Nova Hedwigia 80 3—4 541—545 Stuttgart, May 2005

A morpho-molecular classification of the Anthocerotophyta

(hornworts)
by

Wolfgang Frey and Michael Stech

Institut für Biologie – Systematische Botanik und Pflanzengeographie,
Freie Universität Berlin, Altensteinstr. 6, D-14195 Berlin, Germany

Frey, W. & M. Stech (2005): A morpho-molecular classification of the Anthocerotophyta (hornworts).

- Nova Hedwigia 80: 541-545.

Abstract: Recent molecular analyses and a re-evaluation of morphological and anatomical characters
provided new insights into relationships among hornworts (Anthocerotophyta). Considering these
results, a new suprageneric classification of hornworts is formally proposed. The monospecific
genus Leiosporoceros (Leiosporocerotales) is separated into its own class Leiosporocerotopsida
class. nov., based on its unique combination of molecular and non-molecular characters. Within the
Anthocerotopsida, three orders are distinguished, Anthocerotales (comprising Anthocerotaceae and
Foliocerotaceae), Phaeocerotales (Phaeocerotaceae and Notothyladaceae) and Dendrocerotales
(Dendrocerotaceae). The latter comprises genera Dendroceros, Megaceros and Nothoceros, as well
as some species of Phaeoceros according to the molecular analyses, which are morphologically
intermediate between Megaceros and Phaeoceros. Taxonomic consequences at the generic level
within hornworts are advisable only after further molecular investigations including additional markers
and a broader taxon sampling.

Key words: Anthocerotophyta, suprageneric classification, Leiosporocerotopsida class. nov., mole-

cular data

Introduction

The hornworts (Anthocerotophyta) are a small, isolated and presumably ancient

group of land plants, comprising 11(-12) genera with probably 100-150 species
after generic revisions that are still due for the larger genera. Hornworts are widely
distributed in tropical and temperate zones, with an increasing diversification in the
(sub-)tropical and austral temperate regions, colonizing mostly moist mineral soil of
banks and cliffs, cultivated land and splashed rocks along streams.
Characters separating hornworts from all other land plant lineages comprise, e.g.,
mostly uniplastidic cells with channel thylakoids and pyrenoid apparatus, presence

DOI: 10.1127/0029-5035/2005/0080-0541 0029-5035/05/0080-0541 $ 1.50

© 2005 J. Cramer in der Gebrüder Borntraeger
Verlagsbuchhandlung, D-14129 Berlin · D-70176 Stuttgart
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of a peculiar basal meristem of the sporophyte, non-synchronous development of
spores and pseudoelaters from archesporial tissue, release of mucilage from
nonspecialized cells, as well as characteristic spermatozoid and blepharoplast types
and structure of the gametophyte-sporophyte junction, and the large stem-loop region
P6 of the chloroplast DNA trnL intron. All molecular analyses resolve hornworts as
monophyletic, however, their position in the land plant hierarchy still remains
ambiguous (summarized in Stech et al. 2003).
Relationships within hornworts have been controversially discussed from early studies
to the latest classifications based on morphological and anatomical characters (Hässel
de Menéndez 1988, Schuster 1992, Hyvönen & Piippo 1993, Hasegawa 1994). Recent
molecular analyses based on chloroplast DNA sequences, either rbcL (Duff et al.
2004, results also discussed by Shaw & Renzaglia 2004) or the trnL-trnF region
(Stech et al. 2003), support new and unexpected generic relationships (see below)
and indicate the need of a re-evaluation of traditional characters. A new formal
classification of hornworts considering the latest molecular results is also needed,
especially for a treatment of bryophytes for a new edition of the world-famous
Engler´s Syllabus of Plant Families, which is currently in preparation. Here we
provide a suprageneric taxonomy of Anthocerotophyta that summarizes the current
knowledge of hornwort relationships.

