Salispina intermedia A.L. Jesus, Pires-Zottar. & Marano, sp. nov.
Index Fungorum number: IF 551603, Facesoffungi number: FoF 01277, Fig. 3
Etymology — “intermedia” refers to the presence of intermediate morphological features between S. spinosa and S. lobata.
Holotypus — SP 466378
Mycelium well-developed on PYGs, aerial mycelium scanty, hyaline, irregular, branched, few septate, hyphae 2.5 – 10 μm thick. Zoosporangiophores undifferentiated from the vegetative hyphae, long, simple, bearing one terminal zoosporangium, 6.25 – 12.5 μm (av. 9.8 μm). Zoosporangia of variable morphology, ranging from obovate, obpyriform, globose to elongate, thick-walled, 33 – 197 × 25 – 183 μm (av. 86 × 62 μm); with vacuolated content; smooth to spiny; spines with variable degree of coverage on the zoosporangium, from only one spine at the tip of the zoosporangium to completely spiny; spines (5–)7.5 –35 μm long. (av. 17 μm); basal plug hyaline, 2.5 – 7.5 μm thick (av. 5.5μm). Zoospores discharged through a persistent tube, long or short, 15 – 30 × (7.5–)12.5 –15(–20) μm (av. 23 × 15μm); vesicle absent; encysted zoospores, 6.3 – 12.5 μm diam. (av. 9 μm), germination by one germ tube. Chlamydospores absent. Sexual reproduction unknown.
Culture characteristics — colonies petaloid on PYGs; no growth on Sorghum sp. (L.) seeds.
Material examined — BRAZIL, São Paulo, Cananéia, BParque Estadual da Ilha do Cardoso^ (PEIC), 25°03′05″– 25°18′18″S; 47°53′48″– 48°05′42″W, Perequê river (salinity 2.2 %), from leaves of Rhizophora mangle, 8 Nov 2012, A.L. Jesus, C.L.A. Pires-Zottarelli & A.V. Marano (SP 466378, holotype), ex-holotypes CCIBt 4155, MMBF 12/15; Ibid., Perequê river (salinity 0.8 – 2.8 %), from leaves of R. mangle and Laguncularia racemosa, 30 Aug and 8 Nov 2012, 27 Feb and 5 Jun 2013, A.L. Jesus, C.L.A. Pires-Zottarelli & A.V. Marano (SP 466373, SP 466376, SP 466377, SP466379, paratypes), ex-paratypes CCIBt 4115 =MMBF 07/15, CCIBt 4153 =MMBF 10/15, CCIBt 4154 =MMBF 11/15, CCIBt 4156=MMBF 13/15.
Notes — Salispina intermedia appears as morphologically and phylogenetically intermediate between S. spinosa CBS 591.85 (KT886057) and S. lobata CBS 588.85 (KT886056), Figs. 1, 2 and 3. Its zoosporangial morphology resembles S. spinosa, although their zoosporangia are considerably larger than those observed for the ex-type cultures of S. spinosa (CBS 591.85) and S. lobata (CBS 588.85). Onto PYGs, S. intermedia forms clusters of zoosporangia visible at naked eye. This species was particularly abundant and frequently recovered during spring (Nov) and summer (Feb) samplings, when water temperature was higher (25 – 28 °C) than in the other samplings (18 – 22 °C). We were not able of sequencing the ITS region of Salispina using the primers ITS4 and ITS6 (Cooke et al. 2000) and UN-up18S42 and UN-up28S22 (Robideau et al. 2011).

Fig. 1 Phylogram generated from Bayesian inference analysis (MrBayes 3.2, Ronquist et al. 2012) based on SSU rDNA sequences showing the phylogenetic placement of Salispina gen. nov. in a well-defined clade (indicated in bold). Maximum likelihood (ML) bootstrap support values (PhyML 3.1, Guindon and Gascuel 2003) < 50 % are marked with (-). Clades that do not appear in the ML analysis are indicated with a zero. Bayesian posterior probability values > 0.50 are labelled
numerically. Scale bar indicates the average number of substitutions per site. New taxa are in blue and ex-type strains are in bold.

Fig. 2 Phylogram generated from Bayesian inference analysis (MrBayes 3.2, Ronquist et al. 2012) based on cytochrome oxidase I (COI mtDNA) sequences showing the phylogenetic placement of Salispina gen. nov. in a well-defined clade (indicated in bold). Maximum likelihood (ML) bootstrap support values (PhyML 3.1, Guindon and Gascuel 2003) < 50 % are marked with (-). Clades that do not appear in the ML analysis are indicated with a zero. Bayesian posterior probability values > 0.60 are labelled numerically. Scale bar indicates the average number of substitutions per site. New taxa are in blue and extype strains in bold.

Fig. 3 Salispina intermedia (holotype) a – d Zoosporangia of different morphologies, from smooth (a) to with various degree of spines coverage (b – d) e – i Formation of a persistent tube through which zoospores swim away (no vesicle is formed) f Detail of the zoosporangial basal plug. Bars: a, h = 20 μm, b – g, i = 10 μm.