Philippine Journal of Science 149 (3): 589-602, September 2020 ISSN 0031 - 7683 Date Received: 28 Jan 2020

Microalgae of Pineapple [Ananas comosus (L.) Merr] Phytotelmata from Calauan, ()

Eldrin DLR. Arguelles*

Philippine National Collection of Microorganisms National Institute of Molecular Biology and Biotechnology (BIOTECH) University of the Philippine Los Baños, College, Laguna 4031 Philippines

Phytotelmata are a unique habitat for the existence of new and rare species of microalgae. In the Philippines, no comprehensive study was documented regarding the diversity of microalgae on bromeliad phytotelmata. This study provides the first floristic survey that presents the taxonomy and species composition of phytotelm microalgae associated with bromeliad tanks of pineapple [Ananas comosus (L.) Merr] from Calauan, Laguna. A total of 15 algal taxa were taxonomically identified from the collected samples, of which six species (five genera) are classified as members of Cyanophyceae, four species (three genera) belong to Zygnematophyceae, three species (three genera) to Chlorophyceae, and one species (one genus) each for Euglenophyceae and Klebsormidiophyceae. The study reports an additional of nine species (Phacus monilatus, Monoraphidium lunare, Coelastrum proboscideum, Klebsormidium flaccidum, Cosmarium granatum, Cosmarium contractum var. rotundatum, Phormidium breve, Arthrospira jenneri, and Chroococcus subnudus) as new records in the global inventory list of phytotelm microalgae and 13 new taxa as additional records of microalgae in phytotelm microhabitat of Asia. Also, the occurrence of a rare photosynthetic euglenoid [Phacus monilatus (Stokes) Lemmerman] and a green microalga, [Monoraphidium lunare Nygaard, J. Komárek, J. Kristiansen, & O.M. Skulberg] are described for the first time in the Philippines. Diagnostic descriptions and keys are presented to distinguish the different species of phytotelm algae. The results of this study provide important knowledge regarding the species composition of microalgae in bromeliad phytotelmata found in the Philippines.

Keywords: algae, bromeliaceae, bromeliad tanks, cyanobacteria, phytotelma, taxonomy

INTRODUCTION (such as phytotelmata) remained unexplored. The term phytotelmata is used to describe a volume of water The Philippines is a megadiverse tropic country situated impounded in cavities of plant parts and structures at 116° 40’ and 126° 34’ E longitude and 4° 40’ and 21° that are effective in sustaining micro- and macro-biota 10’ N latitude. It is composed of 7,107 islands with more associated with it (Kitching 2000; Ramos and Moura than 9,000 species of unique plants and animals from 2019). Phytotelmata comes in various forms and are different tropical rain forests in the country. Despite its classified into five major classes: water-filled tree holes, reported biodiversity richness, taxonomic identification plant axils (plants belonging to families Palmae and and characterization of microalgae from unique habitats Pandanaceae), modified leaves (carnivorous plants such *Corresponding Author: [email protected] as those found in Nepenthaceae and Sarraceniaceae),

