Diversity trapped in cages: Revision of Blumenavia Möller (Clathraceae, Basidiomycota) reveals three hidden species

Abstract

Basidiomata of Phallales have a diversified morphology with adhesive gleba that exudes an odor, usually unpleasant that attracts mainly insects, which disperse the basidiospores. The genus Blumenavia belongs to the family Clathraceae and, based on morphological features, only two species are currently recognized: B. rhacodes and B. angolensis. However, the morphological characters adopted in species delimitations within this genus are inconsistent, and molecular data are scarce. The present study aimed to review and identify informative characters that contribute to the delimitation of Blumenavia species. Exsiccates from America and Africa were analyzed morphologically, and molecularly, using ITS, LSU, ATP6, RPB2 and TEF-1α markers for Maximum Parsimony, Bayesian and Maximum likelihood analyses, and also for coalescent based species delimitations (BP&P), as well as for bPTP, PhyloMap, Topo-phylogenetic and Geophylogenetic reconstructions. According to our studies, seven species can be considered in the genus: B. rhacodes and B. angolensis are maintained, B. usambarensis and B. toribiotalpaensis are reassessed, and three new species are proposed, B. baturitensis Melanda, M.P. Martín & Baseia, sp. nov., B. crucis-hellenicae G. Coelho, Sulzbacher, Grebenc & Cortez, sp. nov., and B. heroica Melanda, Baseia & M.P. Martín, sp. nov. Blumenavia rhacodes is typified by selecting a lectotype and an epitype. Macromorphological characters considered informative to segregate and delimit the species through integrative taxonomy include length of the basidiomata, color, width and presence of grooves on each arm as well as the glebifer position and shape. These must be clearly observed while the basidiomata are still fresh. Since most materials are usually analyzed after dehydration and deposit in collections, field techniques and protocols to describe fugacious characters from fresh specimen are demanded, as well as the use of molecular analysis, in order to better assess recognition and delimitation of species in Blumenavia.

Fig 1. Bayesian tree from concatenated matrix (ITS1/5.8S/ITS2/LSU/ATP6/RPB2/TEF-1α).

Numbers above branches indicate maximum parsimony bootstrap (MPbs), posterior probability (PP) and maximum likelihood bootstrap (MLbs) values. Asterisks represents type species. To the right, basidiomata of each species. Scale bars = 10 mm.

Fig 2. Species delimitation for Blumenavia rhacodes, B. heroica sp. nov. and B. baturitensis sp. nov.

(A) Phylogenetic tree obtained by Bayesian analysis on concatenated five loci dataset using MrBayes. (B) bPTP PhyloMap [40] based on Bayesian analysis. (C) Topo-phylogenetic representation [41] of Bayesian analysis. Blumenavia rhacodes specimens: ICN 176968, ICN 177266; B. heroica sp. nov. specimens XAL S. Chacón 5257-A, (paratype), XAL E Gándara 1455 (holotype), XAL D. Jarvio 778 (paratype), B. baturitensis sp. nov. specimens: UFRN-Fungos 1943 (paratype), UFRN-Fungos 2868 (holotype).

Fig 3. Geophylogeny proposed for Blumenavia.

In nodes: pink Blumenavia toribiotalpaensis; orange B. heroica sp. nov.; blue B. baturitensis sp. nov.; red B. rhacodes; purple B. angolensis; brown B. crucis-hellenicae sp. nov.

Fig 4. Blumenavia angolensis.

(A) Fresh basidiomata from São Tomé, reproduced from Degreef et al. [8] Fig 2C, © Publications Scientifiques du Muséum national d’Histoire naturelle, Paris. (B) Fresh basidiomata from São Tomé, reproduced from Desjardin and Perry [11] Fig 6, © Desjardin and Perry. (C) Basionym Laternea angolensis, reproduced from Welwitsch and Currey [5] Tab. 17, Fig 7. (D) Illustration reproduced from Dring [4] Fig 15F, © Board of Trustees of the Royal Botanic Gardens, Kew. (A–D) bar = 10 mm.

Fig 5. Blumenavia baturitensis sp. nov.

(A) Fresh basidiomata, UFRN-Fungos 2868, holotype. (B–I) UFRN-Fungos 1943, paratype: (B) Fresh basidiomata; (C) Dry basidiomata; (D) Wrinkled tentacular projections of glebifer; (E) Glebifer projections forming a larger projection leaving a fusiform space between them; (F, G) Torn and twisted glebifers in arms; (H) Apical exoperidium hyphae in 5% KOH; (I) Apical exoperidium hyphae in 1% Congo red. (J, K) UFRN-Fungos 2868, holotype: (J) Rhizomorph hyphae in 1% Congo red; (K) Basidiospores in Melzer’s reagent. (A, B, D) Arrows indicating membranous wrinkled, tentacled glebifers. (A–C) bar = 10 mm; (D, E) bar = 1 mm; (F, G) bar = 0.5 mm; (H–K) bar = 5 μm.

Fig 6. Blumenavia crucis-hellenicae sp. nov.

