae (all
with sheathing leaves, inflated bracts and falls with a
broad blade and wide wings; T. Hall & A. Seisums,
unpubl. data); I. caucasica ssp. turcica B.Mathew in
this study is genetically more similar to I. caucasica
ssp. caucasica and I. nezahatiae more divergent, but
the relationships of these taxa need further study and
are unresolved in the strict consensus tree. Also
placed in this subclade (C3) are I. persica and both
subspecies of its close relative I. hymenospatha; I. hymenospatha ssp. leptoneura B.Mathew & Wendelbo
appears to be more closely related to other species in
this matrix (BP 75, PP 0.79) than to I. hymenospatha
ssp. hymenospatha, which could mean that this subspecies should be elevated to species rank and confirms the results of morphological studies undertaken
by T. Hall and A. Seisums and cytotaxonomy (A.
Seisums, unpubl. data).
Of particular interest are the positions of Iris postii
and I. edomensis in the C3 subclade. Both are semidesert species (Mathew, 1989), endemics of the Syrian
desert and the Edom region of Jordan, respectively,
which so far cannot be assigned confidently to any of
the western species alliances (T. Hall & A. Seisums,
unpubl. data). Iris postii (from Iraq, eastern Jordan,
eastern Syria and Saudi Arabia) has some morphological features in common with I. aucheri (in subclade C2 here) and its relatives (e.g. a well-developed
stem after flowering; Mathew, 1985). It also has a
similar karyotype (the majority of chromosomes are
large metacentrics; A. Seisums, unpubl. data),
although this may well be coincidental. Therefore, we
added another confidently identified collection of this
species, but this second accession only confirms the
unexpected placement of I. postii. The similarities to
I. aucheri must be parallelisms because both subclades, C2 and C3, are well supported (BP 98 and 93,
PP 0.94 and 0.93, respectively).
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
Iris edomensis, in its foliar characteristics, compressed internodes and membranous bracts, most
resembles I. persica and its close allies, species that
have either one or both bracts noticeably translucent,
especially I. persica and I. hymenospatha. In flower
colour/markings, flower shape, outline of falls and
relatively large standards held well above the horizontal, I. edomensis is certainly divergent. Mathew
(1997) tentatively placed it closer to I. postii, but with
a caveat that the latter species was possibly related to
I. stocksii and other semi-desert species from further
east. Although there is little support for the precise
position of I. postii within this subclade, placement of
I. persica as sister to the Jordanian endemic I. edomensis is supported (BP 75, PP 0.98).
Iris atropatana, one of the most poorly understood
of the western species, is said to come from southern Transcaucasia and northeastern Turkey. Mathew
(1989) considered it a possible synonym of I. caucasica,
whereas Gabrielian (2001) considered these two taxa
distinct. Our studied material corresponds to the
later treatment of I. atropatana and indicates that
it is a member of a subclade (BP 75, PP 0.79) that
includes I. caucasica, I. nezahatiae, I. postii, I. persica,
I. edomensis and I. hymenospatha ssp. leptoneura. It
thus appears close to I. caucasica, but not clearly closer
to this species than to any of the other members of this
highly unresolved group of species.
There is great divergence between the storage roots
and bulbs of some of the western alliance of section
Juno e.g. Iris persica, I. aucheri and I. fosteriana.
However, apart from wingless I. fosteriana, the floral
morphology of western species (clade C), with winged
falls and a slightly raised ridge or prominent, moreor-less entire crest to the midrib of the blade, exhibits
less major variation when compared with those groups
from Central Asia, although certain western taxa have
highly developed petaloid lobes of style arms and
distinctive colour patterns on blades of falls. It is
primarily in stem, foliage and bract characters that
western species have noticeably diverged (T. Hall & A.
Seisums, unpubl. data). Eastern taxa of this section,
however, show much less in the way of foliar and bract
variation, but a much wider diversity of flower shape
and crest characters, and, although the stem is compressed at anthesis in some species, all eastern taxa
have pronounced stem internodes at the fruiting stage.
