Research Article |
Corresponding author: Seung Se Choi ( hepaticae@jbnu.ac.kr ) Academic editor: Matt von Konrat
© 2022 Vadim A. Bakalin, Yulia D. Maltseva, Ksenia G. Klimova, Van Sinh Nguyen, Seung Se Choi, Aleksey V. Troitsky.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Bakalin VA, Maltseva YD, Klimova KG, Nguyen VS, Choi SS, Troitsky AV (2022) The systematic position of puzzling Sino-Himalayan Lophocolea sikkimensis (Lophocoleaceae, Marchantiophyta) is identified. PhytoKeys 206: 1-24. https://doi.org/10.3897/phytokeys.206.84227
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Lophocolea sikkimensis, a little-known Sino-Himalayan species, was collected in North Vietnam and its taxonomic position was identified by molecular genetic techniques. The species is characterized by generally narrowly pointed leaves, which are not seen in other representatives of Lophocoleaceae. We found that it belongs to the recently described genus Cryptolophocolea, although it is clearly morphologically dissimilar to other members of the genus. We propose a corresponding nomenclature combination: Cryptolophocolea sikkimensis comb. nov. This species is the only one in its genus with a predominantly Sino-Himalayan distribution; the vast majority of congeners are distributed in the Southern Hemisphere (mostly in Australasia). Reports of this species in Vietnam further confirm the close phytogeographic relationships of the flora of northern Indochina with those of the Sino-Himalayas and suggest that this species is found in other parts of the Hoang Lien Range and the southern Hengduan Range.
Lophocolea, Lophocoleaceae, molecular phylogenetic, Sino-Himalaya, taxonomy, Vietnam
Lophocolea sikkimensis (Steph.) Herzog & Grolle is a poorly known Lophocoleaceae species but is so different from other known members of the family that it appears to belong to a different genus.
Initially, Lophocolea sikkimensis was placed in the genus Herpocladium as H. sikkimense (
After reviewing the characteristics previously mentioned (plant color, large entire underleaves, dorsally secund leaves, biseriate antheridium stalk, etc.),
In the mid-2000s, important molecular-genetic comparisons were carried out on Lophocoleaceae, providing a new perspective on the old problem. Research by
We analyzed two specimens of Lophocolea sikkimensis in our molecular phylogenetic study using nucleotide sequence data from ribosomal operon of nuclear DNA (ITS1–2) and trnL-F of chloroplast DNA. In addition to Lophocoleaceae, the analysis included sequence data from genetically related Jungermanniales families (
There were too few trnG-intron sequences from Lophocoleaceae in GenBank to construct a reliable phylogenetic tree for this marker that establishes the position of Lophocolea sikkimensis.
Therefore, new trnG-intron sequences were obtained for this taxon but not analyzed properly.
Specimen voucher details, as well as newly identified and previously identified sequences, are listed in Table
Initial species name | Accepted name | Label | GenBank accession number | |
---|---|---|---|---|
ITS1–2 | trnL-F | |||
Bragginsella anomala R.M. Schust. | Bragginsella anomala R.M. Schust. | New Zealand, M. von Konrat & J.J. Engel, L1129 (F) | – | KJ802081 |
Chiloscyphus austrigenus (Hook. f. & Taylor) J.J. Engel & R.M. Schust. | Pachyglossa austrigena (Hook. f. & Taylor) L. Söderstr. | Chile, Hyvönen et al. 5793 (JE) | AM282805 | – |
Chiloscyphus ciliolatus (Nees) J.J. Engel & R.M. Schust. | Cryptolophocolea ciliolata (Nees) L. Söderstr., Crand.-Stotl., Stotler & Váňa | Indonesia, Gradstein, 10327 (GOET) | AM491286 | – |
Chiloscyphus connatus (Sw.) J.J. Engel & R.M. Schust. | Cryptolophocolea connata (Sw.) L. Söderstr. & Váňa | Costa Rica, Gradstein, 9404 (GOET) | AM282806 | – |
Chiloscyphus costatus (Nees) J.J.Engel & R.M.Schust. | Cryptolophocolea costata (Nees) L. Söderstr. | Malaysia, Schäfer-Verwimp & Verwimp, 18724/A (JE) | AM282807 | – |
Chiloscyphus cucullistipulus (Steph.) Hässel | Clasmatocolea cucullistipula (Steph.) Grolle | Chile, Drehwald, 970184 (GOET) | AM491287 | – |
Chiloscyphus cuspidatus (Nees) J.J. Engel & R.M. Schust. | Lophocolea cuspidata (Nees) Limpr. | Germany, Hentschel, Bryo 01411 (JE) | AM491604 | – |
Chiloscyphus fragmentissimus (R.M.Schust.) J.J.Engel & R.M.Schust. | Lophocolea fragmentissima R.M. Schust. | Venezuela, Frahm, 97/5/N (GOET) | AM282809 | – |
