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Rachunia, a new genus of Gesneriaceae from Thailand, is described with a single species, Rachunia cymbiformis. Its relationship to the rest of subtribe Didymocarpinae is investigated through a phylogenetic study based on Bayesian Inference and Parsimony analyses of nuclear ITS and plastid trnL‐trnF (intron‐spacer) sequences. Morphologically, Rachunia differs from the related genera Codonoboea in the large boat‐shaped bracts and orthocarpic vs plagiocarpic fruit; from Microchirita in the bracts, wiry vs fleshy stem, the campanulate vs tubular corolla and the clavate vs chiritoid stigma, and from Henckelia in the clavate vs chiritoid stigma, large boat‐shaped bracts in the inflorescence, free and imbricate sepals, short and campanulate corolla, clavate stigma, and relatively robust orthocarpic fruit.
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NORDIC JOURNAL OF
BOTANY
Nordic Journal of Botany
1
––––––––––––––––––––––––––––––––––––––––
© 2018 e Authors. Nordic Journal of Botany © 2018 Nordic Society Oikos
Subject Editor: John Parnell
Editor-in-Chief: Torbjörn Tyler
Accepted 31 August 2018
2018: e01992
doi: 10.1111/njb.01992
doi: 10.1111/njb.01992 00 1–6
Rachunia, a new genus of Gesneriaceae from ailand, is described with a single
species, Rachunia cymbiformis. Its relationship to the rest of subtribe Didymocarpinae
is investigated through a phylogenetic study based on Bayesian Inference and
Parsimony analyses of nuclear ITS and plastid trnL-trnF (intron-spacer) sequences.
Morphologically, Rachunia differs from the related genera Codonoboea in the large
boat-shaped bracts and orthocarpic vs plagiocarpic fruit; from Microchirita in the
bracts, wiry vs fleshy stem, the campanulate vs tubular corolla and the clavate vs
chiritoid stigma, and from Henckelia in the clavate vs chiritoid stigma, large boat-
shaped bracts in the inflorescence, free and imbricate sepals, short and campanulate
corolla, clavate stigma, and relatively robust orthocarpic fruit.
Keywords: phylogeny, Henckelia, Didymocarpinae
Introduction
In November 2015 a plant in the Gesneriaceae was collected in ong Pha Phum
district in Kanchanaburi province that neither the collectors nor researchers on the
family were able to place to genus. Amongst genera already known from ailand it
shows superficial similarities to Henckelia Spreng. in general appearance, the caulescent
habit (in the ai Henckelia species), opposite leaves (in most Henckelia species) and two
fertile stamens, but differs in the large boat-shaped bracts in the inflorescence, free and
imbricate sepals, short and campanulate corolla, clavate stigma, and relatively robust
orthocarpic fruit. Although under the genus concept adopted by Weber et al. (2011),
most species of Henckelia have a long corolla tube, there are species with a campanulate
corolla, including the type species H. incana Spreng., but no known species has as
short and wide corolla tube as the unknown plant. Also, several species of Henckelia,
including the widespread H. anachoreta (Hance) D.J.Middleton & Mich.Möller and
H. pumila (D.Don) A.Dietr., have orthocarpic or almost orthocarpic fruits but in these
species they are much longer and narrower resulting in a delicate appearance. ese
differences from Henckelia, and the lack of any other genus occurring in ailand,
or indeed Asia as a whole, to which it could be readily compared, led to the need
Rachunia cymbiformis, a new genus and species of Gesneriaceae
from Thailand
D. J.Middleton, G. S.Khew, M.Poopath, M.Möller and C.Puglisi
D. J. Middleton (http://orcid.org/0000-0003-3754-1452) (david_middleton@nparks.gov.sg), G. S. Khew and C. Puglisi, Singapore Botanic Gardens,
National Parks Board, Singapore. – M. Poopath, e Forest Herbarium, Dept of National Parks, Wildlife and Plant Conservation, Bangkok, ailand.
– M. Möller, Royal Botanic Garden Edinburgh, Scotland.
Research
2
for a more detailed study of its generic relationships before
what was quite clearly a new species could be described.
Fortunately, the collectors also collected material in silica gel
and this material was included in a molecular phylogenetic
study to determine the affinities of this plant.
Material and methods
Our phylogenetic study is based on Bayesian Inference
(BI) and Maximum Parsimony (MP) analyses of nuclear
ITS and plastid trnL-trnF (intron-spacer) sequences. e
ingroup encompassed 32 of the 33 known genera in subtribe
Didymocarpinae (sensu Weber et al. 2013), excluding only
Sepikea Schltr., a genus that has tentatively been included in
Cyrtandra J.R.Forst. & G.Forst. (Burtt 2001) and for which
no suitable material for DNA analysis is available. We aimed
to incorporate two or more taxa per genus, including the
type species of each, and excluding ambiguous data when-
ever possible. Following the outcome of preliminary analyses,
we sampled more densely from the genera Codonoboea Ridl.,
Henckelia and Microchirita (C.B.Clarke) Yin Z.Wang. e
outgroup comprised a selection of genera belonging to the sis-
ter subtribes Loxocarpinae and Didissandrinae (Möller et al.
2011a, Middleton and Möller 2012), plus two species of sub-
tribe Streptocarpinae.
Many of the sequences used were published in our previ-
ous studies (Möller et al. 2011a, 2016, Middleton and Möller
2012, Middleton et al. 2015, Puglisi et al. 2016), with the
addition of newly generated data for 12 taxa, including the
unknown plant from ong Pha Phum, Poopath, Sae Wai,
Kheiwbang and Jirakon 1370. A few remaining sequences
were downloaded from GenBank. e final matrices included
82 ingroup and 17 outgroup samples (Appendix 1).
For the newly generated data, we extracted the DNA
with CTAB (Doyle and Doyle 1987) or with the innuPREP
PlantDNA Kit. e PCR protocols follow Middleton et al.