The compressed habit and translucent bracts of some
western species are probably more advanced features.
The final major clade, clade D, consists entirely of
Central Asian species, and all belong to section Juno.
Found in this clade is the morphologically distinct
I. magnifica (Fig. 1) and the other divergent species,
I. narbutii (Fig. 1), which is supported as sister
(BP 62, PP 0.97) to I. warleyensis. Vvedensky (1935)
considered I. warleyensis closer to I. bucharica and
295
I. vicaria (clade B), which our results clearly refute.
In the first subclade, D1 (BP 80, PP 0.92), I. capnoides
is sister (BP 98, PP 0.98) to the remaining species, all
with a Tien-Shan distribution. Some of the members
of this subclade (D1) are morphologically similar to
I. capnoides, although that species has certain features, perhaps reflecting its isolation: leaves that are
invariably sheathing at the base (although leaf
sheaths do appear in some individuals of species that
generally have no sheaths) and seeds with a small,
fragile, aril-like appendage, a swollen seed stalk;
although none of the other taxa in D1 exhibits sheathing leaves, I. graeberiana also has swollen seed
stalks. Previous AFLP studies by A. Seisums (unpubl.
data), limited to those species of subclade D1 in this
study that have a central yellow blotch on the blades
of falls, revealed a closer relationship of I. capnoides
to I. pseudocapnoides than to I. tubergeniana and
I. orchioides, in contrast to these results. Both of the
former species have a raised but undissected/poorly
dissected crest, whereas I. orchioides and the other
yellow-flowered species of subclade D1 have a finely
fimbriate crest (in I. orchioides much of this dissection may occur further down, on the midrib). A.
Seisums (unpubl. data) also indicated that I. tubergeniana s.s. was in a slightly more remote position to all
these other taxa, a position supported by its morphology: it has a compact habit at anthesis and flowers
with relatively contracted wings. A distinct stem and
flowers with wide wings are the norm for I. orchioides
and its close relatives. A morphologically distinct
entity here labelled as I. aff. tubergeniana is well
supported (BP 100) as sister to I. orchioides, which
agrees well with the opinion of T. Hall and A. Seisums
(unpubl. data) of its relationship. In this study it is
genetically distant from I. tubergeniana s.s. and
should be recognized as a distinct species once it has
been further/better characterized. In the same subclade, D1, appear I. graeberiana (synonym Juno
zenaidae Vved.), I. albomarginata and I. willmottiana. Mathew (1997) suggested further investigation
of their relationship, but we recognize their distinct
species status. All three species have flowers within
the blue range, a centrally white blade to the winged
falls and a raised crest that is usually either toothed
or dissected, although in I. willmottiana the wing is
often less expanded and the crest less deeply dissected. Also included are I. kuschakewiczii and its
close relatives, I. inconspicua and I. subdecolorata, all
with a compact habit, wiry brittle roots, flowers with
a dissected crest, and haft of falls either parallel-sided
or only slightly expanded. These three are morphologically and cytologically (A. Seisums, unpubl. data)
extremely similar. The whole I. graeberiana–
I. subdecolorata subclade is well supported (BP 99,
PP 0.99) and also includes I. tubergeniana s.s. and
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
296
N. IKINCI ET AL.
I. pseudocapnoides. However, within this group there
is little molecular divergence; the taxa morphologically little diverged from I. orchioides, I. graeberiana
and I. kuschakewiczii, are intermingled with others
that are morphologically much more divergent. Iris
graeberiana alone stands out as genetically more
divergent (PP 0.95).