589 Philippine Journal of Science Arguelles: Microalgae of Pineapple Phytotelmata Vol. 149 No. 3, September 2020 bamboo internodes, and bromeliad tanks (Ramos and that are considered less common are the synurophyceans Moura 2019). The latter includes the bromeliads, a (Ramos et al. 2018b), xanthophyceans (Sophia 1999; group of monocot flowering plants under the family Brouard et al. 2012), and some cryptophyceans (Duarte Bromeliaceae. The leaves disposition – base of the et al. 2013). Based on these studies, niche characteristics bromeliad plant that appears like an inverted cone with such as having a low water conductivity, low pH, high a spiral arrangement (forming a rosette configuration) water temperature (23.0–29.0 °C), high light intensity, of leaves – of these plants can act as a plant cavity that and organically rich waters favor the occurrence of allows accumulation of rainwater that serves as a special phytotelm microalgae in this habitat. In collation kind of habitat for larvae of insects, nematodes, and with other plant phytotelmata such as the Sarracenia several microorganisms, including the phytotelm algae purpurea (pitcher plant), the pattern of microalgae (Ramos et al. 2018a; Kitching 2000). coenoses shows the dominance of Bacillariophyceae (centric diatoms) and green microalgae (Gebühr et al. Bromeliads possess high ecological value and level of 2006). This observation is caused by differences in niche endemism and give suitable environmental conditions for characteristics of S. purpurea and other bromeliads, such the existence of diverse kinds of flora and fauna. Biota as having a high concentration of dissolved nutrients associated with bromeliad tanks interacts in a processing from detritus present in the phytotelmata of pitcher plant food chain where detritus serves as the ultimate source (Gebühr et al. 2006). of organic matter and energy (Marino et al. 2011). Nevertheless, current studies show that phytotelm algae Floristic studies on phytotelm microalgae in Asia are (autotrophic biomass in phytotelmata) can serve as an scarce. There has been only one report (in Thailand) alternative source of energy. These algae are consumed with few taxa occurring in phytotelmata of bromeliads by micro-metazoans such as crustaceans and other rotifers found in the Queen Sirikit Botanical Garden. This and then later consumed by other higher top-predators taxonomic survey reported a total of 118 taxa classified (e.g. damselfly larvae and chironomids) (Starzomski et to eight major microalgal groups: Chlorophyta (35 taxa), al. 2010). Moreover, the abundance and species richness Euglenophyceae (31 taxa), Cyanobacteria (18 taxa), of phytotelm algae present in bromeliad tanks are highly Bacillariophyceae (17 taxa), Zygnematophyceae (13 reliant on the dissolved nutrients (from detritus) present taxa), Cryptophyceae (two taxa), Synurophyceae (one in such an environment (Marino et al. 2011). Also, taxon), and Xanthophyceae (one taxon). Of these taxa, plant architecture (e.g. number and arrangement of eight species – Aphanothece elabens, Nephrochlamys leaves) of bromeliads can also affect the interaction of subsolitaria, Aphanothece confera, Oedogonium organisms present in bromeliad tanks. More complex leaf curvum, Kirchneriella obtusa, Gyropaigne ukrainica, architecture for bromeliads would mean greater interaction Mougeotia parvula, and Astasia comma – are reported between producers and consumers in the phytotelmata as new records for Thailand (Poniewozik et al. 2020). (Ramos et al. 2018b). In the Philippines, taxonomic and ecological studies of algal flora focus more on phytoplankton and periphyton Bromeliad phytotelmata occur in several continents associated to rocks and aquatic macrophytes in different of the world, even though it is more common in the bodies of water (Arguelles 2020; Arguelles 2019 a, b, c; Neotropical areas because of the high rainfall in the Arguelles and Martinez-Goss 2019; Martinez-Goss et al. region. With the exception of a study done in New 2019; Rañola et al. 1990), but no comprehensive study Zealand by Killick et al. (2014), all other microalgal was documented regarding the species composition of records for bromeliad phytotelmata are reported in the microalgae on phytotelmata. This survey is a pioneering tropical region like those in South America (Brazil) study of phytotelm microalgae in the Philippines and and Asia (Thailand). This is attributable to the endemic presents the first taxonomic survey of the microalgal prevalence of family Bromeliaceae in the neotropical community on pineapple bromeliad tanks. The goal of ecosystems (Ramos and Moura 2019). In general, only a this investigation is to give a quick survey of the algal few studies were documented on the diversity phytotelm species of phytotelmata (in bromeliad tanks of pineapple) microalgae in bromeliad tanks and its ecological in order to improve our understanding on the distribution, significance is still largely unexplored (Marino et al. diversity, and environmental conditions that contribute 2011). Recent taxonomic studies show that the main to the existence of these algae. This taxonomic survey microalgal groups reported from bromeliad phytotelmata of phytotelm algae will highlight morphology-based are chlorophytes (Ramos et al. 2018a), zygnematophytes taxonomic characterization and identification of each (Sophia 1999; Ramos et al. 2017b, 2018c), diatoms algal taxon observed in pineapple bromeliad tanks. (Lyra 1971), cyanobacteria (Ramos et al. 2018b, 2019), dinoflagellates (Ramos et al. 2016), and euglenophytes (Ramos et al. 2017a). Other reported microalgal groups