Chiloscyphus fragrans (Moris & De Not.) J.J.Engel & R.M.Schust. | Lophocolea fragrans (Moris & De Not.) Gottsche, Lindenb. & Nees | Azores, Schwab, 113 (JE) | AM282810 | – |
Chiloscyphus fulvellus (Hooker f. & Taylor) Nees | Clasmatocolea fulvella (Hook. f. & Taylor) Grolle | Chile, Hyvönen, 5313 (GOET) | AM491288 | – |
Chiloscyphus gayanus (Mont.) Gottsche & al. | Clasmatocolea gayana (Mont.) Grolle | Chile, Holz & Franzaring, CH 00-151a (GOET) | AM491289 | – |
Chiloscyphus gottscheoides (Besch. & C.Massal.) J.J.Engel & R.M.Schust. | Pachyglossa gottscheoides (Besch. & C. Massal.) L. Söderstr. | Chile, Drehwald & Mues, 3239 (GOET) | AM282811 | – |
Chiloscyphus guadalupensis (Steph.) J.J.Engel & R.M.Schust. | Cryptolophocolea guadalupensis (Steph.) L. Söderstr. & Váňa | Costa Rica, Gradstein & Mues, 9630 (GOET) | AM282813 | – |
Chiloscyphus helmsianus (Steph.) J.J. Engel & R.M. Schust. | Cryptolophocolea helmsiana (Steph.) L. Söderstr. | New Zealand, Engel & von Konrat, 28439 | – | FJ173297 |
Chiloscyphus humilis (Hook. f. & Taylor) Hässel | Clasmatocolea humilis (Hook.f. et Taylor) Grolle | Chile, Holz & Franzaring, CH 00-44B (GOET) | AM491290 | – |
Chiloscyphus itoanus (Inoue) J.J. Engel & R.M. Schust. | Lophocolea itoana Inoue | China, 1999 Piippo, 60709 | – | AY149868 |
Chiloscyphus japonicus (Steph.) J.J. Engel & R.M. Schust. | Cryptolophocolea compacta (Mitt.) L. Söderstr. | China, 1998 Koponen et al. 50238 | – | AY149869 |
Chiloscyphus lentus (Hook.f. & Taylor) J.J.Engel & R.M.Schust. | Lophocolea lenta (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | New Zealand, Engel & von Konrat, 24002 | – | FJ173298 |
Chiloscyphus leucophyllus (Hook.f. & Taylor) Gottsche, Lindenb. & Nees | Cryptolophocolea leucophylla (Hook. f. & Taylor) L. Söderstr. | New Zealand, Engel & von Konrat, 24319 | – | FJ173299 |
Chiloscyphus liebmannianus (Gottsche) J.J.Engel & R.M.Schust. | Lophocolea liebmanniana Gottsche | Mexico, Burghardt Bryo, 01655 (GOET) | AM282816 | – |
Chiloscyphus martianus (Nees) J.J.Engel & R.M.Schust. | Cryptolophocolea martiana (Nees) L. Söderstr., Crand.-Stotl. & Stotler | Ecuador, Gradstein, 10119 (GOET) | AM282817 | – |
Chiloscyphus massalongoanus Steph. | Cryptolophocolea massalongoana (Schiffn.) L. Söderstr. | Indonesia, Schaefer-Verwimp & Verwimp, S-V 25279 | AM491292 | – |
Chiloscyphus minor (Nees) J.J.Engel & R.M.Schust. | Lophocolea minor Nees | Germany, Hentschel Bryo, 01006 (JE) | AM282818 | – |
Chiloscyphus minor (Nees) J.J.Engel & R.M.Schust. | Lophocolea minor Nees | China, Hunan Province, Zhangjiajie, 1999 Rao, 58428 | – | AY149864 |
Chiloscyphus muricatus (Lehm.) J.J.Engel & R.M.Schust. | Lophocolea muricata (Lehm.) Nees | Australia, Streimann, 51629 (JE) | AM282819 | – |
Chiloscyphus novae-zeelandiae (Lehm. & Lindenb.) J.J.Engel & R.M.Schust. | Lophocolea novae-zeelandiae (Lehm. & Lindenb.) Nees | Australia, Eggers, AUS 3/81 (JE) | AM282820 | – |
Chiloscyphus novae-zeelandiae var. grandistipulus (Schiffn.) J.J.Engel | Lophocolea novae-zeelandiae var. grandistipula (Schiffn.) Váňa | New Zealand, Engel & von Konrat, 24120 | – | FJ173300 |
Chiloscyphus obvolutus (Hook. f. & Taylor) Hässel | Clasmatocolea obvoluta (Hook. f. & Taylor) Grolle | Chile, Hyvoenen, 2827 (GOET) | AM491293 | – |
Chiloscyphus pallescens (Hoffm.) Dumort. | Chiloscyphus pallescens (Hoffm.) Dumort. | Germany, Thuringia, Hentschel Bryo, 01418 (JE) | AM282821 | – |
Chiloscyphus pallescens (Hoffm.) Dumort. | Chiloscyphus pallescens (Hoffm.) Dumort. | Bulgaria, Hentschel Bryo, 0772 (JE) | AM282825 | – |
Chiloscyphus pallescens (Hoffm.) Dumort. | Chiloscyphus pallescens (Hoffm.) Dumort. | Poland, 1993 A. Stenel (W-4) | – | AY149871 |
Chiloscyphus perissodontus (Spruce) J.J.Engel & R.M.Schust. | Cryptolophocolea perissodonta (Spruce) L. Söderstr. | Guyana, Gradstein, 4890 (GOET) | AM282826 | – |
Chiloscyphus perissodontus (Spruce) J.J.Engel & R.M.Schust. | Cryptolophocolea perissodonta (Spruce) L. Söderstr. | Guyana, Gradstein, 5042 (GOET) | AM282827 | – |
Chiloscyphus platensis J.J. Engel & R.M. Schust. | Lophocolea platensis C. Massal. | Bolivia, Churchill et. al., 20950 (JE) | AM491295 | – |
Chiloscyphus platensis J.J. Engel & R.M. Schust. | Lophocolea platensis C. Massal. | Bolivia, Churchill et. al., 22090 (GOET) | AM491294 | – |
Chiloscyphus polyanthos (L.) Corda | Chiloscyphus polyanthos (L.) Corda | Slovakia, Hentschel Bryo, 0318 (JE) | AM282831 | – |
Chiloscyphus polyanthos (L.) Corda | Chiloscyphus polyanthos (L.) Corda | Finland, 2000 He-Nygren & Piippo, 1469 | – | AY149873 |
