Caribbean Journal of Science, Vol. 32, No. 1, 105-110, 1996 Copyright 1996 College of Arts and Sciences University of Puerto Rico, Mayagüez New Records of Water-borne Hyphomycetes for Puerto Rico C A R L O S J . SA N T O S- FL O R E S, C A R L O S B E T A N C O U R T , AND A NGEL M . NI E V E S- RI V E R A Department of Biology, University of Puerto Rico, Mayagüez, Puerto Rico 00680 A BSTRACT . – Conidia of twelve hyphomycete species found in stream foam and leaf litter, and comprising new records for Puerto Rico, are described and illustrated. The genera Chaetendophragmia, Crucispora, Dendrosporium, Dicranidion, Scutisporus, Subulispora and Trinacrium are recorded for the first time in the Island. Conidia of two undescribed species are presented. R ESUMEN . y hojarasca dophragmia, por primers – Se describen e ilustran los conidios de doce especies hifomicetales encontradas en espuma de riachuelos, y que representan nuevos registros para Puerto Rico. Los generos ChaetenCrucispora, Dendrosporium, Dicranidion, Scutisporus, Subulispora y Trinacrium se informan vez en la Isla. Se presentan los conidios de dos especies no descritas. I NTRODUCTION Water-borne hyphomycetes have been studied intensively in Puerto Rico (Betancourt and Justiniano, 1989; Betancourt et al., 1987; Justiniano and Betancourt, 1989; Santos-Flores and Betancourt-Lopez, 1994; Santos-Flores et al., 1995). Many of these aquatic and terrestrial fungi depend on flowing water for dispersal and their conidia are found in streams (Ingold, 1975; Nawawi, 1985). We report herein twelve new records for the water-borne hyphomycete flora of the Island. 1973). Pieces of leaves were stained with cotton blue and mounted on slides. Filters and leaf pieces were examined for loose conidia and conidiophores. R ESULTS AND D ISCUSSION Twelve hyphomycete species representing new records for Puerto Rico were found. The genera Chaetendophragmia, Crucispora, Dendrosporium, Dicranidion, Scutisporus, Subulispora and Trinacrium are recorded for the first time in the Island. Conidia of two undescribed species were also found. M ATERIALS AND M ETHODS Chaetendophragmia triangularis Matsushita 1971 Foam and leaves, collected from several (Figs. 2,3) streams in Puerto Rico (Fig. 1), were placed in sterile polythene bags and transported Conidia conical, pale brown, 60-75 × 6to the laboratory in an ice cooler. Foam was 7 pm. With an apical, 30-35 µm long, filipreserved with clear lactophenol and slides form appendage and the basal cell trunwere prepared to observe loose conidia. cate. With 2-3, 35-55 µm long appendages Each leaf was rinsed in distilled water near middle of axis. and incubated in sterile plastic chambers Conidia common in foam from Rio Anasfor 10-14 days at 21°C. After incubation, co, at Anasco, and Quebrada de Oro, at most of the water in chambers had evap- Mayagüez. orated, leaving the remaining rich in coCrucispora sp. nidia of water-borne hyphomycetes. Co(Fig. 4) nidia were trapped by filtering the water through 8 Mm HA filters (Millipore, Inc.). Conidia hyaline, X-shaped; consisting of Filters were stained with cotton blue and two V-shaped, 40-50 × 2-3 µm, 4-8 septate mounted on slides (Iqbal and Webster, arms. 105 106 C. J. SANTOS-FLORES ET AL. Caribbean Sea F I G. 1. Location of the sampling sites. 