(2017). e sequencing reactions were based on BigDye
Terminator technology, and were run by AIT biotech
(Singapore) on an ABI3730 DNA Analyser. e sequences
were edited in Sequencher ver. 4.7, preliminarily aligned in
Muscle (Edgar 2004) and then manually adjusted in Mesquite
ver. 2.75 (Maddison and Maddison 2011).
e Bayesian inference (BI) phylogenetic analyses were
run in MrBayes ver. 3.2.6 (Ronquist and Huelsenbeck 2003,
Ronquist et al. 2012). We used a likelihood model of DNA
substitution (GTR) with rate variation across sites follow-
ing a gamma distribution, run over 10 000 000 generations
and sampling every 10 000 generations. e heat was set at
0.01 to increase chain swaps. e final standard deviation of
the split sequences was 0.006967 in the combined analysis
( 0.01 in the preliminary single-marker analyses) and the
other output parameters all indicated sufficient sampling,
swaps between chains and convergence. After discarding the
first 25% of the trees, the retained ones were summarised in
the majority-rule consensus tree presented here (Fig. 1), which
was edited in FigTree ver. 1.3.1 (Rambaut and Drummond
2009). e Parsimony analyses were run in PAUP v. 4.0a
(build 161) (Swofford 2002). e heuristic analyses were run
over 100 000 replicates from random stepwise addition, with
MulTrees and TBR with steepest descent on. Statistical sup-
port was inferred by a Parsimony-based bootstrap analysis,
sampled 10 000 times, with random stepwise addition, TBR
on, MulTrees on and steepest descent off.
We analysed the ITS and trnL-trnF independently (not
shown) to assess the combinability of the data. As there were
no hard incongruences (Nishii et al. 2015) between the tree
topologies, we proceeded to analyse the data in a combined
matrix.
Results
e Bayesian analysis of the combined dataset resulted in a
consensus tree with the sample of Poopath et al. 1370 on
the earliest diverging ingroup branch (PP = 0.95), followed
by Codonoboea (low support, PP = 0.69), Microchirita (low
support, PP = 0.55), Henckelia (PP = 0.99), and a polytomy
including all the remaining Didymocarpinae (PP = 1) (Fig. 1).
e 36 trees derived from Parsimony analysis did not have a
robust structure (CI = 0.3827, RI = 0.5809, RC = 0.2223)
and the bootstrap analysis provided high support values
only for the terminal nodes. e resulting polytomy had
most of the genera of Didymocarpinae placed along indi-
vidual branches, and this was the case also for Poopath et al.
1370, Codonoboea, Henckelia and Microchirita. Among the
remaining Didymocarpinae, the few supported relation-
ships between genera matched those obtained by Bayesian
Inference (Fig. 1).
Discussion
e molecular phylogenetic analysis places Poopath et al. 1370
firmly in the Gesneriaceae, subfamily Didymocarpoideae,
tribe Trichosporeae, subtribe Didymocarpinae, by far the
largest subtribe in Asian Gesneriaceae. e outcome of the
analysis presented is similar to those of previous phyloge-
netic studies which extensively covered the Didymocarpinae
(Möller et al. 2009, 2011b, 2016, Middleton et al. 2015).
Phylogenetic relationships among the members of the sub-
tribe remain mostly uncertain, though several clear clades can
be identified among the early diverging branches in the phy-
logeny of the Didymocarpinae. In the present study, at the
base of the subtribe, the collection Poopath et al. 1370 and
the genera Codonoboea, Microchirita, and Henckelia form four
distinct units, followed by the remaining Didymocarpinae
genera on a large polytomy. Codonoboea, Microchirita and
Henckelia have been established as monophyletic and well-
supported (Möller et al. 2011b, Middleton and Möller 2012,
Middleton et al. 2015). Poopath et al. 1370 occupies a sep-
arate branch from the other genera in all the analyses, with
3
0.04
Henckelia urticifolia 3
Didymocarpus antirrhinoides
Allostigma guangxiense
Liebigia barbata
Pseudochirita guangxiensis
Henckelia bifolia
Anna submontana
Raphiocarpus sinicus
Loxostigma glabrifolium
Henckelia pumila 2
Petrocodon scopulorum
Didymocarpus villosus
Microchirita viola
Oreocharis jiangxiensis
Hexatheca fulva
Paraboea clarkei
Henckelia urticifolia 2
Codonoboea malayana
Loxostigma griffithii
Primulina swinglei
Henckelia floccosa
Petrocosmea kerrii
Codonoboea pumila
Damrongia lacunosa
Conandron ramondioides
Microchirita hamosa
Tribounia grandiflora
Petrocosmea nervosa
Henckelia dielsii
Didissandra sp.