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MATERIALS AND METHODS pigmentation, appearance, and color of the mucilaginous sheath; trichomes and filaments characteristics; the existence of specialized cells (akinetes and heterocytes); Sampling of Phytotelm Algae attributes of the cell wall; and features of the chloroplasts The taxonomic survey was done in two planting areas and pyrenoids were noted for each algal taxa (Arguelles of pineapple in Calauan, Laguna: Javier Eco Farm at 2019a; Arguelles et al. 2014). The taxonomic classification Balayhangin (14° 7’ 47.28” N, 121° 18’ 59.04” system done by Desikachary (1959), Prescott (1962), E); and Gapit Fruit Farm in Barangay Imok (14° 6’ 47.16” Velasquez (1962), Martinez (1984), Wehr and Sheath N, 121° 17’ 56.04” E), located in a steep land area along (2003), and Whitton (2002) were used. Taxonomic Calauan – San Pablo highway (Figure 1). Samples were identification of each algal taxa observed was done obtained from water retained in cavities of leaves of the utilizing all accessible information. bromeliad plant (tanks) (Figure 2). The water samples were gathered using rubber tubing connected to a sterile 50-ml syringe. The procedure was repeated three times in each bromeliad tank to make sure that all the biological material present was collected. The samples were then RESULTS placed in a sterile conical specimen tube (Tarson) of 25 Bromeliad tanks are a distinct aquatic microhabitat that mm x 50 mm size (transported by means of a portable contains diverse species of microorganisms. Assemblages cooler) and immediately analyzed in the laboratory in of phytotelm algae in this habitat formed a consortium vivo. Microscopic observation was done using a binocular of microalgae consisted of unicellular and filamentous research microscope (Olympus CX31) provided with species of charophytes, chlorophytes, diatoms, and Infinity X digital camera (Arguelles et al. 2014). cyanobacteria as well as single-celled photosynthetic euglenophytes. A total of 15 algal taxa were taxonomically Morphotaxonomic Enumeration and Identification identified and described from the collected samples from Identification and classification based on morphotaxonomic bromeliad tanks of pineapple, of which six species (five as well as metric characteristics such as cell size as well genera) are classified as members of Cyanophyceae, four as the shape of cells (both specialized and vegetative species (three genera) belong to Zygnematophyceae, cells), features of the filaments, cross wall constrictions, three species (three genera) to Chlorophyceae, and one species (one genus) each to Euglenophyceae and

Figure 1. Location map of the sampling sites in Calauan, Laguna.

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Figure 2. Pineapple bromeliad tanks from sampling areas in Calauan, Laguna.

Klebsormidiophyceae. The predominant group of alga was Family: Selenastraceae Cyanophyceae (40%), followed by Zygnematophyceae Genus: Kirchneriella Schmidle (26.67%), Chlorophyceae (20%), and Euglenophyceae 1. Kirchneriella lunaris (Kirchner) Möbius (6.67%) and Klebsormidiophyceae (6.67%). Taxonomic Pl. I, Fig. 1 characterizations on the basis of the morphology of each Basionym: Rhapidium convolatum var. lunare Kirchner algal isolate – together with current taxonomic names Kirchneriella lunata Schmidle based on Algaebase (Guiry and Guiry 2020), as well as a dichotomous key – are presented in the study. Colonial cells are in clusters of 2-4-8 up to 64 crescent- shaped cells sometimes solitary; cells are cylindrical or sickle-shaped, 2.5–4.0 x 1.5-7.0 µm (length x width); Phylum: CHLOROPHYTA enclosed in a mucilaginous sheath; chloroplasts are Class: Chlorophyceae parietal (with 1–4 pyrenoids per cell) close to the cell wall. Order: Sphaeropleales

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Dichotomous Key

1. Cells without membrane-bound organelles...... 2 1. Cells with membrane-bound organelles...... 7

2. Unicellular and spherical occurring as singly or in clusters of 2–4 cells...... 3 2. Filamentous-type, existing as solitary or in groups of trichomes...... 4