1 = Rio Anasco (this is the correct position for the location given in Santos-Flores et al. 1995, Fig. 1, location 1), 2 = Quebrada de Oro (this is the correct position for the location given in Santos-Flores et al, 1995, Fig. 1, location 3), 3 = Salto Collazo stream, 4 = Dona Juana stream 5 = Rio Valenciano, 6 = Rio Icacos, 7 = Rio Blanco de Naguabo. Isolated from submerged leaves of Roystonea borinquena Cook in Quebrada Dona Juana, at Villalba. Conidia were found in foam from Quebrada de Ore, at Mayaguez. Dendrosporium lobatum Plakidas and Edgerton 1936 (Figs. 5,6) Conidia hyaline to pale green, triangular, 10-12 × -9 µm. With a basal scar and a central septum. Conidia found in foam from Rio Icacos and Rio Blanco de Naguabo, both at Naguabo. dira inermis (Wright) H.B.K. ex D.C. from Quebrada de Oro, at Mayaguez. Diplocladiella appendiculata Nawawi 1987 (Fig. 9) Conidia light brown, 8-celled, Y-shaped, 45-60 µm across (excluding the appendages). With two, 20-45 µm long appendages at the apex of each arm. Two conidia found in a foam sample from Rio Icacos, at Naguabo. This species was known only from Malaysia (Nawawi, 1987). Nawawia nitida Kuth., Liew and Nawawi 1992 (Fig. 10) Dicranidion fissile Ando and Tubaki 1984 Conidia hyaline or subhyaline, round(Figs. 7, 8) tetrahedral to pyramidal, 30-40 µm long, Conidia hyaline, Y-shaped, 7-12 µm long, 18-25 µm at widest point, 3.5-4 µm wide 2.5-4 µm wide at apex, 0.5-1.5 µm wide at at truncate base. With 3-4 thin, hyaline, base, 1-3 septate. With 2-4, lobular, 6-11 35-75 × 1-2 µm filiform appendages aris× 2-3 µm terminal projections. ing from the apical corners. Conidia common in foam samples from Isolated from submerged leaves of A n - WATER-BORNE HYPHOMYCETES 107 FIGS. 2-8. 2, 3. Chaetendophragmia triangularis, 4. Crucispora sp., 5, 6. Dendrosporium lobatum, 7, 8, Dicranidion fissile. (Scale bar= 35 µm for 2, 3; 20 µm for 4; 10 µm for 5-8). Rio Icacos, at Naguabo; Salto Collazo stream, at San Sebastian, and on submerged leaves of Mangifera indica L. in Quebrada de Ore, at Mayagüez. It is similar to N. filiformis (Nawawi) Marvanova, which is smaller (13–18 µm long and 14–18 µm at widest point). Nawawia nitida was known only from Malaysia (Kuthubutheen et al., 1992). Scutisporus brunneus Ando and Tubaki 1985 (Fig. 11) Conidia hyaline to light brown, 4-celled, 12-15 µm wide. Single filiform, 9-20 µm long appendages arise from each corner of the main body. Found in foam from all the surveyed streams. This is one of the most common 108 C. J. SANTOS-FLORES ET AL. FIGS. 9-13. 9. Diplocladiellu appendiculata, 10. Nawawia nitida, 11, Scutisporus brunneus, 12, 13. Speiropsis hy alospora, (Scale bar = 30 µm for 9; 45 µm for 10, 12, 13; 15 µm for 11). species in stream foam in Puerto Rico. It is never found on leaves or other substrates. Speiropsis hyalospora Subramanian and Lodha 1964 (Figs. 12, 13) Conidia consist of one to several branches (up to 65 µm long) of hyaline, 8-11 × 3.5-5 µm cells. Cells joined together by 0.51 µm wide isthmi. Basal cell usually longer (up to 14 µm) than cells in branches. Found on submerged leaves of Syzygium jumbos L. in Quebrada de Oro, at Mayaguez. Unidentified Speiropsis species were reported from Puerto Rico by Padgett (1976) and Betancourt and Justiniano (1989). WATER-BORNE HYPHOMYCETES 109 FIGS. 14-22. 14, 15. Subulispora procurvata, 16, 17. Tricellula inaequalis, 18. Trinacrium sp., 19, 20. Triscelophorus ponapensis, 21, 22. Unknown conidia. (Scale bar = 15 µm for 14, 15; 6 µm for 16, 17; 20 µm for 18, 21, 22; 10 µm for 19, 20). Subulispora procurvata Tubaki and Yokoyama 1971 (Figs. 14, 15) Conidia hyaline, subulate or conoid, truncate at base, 35-60 (including appendage) × 2.5–4 µm, 0-3 septate. With an apical, 8-20 µm long flexible appendage. Isolated from submerged leaves of Sy- zygium jambos L. in Salto Collazo stream, at San Sebastian. Tricellula inaequalis van Beverwijk 1954 (Figs. 16, 17) Conidia hyaline, tricellular. Basal cell 35 × 2-3 µm, oblong to triangular, slightly curved. Apical cell 3–5 × 2-2.5 µm, triangular, pointed and slightly curved. 