Codonoboea leucocodon
Aeschynanthus roseoflorus
Spelaeanthus chinii
Microchirita tubulosa
Cyrtandra pulchella
Agalmyla glabra
Rachunia cymbiformis
Didymostigma obtusum
Codonoboea corrugata
Microchirita mollissima
Ridleyandra quercifolia
Tribounia venosa
Codonoboea racemosa
Henckelia pumila 1
Microchirita sericea
Microchirita caliginosa
Oreocharis urceolata
Petrocodon ainsliifolius
Ornithoboea occulta
Primulina gemella
Dorcoceras geoffrayi
Henckelia grandifolia
Codonoboea floribunda
Henckelia longisepala
Codonoboea codonion
Henckelia_urticifolia 1
Paraboea takensis
Aeschynanthus andersonii
Middletonia regularis
Somrania albiflora
Allocheilos guangxiensis
Lysionotus pauciflorus
Deinost. cyrtocarpa
Metapetrocosmea peltata
Hemiboea follicularis
Codonoboea elata
Oreocharis dasyantha
Deinostigma tamiana
Lysionotus serratus
Primulina luochengensis
Chayamaritia smitinandii
Gyrocheilos lasiocalyx
Ornithoboea flexuosa
Cathayanthe biflora
Glabrella mihieri
Primulina tabacum
Henckelia incana
Streptocarpus rexii
Henckelia anachoreta
Microchirita rupestris
Damrongia purpureolineata
Codonoboea venusta
Didymocarpus kerrii
Microchirita huppatatensis
Petrocodon dealbatus
Agalmyla paucipilosa
Didissandra elongata subsp. minor
Didymostigma trichanthera
Streptocarpus glandulosissimus
Gyrocheilos chorisepalus var. synsepalus
Hemiboea fangii
Codonoboea albomarginata
Cyrtandra pendula
Didissandra frutescens
Aeschynanthus rhododendron
Ridleyandra petiolata
Briggsiopsis delavayi
Lysionotus petelotii
Microchirita involucrata
0.85
1/93
0.55
0.89
1
1/93
1/80
1/100
1/100
1/100
1/100
1/91
0.95
1/100
1/100
1/100
1/100
1/100
0.8
1/100
0.77
1/85
0.69
1/85
0.57
1/98
1/93
1
0.92
1
0.99
1/100
1
0.7
1
1/100
0.9
1/94
0.98
0.98
1
0.92
1/98
1/78
1/100
0.97
0.61
0.69
1/99
1/88
0.99
1/100
1/100
1/93
1/99
0.55
0.65
1/100
1/100
1/100
1/94
1/97
1/99
1/96
1/100
1
0.94
1/99
0.74
0.93
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1/94
0.51
1/100
1/97
0.88
1
1/99
0.99
0.72
1/100
1/95
Ingroup
All other Didymocarpinae
Codonoboea
Microchirita
Henckelia
Figure 1. Bayesian Inference 50% majority rule consensus tree from the analysis of ITS and trnL-trnF. All the posterior probabilities and
only the bootstrap values 75% are added at the nodes.
4
no immediate sister taxon. Morphologically, it differs from
Codonoboea in the large boat-shaped bracts and orthocarpic
vs plagiocarpic fruit; from Microchirita in the bracts, wiry vs
fleshy stem, the campanulate vs tubular corolla and the cla-
vate vs chiritoid stigma (a stigma in which the upper lip of
the stigma is absent and the lower lip is expanded and often
bilobed), and from Henckelia in the characters noted in the
introduction and particularly in the clavate vs chiritoid stigma.
e other genera in this subtribe are in a clade well-sup-
ported by the Baysian Inference (PP = 1), and are more dis-
tantly related to Poopath et al. 1370. Morphologically, the
genera which share a caulescent habit and two fertile stamens
with Poopath et al. 1370 are Allostigma W.T.Wang, Cyrtandra
J.R.Forst. & G.Forst., Deinostigma W.T.Wang, most
Didymocarpus Wall., Didymostigma W.T.Wang, Hemiboea
C.B.Clarke, Liebigia Endl. and Pseudochirita W.T.Wang.
Poopath et al. 1370 differs from all of them in the presence
of large boat-shaped bracts, and additionally from Allostigma
in the parietal (Poopath et al. 1370) vs axile (Allostigma) pla-
centation, from Cyrtandra in dehiscent vs indehiscent fruit,
from Deinostigma in opposite vs alternate leaves and clavate
vs chiritoid stigma, from Didymocarpus in the imbricate
vs tubular or widely separated sepals and having a shorter
corolla tube, from Didymostigma in the insertion of stamens
being basal vs distal in the corolla tube and the clavate vs bi-
lobed stigma, from Hemiboea in the unilocular ovary with
2 equal parietal placentae vs bi-locular ovary with only 1
axile placenta developing, from Liebigia in the campanulate
vs tubular corolla and clavate vs chiritoid stigma, and from
Pseudochirita in the free and imbricate vs tubular calyx and
in the clavate vs bi-lobed and unequal-lobed stigma shape.
Based on the results of the molecular phylogenetic study
and the morphological distinctness of the plant, we hereby
describe a new species in a new genus which we name
Rachunia cymbiformis.
Rachunia D.J.Middleton & C.Puglisi gen. nov.
Type species: Rachunia cymbiformis D.J.Middleton.
Etymology
e genus is named in honour of the ai botanist Dr Rachun
Pooma of the Forest Herbarium Bangkok (BKF) to recognise
his great contribution to our understanding of plant diversity
in ailand and the wider region.
Description
Caulescent herb; stems not fleshy. Leaves opposite, petiolate;
blades entire to serrulate at margin, with pinnate venation.
Inflorescences axillary, dichasial, 6–14 pair-flowers, similar in
length to subtending leaves, with large boat-shaped bracts.
Calyx 5-lobed; lobes free to base, strongly imbricate; margins
entire. Corolla tube short, campanulate; limb 2-lipped, with
upper lip 2-lobed and lower lip 3-lobed, all lobes orbicular.
Fertile stamens 2, inserted near base of corolla tube; filaments
narrow at base, thickened above, strongly curved with the
anther apex pointing towards dorsal side of corolla; anthers
coherent at apices, held face to face; staminodes 3, medial one
smaller than 2 lateral ones. Disc annular. Ovary uni-locular
with 2 equal-sized parietal placentae; style narrow; stigma
clavate. Fruit a long and narrow capsule, orthocarpic (only
seen immature but likely to be loculicidal as in most other
Didymocarpinae with dehiscent fruit); seeds tiny, globose.
Distribution
ailand, Kanchanaburi. e genus currently only has one
species which is only known from the type collection. Given
the collection locality on the ai-Myanmar border it almost
certainly also occurs in Myanmar.
Rachunia cymbiformis D.J.Middleton sp. nov. (Fig. 2)
Type: Kanchanaburi, ong Pha Phum, Ban E Tong,
near ai-Myanmar border, 900 m a.s.l., 3 Nov 2015,
M. Poopath, J. Sae Wai, W. Kheiwbang and S. Jirakon 1370
(holotype: BKF, isotype: SING).