Systematic arrangement of taxa

According to the rbcL analysis of representatives of seven hornwort genera by Duff
et al. (2004), Leiosporoceros occupies the basalmost position among hornworts. The
remaining genera are divided into two clades, the first comprising Anthoceros and
Folioceros and the second Phaeoceros, Notothylas, Dendroceros, Megaceros and
Nothoceros. Both the analyses of Stech et al. (2003) and Duff et al. (2004) support
polyphyly of Phaeoceros as well as close relationships of Notothylas and Phaeoceros
p.p. on the one hand and Dendroceros, Megaceros and Phaeoceros p.p. on the other
hand. The following classification considers these new molecular results. Numbers
of species within genera are provisional pending generic taxonomic revisions based
on molecular and morphological characters.

1. Class Leiosporocerotopsida W.Frey & Stech, classis nova

Differt ab classe Anthocerotopsida tetradibus sporarum isobilateralibus, sporis ovoideis pellucidis
laevibus, capsulis sutura profunda in longitudinem sulcatum, anatomia chloroplastorum et sequentia
rbcL.
Thalli solid, without mucilage clefts. Spore tetrads isobilateral. Spores minute, ovoid,
thin-walled, smooth; monolete. Pseudoelaters long, consisting of 1(-2?) cells, thick-
walled. Capsule with stomata, suture highly differentiated and visible as a deep
longitudinal groove. Chloroplasts with peripheral starch and central grana, pyrenoid
lacking. Sister group to all other hornworts according to rbcL sequences. Genetically
isolated by apparent lack of RNA editing of the rbcL transcript (Duff et al. 2004).
Only one species.

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Order Leiosporocerotales Hässel
Fam. Leiosporocerotaceae Hässel
Leiosporoceros Hässel (1). L. dussii (Steph.) Hässel, trop., Mexico, Antilles, Panama,
Ecuador.

2. Class Anthocerotopsida Engel

Thalli cavernous or solid. Spore tetrads tetrahedral. Spores trilete, ill-defined only
in some Folioceros species, spinose except for some Notothylas species. Pseudoelaters
short, thin-walled or long, thick-walled; helical bands partly present.

1. Order Anthocerotales Limpr.

More than two antheridia per antheridial chamber. Antheridial jacket consisting of 4
cell tiers. Stomata present. Spores dark brown or blackish.
Fam. Anthocerotaceae Dumort.
Thalli cavernous, with mucilage-filled or Nostoc containing cavities. Pseudoelaters
short, thin-walled. Two genera with about 80 species: (i) Anthoceros L. (c. 80),
temp.-trop. Thalli variously lobed. Spores with trilete mark. A. neesii Prosk. is
endemic to Europe. (ii) Sphaerosporoceros Hässel (2), temp.-subtrop., North America,
Colombia. Similar to Anthoceros. Spores spherical, trilete mark reduced.
Pseudoelaters break up into their individual short cells.
Fam. Foliocerotaceae Hässel
Thalli dissected into variously shaped lobes, with large mucilagenous cavities. Spores
with reduced trilete mark. Pseudoelaters 4 cells long, with a dark thickened peripheral
wall; lumens as narrow slits. One genus, Folioceros D.C.Bharadwaj with 17 species,
subtrop.-trop., mainly Asiatic, in South America only F. apiahynus (Steph.) Hässel,
two species in Africa.

2. Order Phaeocerotales Prosk.

More than two antheridia per antheridial chamber. Antheridial jacket consisting of
irregularly arranged cells. Stomata present or absent. Spores yellow to blackish.
Fam. Phaeocerotaceae D.C.Bharadwaj
Thalli solid, lacking cavities, lobed; with continous growth after fertilization.
Sporophytes long. One genus, Phaeoceros Prosk. originally with about 40 species,
but polyphyletic according to the molecular data, with some species closely related
to Megaceros (Dendrocerotaceae), temp.-trop. Pseudoelaters short, thin-walled. Spores
yellow. P. carolinianus (Michx.) Prosk., nearly cosmopolitan.
Fam. Notothyladaceae (Milde) Müll.Frib.
Thalli solid [except N. orbicularis (Schwein.) Sull., scanty cavernous], only up to
5(-7) mm in Ø; growth stops at the apices after fertilization. 1(-3) chloroplasts per

