Onodrimia javanensis Jahodářová, Dvořák et Hašler, 2017

Jahodářová, Eva, Dvořák, Petr, Hašler, Petr & Poulíčková, Aloisie, 2017, Revealing hidden diversity among tropical cyanobacteria the new genus Onodrimia (Synechococcales, Cyanobacteria) described using the polyphasic approach, Phytotaxa 326 (1), pp. 28-40 : 30-34

Onodrimia javanensis Jahodářová, Dvořák et Hašler , sp. nov. ( Figs. 1–3 )

Thallus usually macroscopic, in mats, occasionally creeping, bright green to grey-green. Filaments straight to bent, occasionally coiled or entangled together, frequently false branched. Sheath facultative, thin, colorless, distinct, roundly closed at the ends or opened after hormogonia release, exceeding trichome or with trichome protruding from sheath. Trichomes, bright green, grey-green to yellowish, slightly constricted at cross-walls, narrowed at the ends, immotile, sometimes in tight loops. Cells usually rectangular, isodiametric to longer then wide, 1.39–2.55 μm (average 1.9 μm) wide, 1.5–8.35 μm (average 3.6 μm) long, with visible parietal chromatoplasm and inner pale centroplasm, with distinct storage granules. Apical cells usually longer than wide, rounded or conical, without calyptra, sometime pale with orange granules (probably dying cells). Reproduction into short hormogonia or hormocytes by help of necridic cells. Both hormogonia and hormocytes frequently form groups or tree-like tufts and attach via sheath to other filaments.

Type: — INDONESIA. Java, near the Tamanjaya village, Ujung Kulon NP, 6° 47’0.81’’S’, 105°30’9.14’’E, 49 m a.s.l., submersed bark of tree branches which fell into the hot water spring in rainforest, coll. L. Majeský, 25 February 2012 (holotype: OLM! Botany 24: Lichens and others No. 9224 dried sample is deposited in Regional Museum in Olomouc, Czech Republic. Reference strain: UPOC E28/2016, deposited at the culture collection of Department of Botany, Palacký University Olomouc, Czech Republic).

Habitat: —Submersed bark of tree branches which fell into the hot water spring in rainforest.

Etomology: —The species name was established based on the first place of discovery of Onodrimia .

Phylogeny: —All 16S rRNA sequences of Onodrimia were 100% identical. Bayesian inference based on a partial sequence 16S rRNA phylogeny revealed a monophyly of Onodrimia lineage among other cyanobacterial genera – most related were Phormidesmis Turicchia, Ventura, Komárková & Komárek (2009: 179) , Stenomitos , Neosynechococcus Dvořák, Hindák, Hašler & Hindáková in Dvořák et al. (2014b: 26), Pantanalinema Vaz et al. (2015: 301) and some unspecified Leptolyngbyaceae species. Other compact and highly supported genera were Nodosilinea , Spirulina Turpin ex Gomont (1893: 249) , Symploca Kützing ex Gomont (1893: 104) , Oculatella , Coleofasciculus Siegesmund, Johansen & Friedl in Siegesmund et al. (2008: 1575), Wilmottia Strunecký, Elster et Komárek (2011: 62) ( Fig. 4 ). Onodrimia´s cluster was strongly supported by both posterior probability and bootstrap values. Uncultured bacterium TG-102 ( JQ 769612.1) forms a sister lineage to Onodrimia , but the support is not significant. In our phylogenetic tree, Leptolyngbya appeared in 10 separate lineages. Thus, we show that polyphyly of Leptolyngbya is larger than previously shown (see Fig 2 ).

The closest relative to Onodrimia based on a BLAST search (28 th of July 2015) was an uncultured bacterium TG-102 ( JQ769612.1 ; with sequence similarity 96.2%) (Table S 1), which was found in biological soil crust of a copper mine tailings wasteland .