Etymology
e specific epithet ‘cymbiformis’ refers to the boat-shaped
bracts in the inflorescence.
Figure 2. Rachunia cymbiformis D.J.Middleton. (A) upper leaves,
inflorescence and infructescence, (B) close-up of bracts and flower.
Photos by Manop Poopath.
5
Description
Terrestrial herb, 20–40 cm tall, decumbent at base and root-
ing along stems; stems densely covered in long dark brown
hairs to 3 mm long. Leaves opposite; petioles 13–19 mm
long, densely covered in long golden brown hairs; blade
elliptic to obovate, symmetrical or slightly falcate, 6.0–13.5 ×
2.7–5.3 cm, 2.1–3.6 times as long as wide, narrowed at base
but then ultimately rounded, sometimes inserted on pedicel
at slightly different levels on each side, acuminate at apex;
margin entire at base to serrulate in upper half; secondary
veins 7–9 on each side; tertiary venation reticulate, sparsely
covered with golden brown hairs above and beneath, beneath
more densely so on venation. Inflorescence 5–15 cm long, as
a simple dichasium with 6 pair-flowers or branched one more
time and then with up to 14 flowers; peduncle 3.0–10.1 cm
long, sparsely to densely covered in long golden brown hairs
which are 1.2–2.5 mm long; bracts boat-shaped, green tinged
purplish, ovate to elliptic, 9–11 × 4.5–8.0 mm, acuminate
at apex, sparsely to densely covered in long golden brown
hairs; pedicels 8.0–13.5 mm long, glabrous. Calyx purplish,
of 5 lobes which are free to base; lobes elliptic, 4.8–5.5 ×
1.4–2.2 mm, acute at apex, with entire margin, glabrous.
Corolla with a short campanulate tube and spreading limb;
limb 2-lipped, with all lobes orbicular; tube whitish, its lobes
pale purple, whitish at base of upper lip, glabrous outside,
with sparse long hairs in lower half of tube inside; tube
4.5 mm long; upper lip 5 mm long, its lobes 4 × 5 mm; lower
lip 6 mm long, its lateral lobes 4.5 × 5.0 mm and medial
lobe 4.5 × 4.5 mm. Fertile stamens 2, inserted at 1.2 mm
from base; filaments white, narrow at base, thickened above,
strongly curved with the anther apex pointing towards dor-
sal side of corolla, glabrous; anthers coherent at apices, held
face to face, ca 1.1 × 2.7 mm, glabrous; lateral staminodes
0.6 mm long, medial 0.3 mm long. Disc annular, 0.2 mm
long. Pistil white; ovary 3 mm long, glabrous, unilocular with
2 equal-sized parietal placentae; style 7 mm long, glabrous;
stigma clavate. Fruit orthocarpic, 4–6 cm long, 1.2–1.5 mm
wide; seeds globose, 0.2 mm in diameter.
Distribution and habitat
Rachunia cymbiformis is currently only known from the type
collection from ailand, Kanchanaburi province, ong
Pha Phum district, Ban E Tong, near the ai-Myanmar
border at 900 m a.s.l. Given the collection locality on the
ai-Myanmar border it almost certainly also occurs in
Myanmar. e habitat is in moist evergreen forest on a slope
in shade. Although not recorded on the specimen label, the
underlying bedrock is granite and the soil is loam.
Conservation status
As Rachunia cymbiformis is only known from the type
collection and its distribution and population size are
unknown, it must currently be categorised as ‘Data
Deficient’ (DD). On the ai side of the border the forest
is rather degraded but on the Myanmar side the forest is
currently rather extensive.
Acknowledgements – We thank the additional collectors of the
plant, J. Sae Wai, W. Kheiwbang and S. Jirakon, M. Niissalo,
R. Goh and Khoo-Woon M.H. for their assistance in the Singapore
Botanic Gardens molecular laboratory. e Royal Botanic Garden
Edinburgh is supported by the Rural and Environment Science and
Analytical Services division (RESAS) in the Scottish Government.
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6
Appendix 1. The following is a list of the accessions
used, ordered alphabetically by taxon, with the
GenBank codes for the ITS and trnL-trnF sequences
respectively. The taxa whose sequences are newly
published are listed with the complete voucher
information.
Aeschynanthhus andersonii, Myanmar, Shan State, 19 Sep
2015, Y. Baba et al. FOMIC103456 (SING); Aeschynanthus
rhododendron HQ632993, HQ632895; Aeschynanthus roseo-
florus FJ501333, HQ632896; Agalmyla glabra HQ632989,
HQ632892; Agalmyla paucipilosa HQ632990, HQ632893;
Allocheilos guangxiensis HQ632994, HQ632897; Allostigma
guangxiense HQ632977, HQ632880; Anna submontana
FJ501362, FJ501542; Briggsiopsis delavayi HQ632976,
HQ632879; Cathayanthe biflora HQ632996, HQ632899;
Chayamaritia smitinandii KP325425, KP325432; Codonoboea
albomarginata HQ632961, AJ492297; Codonoboea codonion
JF912565, JF912538; Codonoboea corrugata HQ632962,
FJ501484; Codonoboea elata JF912550, JF912523;
Codonoboea floribunda JF912566, JF912539; Codonoboea
leucocodon JF912567, JF912540; Codonoboea malayana
JF912568, JF912541; Codonoboea pumila JF912570,
JF912543; Codonoboea racemosa JF912571, JF912544;
Codonoboea venusta JF912572, JF912545; Conandron
ramondioides FJ501340, FJ501515; Cyrtandra pendula
FJ501354, FJ501530; Cyrtandra pulchella EU919941,
HQ632906; Damrongia lacunosa KU203801, KU203896;
Damrongia purpureolineata KU203798, KU203893;
Deinostigma cyrtocarpa JX506885, JX506777; Deinostigma
tamiana, Vietnam, Quang Ngai, 2 Apr 2017, Lý Ngc Sâm
Lý 882 (VNM); Didissandra elongata ssp. minor KP325420,
KP325427; Didissandra frutescens JN934793, FJ501522;
Didissandra sp. KP325422, KP325429; Didymocarpus
antirrhinoides DQ912671, FJ501513; Didymocarpus kerrii,
ailand, Mae Hong Son, 21 Oct 2014, D.J. Middleton, C.
