Penicillium vaccaeorum Quintanilla, Mycopathol. 80: 74. 1982.

Torres-Garcia, Daniel, Gene, Josepa & Garcia, Dania, 2022, New and interesting species of Penicillium (Eurotiomycetes, Aspergillaceae) in freshwater sediments from Spain, MycoKeys 86, pp. 103-145 : 103

Penicillium vaccaeorum Quintanilla, Mycopathol. 80: 74. 1982.

Figure 16

Penicillium lacussarmientei = Penicillium lacussarmientei Ramírez, Mycopathol. 96: 29. 1986.

Type.

Spain, Valladolid, San Miguel del Arroyo , from sandy soil under pine tree; J.A. Quintanilla (holotype CBS H-148.83, cultures ex-type CBS 148.83, DTO 9E2, CECT 2753) .

Subgeneric classification.

Subgenus Penicillium Aspergilloides , Penicillium section Citrina , series Penicillium Roseopurpurea

Description.

Mycelium superficial and immersed composed of septate, smooth-walled, hyaline hyphae of 1.5-2.5 μm wide. Conidiophores monoverticillate, rarely biverticillate and divaricate; stipes smooth-walled, vesiculate, 22.5-103 × 1.5-2.5 μm (vesicle up to 4.5 µm); metulae divergent 2-3, unequal in length, 7-37 × 1.5-3 µm; phialides 2-5 per stipe/metula, ampulliform, 6-8.5 × 2-2.5 μm; conidia smooth- or finely roughened, globose, 2-2.5 × 2-2.5 μm.

Culture characteristics (7 d at 25 °C).

Colonies on CYA, 20-22 mm diam., slightly raised, velvety, radially sulcate, dull red (8C3) at center to light yellow (4A5) and white (1A1) towards periphery, margins slightly undulate, sporulation sparse, conidial masses grayish green (28B3); reverse brownish orange (5C6); with reddish soluble pigment. On MEA, 24-27 mm diam., slightly elevated, velvety, light yellow (4A5) and pale orange (5A2) at periphery, margins low and entire, sporulation sparse, conidial masses grayish green (27C4); reverse golden yellow (5B7) and reddish-yellow (4A6) at periphery; soluble pigment absent. On YES, 30-32 mm diam., slightly raised at center, velvety, radially sulcate, pale yellow (3A3) to white (1A1) and brownish orange (5C3) towards periphery, margins slightly undulate, sporulation abundant, conidial masses grayish green (28B3); reverse brownish yellow (5C8); soluble pigment absent. On OA, 18-20 mm diam., flattened, velvety, dark green (28F4) to light gray (25D1) and white (1A1) towards periphery, margins low and entire, sporulation abundant, conidial masses dull green (25D3); reverse brown (6E4) and yellowish gray (4B2) at periphery; soluble pigment absent. On DG18, 12-13 mm, slightly raised at center, velvety, radially sulcate, white (1A1) and yellowish white (1A2) towards periphery, margins regular, sporulation sparse, conidial masses grayish green (27C3); reverse light yellow (4A5) and white (1A1) at periphery; soluble pigment absent. On CREA, 9-11 mm diam., flattened, floccose, yellowish green (29B7) and white (1A1) towards periphery, margins entire, sporulation sparse, conidial masses grayish green (28B3); reverse dark gray (1F1); soluble pigment and production of acid absent.

Colony diameter on CYA after 7d (mm).

On CYA: 5 °C no growth, 15 °C 13-14, 20 °C 16-17, 30 °C 18-20, 35 °C 6-11, 37 °C no growth, 40 °C no growth.

Specimens examined.

Spain, Catalonia, Fogars de Montclús, La Costa de l'Infern , from stream sediments, Oct 2018, D. Torres ( FMR 17531 ) ; Fogars de Montclús, La Costa de l'Infern , from stream sediments, Oct 2018, D. Torres ( FMR 17534 ) ; Aitona, Segre River , from sediments, Dec 2020, D. Torres & J. Gené ( FMR 18100 ) ; La Granja d’Escarp, Segre River , from sediments, Dec 2020, D. Torres & J. Gené ( FMR 18123 ) ; Balearic Islands, Mallorca, Serra de Tramontana , from stream sediments, Dec 2018, J. F. Cano ( FMR 17616 ) ; Basque Country, from stream sediments, Aug 2019, J. Gené ( FMR 17967 ) .

Distribution.

Argentina, Canada, Chile, Spain, The Netherlands and Turkey.

Notes.

Penicillium vaccaeorum and P. lacussarmientei , two species described from sandy soils in Spain and Chile ( Quintanilla 1982; Ramírez 1986), respectively, were considered synonyms of P. roseopurpureum by Frisvad et al. (1990), noting that both species were fast growing variants of P. roseopurpureum . Later on, based on that criterion and the lack of morphological differences, Houbraken et al. (2011) considered the two former species synonyms of P. sanguifluum despite some sequence variation where P. vaccaeorum and P. lacussarmientei clustered together in a clade sister to that of P. sanguifluum . Our phylogeny correlates with Houbraken et al. (2011) who found the same topology. Having the opportunity to examine specimens from both monophyletic sister clades (Fig. 8 ), we observed consistent phenotypic features to distinguish them. For instance, isolates of P. vaccaeorum had longer stipes (up to 103 μm; up to 120 μm in the protologue of the species) ( Quintanilla 1982), they were able to grow on CYA at 35 °C (6-11 mm diam. after 7 d), had good sporulation and faster growth on YES agar (30-32 mm diam. 7 d) and more restricted on DG18 (12-13 mm diam. 7 d). In contrast, isolates of the P. sanguifluum clade showed considerably shorter conidiophores (15-50 µm long), they were unable to grow above 30 °C, and the colonies on YES and DG18 showed sparsely or absent sporulation and attained 18-28 mm and 16-22 mm diam., respectively ( Houbraken et al. 2011). Hence, genetic and phenotypic differences support the reinstatement of P. vaccaeorum as an accepted species, with P. lacussarmientei considered synonym. This species together with P. sanguifluum and P. roseopurpureum are classified in series Roseopurpurea , which differs from almost all series of the section Citrina by species’ monoverticillate conidiophores. The only other series in the section with monoverticillate conidiophores is Gallaica , represented exclusively by P. gallaicum , which differs from the former series mainly by the production of sclerotia ( Houbraken et al. 2020).

According to the revised data, P. vaccaeorum occurs worldwide, and is commonly isolated from sandy soils of beaches and forests, and even associated with ants (Table 1 ).

Figures

Figure 8. Phylogenetic tree of Penicillium section Citrina based on ML analysis obtained by RAxML inferred from the combined tub 2, cmdA, and ITS loci. Branch lengths are proportional to phylogenetic distance. Bootstrap support values / Bayesian posterior probability scores above 70 % / 0.95 are indicated on the nodes. Bold branches indicate bs / pp values 100 / 1. The tree is rooted to P. cainii DAOM 239914 and P. jacksonii CCFC 239937. The name in green is the resurrected species P. vaccaeorum included in this study. T = Ex-type strain.

Figure 16. Morphological characters of Penicillium vaccaeorum (FMR 17967). A colonies from left to right (top row) CYA, MEA, YES, and OA; (bottom row) CYA reverse, MEA reverse, DG 18, and CREA B-E conidiophores on MEA F conidia. Scale Bars: 25 μm (B), 10 μm (C-F).