241
Balanopho a xin eniae (Balanopho aceae), a new species om
Xizang, China
Chen-Long Fu1, Jia-Ning Zhou1, Wei-Hua Liao1, Tong Zhang2, Bo Xu3, Meng Li1
1 Co-Inno a ion Cen e o he Sus ainable Fo es y in Sou he n China, College o li e sciences, Nanjing Fo es y Uni e si y, Nanjing 210037, Jiangsu, China
2 Cen al Sou h In en o y and Planning Ins i u e o Na ional Fo es y and G assland Adminis a ion, Changsha, 410014, China
3 CAS Key Labo a o y o Moun ain Ecological Res o a ion and Bio esou ce U iliza ion & Ecological Res o a ion and Biodi e si y Conse a ion Key Labo a o y o
Sichuan P o ince, Chengdu Ins i u e o Biology, Chinese Academy o Sciences, Chengdu 610213, Sichuan, China
Co esponding au ho s: Bo Xu ([email p o ec ed]); Meng Li ([email p o ec ed])
Copy igh : © Chen-Long Fu e al.
This is an open access a icle dis ibu ed unde
e ms o he C ea i e Commons A ibu ion
License (A ibu ion 4.0 In e na ional – CC BY 4.0).
Resea ch A icle
Abs ac
Balanopho a xin eniae C.L.Fu, M.Li & B.Xu, a new species disco e ed in Xizang, China,
is desc ibed and illus a ed he e. Molecula phylogene ic analyses and mo phological
compa isons s ongly suppo B. xin eniae as a new species wi hin he genus Balanopho a.
The mos dis inc i e cha ac e is ic o he new species is i s dioecious sexual sys em, wi h
male lowe s ha ing a 3-lobed pe ian h, opposi e lea es wi h se a ed ips, and yellow
scapes. Al hough i sha es mo phological simila i ies wi h B. hen yi, B. xin eniae can be dis-
inguished by i s la sphe oid ube s wi hou s ella e len icels, yellow scape, p ominen ly
se a ed apical lea ma gins, and nea ly sphe ical o o oid-ellipsoid emale in lo escences.
Key wo ds: Balanopho a xin eniae, China, holopa asi ic plan s, phylogeny, axonomy
In oduc ion
Balanopho a J. R. Fo s . & G. Fo s . is a genus o holopa asi ic plan s con-
sis ing o abou 25 species dis ibu ed mainly in opical Sou heas and Eas
Asia (Hansen 1972; Mu inda e al. 2024). In adi ional medicine, species o
Balanopho a a e o en used o ea hemo hoids, s omach aches, and li e ail-
men s and can also be used as an ipy e ics (Teng e al. 2000; Li e al. 2006;
T akulsomboon e al. 2006; Mu inda e al. 2024). Rega ding hei li es yle,
Balanopho a comp ises highly specialized oo holopa asi es cha ac e ized by
unde g ound connec ions wi h hei hos s. O e an ex ended pe iod o adap-
i e e olu ion, hese plan s ha e expe ienced signi ican gene loss om hei
genomes (Su e al. 2019; Chen e al. 2023) and ex ensi e mo phological educ-
ion, lea ing only a limi ed numbe o ai s a ailable o axonomic classi ica-
ion (Job and Be el 2015; Yu e al. 2022).