Hemrat, P. Karaket, C. Puglisi and S. Suddee 5812 (SING);
Didymocarpus villosus HQ633001, HQ63290; Didymostigma
obtusum HQ632971, HQ632875; Didymostigma trichan-
thera HQ632972, HQ632876; Dorcoceras geoffrayi, ailand,
Sukhothai, 24 Oct 2014, D.J. Middleton, C. Hemrat, P.
Karaket, C. Puglisi and S. Suddee 5833 (SING); Glabrella
mihieri FJ501363, FJ501544; Gyrocheilos chorisepalus var.
synsepalus HQ632997, HQ632900; Gyrocheilos lasiocalyx
HQ632998, HQ632901; Hemiboea fangii HQ632979,
HQ632882; Hemiboea follicularis HQ632982, HQ632885;
Henckelia anachoreta 1 HQ632966, HQ632870; Henckelia
bifolia JF912549, JF912522; Henckelia dielsii HQ632967,
HQ632871; Henckelia floccosa HQ632964, FJ501486;
Henckelia grandifolia JF912554, JF912527; Henckelia incana
HQ632965, HQ632869; Henckelia longisepala HQ632963,
HQ632868; Henckelia pumila 1 JF912556, JF912529;
Henckelia pumila 2 FJ501327, FJ501491; Henckelia urticifolia
1 DQ872835, DQ872821; Henckelia urticifolia 2 JF912559,
JF912532; Henckelia urticifolia 3 FJ501328, FJ501492;
Hexatheca fulva HQ632969, HQ632873; Liebigia barbata
DQ912668, FJ501538; Loxostigma glabrifolium HQ633006,
HQ632910; Loxostigma griffithii FJ501338, FJ501508;
Lysionotus pauciflorus FJ501331, FJ501497; Lysionotus petel-
otii HQ632974, FJ501496; Lysionotus serratus, Myanmar,
Shan State, 13 Sep 2015, Y. Baba et al. FOMIC103132
(SING); Metapetrocosmea peltata HQ632968, HQ632872;
Microchirita caliginosa FJ501325, FJ501488; Microchirita
hamosa, ailand, Tak, 17 Oct 2014, D.J. Middleton, C.
Hemrat, P. Karaket, C. Puglisi and S. Suddee 5762 (BKF);
Microchirita huppatatensis, ailand, Uthai ani, 14 Oct
2014, D.J. Middleton, C. Hemrat, P. Karaket, C. Puglisi
and S. Suddee 5689 (BKF); Microchirita involucrata 2
JF912553, JF912526; Microchirita mollissima JF912555,
JF912528; Microchirita rupestris, ailand, Kanchanaburi,
28 Oct 2009, D.J. Middleton and P. Triboun 5204 (E);
Microchirita sericea JF912548, JF912521; Microchirita tubu-
losa JF912558, JF912531; Microchirita viola JF912560,
JF912533; Middletonia regularis KU203789, KU203884;
Oreocharis dasyantha HQ633014, HQ632918; Oreocharis
jiangxiensis HQ633029, HQ632933; Oreocharis urceolata
HQ633018, HQ632922; Ornithoboea flexuosa KU203836,
KU203931; Ornithoboea occulta, ailand, Tak, 15 Oct
2014, D.J. Middleton, C. Hemrat, P. Karaket, C. Puglisi
and S. Suddee 5702 (SING); Paraboea clarkei JN934757,
JN934715; Paraboea takensis, ailand, Tak, 16 Oct 2014,
D.J. Middleton, C. Hemrat, P. Karaket, C. Puglisi and S.
Suddee 5706 (SING); Petrocodon ainsliifolius HQ633038,
HQ632941; Petrocodon dealbatus FJ501358, FJ501537;
Petrocodon scopulorum HQ633044, HQ632947; Petrocosmea
kerrii FJ501334, FJ501502; Petrocosmea nervosa FJ501335,
AJ492299; Primulina gemella FJ501345, FJ501523;
Primulina luochengensis HQ633046, HQ632949; Primulina
swinglei, Vietnam, cultivated at Singapore Botanic Gardens,
27 Feb 2017, J. Leong-Škorničková JLS 3188 (SING);
Primulina tabacum FJ501352, AJ492300; Pseudochirita
guangxiensis HQ633003, HQ632908; Rachunia cymbifor-
mis, ailand, Kanchanaburi, 3 Nov 2015, M. Poopath,
J. Sae Wai, W. Kheiwbang and S. Jirakon 1370 (BKF);
Raphiocarpus sinicus HQ632973, HQ632877; Ridleyandra
petiolata HQ633032, HQ632935; Ridleyandra quercifolia
HQ633033, HQ632936; Somrania albiflora KU203792,
KU203887; Spelaeanthus chinii FJ501307, FJ501457;
Streptocarpus glandulosissimus AF316918, KR703972;
Streptocarpus rexii AF316979; AJ492305; Tribounia gran-
diflora JX839280, JX839281; Tribounia venosa JX839283,
JX839282.
... B.Clarke) B.L.Burtt [85]. On the other hand, phylogenetic studies of materials sampled from further botanical exploration also revealed lineages that are distinct and worthy of generic recognition (e.g., Actinostephanus F. Wen [66], Middletonia C.Puglisi [89], Rachunia D.J. Middleton & C.Puglisi [90], etc.). These recent taxonomic changes and novelties of the didymocarpoid Gesneriaceae underscore the importance of incorporating molecular information in achieving a stable classification of Asian Gesneriaceae. ...