Chiloscyphus polychaetus (Spruce) J.J. Engel & R.M. Schust. | Heteroscyphus polychaetus (Spruce) Hentschel & Heinrichs | Ecuador, Gradstein & Mandl, 10139 (GOET) | AM491296 | – |
Chiloscyphus profundus (Nees) J.J.Engel & R.M.Schust. | Lophocolea profunda Nees | Germany, Hentschel Bryo, 01414 (JE) | AM282832 | – |
Chiloscyphus profundus (Nees) J.J.Engel & R.M.Schust. | Lophocolea profunda Nees | Finland, 2000 & Piippo, 1470 | – | AY149874 |
Chiloscyphus randii (S.W.Arnell) J.J.Engel & R.M.Schust. | Lophocolea randii S.W. Arnell | Prince Edward Isles, Gremmen, 98-63 (JE) | AM282833 | – |
Chiloscyphus sabuletorum (Hook.f. & Taylor) J.J.Engel & R.M.Schust. | Lophocolea sabuletorum (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Argentina, Hyvönen, 3233 (JE) | AM282834 | – |
Chiloscyphus sabuletorum (Hook.f. & Taylor) J.J.Engel & R.M.Schust. | Lophocolea sabuletorum (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Chile, Busch et al. Bryo, 01396 (JE) | AM282835 | – |
Chiloscyphus semiteres (Lehm.) Lehm. & Lindenb. | Lophocolea semiteres (Lehm.) Mitt. | Australia, Streimann, 58464 (GOET) | AM282836 | – |
Chiloscyphus semiteres (Lehm.) Lehm. & Lindenb. | Lophocolea semiteres (Lehm.) Mitt. | The Netherlands, Stieperaere, 8611 (JE) | AM282837 | – |
Chiloscyphus semiteres (Lehm.) Lehm. & Lindenb. | Lophocolea semiteres (Lehm.) Mitt. | New Zealand, Engel & von Konrat, 27982 | – | FJ173301 |
Chiloscyphus spinifer (Hook.f. & Taylor) J.J.Engel & R.M.Schust. | Cryptolophocolea spinifera (Hook. f. & Taylor) L. Söderstr. | New Zealand, Schäfer-Verwimp & Verwimp, 13808 (JE) | AM282838 | – |
Chiloscyphus spinifer (Hook.f. & Taylor) J.J.Engel & R.M.Schust. | Cryptolophocolea spinifera (Hook. f. & Taylor) L. Söderstr. | New Zealand, Engel & von Konrat, 28452 | – | FJ173302 |
Chiloscyphus trachyopus (Hook. f. & Taylor) Hässel | Clasmatocolea trachyopa (Hook. f. & Taylor) Grolle | Chile, Hyvoenen, 5933 (GOET) | AM491298 | – |
Chiloscyphus vermicularis (Lehm.) Hässel | Clasmatocolea vermicularis (Lehm.) Grolle | Ecuador, Sauer & Gradstein, MS-E065 (GOET) | AM491299 | – |
Clasmatocolea ctenophylla (Schiffn.) Grolle | Clasmatocolea ctenophylla (Schiffn.) Grolle | Chile, Engel, 25779 | – | FJ173304 |
Clasmatocolea humilis (Hook.f. & Taylor) Grolle | Clasmatocolea humilis (Hook.f. & Taylor) Grolle | Chile, Engel, 25274 | – | FJ173305 |
Clasmatocolea obvoluta (Hook.f. & Taylor) Grolle | Clasmatocolea obvoluta (Hook.f. & Taylor) Grolle | Chile, Engel, 25696 | – | FJ173306 |
Cyanolophocolea echinella (Lindenb. & Gottsche) R.M. Schust. | Heteroscyphus echinellus (Lindenb. & Gottsche) J.J. Engel & X.L. He | New Zealand, Lewington, 1140 (H) | – | FJ919297 |
Cyanolophocolea echinella (Lindenb. & Gottsche) R.M. Schust. | Heteroscyphus echinellus (Lindenb. & Gottsche) J.J. Engel & X.L. He | New Zealand, Engel, 27818 (F) | – | FJ919304 |
Herbertus dicranus (Taylor ex Gottsche, Lindenb. & Nees) Trevis. | Herbertus dicranus (Taylor ex Gottsche, Lindenb. & Nees) Trevis. | H3230549 (H) | KU523784 | KU523718 |
Heteroscyphus argutus (Reinw., Blume & Nees) Schiffn. | Heteroscyphus argutus (Reinw., Blume & Nees) Schiffn. | Nepal, D.G. Long, 30333 (JE) | – | AY149861 |
Heteroscyphus aselliformis (Reinw., Blume & Nees) Schiffn. | Heteroscyphus aselliformis (Reinw., Blume & Nees) Schiffn. | Indonesia, Gradstein, 10240 (GOET) | AM180588 | – |
Heteroscyphus biciliatus (Hook. f. & Taylor) J.J. Engel | Heteroscyphus biciliatus (Hook. f. & Taylor) J.J. Engel | New Zealand, Frahm, 20-6 (GOET) | AM491300 | – |
Heteroscyphus coalitus J.J. Engel | Heteroscyphus coalitus J.J. Engel | Nepal, D.G. Long, 17402 (JE) | AM282839 | – |
Heteroscyphus coalitus J.J. Engel | Heteroscyphus coalitus J.J. Engel | Nepal, D.G. Long, 30316 (JE) | – | AY149865 |
Heteroscyphus cuneistipulus (Steph.) Schiffn. | Heteroscyphus cuneistipulus (Steph.) Schiffn. | New Zealand, Frahm, 9-15 (GOET) | AM282840 | – |
Heteroscyphus fissistipus (Hook.f. & Taylor) Schiffn. | Heteroscyphus fissistipus (Hook.f. & Taylor) Schiffn. | Ireland, D.G. Long, H4064 (JE) | AM282841 | – |
Heteroscyphus inflatus (Steph.) S.C. Srivast. & A. Srivast. | Heteroscyphus inflatus (Steph.) S.C. Srivast. & A. Srivast. | Nepal, D.G. Long, 30457 (JE) | – | AY149875 |
Heteroscyphus planus (Mitt.) Schiffn. | Heteroscyphus planus (Mitt.) Schiffn. | Japan, 1992 Mizutani, 15828 | – | AY149872 |