Mo phologically, Balanopho a can be eadily di ided in o wo subgene a:
subgenus Balanopho a (4- o 6-me ous male lowe s) and subgenus Balania
(3-me ous male lowe s), a esul ha is also suppo ed by molecula sys-
ema ics (Yu e al. 2022). Howe e , esol ing ela ionships among hese subge-
ne a a he species le el con inues o be a challenging ask. DNA egions wi h
Academic edi o :
Ma cos A. Ca aballo-O iz
Recei ed:
1 Janua y 2025
Accep ed:
5 No embe 2025
Published:
19 No embe 2025
Ci a ion: Fu C-L, Zhou J-N,
Liao W-H, Zhang T, Xu B, Li M
(2025) Balanopho a xin eniae
(Balanopho aceae), a new species
om Xizang, China. Phy oKeys 266:
241–252. h ps://doi.o g/10.3897/
phy okeys.266.147400
Phy oKeys 266: 241–252 (2025)
DOI: 10.3897/phy okeys.266.147400
242
Phy oKeys 266: 241–252 (2025), DOI: 10.3897/phy okeys.266.147400
Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
high subs i u ion a es p o ide e ec i e ools o e alua e he monophyly o
Balanopho a species de ined by mo phological ai s. Recen s udies ha e im-
p o ed ou unde s anding o species bounda ies wi hin his genus by compa -
ing classi ica ions based on mo phological cha ac e s wi h esul s om molec-
ula phylogene ic analyses (Hsiao e al. 2010; Su e al. 2012; Yu e al. 2021). Fo
ins ance, Hansen (1972) conside ed he mo phological di e ences among se -
e al species as in aspeci ic a ia ion, synonymizing hem unde B. ha landii. In
con as , a mo e ecen s udy based p ima ily on molecula phylogene ic anal-
ysis p o ided suppo o ecognizing h ee dis inc axa—B. ha landii, B. hen yi,
and B. kawakamii—wi h clades co esponding o hese axa being s ongly sup-
po ed by molecula da a (Yu e al. 2022).
Du ing ou ieldwo k om Ap il o May 2024 in he o es s nea Gelin Village,
Mo uo Coun y, Xizang (China), we disco e ed a Balanopho a ha di e ed om all
p e iously desc ibed species epo ed in China, India, Nepal, and o he coun ies.
Th ough mo phological obse a ions and he cons uc ion o a phylogene ic ee
using nuclea ibosomal in e nal ansc ibed space (ITS) sequences, we con-
cluded ha his axon ep esen s an undesc ibed species o Balanopho a.
Ma e ials and me hods
Mo phological s udy
Mo phological obse a ions we e conduc ed based on li ing plan s in he ield.
Measu emen s we e pe o med manually wi h ule s in he ield o wi h he so -
wa e ImageJ in he labo a o y using pho og aphs. We es ic ed mo phological
compa isons o h ee closely ela ed species. A he same ime, all images o
specimens po en ially ela ed o he new species om China, India, Nepal, and
o he coun ies deposi ed in he he ba ia A, CDBI, CDCM, GAC, HHBG, IBSC, IBK,
LBG, K, KUN, NAS, P, PE, SHM, and ZM we e obse ed and s udied (ac onyms
ollow Thie s 2024). The e minology used in he mo phological desc ip ion ol-
lows he Flo a o China (Huang and Mu a a 2003). Obse a ions o lowe s and
cla i o m bodies we e ca ied ou using an Olympus SZX10 s e eomic oscope.
Phylogene ic s udy
DNA ex ac ions, PCR p ocedu es, and p ime s ollowed Yu e al. (2021). The
ITS egion o he new species was sequenced using he Sange me hod. In ad-
di ion, we downloaded ITS sequences o o he Balanopho a species a ailable in
he Nucleo ide da abase o NCBI (h ps://www.ncbi.nlm.nih.go ). Langsdo ia
hypogaea was selec ed as he ou g oup o his s udy based on he esul s p e-
sen ed by Sanchez-Pue a e al. (2023). A o al o 54 samples we e used in his
s udy, including mul iple accessions o some species (Table 1). All sequences
we e aligned using MAFFT 7.526 ollowing de aul pa ame e s (Ka oh e al.
2002; Ka oh and S andley 2013), and he ends we e manually immed. The
model TPM3u+F+I+R2 o ML analysis and TPM3u +I+G o BI analysis was
selec ed based on he Bayesian In o ma ion C i e ion (BIC) using jModelTes
2.1.10 (Da iba e al. 2012). Maximum likelihood (ML) analyses we e pe -
o med using IQ-TREE 2.3.6 wi h 10,000 ul a as boo s ap app oxima ions
(Minh e al. 2020). A Bayesian in e ence (BI) analysis was pe o med using
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Phy oKeys 266: 241–252 (2025), DOI: 10.3897/phy okeys.266.147400
Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
M Bayes 3.2.7 (Ronquis e al. 2012). A Ma ko chain Mon e Ca lo analysis
was conduc ed o 20,000,000 gene a ions wi h one cold and h ee ho chains,
each s a ing om a andom ee and sampled e e y 1,000 gene a ions. The
i s 25% o he ees we e disca ded as bu n-in, and he emaining ees we e
used o cons uc majo i y- ule consensus ees. The inal ees ob ained om
he ML and BI analyses we e isualized using FigT ee 1.4.2 (Rambau 2009).