... Within the subtribe Didymocarpinae, Rachunia, Codonoboea, Microchirita, and Henckelia formed successive sister groups to an unresolved clade composed of all other genera of the subtribe (i.e., 'core' Didymocarpinae), congruent with previous studies [66,90]. As shown in previous phylogenetic studies of Didymocarpinae using ITS and trnL-F sequences, relationships within the 'core' Didymocarpinae remained poorly resolved [75,77,79,86,87,90]. ...
... Within the subtribe Didymocarpinae, Rachunia, Codonoboea, Microchirita, and Henckelia formed successive sister groups to an unresolved clade composed of all other genera of the subtribe (i.e., 'core' Didymocarpinae), congruent with previous studies [66,90]. As shown in previous phylogenetic studies of Didymocarpinae using ITS and trnL-F sequences, relationships within the 'core' Didymocarpinae remained poorly resolved [75,77,79,86,87,90]. The employment of genomic data such as whole plastome sequences [96] and target capture [101] will be essential to clarify and understand this apparent radiation of Asian gesneriad clade. ...
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Situated in the southern end of the Annamite Mountain Range, Langbiang Plateau is a major biodiversity hotspot of southern Vietnam known for high species diversity and endemicity. To achieve effective conservation, parts of the plateau were designated as the Langbiang Biosphere Reserve, an UNESCO World Network aiming to improve relationships between inhabitants and their environments. Amongst the rich endemic flora of the plateau are three gesneriads ascribed to Primulina, a calciphilous genus with high species diversity in the vast limestone karsts stretching from southern China to northern Vietnam. However, a recent phylogenetic study questioned the generic placement of the Langbiang Primulina, corroborating with observations on the geographical distribution, habitat preference, and phyllotaxy of the three species. Based on phylogenetic analyses of nuclear ITS and plastid trnL-F DNA sequences of a comprehensive sampling covering nearly all genera of the Old World Gesneriaceae, we demonstrate that the three Langbiang Primulina species form a fully supported clade distantly related to other Primulina. As this clade is biogeographically, ecologically, morphologically, and phylogenetically distinct worthy of generic recognition, we propose to name it Langbiangia gen. nov. to highlight the rich and unique biodiversity of the Langbiang Plateau. By means of this taxonomic endeavor, we are hoping to raise the conservation awareness of this biodiversity heritage of southern Vietnam and promote the importance of Langbiang Biosphere Reserve that is crucial for achieving action-oriented global targets of the post-2020 global biodiversity framework (GBF) of the UN Convention on Biological Diversity (CBD)—effective conservation and management of at least 30% of biodiverse terrestrial, inland water, and costal and marine areas by 2030—that has been agreed at the COP15 in Montréal in December 2022.
... In total, 61 f b (C) scores were significantly elevated at a Bonferronicorrected P < 0.05, and 10 of the 40 branches in the phylogeny showed significant excess allele sharing with at least one other species C (Fig. S10) Table S2), and the chloroplast phylogenetic tree was estimated by IQ-TREE based on matrix_1 (see Table S2) Species-level relationships within the redefined Henckelia have not been resolved, with only a few studies including very limited Henckelia species and a small number of gene regions (e.g. ITS and trnL-trnF; Weber et al., 2011;Roalson and Roberts, 2016;Middleton et al., 2018;Li et al., 2022). Here, we used WGS sequencing to generate massive amounts of genomic data to reconstruct the phylogenetic relationships among all Chinese Henckelia species. ...
... Using these datasets, we verified the monophyly of most current morphological species and generally obtained highly supported relationships (LPP > 0.95 or UFBoot > 95). Specifically, our phylogenies revealed that H. longisepala is sister to the rest of the genus, consistent with some previous results (Roalson and Roberts, 2016;Middleton et al., 2018) but in conflict with the results of Weber et al. (2011). The early divergence of H. longisepala in Henckelia is consistent with its distinct morphological characteristics (i.e. ...
Article
Background and Aims Hybridization has long been recognized as an important process for plant evolution and is often accompanied by polyploidization, another prominent force in generating biodiversity. Despite its pivotal importance in evolution, the actual prevalence and distribution of hybridization across the tree of life remain unclear. Methods We used whole-genome shotgun (WGS) sequencing and cytological data to investigate the evolutionary history of Henckelia, a large genus in the family Gesneriaceae with a high frequency of suspected hybridization and polyploidization events. We generated WGS sequencing data at about 10× coverage for 26 Chinese Henckelia species plus one Sri Lanka species. To untangle the hybridization history, we separately extracted whole plastomes and thousands of single-copy nuclear genes from the sequencing data, and reconstructed phylogenies based on both nuclear and plastid data. We also explored sources of both genealogical and cytonuclear conflicts and identified signals of hybridization and introgression within our phylogenomic dataset using several statistical methods. Additionally, to test the polyploidization history, we evaluated chromosome counts for 45 populations of the studied 27 Henckelia species. Key Results We obtained well-supported phylogenetic relationships using both concatenation and coalescent-based methods. However, the nuclear phylogenies were highly inconsistent with the plastid phylogeny, and we observed intensive discordance among nuclear gene trees. Further analyses suggested that both incomplete lineage sorting (ILS) and gene flow contributed to the observed cytonuclear and genealogical discordance. Our analyses of introgression and phylogenetic networks revealed a complex history of hybridization within the genus Henckelia. In addition, based on chromosome counts for 27 Henckelia species, we found independent polyploidization events occurred within Henckelia after different hybridization events. Conclusions Our findings demonstrated that hybridization and polyploidization are common in Henckelia. Furthermore, our results revealed that H. oblongifolia is not a member of the redefined Henckelia and suggested several other taxonomic treatments in this genus.