In the final, strongly supported subclade, D2
(BP 100, PP 0.97), the highly morphologically divergent I. magnifica, with a well-developed stem, markedly swollen, pencil-like roots, flowers with wide
wings and an entire raised crest, for which affinities
previously have never been discussed, falls into a
basal polytomy with several other species, including
I. parvula, I. linifolia, I. tadshikorum and I. narynensis. The D2 subclade primarily has a Pamir-Alai distribution, with the more widespread I. narbutii
extending into the foothills of the Tien-Shan, I. linifolia into the Kurama range of the southwestern
Tien-Shan and I. narynensis restricted to the southwestern Tien-Shan. The I. parvula alliance (which
also includes I. linifolia, I. tadshikorum and I. narynensis) consists of plants with roots swollen above and
tapering abruptly below, sheathing leaves (except in
I. narynensis), seeds usually with a small, fragile
aril-like appendage (but, as with I. capnoides and
I. graeberiana of D1, this swollen seed-stalk is unlike
the prominent aril found in species of section Physocaulon) and flowers in which the falls have a moreor-less parallel-sided haft and a toothed to markedly
fimbriate crest. In some trees (like the one illustrated; Fig. 3), the species of the I. parvula alliance
are placed together in subclade D2, but not in all
trees. Iris narynensis, endemic to the low mountains
of the Fergana Valley (which separates the TienShan from the Pamir-Alai), has flowers vaguely
reminiscent of I. kuschakewiczii and its allies (all
with a Tien-Shan foothills distribution), but is placed
here (BP 62, PP 0.97) with the I. parvula alliance
(plants primarily of the Pamir-Alai but extending
into the Tien-Shan). Its root and flower features
(including a yellow blotch in the crest area of the
falls) match those of the other taxa in this natural
grouping, and Hall (2007) observed a similar small
aril-like appendage on the seeds of I. narynensis,
although this soon becomes detached; this species
deviates from the rest of the I. parvula group in
lacking leaf sheaths. Iris aff. parvula is a distinct
entity with long-tapering, dark-green foliage different from the linear, mid-green leaves, abruptly
pointed, of I. parvula s.s. and is shown in this study
to be unresolved with respect to other members of
the alliance. It could represent a new species and
should be studied further.
In this final subclade, I. warleyensis and I. narbutii
are positioned with a trio of morphologically similar
species, I. maracandica, I. svetlanae and I. hippolyti,
with moderate/strong support (BP 80, PP 0.96). These
last three species have roots that are markedly
swollen above, narrowing abruptly below (like I. narbutii and I. parvula and its relatives), but widely
winged falls with an undissected prominent crest
(similar to I. magnifica, also in subclade D2).
The placement of I. warleyensis here is far removed
from I. bucharica and I. vicaria (see clade B); all three
species were considered to be closely related by
Vvedensky (1935), a relationship supported by both
morphological and cytological data (A. Seisums,
unpubl. data), but these two species of clade B are
distantly related to I. warleyensis in our analyses.
They all have long, fleshy roots (admittedly more
slender in I. warleyensis, a feature in junos indicative
of a more arid environment), a well-developed stem
with long internodes, flowers with wingless falls and
a prominently raised, undissected crest. Iris warleyensis is strongly supported (BP 100, PP 0.97) to be
within a minor subclade (but with low bootstrap
support at several nodes) of the main eastern (Central
Asian) alliance and paired specifically (BP 62,
PP 0.97) with the morphologically distinct I. narbutii,
a compact species with somewhat similarly shaped
and coloured falls, but with large, purple, paddleshaped standards and almost radish-like roots. This
result caused us to sequence an additional accession
of both I. warleyensis and I. narbutii, but these additional accessions confirmed the relationships illustrated here (results not shown). The placement of
I. warleyensis leaves us with a conundrum; there are
two possible explanations, hybridization between an
I. vicaria-like ancestor (also from the Pamir-Alai) and
I. narbutii (with the latter being the seed parent,
which would explain why I. warleyensis is placed so
close to I. narbutii) or parallelism in the apparently
shared traits of I. warleyensis and I. vicaria. The
chromosome morphology of I. narbutii and one of the
entities within I. vicaria is similar (A. Seisums,
unpubl. data), although this may be coincidental. It
is likely that this mystery could be resolved if we
developed low-copy nuclear DNA loci for subgenus
Scorpiris.