Resul s and discussion
The ci cumsc ip ion o B. la a, B. a gesii, and B. in oluc a a emains a pe sis en
axonomic con en ion. Geng (2016) p oposed ha B. a gesii and B. in oluc a a
a e sis e g oups wi h dis inc clades based on ITS sequences. In con as , Lidén
(2019) sugges ed ha B. a gesii is synonymous wi h B. in oluc a a, while Yu e al.
(2022) posi ed ha B. la a and B. a gesii a e synonymous wi h B. in oluc a a. In
his s udy, hese species a e conside ed pa o he B. in oluc a a complex, and we
a e no add essing axonomic di isions wi hin his g oup he e. Ins ead, o cla i y
and o acili a e mo phological compa isons, we e e o he dioecious indi iduals
wi hin he complex as B. la a and he monoecious indi iduals as B. in oluc a a.
Table 1. Taxa used in he phylogene ic analyses wi h hei GenBank accession numbe s.
Species Accession numbe Species Accession numbe
B. app essa MW414662 B. laxi lo a JN392888
B. app essa MW414670 B. laxi lo a JN392889
B. la a OM103570 B. laxi lo a MW414661
B. la a OM103567 B. laxi lo a MW414659
B. ungosa JN392881 B. laxi lo a JN392890
B. ha landii OM103582 B. laxi lo a JN392891
B. ha landii OM103583 B. laxi lo a JN392892
B. ha landii OM103585 B. laxi lo a JN392893
B. hen yi OM103573 B. pa ajaponica MT482093
B. hen yi OM103574 B. pa ajaponica MT498081
B. hen yi OM103575 B. pa ajaponica MW414666
B. hen yi OM103576 B. pa ajaponica MW414667
B. hen yi OM103577 B. e lexa EU598798
B. indica JN392880 B. e lexa KT763380
B. in oluc a a OM103568 B. subcupula is OM103584
B. in oluc a a OM103569 B. subcupula is OM418783
B. in oluc a a OM103571 B. subcupula is OM418784
B. in oluc a a OM103572 B. obi acola JN392901
B. japonica JN392882 B. obi acola OM103578
B. japonica MW414664 B. xin eniae PQ640032
B. japonica MW414665 B. xin eniae PQ640033
B. kawakamii JN392900 B. yakushimensis JN392894
B. kawakamii OM103579 B. yakushimensis MW414660
B. kawakamii OM103580 B. yakushimensis MW414663
B. kawakamii OM103581 B. yakushimensis MW414668
B. la isepala OM418779 B. yakushimensis MW414669
B. la isepala OP120928 Langsdo ia hypogaea KT709672
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Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
The opologies o he phylogene ic ees cons uc ed by he ML and BI me h-
ods a e highly consis en (Suppl. ma e ials 1, 2). Ou esul s showed ha wo
samples o he new species we e well esol ed as a dis inc clade, which is he
sis e g oup o he B. in oluc a a complex (BS = 1; PP = 98%) (Fig. 1).
Figu e 1. The cladog am o Balanopho a cons uc ed using maximum likelihood (le ) and Bayesian in e ence ( igh )
based on ITS sequences. Numbe s a nodes indica e pos e io p obabili ies (PP) o boo s ap pe cen ages (BS). B anches
highligh ed in ed indica e opological incong uence be ween he wo ees. Labels o conco dan opologies a e no
displayed on he BI ee.