110 C. J. SANTOS-FLORES ET AL. Isolated from foam and submerged leaves of Mangifera indica L. from Rio Icacos, at Naguabo. This species is similar to T. curvatis Haskins in conidial size and shape, but on leaves T. inaequalis has apical cell of conidium in line with basal cell, while T . curvatis does not (Haskins, 1958). Trinacrium sp. (Fig. 18) Conidia pale brown, triradiate. Appendages 25-35 × 2-3.5 µm, 3-5 septate. Found as loose germinating conidia in foam from Rio Icacos, at Naguabo, and Rio Valenciano, at Juncos. Found on submerged leaves of Syzygium jambos L. from Salto Collazo stream, at San Sebastian. Conidia resemble a form isolated in Thailand by Tubaki et al. (1983; Fig. 30). Triscelophorus ponapensis Matsushita 1981 (Figs. 19, 20) Conidial axis hyaline or subhyaline, subulate, 2–4 septate, 15–30 × 4-6 µm. With 0-3, 8-15 × 3.5-5 µm, 2-4 septate appendages. Found on submerged leaves of Syzygium jambos from Salto Collazo stream, San Sebastian. Unknown 1 (Fig. 21) Conidia hyaline, triradiate, unicellular, 10-12 µm across, with a large central vacuole. With 3, lobulate, 3–6 µm long projections. Found in foam from Rio Icacos and Rio Blanco de Naguabo, both at Naguabo. A very similar conidium was illustrated from Malaysia by Nawawi (1985; Fig. 71; P1. 9,E). Similar conidia but with 3 single, apical projections were found in South Africa (Webster et al., 1994; Fig. 2K, unknown 4). Unknown 2 (Fig. 22) Conidia hyaline, tetradiate, unicellular, 11-14 µm across, with large central vacu- ole. With 4, rounded 3–6 µm long projections. Found in foam from Rio Icacos, at Naguabo. LITERATURE C ITED Betancourt, C., and J. Justiniano. 1989. Hongos ingoldianos presentes en el arroyo de Ore, Mayagüez, Puerto Rico. Int. J. Mycol. Lichenol. 4(1-2): 1-8. —, J. Cruz, and J. Garcia. 1987, Los hifomicetos acuaticos de la Quebrada Dona Juana en el Bosque Estatal de Toro Negro, Villalba, Puerto Rico. Carib. J. Sci. 23(2):278-284. Haskins, R. H. 1958. Hyphomycetous fungi: Volucrispora aurantiaca n. gen., n. sp.; V. ornithomorpha (Trotter) n. comb.; Tricellula curvatis n. sp., with the genus Tricellula emended. Can. J. Microbiol. 4:273285. Ingold, C. T. 1975. An illustrated guide to aquatic and waterborne Hyphomycetes (Fungi Imperfect) with notes on their biology. Freshwater Biol. Assoc. Sci. Publ. 30:1-96. Iqbal, S. H., and J. Webster. 1973. Aquatic hyphomycete spora of the River Exe and its tributaries. Trans. Br. Mycol. Soc. 61:331-346. Justiniano, J., and C. Betancourt. 1989. Hongos ingoldianos presentes en el Rio Maricao, Puerto Rico, Carib. J. Sci. 25(3-4):111-114. Kuthubutheen, A. J., G. M. Liew, and A. Nawawi. 1992. Nawawia nitida anam. sp. nov. (Hyphomycetes) and further records of Nawawia filiformis from Malaysia. Can. J. Bet. 70:96-100. Nawawi, A. 1985. Aquatic hyphomycetes and other water-borne fungi from Malaysia. Malay. Nat. J. 39:75-134. —. 1987. Diplocladiella appendiculata sp. nov. a new aero-aquatic hyphomycete. Mycotaxon 28(2): 297-302. Padgett, D. E. 1976. Leaf Decomposition by Fungi in a Tropical Rainforest Stream. Biotropica 8(3): 166-178. Santos-Flores, C., and C. Betancourt-Lopez. 1994. Aquatic Hyphomycetes (Deuteromycotina) from Rio Loco at Susua State Forest, Puerto Rico. Carib. J. Sci. 30(3-4):262-267. —, A. M. Nieves-Rivers, and C. Betancourt. 1995. Hyphomycetes with multiradiate conidia in stream foam from Puerto Rico. Carib. J. Sci. 31(1-2):4956. Tubaki, K., K. Watanabe, and L. Manoch. 1983. Aquatic hyphomycetes from Thailand, Trans. Mycol. Soc. Jpn. 24:451-457. Webster, J., L. Marvanova and A. Eicker. 1994. Spores from foam from South African rivers. Nova Hedwigia 57(3-4):379-398.