We note here that features such as winged falls vs.
parallel-sided haft, specific colour patterns (guides
for pollinators) on the blade of falls and even an
entire crest vs. a fimbriate crest appear to have
arisen independently within some of these eastern
subclades, and these are therefore not such reliable
features for species groupings as we have previously
assumed. How such traits function during pollination is unknown, but it is possible that there is
strong selection operating on these floral features,
which could make them unreliable indicators of
relationships.
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300
MOLECULAR PHYLOGENETICS OF THE JUNO IRISES
CONCLUSIONS
This study confirms that subgenus Scorpiris is a
discrete, monophyletic taxonomic group within Iris. It
is shown that the primarily Afghan section Physocaulon (clade A) is sister to all other sampled juno irises
(clades B, C and D), which agrees with its separation
on bulb morphology grounds. The highly divergent
I. leptorrhiza, assigned by Rodionenko (1994) to
section Juno, is placed firmly here in section Physocaulon (clade A). The two strongly supported major
clades C and D have, on the whole, good geographical
cohesion, evolving either northwards through the
Pamir-Alai and Tien-Shan ranges of Central Asia
(clade D) or westwards via western Asia and extending as far as North Africa and the western Mediterranean (clade C). Although the precise position of the
species of minor clade B (Pamir-Alai) is unresolved,
they are possibly associated with clade D (species
entirely Central Asian in distribution), perhaps as
sister to subclade D2, also consisting of species
mainly from the Pamir-Alai. Subclade D1 contains
species of Tien-Shan origin.
Divergent members of section Juno with, at least in
part, an Afghan distribution are more closely related
to species of the western alliance in section Juno than
to the Central Asian species; the minor clade C1 consists of three of these, although a fourth (I. microglossa) appears highly isolated and unresolved in this
study. It has been demonstrated that the two species
of the Acanthospora group, separated by Rodionenko
(1961, 1994) from section Juno on account of their
unique pollen architecture, should not be considered
distinct and here are placed firmly within C2, one of
the two western subclades. Similarly there is no justification for the creation of a section exclusively for
I. cycloglossa (Rodionenko, 1994). The well-supported
position of I. galatica, in subclade C2 with the morphologically distinct I. aucheri and its close relatives,
is perplexing but may be explained by either parallelism or hybrid origin. Of particular interest is confirmation of the placement of divergent I. fosteriana
within the second western subclade C3, which makes
it the only wingless species within the whole western
alliance. From this study it is clear that both I. galatica and I. stenophylla (elements of the latter species
complex that were sampled for this study, at least)
are not closely related to I. persica, as had been
previously considered, and that support is strong for
the minor subclade that links southwestern Turkish
I. stenophylla with the two species of the Mediterranean Acanthospora group.
Individual Central Asian species of section Juno are
also unexpectedly placed with high support. The two
most divergent eastern alliance junos, Iris magnifica
and I. narbutii, are firmly embedded within sub-
297
clade D2, supported to some extent by their morphology and geography. The surprising placement of
I. warleyensis (until now considered most closely
related to the species of clade B) may be attributed to
hybridization.
However, within some subclades, a lack of molecular divergence has probably contributed to poor resolution at lower taxonomic levels within species groups
(e.g. Iris persica and its closest allies in C3) and
between species groups that appear morphologically
distinct from each other (most notably between I. orchioides, I. kuschakewiczii and their respective allies
in D1). There are likewise examples of well-supported
clades in which similar floral or vegetative features
appear more than once and have therefore evolved
independently. In several cases, the adaptive significance of such traits is imperfectly understood, so it is
unclear how reliable they might be as taxonomic
characters.