B. laxi lo a JN392891
B. hen yi OM103573
B. japonica MW414664
B. laxi lo a JN392889
B. la isepala OP120928
B. japonica MW414665
B. la a OM103567
B. subcupula is OM103584
B. kawakamii OM103579
B. la isepala OM418779
B. laxi lo a JN392892
B. japonica JN392882
B. la a OM103570
B. yakushimensis JN392894
B. laxi lo a JN392893
B. ha landii OM103585
B. pa ajaponica MT482093
B. obi acola JN392901
B. app essa MW414670
B. app essa MW414662
B. kawakamii OM103580
B. hen yi OM103577
B. in oluc a a OM103569
B. hen yi OM103574
B. laxi lo a MW414659
B. in oluc a a OM103568
B. yakushimensis MW414668
B. in oluc a a OM103571
B. yakushimensis MW414660
B. pa ajaponica MW414666
Langsdo ia hypogaea KT709672
B. in oluc a a OM103572.
B. laxi lo a MW414661
B. pa ajaponica MW414667
B. ha landii OM103583
B. subcupula is OM418784
B. subcupula is OM418783
B. yakushimensis MW414669
B. e lexa KT763380
B. kawakamii OM103581
B. kawakamii JN392900
B. yakushimensis MW414663
B. xin eniae PQ640032
B. hen yi OM103576
B. laxi lo a JN392888
B. e lexa EU598798
B. xin eniae PQ640033
B. hen yi OM103575
B. pa ajaponica MT498081
B. ungosa JN392881
B. indica JN392880
B. obi acola OM103578
B. laxi lo a JN392890
B. ha landii OM103582
99
98
100
97
92
97
100
19
36
100
84
71
98
49
100
100
100
100
100
100
100
67
95
80
100
53
100
59
100
99
57
100
100
100
100
91
100
100
100
56
58
100
98
100
0.98
0.34
1
0.87
0.35
1
OM418783
OM418784
OM103584
JN392889
JN392893
JN392892
JN392890
JN392891
0.38
0.38
1
1
0.32
1
1
0.92
1
0.45
0.78
0.81
1
0.39
0.08
0.12
0.12
1
1
1
1
1
1
1
0.94
0.89
0.33
1
1
0.97
0.73
1
1
1
0.69
0.18
0.18
OM103574
OM103577
OM103575
OM103576
1
0.40
0.87
0.43
1
0.98
0.86
0.45
ML ee BI ee
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Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
The new species desc ibed in his s udy is mo phologically mos simila o
B. hen yi, along wi h he closely ela ed species B. in oluc a a and B. la a. In
he phylogeny, he new species was placed in a clade con aining hese h ee
species (Fig. 1). We he e o e ocused ou compa ison o mo phological cha -
ac e s on hese ou species (Table 2; Fig. 2). The mos dis inc i e cha ac e -
is ics o he new species a e i s dioecious sexual sys em ( s. monoecious in
B. in oluc a a), opposi e lea es ( s. conna e in o a single shea h-like who l in
B. la a and B. in oluc a a) wi h se a ed ma gins ( s. c ena e in B. la a and B.
in oluc a a and en i e in B. hen yi), and yellow scapes ( s. ed in B. in oluc a a
and ed o ed o yellow in B. hen yi).
Based on he esul s o molecula and mo phological analyses, we belie e
ha he e is su icien e idence o suppo he alidi y o his axon as dis inc ly
di e en om B. hen yi, B. in oluc a a, and B. la a. The e o e, we ecommend
ha i be ea ed as a new species.
Figu e 2. Mo phology o Balanopho a xin eniae (A–C), B. hen yi (D–F), B. in oluc a a (G–I), and B. la a (J–M). A, D, G, J, K. Male
and emale indi iduals; B, E, H, L. Lea es; C, F, I, M. Tube s. Pho os by Meng Li and Wei-Hua Liao.
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Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
Table 2. Compa ison o mo phological ea u es among ou ela ed species o Balanopho a.