Heteroscyphus splendens (Lehm. & Lindenb.) Grolle | Heteroscyphus splendens (Lehm. & Lindenb.) Grolle | Papua New Guinea, 1989 Hoffmann, 89-749 | – | AY149876 |
Heteroscyphus zollingeri (Gottsche) Schiffn. | Heteroscyphus zollingeri (Gottsche) Schiffn. | China, 1998 Koponen et al. 57927 | – | AY149879 |
Hygrolembidium acrocladum (Berggr.) R.M. Schust. | Hygrolembidium acrocladum (Berggr.) R.M. Schust. | Australia, Curnow, 5587 | – | AY463560 |
Leiomitra lanata (Hook.) R.M. Schust. | Leiomitra lanata (Hook.) R.M. Schust. | New Zealand, Glenny s.n., 2001 | – | AY463565 |
Lepicolea attenuata (Mitt.) Steph. | Lepicolea attenuata (Mitt.) Steph. | New Zealand, South Island, Stotler & Crandall-Stotler, 4586 (ABSH) | – | JF316578, AY507540 |
Lepicolea ochroleuca (Spreng.) Spruce | Lepicolea ochroleuca (Spreng.) Spruce | Chile, Hyvonen, 2938 | – | AY463566 |
Lepicolea scolopendra (Hook.) Dumort. ex Trevis. | Lepicolea scolopendra (Hook.) Dumort. ex Trevis. | Australia, Streimann, 55445 | – | AY463568 |
Leptophyllopsis laxa (Mitt.) Hamlin | Leptophyllopsis laxa (Mitt.) Hamlin | Australia, Streimann, 43810 (JE) | AM491301 | – |
Leptoscyphus amphibolius (Nees) Grolle | Leptoscyphus amphibolius (Nees) Grolle | Brazil, Schafer-Verwimp, Schafer 14748 | – | EU350474 |
Leptoscyphus gibbosus (Taylor) Mitt. | Leptoscyphus gibbosus (Taylor) Mitt. | Dominican Republic, Schafer-Verwimp (herb. Schafer), 17647 | – | DQ176702 |
Leptoscyphus gibbosus (Taylor) Mitt. | Leptoscyphus gibbosus (Taylor) Mitt. | Costa Rica, Herbarium Schafer-Verwimp, SV/H-0364 | – | EU350480 |
Leptoscyphus porphyrius (Nees) Grolle | Leptoscyphus porphyrius (Nees) Grolle | Ecuador, Schafer-Verwimp (herb. Schafer), 23229/a | – | DQ176707 |
Leptoscyphus porphyrius (Nees) Grolle | Leptoscyphus porphyrius (Nees) Grolle | Ecuador, Herbarium Schafer-Verwimp, Schafer 24214/a | – | EU350481 |
Lophocolea bidentata (L.) Dumort. | Lophocolea bidentata (L.) Dumort. | Poland: Silesian upland, K. Jedrzejko & A. Stenel (W-58) | – | AY149862 |
Lophocolea cuspidata (Nees) Limpr. | Lophocolea bidentata (L.) Dumort. | China, Hunan Province, Sang-Zhi Co., Koponen et al. 48430 | – | AY149866 |
Lophocolea heterophylla (Schrad.) Dumort. | Lophocolea heterophylla (Schrad.) Dumort. | USA, Indiana, ML Sargent’s culture collection, #481 | – | AF231899 |
Lophocolea martiana Nees | Cryptolophocolea martiana (Nees) L. Söderstr., Crand.-Stotl. & Stotler | French Guiana, Kourou, Gradstein, 6265 | – | AY149870 |
*Lophocolea sikkimensis (Steph.) Herzog & Grolle | *Cryptolophocolea sikkimensis (Steph.) Bakalin et Maltseva | Vietnam, Lao Cai Province, V.A. Bakalin & K.G. Klimova, V-12-17-17 (VBGI) | OK523503 | OK562106 |
**Lophocolea sikkimensis (Steph.) Herzog & Grolle | **Cryptolophocolea sikkimensis (Steph.) Bakalin et Maltseva | Vietnam, Lao Cai Province, V.A. Bakalin, V-3-86-16 (VBGI) | OK523504 | – |
Mastigophora woodsii (Hook.) Nees | Mastigophora woodsii (Hook.) Nees | China, D.Long, 33696 (E) | – | JF316581 |
Mastigophora woodsii (Hook.) Nees | Mastigophora woodsii (Hook.) Nees | Australia, Frahm, CANB639918, 2000 | – | AY463574 |
Pachyglossa tenacifolia (Hook. f. & Taylor) Herzog & Grolle | Pachyglossa tenacifolia (Hook. f. & Taylor) Herzog & Grolle | New Zealand, Bartlett 196 (JE) | AM491297 | – |
Pedinophyllum interruptum (Nees) Kaal. | Pedinophyllum interruptum (Nees) Kaal. | Germany, Schaefer-Verwimp, 35485 (M) | KT992498 | – |
Pedinophyllum interruptum (Nees) Kaal. | Pedinophyllum interruptum (Nees) Kaal. | Russia, N.A. Konstantinova & A.N. Savchenko, k508/7-07 (F) | – | KJ802073 |
Plagiochila alternans Lindenb. & Gottsche | Plagiochila alternans Lindenb. & Gottsche | Bolivia, Heinrichs et al. GP 16 (GOET) | AY550130 | – |
Plagiochila asplenioides (L.) Dumort. | Plagiochila asplenioides (L.) Dumort. | Finland, Nuuksio National Park, He-Nygren and Piippo 1467 | – | AY149858 |
Plagiochila fruticella (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Plagiochila fruticella (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | New Zealand, Engel & von Konrat, 23943 (GOET) | AM180613 | – |
Plagiochila pleurata (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Plagiochila pleurata (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | New Zealand, Schaefer-Verwimp & Verwimp, 13777 (GOET) | AM180615 | – |
Plagiochila porelloides (Torr. ex Nees) Lindenb. | Plagiochila porelloides (Torr. ex Nees) Lindenb. | Germany, Schaefer-Verwimp, 31077 (M) | KX896587 | – |