... This has entailed a deep reconstruction of the subfamily that is still in process, allowing more homogeneous species classification [1,[13][14][15]. Some genera have been newly established (Billolivia, Michaelmoelleria, Chayamaritia, Glabrella, Microchirita, Middletonia, Rachunia, Somrania) or recovered (Dorcoceras, Loxocarpus), while others have gained species from other genera (Damrongia, Oreocharis, Loxostigma, Deinostigma, Paraboea, Primulina, Streptocarpus), lost species by relocation to other genera (Boea), or lost species by synonymization (Acanthonema, Hovanella, Colpogyne, Linnaeopsis, Nodonema, Schizoboea, Briggsia) [13,[16][17][18][19][20][21][22][23][24][25][26][27][28]. ...
Article
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Gesneriaceae is a pantropical family of plants that, thanks to their lithophytic and epiphytic growth forms, have developed different strategies for overcoming water scarcity. Desiccation tolerance or “resurrection” ability is one of them: a rare phenomenon among angiosperms that involves surviving with very little relative water content in their tissues until water is again available. Physiological responses of desiccation tolerance are also activated during freezing temperatures, a stress that many of the resurrection gesneriads suffer due to their mountainous habitat. Therefore, research on desiccation- and freezing-tolerant gesneriads is a great opportunity for crop improvement, and some of them have become reference resurrection angiosperms (Dorcoceras hygrometrica, Haberlea rhodopensis and Ramonda myconi). However, their difficult indoor cultivation and outdoor accessibility are major obstacles for their study. Therefore, this review aims to identify phylogenetic, geoclimatic, habitat, and morphological features in order to propose new tentative resurrection gesneriads as a way of making them more reachable to the scientific community. Additionally, shared and species-specific physiological responses to desiccation and freezing stress have been gathered as a stress response metabolic basis of the family.
... Gesneriaceae (Lamiales) consists of ca. 150 genera and around 3500 species of perennial herbs, shrubs or small trees, with the main distribution in the tropics and subtropics Möller et al. 2016a;Middleton et al. 2018). In China, there are >600 species in 44 genera (Möller et al. 2016a, b;Xu et al. 2017). ...
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Four new species of Gesneriaceae from Yunnan, southwest China, are described and illustrated. They are Petrocosmea rhombifolia , Petrocosmea tsaii , Didymocarpus brevipedunculatus , and Henckelia xinpingensis . Diagnostic characters between the new species and their morphologically close relatives are provided. Their distribution, ecology, phenology, and conservation status are also described.
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Based on an updated taxonomy of Gesneriaceae, the biogeography and evolution of the Asian Gesneriaceae are outlined and discussed. Most of the Asian Gesneriaceae belongs to Didymocarpoideae, except Titanotrichum was recently moved into Gesnerioideae. Most basal taxa of the Asian Gesneriaceae are found in the Indian subcontinent and Indo-China Peninsula, suggesting Didymocarpoideae might originate in these regions. Four species diversification centers were recognized, i.e. Sino-Vietnam regions, Malay Peninsula, North Borneo and Northwest Yunnan (Hengduan Mountains). The first three regions are dominated by limestone landscapes, while the Northwest Yunnan is well-known for its numerous deep gorges and high mountains. The places with at least 25% species are neoendemics (newly evolved and narrowly endemic) which were determined as evolutionary hotspots, including Hengduan Mountains, boundary areas of Yunnan-Guizhou-Guangxi in Southwest China, North Borneo, Pahang and Terengganu in Malay Peninsula, and mountainous areas in North Thailand, North Sulawesi Island. Finally, the underlying mechanisms for biogeographical patterns and species diversification of the Asian Gesneriaceae are discussed.
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The new species Ornithoboea grandiflora D.J. Middleton and new variety Ornithoboea maxwellii var. minutiflora D.J. Middleton are described.
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Based on molecular, morphological and cytological studies the previously monotypic genus Deinostigma W.T.Wang & Z.Y.Li has been expanded to include several species previously ascribed to Primulina Hance. Deinostigma now comprises seven species, including one previously placed in synonymy. The new combinations Deinostigma cicatricosa are made. Deinostigma eberhardtii is lectotypified. The genus is defined by a combination of an alternate leaf arrangement, hooked hairs on many plant parts, flowers with the pedicel inserted at an angle and off-centre on the receptacle, and, where known, a somatic chromosome number (2n) of < 36. This new circumscription of the genus expands its distribution from Vietnam into South China.
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The Loxocarpinae, also known as the “Boea group”, are the subtribe of Gesneriaceae which includes Boea and a number of segregated genera and close relatives. This group currently comprises over 200 species in 15 genera. Here we present the most up-to-date phylogeny, covering all the genera known to belong to the group, based on Bayesian inference and parsimony of the nuclear ITS and the plastid regions trnL-trnF (intron and spacer) and ndhF-trnL UAG(spacers). The results show discrepancies between the current generic delimitation in the subtribe and the clades delineated by the phylogeny. As a result Boea, Damrongia, Paraboea and Streptocarpus are recircumscribed in an attempt to establish a more natural classification and new combinations are made. The new genus Middletonia is described.
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Based on a phylogenetic analysis of Asian Gesneriaceae with the most comprehensive coverage at the genus level to date, the new genus Chayamaritia is established and described in subfamily Didymocarpoideae, tribe Trichosporeae, subtribe Didymocarpinae. It contains two species, of which one, Chayamaritia smitinandii (B.L.Burtt) D.J.Middleton, was formerly placed in the genera Chirita and Henckelia. The other, Chayamaritia banksiae D.J.Middleton, is newly described. The exclusion of Chayamaritia smitinandii from Henckelia further clarifies the taxonomic and biogeographic limits of Henckelia following its considerable recircumscription during the recent remodelling and synonymisation of Chirita.