3. Cells are 3.0–5.0 μm in diameter...... Chroococcus minutus 3. Cells are 25.0–32.0 μm in diameter...... Chroococcus subnudus

4. Trichomes are cylindrical and spirally coiled...... Arthrospira jenneri 4. Trichomes are straight, solitary, and scattered...... 5

5. Trichomes with observable constrictions at the cross walls...... Pseudanabaena catenata 5. Trichomes without observable constrictions at the cross walls...... 6

6. Cells with granulated protoplasm...... Oscillatoria subbrevis 6. Cells without granulated protoplasm...... Phormidium breve

7. Cells with red eyespot and proteinacious pellicle...... Phacus monilatus 7. Cells without red eyespot and proteinacious pellicle...... 8

8. Filamentous, prone to fragmentation...... Klebsormidium flaccidum 8. Unicellular occuring as solitary cells...... 9

9. Cells are long, 365.0–520.0 μm (length) and 40.0–47.0 μm (width)...... Pleurotaenium trabecula 9. Cells are short, 2.5–400.0 μm (length) and 1.5–28.5 μm (width)...... 10

10. Cells capable of semicell formation...... 11 10. Cells not capable of semicell formation...... 12

11. Semicells pyramidate with round concave lateral margins...... Cosmarium granulatum 11. Semicells are globose that are connected by an isthmus...... Cosmarium contractum var. rotundatum

12. Cells unicellular, cylindrical, and narrowly fusiform towards the apex...... Closterium acerosum 12. Cells are ovoid, spindle, or crescent-shaped...... 13

13. Cells capable of forming coenobia (tetrahedrical) ...... Coelastrum proboscideum 13. Cells not capable of forming coenobia...... 14

14. Capable of colony formation with lunate or crescent-shaped cells...... Kirchneriella lunaris 14. Cells are solitary and spindle-shaped...... Monoraphidium lunare

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Plate I. Photomicrographs of (1) Kirchneriella lunaris (Kirchner) Möbius; (2) Monoraphidium lunare Nygaard, J. Komárek, J. Kristiansen, & O.M. Skulberg; (3) Coelastrum proboscideum Bohlin; (4) Klebsormidium flaccidum (Kützing) P.C. Silva, K.R. Mattox, & W.H. Blackwell; (5) Pleurotaenium trabecula Nägeli; (6) Cosmarium granatum Brébisson ex Ralfs; (7) Cosmarium contractum var. rotundatum Borge; (8) Closterium acerosum Ehrenberg ex Ralfs; and (9) Chroococcus subnudus (Hansgirg) G. Cronberg & J. Komárek. All scale bars = 10 μm.

Specimen: , Laguna, Calauan (Brgy. Balayhangin, Kristiansen, & O.M. Skulberg Pl. I, Fig. 2 Javier Eco Farm), E. DLR. Arguelles s.n. Photomicrograph prepared from the mounted specimen. Cells uninucleated and are spindle-shaped (to broadly fusiform), slightly curved or arcuate, with pointed ends, Reference: John and Tsarenko 2011, Phylum Chlorophyta 8.0–16.0 μm long, 1.0–3.0 μm wide, chloroplast is parietal (Green Algae) In: The Freshwater Algal Flora of the and is not extending throughout the cytoplasm; cell British Isles: An Identification Guide to Freshwater and division produces 2–4 or 8 autospores arranged in a series Terrestrial Algae, 361p., pl. 98I. set free by lengthwise rupture of the mother cell wall. Class: Chlorophyceae A new record for the Philippines. Order: Sphaeropleales Specimen: LUZON, Laguna, Calauan (Brgy. Balayhangin, Family: Selenastraceae Javier Eco Farm), E. DLR. Arguelles s.n. Photomicrograph Genus: Monoraphidium Komárková-Legnerová prepared from the mounted specimen. 1. Monoraphidium lunare Nygaard, J. Komárek, J.