Plagiochila porelloides (Torr. ex Nees) Lindenb. | Plagiochila porelloides (Torr. ex Nees) Lindenb. | USA, Alaska, B. Shaw, F955/1 (DUKE) | – | KF943056 |
Plagiochila sichotensis Bakalin & Vilnet | Plagiochila sichotensis Bakalin & Vilnet | Russia, Russian Far East, Primorsky Territory, V.A. Bakalin & G.A. Arutinov, Arutinov 1-25-13 (VBGI) | MF947695 | MF947697 |
Plagiochila xerophila Bakalin & Vilnet | Plagiochila xerophila Bakalin & Vilnet | China, Sichuan Province, V.A. Bakalin & K.G. Klimova, China-46-2-17 (VBGI) | – | MK123266 |
Tetracymbaliella cymbalifera (Hook. f. & Taylor) Grolle | Tetracymbaliella cymbalifera (Hook. f. & Taylor) Grolle | New Zealand, M.A.M. Renner, 6139 (NSW) | KT992470 | – |
Tetracymbaliella cymbalifera (Hook. f. & Taylor) Grolle | Tetracymbaliella cymbalifera (Hook. f. & Taylor) Grolle | New Zealand, Frahm 1-23 (MO-5131915) | – | DQ026625 |
Triandrophyllum subtrifidum (Hook. f. & Taylor) Fulford & Hatcher | Triandrophyllum subtrifidum (Hook. f. & Taylor) Fulford & Hatcher | Bolivia, Churchill et al. 22800 | AJ972455 | – |
Triandrophyllum subtrifidum (Hook. f. & Taylor) Fulford & Hatcher | Triandrophyllum subtrifidum (Hook. f. & Taylor) Fulford & Hatcher | Venezuela, Ricardi, 9730/T | – | JF316580 |
Trichocolea tomentella (Ehrh.) Dumort. | Trichocolea tomentella (Ehrh.) Dumort. | China, He-Nygren, 1137 | – | AY463590 |
Trichotemnoma corrugatum (Steph.) R.M. Schust. | Trichotemnoma corrugatum (Steph.) R.M. Schust. | New Zealand, Glenny 8426 | – | AY463591 |
Zoopsis argentea (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Zoopsis argentea (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Australia, Streimann, 51704 | – | AY463595 |
Zoopsis argentea (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | Zoopsis argentea (Hook. f. & Taylor) Gottsche, Lindenb. & Nees | New Zealand, J.J.Engel, 23962 | – | JF316577 |
DNA was extracted from dried liverwort tissues using the NucleoSpin Plant II Kit (Macherey-Nagel, Germany). Amplification of ITS1–2, trnL-F, and the trnG-intron was performed using an Encyclo Plus PCR kit (Evrogen, Moscow, Russia) with the primers listed in Table
Locus | Sequence (5’-3’) | Direction | Annealing temperature (°C) | Reference |
---|---|---|---|---|
ITS 1–2 nrDNA | CGTTGTGAGAAGTTCATTAAACC | forward | 64 |
|
ITS 1–2 nrDNA | GATATGCTTAAACTCAGCGG | reverse | 58 |
|
trnL-F cpDNA | CGAAATTGGTAGACGCTGCG | forward | 62 |
|
trnL-F cpDNA | ATTTGAACTGGTGACACGAG | reverse | 58 |
|
trnG-intron cpDNA | ACCCGCATCGTTAGCTTG | forward | 56 |
|
trnG-intron cpDNA | GCGGGTATAGTTTAGTGG | reverse | 54 |
|
The polymerase chain reaction was performed in a total volume of 20 µl, including 1 µl of template DNA, 0.4 µl of Encyclo polymerase, 5 µl of Encyclo buffer, 0.4 µl of dNTP-mixture (included in Encyclo Plus PCR Kit), 13.4 µl (for trnL-F and the trnG-intron)/12.4 µl (for ITS1–2) of double-distilled water (Evrogen, Moscow, Russia), 1 µl of dimethylsulfoxide/DMSO (for ITS1–2) and 0.4 µl of each primer (forward and reverse, at a concentration of 5 pmol/µl). Polymerase chain reactions were carried out using the following program: 180 s initial denaturation at 95 °C, followed by 30–40 cycles of 30 s denaturation at 94 °C, 20 (for trnL-F) – 30 s (for ITS1–2, trnG-intron) annealing at 56 °C (trnG-intron) or 58 °C (trnL-F and ITS1–2), and 30 s elongation at 72 °C. Final elongation was carried out in one 5-min step at 72 °C. Amplified fragments were visualized on 1% agarose TAE gels by EthBr staining and purified using the Cleanup Mini Kit (Evrogen, Moscow, Russia). The DNA was sequenced using the BigDye Terminator v. 3.1 Cycle Sequencing Kit (Applied Biosystems, USA) with further analysis of the reaction products following the standard protocol on an ABI Prism 3100-Avant Genetic Analyser (Applied Biosystems, USA) in the Genome Center (Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow).
The datasets were produced for the ITS1–2 and trnL-F loci. Both datasets were aligned using MAFFT (
Phylogenies were reconstructed under three criteria: maximum parsimony (MP) with Mega X (
MP analysis for both datasets included 1,000 bootstrap replicates, default settings for all other parameters, and treated gaps as partial deletions with a site coverage cut-off of 95%.
For the ML analysis, the best fitting evolutionary model of nucleotide substitutions according to the BIC value was TIM3+F+I+G4 for the ITS dataset and TVM+F+I+G4 for the trnL-F dataset as determined by IQ-tree. Consensus trees were constructed with 1000 bootstrap replicates.