Secondary structure of 16S-23S ITS: —All three strains (E27, E28, E30) possessed a single, identical operon containing genes for tRNAIle and tRNAAla (490 nt total). The D1-D1’ helix (105 nt) and Box-B (46 nt) was the same between all strains ( Fig. 5 ). The base of the stem always consisted of 5 bp clamp followed by a 3’ asymmetric loop of 9 nt. The middle part contains three loops. The first loop consisted of 6 nt, with the second largest loop (14 nt) and the third loop the smallest (5 nt). The terminal loop of the D1-D1´consisted of 10 nt. The Box-B helix was conserved in all strains. Many cyanobacteria have a V 2 helix between the two tRNA genes but all of our strains lack this structure, having only 9 nt between the tRNA genes. We compared D1-D1’ helix and Box-B of Onodrimia with secondary structures of Nodosilinea nodulosa (Z.Li & J.Brand) Perkerson & Casamatta in Perkerson et al. (2011: 1405) UTEX 2910 ( KF 307598), Oculatella subterranean Zammit, Billi & Albertano (2012: 352) , Leptolyngbya boryana (Gomont) Anagnostidis & Komárek (1988: 391) UTEX B 485 ( EF 429291), Leptolyngbya appalachiana Johansen & Olsen in Johansen et al. (2008: 24) GSM-SFF-MF60 ( EF 429286), Phormidesmis sp. WJT36- NPBG 20 ( KJ 939034), Phormidesmis sp. WJT67- NPBG 4A ( KJ 939043), Stenomitos rutilans Miscoe & Johansen in Miscoe et al. (2016: 85) HA7619-LM2 ( KF 417430) and Neosynechococcus sphagnicola CAUP A 1101 ( KJ 469130). We found only little resemblance in a shape of base composition except the fact that L. appalachiana also showed an exceptionally long D1-D1’ helix similar to Onodrimia ( Onodrimia 105 nt and L. appalachiana 115 nt), but they differed in overall shape. Moreover, L. appalachiana exhibited also an exceptionally long Box-B (74 nt), while Onodrimia had only 46 nt.

Figures

FIGURE 1. Morphological variability of Onodrimia javanensis sp. nov. (A–B) arrangement of trichomes in colony and old sheaths. (D, F, J) trichomes with necridic cells (narrow arrow). (C, E, H) trichomes with exceeding sheath (bold arrow). (I, G) appearance of apical cells. (B, J) formation of hormogonia (asterisk). Scale bar 10 μm.

FIGURE 2. Variability in reproduction of Onodrimia javanensis sp. nov. Arrow=pseudobranching, asterisk=attachment of sheath to trichome, cross=clusters of hormogonia and hormocytes, double cross=growing trichomes from hormogonia and hormocytes, circle=pale apical cell with orange granules (probably dying cell), plus=trichome protruding from sheath, double plus=empty sheaths suggesting branching of trichomes. Scale bar 10 μm.

FIGURE 3. Iconotype of Onodrimia javanensis. A–F filaments with close sheath. G–K filaments with open sheath. In the lower part are drawn hormogonia, hormocytes and tree-like tufted hormogonial production (asterisk).

FIGURE 4. A phylogenetic reconstruction of 146 taxa based on 16S rRNA using Bayesian inference. Studied strains Onodrimia javanensis and Leptolyngbya sensu stricto are in bold. Supports at the nodes (Bayesian inference/maximum likelihood/maximum parsimony) represent only posterior probabilities>90 and bootstrap values>50, symbol of asterisk on the nodes represent 100 posterior probabilities and bootstrap values. Lines on site of the tree represent separate lineages of Leptonygbya sensu stricto and other Leptolyngbyaceae genera including Onodrimia.

FIGURE 5. Estimated 16S-23S ITS secondary structures D1-D1´and Box-B helices of (A, J) Onodrimia javanensis E27, E28, E30, (B, K) Leptolyngbya appalachiana GSM-SFF-MF60 (EF429286), (C, L) Phormidesmis sp. WJT36-NPBG20 (KJ939034), (D, M) Phormidesmis sp. WJT67-NPBG4A (KJ939043), (E, N) Stenomitos rutilans HA7619-LM2 (KF417430), (F, O) Neosynechococcus sphagnicola sy1 (KJ469130), (G, P) Nodosilinea nodulosa UTEX 2910 (KF307598), (H, Q) Oculatella subterranea, (I, R) Leptolyngbya boryana UTEX B 485 (EF429291).