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cell. Sporophytes in pairs, fusiform to ellipsoidal, horizontal on the thallus, enclosed
to maturity in an involucre, cleistocarpous to tardily dehiscing by 1-2 lines. Mature
sporophytes only slightly exceeding the involucre. Columella poorly developed or
lacking. Stomata normally lacking. DNA studies support a sistergroup relationship
of Notothylas and Phaeoceros. One genus, Notothylas Sull. ex A.Gray with 22 species,
cosmopolitan in warm climates, with a centre of diversification on the Indian
subcontinent and East Asia. Spores yellowish to blackish. Pseudoelaters break up
into individual short cells with irregular thickenings. N. orbicularis (Schwein.) Sull.,
western Europe, eastern North America, Korea, Japan.

3. Order Dendrocerotales Hässel

Mainly 1 antheridium per antheridial chamber. Antheridial jacket consisting of
irregularly arranged cells. Stomata lacking. Spores transparent. Pseudoelaters with
helical thickenings.
Fam. Dendrocerotaceae (Milde) Hässel
With the characters of the order. Three genera with about 50 species. Gondwanalandic-
trop. (i) Dendroceros Nees (c. 30), trop.-subtrop. Thalli solid or cavernous, with
midrib and lateral unistratose wings. Pyrenoid present. Columella slight. Spores
multicellular. Germination endosporous. Epiphytic, sometimes epiphyllous. D.
javanicus (Nees) Nees, trop.-subtrop. Asia, S Pacific, Tanzania. (ii) Megaceros Campb.
(20), trop.-Stemp., widely distributed in the Southern Hemisphere. Thalli flat, solid.
1–4(12) chloroplasts per cell. Pyrenoid lacking. Columella massive. Spores unicellular.
M. flagellaris (Mitt.) Steph., widely distributed in tropical and subtropical Asia, S
Pacific, Australia, New Zealand, Himalayas, Africa. (iii) Nothoceros (R.M.Schust.)
J.Haseg. [Megaceros subg. Nothoceros sensu R.M.Schust., subg. Australoceros sensu
J.Haseg.] (3), palaeoaustral, subantarctic. Thalli solid, remotely furcately branched,
with lingulate segments (margins unistratose, laciniate, crispulate). >2 chloroplasts
per cell. Pyrenoid lacking. Spores pale yellow. N. giganteus (Lehm. & Lindenb.)
J. Haseg., S New Zealand, probably conspecific with N. endiviaefolius (Mont.) J.
Haseg., Tierra del Fuego and southern Patagonia.

Future considerations

The systematic positions of Hattorioceros (J.Haseg.) J.Haseg., Mesoceros Piippo

and Sphaerosporoceros Hässel remain to be tested by molecular analyses. Whereas
the latter is morphologically close to Anthoceros, relationships of Hattorioceros and
Mesoceros are ambiguous also at the morphological level. Hattorioceros striatisporus
(J.Haseg.) J.Haseg. was originally described as Phaeoceros striatisporus J.Haseg.,
but separated into its own monospecific genus because of basically ovoid to spheroid
spores with a triradiate mark, a unique spore type in Anthocerotophyta. Mesoceros
comprises two species that are intermediate between Anthoceros and Phaeoceros.
The polyphyly of Megaceros and Phaeoceros, as indicated by rbcL and trnL intron
sequences (Stech et al. 2003, Duff et al. 2004), will finally require taxonomic

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consequences after investigation of additional species. So far, three Phaeoceros species,
P. chiloensis (Steph.) Hässel, P. coriaceus (Steph.) E.O.Campb. and P. hirticalyx
(Steph.) J.Haseg. have been revealed as closely related to Megaceros and should,
consequently, be transferred to Dendrocerotaceae. These species have one androecium
per cavity and multiple chloroplasts lacking pyrenoids, diagnostic characters of
Megaceros. In contrast, lack of helical pseudoelaters, presence of stomata and yellow
spores are characters that define Phaeoceros. Taxa with these character combination
may be grouped into a new genus, and Megaceros may be separated into two genera.

Acknowledgement

Sincere thanks are due to Dr D. Quandt (Dresden) for critical reading of the manuscript.

References

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Received 15 December 2004, accepted in revised form 10 February 2005.

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