Although most of the clades appear to be natural
groupings, and the placement of many of the divergent species has strong support, more data (especially
for nuclear DNA) are required before some of the
internally unresolved clades and unexpectedly placed
species are better understood. It is also apparent that
some species complexes require further study. In a
few instances, taxonomic changes are likely to be
needed. Finally, an accurate picture of the infrasectional groupings within section Physocaulon is impossible without DNA samples of the poorly known
Afghan species currently not in cultivation, but tentatively placed within that section (see below).
SPECIES NOT INCLUDED IN THIS STUDY
Iris drepanophylla is undoubtedly closely related to
I. kopetdagensis, but only the latter was included in
this study. Although the recent regional treatment by
Nikitin & Geldihanov (1988) considered them conspecific, we follow Wendelbo & Mathew (1975), who recognized their distinct status. The primary
morphological difference between them is found in
their falls, with downturned margins to the haft of
the former and a gutter-like haft to the latter. These
characters are difficult to assess in herbaria, resulting
in frequent misidentification (see the Physocaulon
subgroup, under Discussion above). Iris xanthochlora
Wendelbo and I. porphyrochrysa Wendelbo were considered closely related to I. kopetdagensis by Wendelbo & Mathew (1975) and Mathew (1989); however,
Rodionenko (1994) placed them with species of the
Rosenbachianae subgroup. Shared features (a similar
flower shape, reduced standards, firm-textured green
bracts, visible internodes at the fruiting stage, etc.)
suggest their close affinity with I. kopetdagensis
and their inclusion in the Physocaulon subgroup.
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N. IKINCI ET AL.
Wendelbo & Mathew (1975) were less certain about
the relationships of I. wendelboi Grey-Wilson &
B.Mathew and the related I. carterorum B.Mathew &
Wendelbo, although both probably belong in section
Physocaulon; neither the morphology of their bulbs
nor the seeds of the former have ever been studied,
although I. wendelboi has rather short, swollen roots
and I. carterorum has arillate seeds (Wendelbo &
Mathew, 1975). Rodionenko (1994) placed them in the
Rosenbachianae subgroup, primarily on account of
their reportedly clypeate pollen.
Iris cabulica Gilli certainly fits morphologically into
the Rosenbachianae subgroup, and I. doabensis
B.Mathew possibly belongs here also. The most morphologically isolated of the eastern Afghan/adjacent
Pakistan juno species is perhaps I. platyptera
B.Mathew & Wendelbo, a distinctly arillate species
that also has short, thick roots. Although it is not
recorded if this species has a swollen stem base,
several herbarium specimens clearly suggest that
one is present (T. Hall & A. Seisums, unpubl. data).
B.Mathew & Wendelbo tentatively placed it in section
Physocaulon. However, a combination of characters,
such as membranous bracts, distinctive foliage and,
especially, flowers that are prominently winged,
makes it morphologically unique within this section.
Rodionenko (1994) placed the above three species in
the Rosenbachianae subgroup, but our knowledge of
their morphology is fragmentary, so the later opinion
would appear premature.
Neither Iris drepanophylla (from eastern Iran,
Turkmenistan and northern Afghanistan) nor any of
the other species listed above (mostly Afghanistan
endemics) are cultivated in the UK, so they could not
be sampled for this study. It may be that once material becomes available of all these, in some cases
poorly understood, junos, then the composition of
section Physocaulon may change slightly or even
radically.
Iris almaatensis Pavlov is morphologically similar
to I. kuschakewiczii, I. inconspicua and I. subdecolorata. It was considered by Vvedensky (1971) and
Mathew (1989) to be synonymous with I. subdecolorata, whereas Pavlov & Poliakov (1958), having
compared I. almaatensis and I. subdecolorata in
cultivation, considered them distinct species. Iris
almaatensis was described from cultivated material
originating from the outskirts of Tschemalgan village
(near Almaaty) in southern Kazakhstan, a population
disjunct from the general area of I. subdecolorata. We
were unable to include material from this population.