Indels for both datasets were coded with FastGap ver. 1.2 (
The infrageneric and infraspecific variability of ITS1–2 and trnL-F were quantified as the average pairwise p-distances calculated in Mega X (
Five new sequences from Lophocolea sikkimensis specimens were deposited in GenBank: two for ITS1–2, one for trnL-F and two for trnG-intron cpDNA. ITS1–2 alignment of the 55 specimens consisted of 955 character sites, and the trnL-F alignment of 53 specimens consisted of 612 character sites. The parameters of the tested alignments are shown in Table
The characteristics of ITS1–2, and trnL-F nucleotide sequence alignments.
Locus | Total sites | Conservative sites | Variable sites | Parsimony-informative sites | |||
---|---|---|---|---|---|---|---|
base pairs | % | base pairs | % | base pairs | % | ||
ITS1–2 | 955 | 376 | 39.37 | 376 | 39.37 | 431 | 45.13 |
trnL-F | 612 | 325 | 53.11 | 325 | 53.11 | 208 | 33.99 |
The MP analysis for ITS1–2 yielded a single parsimonious tree with CI = 0.364388 and RI = 0.619946. The ML criterion recovered a bootstrap consensus tree with a log-likelihood = -10978.46. The arithmetic means of the log likelihoods in Bayesian analysis for each sampling run were -11026.6 and -11028.06.
The MP analysis for trnL-F yielded five equally parsimonious trees with CI = 0.480315 and RI = 0.697248. The ML criterion recovered a bootstrap consensus tree with a log-likelihood = -4951.09. The arithmetic means of log likelihoods in the Bayesian analysis for each sampling run were -4993.95 and -4989.08.
The trees constructed for each dataset by the three different methods appeared highly congruent. Fig.
Phylogram obtained in a Bayesian analysis for the genus Cryptolophocolea and related taxa based on ITS1–2 dataset. The values of Bayesian posterior probabilities and bootstrap support from the MP and ML analyses greater than 0.50 (50%) are indicated. Taxon names and GenBank accession numbers are provided. Newly studied specimens are marked in bold 1 family Lophocoleaceae 2 family Brevianthaceae 3 family Plagiochilaceae.
The topologies obtained here are quite similar to previously published phylogenies in the reinstatement of Lophocoleaceae (
Phylogram obtained in a Bayesian analysis for the genus Cryptolophocolea and related taxa based on trnL-F dataset. The values of Bayesian posterior probabilities and bootstrap support from the MP and ML analyses greater than 0.50 (50%) are indicated. Specimen names and GenBank accession numbers are provided. Newly studied specimens are marked in bold 1 family Lophocoleaceae 2 family Brevianthaceae 3 family Plagiochilaceae.
The intergroup average distance between Lophocolea sikkimensis and Cryptolophocolea (Table
Inter- and Infrageneric p-distances, ITS1–2 and trnL-F. The number of base differences per site from averaging over all sequence pairs within and between each group are shown. The upper triangle – data for ITS 1–2, the lower triangle – data for trnL-F; — – data are absent.
trnL-F, % | Taxon | Lophocolea sikkimensis+ Cryptolophocolea | Lophocolea sikkimensis | Cryptolophocolea | Lophocolea | Clasmatocolea | Chiloscyphus | Leptoscyphus | Heteroscyphus | Pachyglossa | ITS 1–2, % |
---|---|---|---|---|---|---|---|---|---|---|---|
0.061 | Lophocolea sikkimensis+Cryptolophocolea | – | – | 0.164 | 0.150 | 0.153 | – | 0.167 | 0.156 | – | |
0 | Lophocolea sikkimensis | – | 0.128 | 0.142 | 0.129 | 0.130 | – | 0.154 | 0.138 | 0.011 | |
0.070 | Cryptolophocolea | – | 0.058 | 0.168 | 0.155 | 0.158 | – | 0.170 | 0.161 | 0.135 | |
0.040 | Lophocolea | 0.092 | 0.085 | 0.094 | 0.088 | 0.096 | – | 0.163 | 0.100 | 0.074 | |
0.058 | Clasmatocolea | 0.082 | 0.069 | 0.088 | 0.068 | 0.074 | – | 0.146 | 0.075 | 0.063 | |
0.037 | Chiloscyphus | 0.081 | 0.069 | 0.087 | 0.062 | 0.056 | – | 0.147 | 0.079 | 0.013 | |
0.021 | Leptoscyphus | 0.086 | 0.081 | 0.087 | 0.094 | 0.086 | 0.090 | – | – | – | |
0.055 | Heteroscyphus | 0.088 | 0.080 | 0.091 | 0.091 | 0.085 | 0.083 | 0.074 | 0.151 | 0.135 | |
– | Pachyglossa | – | – | – | – | – | – | – | – | 0.044 |
Therefore, according to the estimated phylogenetic relationships and level of genetic differences, Lophocolea sikkimensis should be transferred to the genus Cryptolophocolea.