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A new formal classification of Gesneriaceae is proposed. It is the first detailed and overall classification of the family that is essentially based on molecular phylogenetic studies. Three subfamilies are recognized: Sanangoideae (monospecific with Sanango racemosum), Gesnerioideae and Didymocarpoideae. As to recent molecular data, Sanango/Sanangoideae (New World) is sister to Gesnerioideae + Didymocarpoideae. Its inclusion in the Gesneriaceae amends the traditional concept of the family and makes the family distinctly older. Subfam. Gesnerioideae (New World, if not stated otherwise with the tribes) is subdivided into five tribes: Titanotricheae (monospecific, East Asia), Napeantheae (monogeneric), Beslerieae (with two subtribes: Besleriinae and Anetanthinae), Coronanthereae (with three subtribes: Coronantherinae, Mitrariinae and Negriinae; southern hemisphere), and Gesnerieae [with five subtribes: Gesneriinae, Gloxiniinae, Columneinae (5the traditional Episcieae), Sphaerorrhizinae (5the traditional Sphaerorhizeae, monogeneric), and Ligeriinae (5the traditional Sinningieae)]. In the Didymocarpoideae (almost exclusively Old World, especially E and SE Asia/Malesia) two tribes are recognized: Epithemateae [with four small, but morphologically and genetically very distinctive subtribes: Loxotidinae (monogeneric with Rhynchoglossum), Monophyllaeinae, Loxoniinae and Epithematinae (monogeneric)] and Trichosporeae (the earliest name at tribal rank for the ‘‘Didymocarpoid Gesneriaceae’’). The last is subdivided into ten subtribes: Jerdoniinae (monospecific), Corallodiscinae (monogeneric), Tetraphyllinae (monogeneric), Leptoboeinae, Ramondinae (Europe), Litostigminae (monogeneric), Streptocarpinae (Africa and Madagascar), Didissandrinae, Loxocarpinae and Didymocarpinae. Didymocarpinae is the largest subtribe (ca. 30 genera and .1600 species) and still requires intensive study. It includes the most speciose genera such as Cyrtandra, Aeschynanthus, Agalmyla, Didymocarpus, Henckelia, Codonoboea, Oreocharis and Primulina and the types of the traditional tribes Didymocarpeae, Trichosporeae and Cyrtandreae.
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Based on molecular data and a morphological evaluation, evidence is provided that the species of eleven, mostly small-sized and monotypic genera of Chinese Gesneriaceae (Ancylostemon, Bournea, Briggsia s.str., Dayaoshania, Deinocheilos, Isometrum, Opithandra, Oreocharis, Paraisometrum, Thamnocharis, Tremacron) form a highly-supported group in which the species interrelationships run across traditional generic boundaries. The data confirm previous doubts on the naturalness of some of these genera and, after a detailed discussion of the particular genera, the conclusion is reached that the whole group is best regarded as a single genus, Oreocharis, which is thus expanded to comprise over 80 species. A list of the species is given and the necessary transfers are made. The new delimitation provides a framework for studying the species relationships and working out an infrageneric classification. Oreocharis provides an excellent example of a major monophyletic group that has experienced a rapid radiation early in its evolution and shows manifold convergences in floral characters (corolla form and coloration, fertility of stamens, anther shape and dehiscence mode), apparently reflecting different pollination strategies, but has little variation in vegetative habit and fruit structure.
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The new genus Tribounia is described with two species, Tribounia venosa (Barnett) D.J. Middleton, a new combintion, and Tribounia grandiflora D.J. Middleton, a new species. A key to the species and conservation assessments are provided.
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The limits of Cyrtandra, its synonyms and the characters used for its division into subgenera and sections are surveyed chronologically. Additional features, known as yet for only a few groups, are discussed.
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The taxonomy of the African, Madagascan and Comoro Island (Afro-Malagasy) Gesneriaceae attracts a large amount of interest given the horticultural importance of Cape Primroses (Streptocarpus) and African Violets (Saintpaulia). Earlier studies indicated that the Afro-Malagasy genera form a strongly supported clade, and recent classifications have included some of the genera within an expanded Streptocarpus. Given the global importance of this group, we carried out a comprehensive molecular phylogenetic analysis of all Afro-Malagasy genera in subfamily Didymocarpoideae, tribe Trichosporeae, subtribe Streptocarpinae, to investigate species relationships in these genera as the basis for a new classification. Phylogenetic analyses of the nuclear ribosomal spacer (ITS, 5S NTS) and chloroplast intron and spacer regions (rpl20-rps12 spacer, trnL intron, trnLF spacer) of 226 samples were performed, including all Streptocarpinae genera, except the monotypic Nodonema. The molecular phylogenies demonstrate that the genera with non-twisted fruits are nested within Streptocarpus which has twisted fruits. Two main clades were found, one comprising herbaceous caulescent Streptocarpus that also included Saintpaulia, the caulescents Hovanella and Schizoboea, and the unifoliates Acanthonema and Trachystigma. The second clade comprises the woody caulescents and acaulescent Streptocarpus, Colpogyne and Linnaeopsis. Altogether, twelve well-supported subclades can be recognized, each with a combination of distinct morphological characteristics. A new classification of tribe Streptocarpinae, de facto Streptocarpus, is presented, retaining the two subgenera, Streptocarpus and Streptocarpella, and dividing them into five and seven sections respectively. Nodonema is attributed to subg. Streptocarpus for morphological reasons. The former genus Saintpaulia is classified as Streptocarpus subg. Streptocarpella sect. Saintpaulia with ten species recognized.
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— We studied sequence variation in 16S rDNA in 204 individuals from 37 populations of the land snail Candidula unifasciata (Poiret 1801) across the core species range in France, Switzerland, and Germany. Phylogeographic, nested clade, and coalescence analyses were used to elucidate the species evolutionary history. The study revealed the presence of two major evolutionary lineages that evolved in separate refuges in southeast France as result of previous fragmentation during the Pleistocene. Applying a recent extension of the nested clade analysis (Templeton 2001), we inferred that range expansions along river valleys in independent corridors to the north led eventually to a secondary contact zone of the major clades around the Geneva Basin. There is evidence supporting the idea that the formation of the secondary contact zone and the colonization of Germany might be postglacial events. The phylogeographic history inferred for C. unifasciata differs from general biogeographic patterns of postglacial colonization previously identified for other taxa, and it might represent a common model for species with restricted dispersal.