Ivaschenko (2005) stated that much of its already
restricted natural habitat has been lost as a result of
agricultural practices, and no representative of I. almaatensis has been found in the last 15 years. It may
be extinct.
The final juno not included in this study is I. odontostyla B.Mathew & Wendelbo, seemingly a close
Afghanistan relative of I. stocksii. Rodionenko (1961)
considered the latter species (recently reintroduced to
the UK) to be part of section Juno, and Wendelbo &
Mathew (1975) placed both species in the same
section. Rodionenko (1994) has since placed them
both, under genus Juno, in section Juno. They have
some morphological characters in common with I. persica and its allies, although the seeds of I. odontostyla
are unknown. Mathew (1997) conjectured that they
are both possibly associated with the equally semidesert Middle Eastern I. postii, a hypothesis not supported by this study, at least in regard to I. stocksii,
which falls with I. cycloglossa, the pair placed with
the western alliance, which includes both I. persica
and I. postii.
Certain elements of species complexes such as
I. persica, I. stenophylla, I. aucheri, I. pseudocaucasica and I. bucharica were also not included, primarily because it was realized early on that resolution of
such species complexes would not be possible in the
present study utilizing only plastid markers. These
sorts of questions are better addressed with DNA
fingerprinting techniques or highly variable low-copy
nuclear genes.
ACKNOWLEDGEMENTS
This research received support from the SYNTHESYS Project (GB-TAF-158) http://www.synthesys.
info/, which is financed by European Community
Research Infrastructure Action under the FP7
‘Capacities’ Program, and N.İ. was also supported by
The Scientific and Technological Research Council of
Turkey (TUBITAK-BIDEB-2219). Margaret Johnson
was helpful in setting up the visit by N.İ. to the Royal
Botanic Gardens, Kew, and in providing advice on the
plants, for which we thank her. Cultivation of the
material analyses here and the laboratory work was
supported by the Royal Botanic Gardens, Kew.
Maarten Christenhusz is thanked for his help in
preparing the figures.
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APPENDIX
NOMENCLATURAL
NOTES
Several species originally described as species of Juno
Tratt. do not have combinations in Iris L.; these are
provided below.
Iris capnoides (Vved.) T.Hall & Seisums, comb.
nov.
Basionym: Juno capnoides Vved., Opred. Rast. Sred.
Azii 2: 321 (1971).
Iris inconspicua (Vved.) T.Hall & Seisums, comb.
nov.
Basionym: Juno inconspicua Vved., Opred. Rast.
Sred. Azii 2: 321 (1971).
Iris svetlanae (Vved.) T.Hall & Seisums, comb. nov.
Basionym: Juno svetlanae Vved., Opred. Rast. Sred.
Azii 2: 322 (1971).
Iris zaprjagajevii (N.V.Abramov) T.Hall & Seisums,
comb. nov.
Basionym: Juno zaprjagajevii N.V.Abramov, Novosti
Sist. Vyssh. Rast. 8: 115 (1971).
The names Iris parvula Vved. and Iris vicaria Vved.
published by Vvedensky (1935) were accompanied by
a Russian but no Latin description or diagnosis; new
combinations based on validly published Juno
parvula Vved. and Juno vicaria Vved. by Vvedensky
(1963) are needed.
Iris parvula (Vved.) T.Hall & Seisums, comb. nov.
Basionym: Juno parvula Vved., Fl. Tadjiksk. SSR. 2:
425 & 388 (1963) ≡ Iris parvula Vved. in Kom. (ed.),
Fl. SSSR, 4: 563 (1935), descr. ross., nom. invalid.
Iris vicaria (Vved.) T.Hall & Seisums, comb. nov.
Basionym: Juno vicaria Vved., Fl. Tadjiksk. SSR, 2:
425 & 390 (1963) ≡ Iris vicaria Vved. in Kom. (ed.), Fl.
SSSR, 4: 569 (1935), descr. ross., nom. invalid.
© 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 281–300