Due to the obvious position of the studied specimens in the Cryptolophocolea clade, we provide the corresponding new combination for Lophocolea sikkimensis:
Herpocladium sikkimense Steph., Sp. Hepat. (Stephani) 6: 349, 1922 (=Lophocolea sikkimensis (Steph.) Herzog & Grolle, Rev. Bryol. Lichénol. 27 (3/4): 164, 1958 [1959])
However, the morphology of Cryptolophocolea sikkimensis is highly variable, and along with well-developed plants with ovate leaves and entire underleaves, modifications with shortly bilobed to bidentate leaves and underleaves may be observed. Indeed,
Notably, the Vietnamese populations, located between the extreme flanks of the species range, sometimes exhibit an intermediate morphology: plants with bilobed leaves and emarginate underleaves are often found. However, these plants are usually smaller than the well-developed individuals and characterized by distanced leaves and underleaves; they generally provide an impression of weakly developed or “suppressed” shoots. This intermediate morphology corresponds to the observations by
Plants yellowish green, greenish and whitish yellowish to yellowish brownish, sometimes grading to grayish brownish in the herbarium, gentle, very fragile and glistening when dry, forming loose pure patches over other bryophytes or rarely intermixed with Riccardia sp., Herbertus dicranus, Plicanthus, Scapania ciliatospinosa, Mnioloma fuscum; creeping to loosely ascending (very rarely suberect in dense patches); normally developed shoots 1.1–2.0 mm wide (narrower, depauperate plants are commonly occurring) and 8–20(–30) µm long. Rhizoids regular, in erect or upraise spreading fascicles, originating from a small-celled initial zone of the underleaf adjacent to the stem in the axial part of the underleaf, fascicles 0.1–0.5 mm long. Stem rarely intercalary (lateral, from the middle part of the sinus) branched; cross section slightly transversely ellipsoidal, ca. 170–200 × 220–250 μm, external wall distinctly thickened, cell in 1(–2) marginal rows thick-walled, with large (sometimes loosely confluent) concave trigones, 17–27 μm in diameter, inner cells thin-walled, trigones moderate to large, concave, 23–27 μm in diameter. Leaves contiguous to distant in depauperate shoots, obliquely spreading, very obliquely to obliquely inserted (insertion line 20–45° with stem axis), barely decurrent dorsally, very ventral end of the insertion line subtransverse, dorsally leaves alternate to subopposite with a somewhat adjacent one to another dorsal bases, ventrally widely connate with underleaves; in general outline slightly convex to concave (never flat), with leaf apex commonly turned to the apical part of the shoot, when flattened in the slide widely ovate to obliquely ovate and widely ovate-triangular, widest very near to the base, apex acute to apiculate, rarely shortly bilobed with unequal to subequal lobes (bilobed apex mostly present in small shoots), normally developed leaves 900–1200 × 950–1100 μm. Underleaves loosely canaliculate, if looking from the ventral side, widely connate with leaves in both sides, transversely ellipsoidal, with apex entire, rarely emarginate to shortly bilobed (with sinus semicrescentic), insertion line arcuate (sinuate), 400–600 × 700–850 μm. Midleaf cells subisodiametric, 22–33 μm in diameter or shortly oblong, to 38 μm long, thin-walled, trigones large, mostly triangle to slightly concave or slightly convex, cuticle virtually smooth; cells along leaf margin subisodiametric (subquadrate), 21–25 μm in diameter to slightly elongate along the margin, to 25–27 μm long; oil bodies in the midleaf cells 2–5 per cell, finely granulate, irregularly oblong, ellipsoidal to shortly fusiform, 8–17 × 5–7(–8) μm, grayish (Figs
Cryptolophocolea sikkimensis (Steph.) Bakalin & Maltseva, comb. nov. A oil bodies in apical part of the leaf B mat C oil bodies in leaf margin cells D shoots, fragment, dorsal view E oil bodies in midleaf cells F shoot, fragment, ventral view. Scale bars: 100 µm (A, C, E); 5 mm (B); 2 mm (D, F). All from V-8-54-17 (VBGI).
Specimens examined (North Vietnam). Vietnam • Lao Cai Province, Sa Pa District, San Sa Ho Commune, Hoang Lien Range, Hoang Lien National Park, one of the ways to the Phan Xi Pan Peak; 22°18.8'N, 103°45.933'E; 2727 m a.s.l.; 3 Apr. 2018; V.A. Bakalin & K.G. Klimova leg.; thickets of Sinobambusa with many rocky outcrops and Rhododendron trees, partly shaded moist cliff, over Sphagnum mat; VBGI V-16-6-18 • same collection data as for preceding; 22°19.2'N, 103°46.183'E; 2610 m a.s.l.; 22 Apr. 2017; V.A. Bakalin & K.G. Klimova leg.; evergreen south subtropical mountain forest with bamboo thickets and many rocky outcrops, open moist cliff; VBGI V-12-17-17 • same collection data as for preceding; 22°18.45'N, 103°46.567'E; 2900 m a.s.l.; 20 Apr. 2017; V.A. Bakalin & K.G. Klimova leg.; Rhododendron dominated forest with bamboo thickets and many rocky outcrops, moist cliff in part shade; VBGI V-9-22-17; • same collection data as for preceding; 22°18.25'N, 103°46.5'E; 3050 m a.s.l.; 20 Apr. 2017; V.A. Bakalin & K.G. Klimova leg.; Rhododendron dominated forest with bamboo thickets and many rocky outcrops, moist open cliffs; VBGI V-8-14-17, V-8-29-17, V-8-32-17, V-8-52-17, V-8-53-17, V-8-54-17, V-8-62-17, V-8-73-17, V-8-13-17 • same collection data as for preceding; 22°18.183'N, 103°46.517'E; 3100 m a.s.l.; 17 Mar. 2016; V.A. Bakalin leg.; evergreen south subtropical mountain forest over the peak, partly shaded moist cliffs; VBGI V-3-86-16, V-3-91-16, V-3-61-16, V-3-81-16 • same collection data as for preceding; Lai Châu Province, Ta Leng Commune, Pu Ta Leng Mt. summit; 22°25.367'N, 103°36.233'E; 3050 m a.s.l.; 30 Mar. 2018; V.A. Bakalin & K.G. Klimova leg.; rhododendron trees with a dense bamboo understory, partly shaded moist decaying decorticated fallen tree trunk; VBGI V-11-45-18, V-11-16-18 • same collection data as for preceding; partly shaded mesic trunk of a living tree; VBGI V-11-36-18.
Cryptolophocolea sikkimensis has a pronounced Sino-Himalayan distribution. Its range stretches from Nepal to Taiwan and Borneo. Specifically, the species is found in China (Yunnan and Taiwan Provinces), North Borneo, Bhutan, Nepal, India (Sikkim, Darjeeling), North Thailand, and Vietnam (
As described in the following section, the distribution of Cryptolophocolea sikkimensis is quite unusual within the genus. The vast majority of taxa principally exhibit a different distribution pattern. The phylogenetic tree shows that Cryptolophocolea sikkimensis forms a sister branch to all other taxa widespread in Southeast Asia (widely irrigated to Melanesia) and one pantropical species (Cryptolophocolea connata). This somewhat correlates with the distinctly different distribution and unique morphology of C. sikkimensis. The available data are insufficient for determining the morphological evolution pathways and distribution history within the genus. However, C. sikkimensis is assumed to belong to an isolated and morphologically specialized branch. The taxon probably had a wide range in the past that is now disjunctively distributed; in fact, the species is ‘locked’ in the mountainous regions from the Sino-Himalaya to Borneo, considering its ecological preferences.