B. xin eniae B. hen yi B. in oluc a a B. la a
Rep oduc ion Dioecious Dioecious Monoecious Dioecious
Tube b anching
shape
Fla sphe oid o
subglobose
I egula ly sphe ical
o obla e
U ceola e, sligh ly
elonga ed
U ceola e
Tube su ace Densely co e ed
wi h g anula wa s
Densely co e ed
wi h g anula wa s
Densely co e ed wi h
g anula wa s and
yellowish s ella e len icels
Densely co e ed wi h
g anula wa s and
yellowish s ella e len icels
Colo o scapes Yellow Red o ed o yellow Red Yellow
Lea a angemen Opposi e Opposi e Conna e in o a single
shea h-like who l
Conna e in o a single
shea h-like who l
Lea es apex Se a ed En i e C ena e C ena e
Shape o emale
in lo escence
Nea ly sphe ical o
o oid-ellipsoid
B oadly o oid O oid o sho ellipsoid O oid o sho ellipsoid
Flowe ing pe iod Ap il o May Sep embe o
No embe
July o Augus July o Augus
Taxonomic ea men
Balanopho a xin eniae C.L.Fu, M.Li & B.Xu, sp. no .
u n:lsid:ipni.o g:names:77372106-1
Figs 3, 4
Type. China • Xizang Au onomous Region: Mo uo Coun y (Mêdog), Gelin Vil-
lage, 29.210161, 95.191856, al . 1650 m, 29 Ap 2024 ( l.), Meng Li, C.L. Fu &
J.N. Zhou YLZB11691-A (holo ype: CDBI! Ba code numbe : CDBI0298364, ♀;
iso ypes: NAS! Ba code numbe : NAS00708465; NF!).
Diagnosis. Balanopho a xin eniae is mo phologically simila o B. hen yi, bu he
wo species can be dis inguished by se e al ea u es. The ube b anching shape
o B. xin eniae is cha ac e ized as la sphe oid o subglobose, whe eas ha o B.
hen yi is i egula ly sphe ical o obla e. The scapes o B. xin eniae a e yellow, in
con as o he ed o ed- o-yellow scapes o B. hen yi. In B. xin eniae, he apical
ma gins o he lea es a e p ominen ly se a e, whe eas hose o B. hen yi a e en-
i e. The emale in lo escence o B. xin eniae is nea ly sphe ical o o oid-ellipsoid,
while ha o B. hen yi is b oadly o oid. The lowe ing pe iod o B. xin eniae is om
Ap il o May, compa ed o Sep embe o No embe o B. hen yi (Table 2; Fig. 2).
Desc ip ion. Plan s dioecious. Tube yellowish b own, b anched o unb anched,
su ace scab ous, densely co e ed wi h g anula wa s; ube b anches la sphe -
oid o subglobose, 0.8–2.0 cm in diam. Scapes yellow, 2–12 × 0.5–1.2 cm. Lea es
6–10, yellow o eddish, opposi e, scaly, oblong-o a e, apical ma gins p ominen ly
and i egula ly se a e, wi h se a ions a anged app oxima ely pa allel o he lea
ma gin, 1.2–3.5 × 0.3–1.8 cm. Male in lo escences subsphe oid o o oid-ellip-
soid, 1.8–2.5 × 1.5–2.0 cm. B ac s unca e wi h expanded liplike ma gin, used
side by side in o a hexagonal al eolus. Male lowe s: pedicella e, inse ed basally
in al eolus, 3-me ous, 2–4 mm in diam. Pe ian h lobes b oadly del oid. Synand ia
subdiscoid; an he s 3, ans e sely dehiscen . Female in lo escence nea ly sphe -
ical o o oid-ellipsoid. Cla i o m bodies obo oid, sho ly s iped, apically unca e;
cu icula idges o apical cells laby in h-like. Female lowe s: bo h in he lowe
pa s o he cla i o m bodies and on he main axis o he in lo escence.
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Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
Dis ibu ion and habi a . The new species is cu en ly known only om
he ype locali y in Gelin Village, Mo uo Coun y, Xizang Au onomous Region
(Fig. 5). I g ows in shaded o es habi a s on mois moun ain slopes. The
hos s a e usually species o Fagaceae, such as Que cus lamellosa and
Cas anopsis indica.