(A, B) Fresh basidiomata, SMDB 18116, holotype. (C) Dry basidiomata, ICN 177268, paratype. (D, E) Glebifer as a cross-shaped pyramidal structure: (D) Fresh, SMDB 18116, holotype; (E) Dry stuck in one arm, ICN 177268, paratype. (F, G) Glebifers like teeth along the extension of the arms: (F) Fresh, SMDB 18116, holotype; (G) Dry, ICN 177268, paratype. (H–K) ICN 177268, paratype: (H) Basidiospores in Melzer’s reagent; (I) Rhizomorph hyphae in 1% Congo red; (J) Apical exoperidium hyphae in 1% Congo red; (K) Apical exoperidium hyphae in 5% KOH. (A) Arrows indicating the longitudinal outer and inner grooves. (A–E) bar = 10 mm; (F, G) bar = 5 mm; (H–K) bar = 5 μm.

Fig 7. Blumenavia heroica sp. nov.

(A–B) Dry basidiomata: (A) S. Chacón 5257-A, paratype; (B) XAL R. Medel 672, paratype. (C) Groove in the outer face of the arm, XAL D. Jarvio 370, paratype. (D) Glebifers forming by parallel lines curving inward, S. Chacón 5257-A, paratype. (E, F) XAL R. Medel 672, paratype: (E) Glebifers forming by parallel lines curving inward not sparse in the upper half of the basidiomata; (F) Spaced glebifers. (G, H) XAL E. Gándara 1455, holotype: (G) Apical exoperidium hyphae in 5% KOH; (H) Apical exoperidium hyphae in 1% Congo red. (I) Rhizomorph hyphae in 1% Congo red, XAL S. Chacón 4475, paratype. (J) Basidiospores in 5% KOH, XAL E. Gándara 1455, holotype. (A, D, E, F) Arrows indicating glebifers. (B, C) Arrows indicating longitudinal outer groove. (A, B) bar = 10 mm; (C, E, F) bar = 1 mm; (D) bar = 5 mm; (G–J) bar = 5 μm.

Fig 8. Blumenavia rhacodes photographs.

(A–C) Reproduced from Möller [3], Tab. III: (A) Fig 1A sintype; (B) Fig 2 sintype; (C) Fig 3, lent from B Herbarium B700016469, sintype. (D) Lent from B Herbarium, B700016470. (A–D) bar = 10 mm.

Fig 9. Blumenavia rhacodes.

(A) Peridium layers, ICN177267: Ex = exoperidium, M = mesoperidium, E = endoperidium. (B–C) ICN 177266, epitype: (B) Fresh basidiomata; (C) Dry basidiomata. (D) Basidiomata preserved in alcohol, HBG 024640. (E–G) B700016478, lectotype: (E) Dry basidiomata; (F) Groove in the arm. (G) Dry glebifer. (H–K) ICN 177266, epitype. (H) Dry glebifer; (I) Fresh glebifer; (J–K) ICN 177266, epitype: (J) Apical exoperidium hyphae in 5% KOH; (K) Apical exoperidium hyphae in 1% Congo red. (L) Rhizomorph hyphae in 1% Congo red, PACA 12552. (M) Basidiospores in 5% KOH, PACA 12550. (B, C, G, H, I) Arrows indicating glebifers. (F) Arrow indicating the longitudinal outer groove. (A) bar = 0.2 mm; (B, C, E) bar = 10 mm; (F–I) bar = 1mm; (J–M) bar = 5 μm. Photographs: (B, I) sent by Fazolino E. P.; (D) sent by Matthias Schultz.

Fig 10. Blumenavia toribiotalpaensis.

(A) Fresh basidiomata, black and white image reproduced Vargas-Rodriguez and Vázquez-García [7] Fig 3, © Vargas-Rodriguez and Vázquez-García. (B, C) Dry basidiomata, BPI 870955, holotype. (D) Dry basidiomata, IBUG 456, paratype. (E) Top view of fresh basidiomata show groove in the arm, black and white image reproduced from Vázquez-García [7] Fig 5, © Vargas-Rodriguez and Vázquez-García. (F) Lacerated projections of glebifer, BPI 870955, holotype. (G, H) Torn and twisted points of glebifer, BPI 870955, holotype. (I, J) IBUG 454, paratype: (I) Apical exoperidium hyphae in 5% KOH; (J) Apical exoperidium hyphae in 1% Congo red. (K) Rhizomorph hyphae in 1% Congo red, IBUG 240a, isotype. (L) Basidiospores in 5% KOH, IBUG 456, paratype. (A–B) bar = 10 mm; (F–H) bar = 1 mm; (I–L) = 5 μm.

Fig 11. Blumenavia usambarensis, B 700016468, holotype.

(A, B) Dry basidiomata. (C) Arm edge and glebifer. (D) Rhizomorph hyphae in 1% Congo red. (E) Basidiospores in 5% KOH. (A, B) bar = 10 mm; (C) bar = 1 mm; (D–E) = 5 μm.