Within Vietnam, the distribution of the species is limited to the peak surroundings of Phan Xi Pan Mt., a refugium containing a number of Sino-Himalayan species (
Cryptolophocolea includes 32 species (including the newly transferred C. sikkimensis), of which the species status is questionable for eight (one star in the World Liverwort Checklist,
Australasia and New Zealand contain 10 species, including four restricted to New Zealand and adjacent islands (Cryptolophocolea aculeata (Mitt.) L. Söderstr., C. helmsiana (Steph.) L. Söderstr., C. spinifera (Hook.f. & Taylor) L. Söderstr., C. tuberculata (J.J. Engel) L. Söderstr.), one taxon restricted to Tasmania (C. connatifolia (J.J. Engel) L. Söderstr.), three restricted to Southeast Australia and New Zealand (C. trialata (Gottsche) L. Söderstr., C. subopposita (J.J. Engel) L. Söderstr., C. pallida (Mitt.) L. Söderstr.), and two restricted to Tasmania, New Zealand, Antipodean Islands and some other small adjacent islands (C. leucophylla (Hook.f. & Taylor) L. Söderstr., C. mitteniana (Colenso) L. Söderstr.).
Cryptolophocolea chiloscyphoidea (Lindenb.) L. Söderstr. & Crand.-Stotl. is broadly distributed in Australasia, South America, and the subantarctic islands (and also recorded in India, but that record may be doubted for phytogeographic reasons). South America contains four taxa (in addition to the one mentioned above): C. fleischeri (Steph.) L. Söderstr. (also in Mexico), C. proteus (Herzog) L. Söderstr., C. pycnophylla (Spruce) L. Söderstr., C. tricorata (Hässel) Crand.-Stotl. & Stotler.
Cryptolophocolea connata (Sw.) L. Söderstr. & Váňa is broadly distributed from Africa to Malesia, Australasia, the Neotropics and Polynesia (Hawaii). Africa and South America contain two species that extend beyond this large region: C. martiana (Nees) L. Söderstr. (also in the southern part of the U.S.A.) and C. pallidovirens (Hook.f. & Taylor) L. Söderstr. (also circumsubantarctic by subantarctic island). Africa has a restricted distribution of C. lilliena (Steph.) L. Söderstr. (Kenya only) and C. regularis (Steph.) L. Söderstr. (Madagascar, Réunion, and Mauritius). South Asia contains C. fleischeri (Steph.) L. Söderstr. (Sri Lanka only). C. compacta (Mitt.) L. Söderstr. is strictly found in temperate East Asia (East China, Korea, Japan, also a questionable record from Thailand).
The large region stretching from Southeast Asia (Indochina) to Melanesia contains eight species, with three species distributed across this large area: C. ciliolata (Nees) L. Söderstr., Crand.-Stotl., Stotler & Váňa (also in southeast China (Hainan, Taiwan), Sri Lanka in south Asia and Hawaii in Polynesia), C. costata (Nees) L. Söderstr. (also in Taiwan) and C. edentata (J.J. Engel) L. Söderstr. (also in Taiwan). Melanesia has a restricted distribution of C. explanata (Mitt.) Váňa & Crand.-Stotl. (New Caledonia and Samoa). Malesia and Melanesia have a restricted distribution of C. levieri (Schiffn.) L. Söderstr. Malesia additionally contains three species: C. massalongoana (Schiffn.) L. Söderstr., C. stephanii (Schiffn.) L. Söderstr. (Java only), C. thermarum (Schiffn.) L. Söderstr. (Java only). Finally, Polynesia contains C. whittieriana (Inoue & H.A.Mill.) L. Söderstr. (Caroline Island only).
Therefore, the highest taxonomic diversity is found in New Zealand and adjacent islands (and, to some extent, Tasmania); a less prominent taxonomic ‘peak’ can be found in the southern part of South America, and the third-most taxonomically diverse area is Malesia to Melanesia. The Indochina Peninsula (north Thailand only) contains four species. Northwards of Indochina, the distinctly East Asian Cryptolophocolea compacta and predominantly Paleotropical C. ciliolata (reaching Yunnan Province in China) are found. None of the species listed are referred to as Sino-Himalayan floral elements. Therefore, C. sikkimensis is the first known species whose area core is distinctly Sino-Himalayan.
Cryptolophocolea sikkimensis possesses generally narrowly pointed ovate leaves that are unique in the genus. Its phylogenetic affinity could not be clearly identified without molecular genetic investigations. In the present study, such bright and easily noted leaf features were the only possible variants of morphological pathways that occurred in the genus, whereas the underleaves (widely connate with the leaves), and biseriate antheridium stalk show much stronger taxonomic value. The species’ atavistic traits are generally typically evidenced by depauperate plants with bidentate leaves and underleaves. The unique morphology of C. sikkimensis is associated with its unique distribution – the species has the only predominantly Sino-Himalayan distribution in the genus.
The line art pictures were prepared by Mr. Matvei Bakalin, to whom the authors are sincerely grateful. The research was partially performed using the large-scale research facilities “Herbarium of the Polar-Alpine Botanical Garden-Institute (KPABG)”, reg. No. 499397. The work of VB, YM, and KK was partially supported by the Russian Foundation for Basic Research (grant no. 20-04-00278) and is within the frame of the institutional research project “Cryptogamic Biota of Pacific Asia” (no. 1021043000529-9). The work was partially supported by the Vietnam Academy of Science and Technology (grant no. KHCBTD.02/21-23) for VB, KK and NVS. Also, the work of SC was partially supported by a grant from the National Ecosystem Survey of the National Institute of Ecology (NIE-A-2022-01).