Figu e 3. Balanopho a xin eniae C.L. Fu, M. Li & B. Xu. A, B. Habi a s (bo h male and emale; a ows poin o emale
indi iduals); C. Female indi idual (uppe ) and male indi idual (lowe ); D. Male indi idual; E. Female indi idual; F. Lea es
and scape; G. Lea es; H. Tube ; I. Male in lo escence; J. Female in lo escence; K. Female lowe and cla i o m body.
Pho os by Meng Li and Chen-Long Fu.
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Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
Figu e 4. Line illus a ion o Balanopho a xin eniae. A. Male indi idual; B. Male lowe ; C. Female indi idual; D. Female lowe s
su ounding one cla i o m body. D awn by Zi-Heng Yu based on YLZB11691-A and YLZB11691-B specimens s o ed in CDBI.
Figu e 5. Dis ibu ion o Balanopho a xin eniae (Balanopho aceae).
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Chen-Long Fu e al.: Balanopho a xin eniae (Balanopho aceae) om China
Phenology. The species has been obse ed lowe ing om Ap il o May.
E ymology. The species epi he xin eniae hono s P o . Xin-Fen Gao ( o me
cu a o o he CDBI He ba ium), a dis inguished bo anis who made signi ican
con ibu ions o he axonomy o seed plan s, lycopods, and p e idophy es.
Ve nacula name. Chinese Manda in: Xìn ēn shé gū (信芬蛇菰).
Addi ional specimens examined. China • Xizang Au onomous Region:
Mo uo Coun y, Gelin Village, 29.210161°N, 95.191856°E, al . 1650 m, 29 Ap
2024 ( l.), Meng Li, C.L. Fu & J.N. Zhou YLZB11691-B (pa a ype: CDBI! Ba -
code numbe : CDBI0298363, ♂; NAS! Ba code numbe : NAS00708510, ♂; NF!,
♂); • Gelin Village, 29.220615°N, 95.189094°E, al . 1602 m, 1 May 2024 ( l.),
Meng Li MD2405013-B (pa a ype: CDBI! Ba code numbe : CDBI0298362, ♂);
• Gelin Village, al . 1700 m, 24 May 1983 ( l.), B.S. Li, S.Z. Cheng & Z.C. Nie
03761 (PE! Ba code numbe : PE01491372; PE01801408); • Beibeng Town-
ship, al . 1650 m, 11 May 1983 ( l.), S.Z. Cheng, Z.C. Nie & B.S. Li 04844 (PE!
Ba code numbe : PE01491371).
Key o he new species o Balanopho a and h ee mo phologically
simila axa
1 Lea es conna e in o a single shea h-like who l ...........................................2
– Lea es opposi e ...........................................................................................3
2 Plan s monoecious, male lowe s clus e ed basally on in lo escence ........
................................................................................. Balanopho a in oluc a a
– Plan s dioecious ................................................................Balanopho a la a
3 Scapes yellow, apical lea ma gins p ominen ly and i egula ly se a e, emale
in lo escence nea ly sphe ical o o oid-ellipsoid .......... Balanopho a xin eniae
– Scapes ed o ed o yellow, lea ma gins en i e, emale in lo escence
b oadly o oid .................................................................. Balanopho a hen yi
Acknowledgemen s
We since ely hank he e iewe s and edi o s o hei aluable commen s. We
a e g a e ul o P o . Yong Yang o his guidance on specimen p epa a ion and o
Xiao-Xia Ding and Xiao-Yu Dong o p epa ing he ype specimens. We also hank
Yan-Feng Song o assis ance in examining he specimens s o ed in PE and ex-
end special hanks o Zi-Heng Yu o d awing he line illus a ions. We app ecia e
he suppo o he Fo es y and G assland Bu eau o Linzhi Ci y and he Fo es y
and G assland Bu eau o Mo uo Coun y o issuing he collec ion pe mi s.
Addi ional in o ma ion
Con lic o in e es
The au ho s ha e decla ed ha no compe ing in e es s exis .
E hical s a emen
No e hical s a emen was epo ed.
Use o AI
No use o AI was epo ed.