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Reference: González and Krienitz 1997, Algological characterized by having a truncated apex and a basal Studies, 31p., Fig. 7. swelling. Apex is smooth (without spines or conical warts) and the cell is 25.0–33.0 μm wide while the isthmus Class: Chlorophyceae (containing the nucleus) is 28.0–35.0 μm wide. Semicell Order: Sphaeropleales apex usually has a vacuole that holds on tiny granules Family: Scenedesmaceae (possibly made up of calcium sulfate crystals). The lateral Genus: Coelastrum Nágeli margins of the cells are parallel to each other with clear, 1. Coelastrum proboscideum Bohlin Pl. I, Fig. 3 hyaline cell walls. Chloroplasts are parietal and ribbon- Cells are ovoid and hexagonal at lateral view and triangular like and are scattered in each of the semicell; several in polar view; coenobia tetrahedrical, cubic, or spherical pyrenoids are present within the protoplasm. composed of 4–8 cells, 6.0–18.0 μm in diameter, with Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit small and regular intercellular spaces between the cells Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph held together intercellularly with cell wall outgrowths. prepared from the mounted specimen. Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit References: Ramos et al. 2018c, Phytotaxa 346(1): 69, Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph Fig. 97–100; Gerrath 2003, Conjugating Green Algae prepared from the mounted specimen. and Desmids, In: Freshwater Algae of North America: Reference: Comas and Krienitz 1997, Algological Studies. Ecology and Classification, 377p. 89p., Fig. 10–14. Class: Zygnematophyceae Order: Desmidiales Phylum: CHAROPHYTA Family: Desmidiaceae Class: Klebsormidiophyceae Genus: Cosmarium Corda ex Ralfs Order: Klebsormidiales 1. Cosmarium granatum Brébisson ex Ralfs Family: Klebsormidiaceae Pl. I, Fig. 6 Genus: Klebsormidium P.C.Silva, Mattox & W.H. Cells are 1.2–1.5 times longer than broad, 30.0–39.8 μm in Blackwell length and 20.0–28.5 μm in width; semicells pyramidate with 1. Klebsormidium flaccidum (Kützing) P.C. Silva, K.R. round angles and concave lateral margins, apex of both the Mattox &, W.H. Blackwell Pl. I, Fig. 4 semicells are smooth and rounded; basal and apex angles are Basionym: Ulothrix flaccida Kützing rounded with slight thickening in the apical part of the cell; Filaments are long and not constricted, prone to isthmus (6.0 –10.0 μm) is closed with narrow sinus; cells fragmentation into small pieces; cells are usually with deep median constriction and cell wall is punctuate; cylindrical not attenuated (smooth and curved ends), 16.0 semicells each with chloroplasts containing single pyrenoid. μm long and 6.0 μm wide with slightly firm cell wall; Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit parietal-band shaped chloroplasts (with a single pyrenoid) Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph covering half of the cell. prepared from the mounted specimen. Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit Reference: Nandi et al. 2019, Phytomorphology, 43p., Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph Fig. 1L. prepared from the mounted specimen. 2. Cosmarium contractum var. rotundatum Borge References: Arguelles and Monsalud 2017, Philippine Pl. I, Fig. 7 Journal of Systematic Biology, 33p., Pl. II, Fig.5; Mikhai- lyuk et al. 2015, Journal of Phycology, 755p., Fig. 2a–f; Cells 1.5 – 1.8 times longer than broad, small and globose, Ortega-Calvo et al. 1993, Nova Hedwigia, 246p., Pl. 3, Fig. 24.0–29.0 μm long and 11.5–13.0 μm wide; semicells are 32; Pantastico 1977, Taxonomy of the Freshwater Algae of globose to subcircular that are connected by an isthmus, and Vicinity, 143p., Pl. XI, Fig. 1. lateral margins of the semicells are convex with smooth Class: Zygnematophyceae and rounded apical margin; isthmus is 3.5 –5.5 μm in Order: Desmidiales length; cells with deep median constriction and acute, open Family: Desmidiaceae median sinus; semicells each having a single chloroplast Genus: Pleurotaenium Nägeli containing a pyrenoid. 1. Pleurotaenium trabecula Nägeli Pl. I, Fig. 5 Specimen: LUZON, Laguna, Calauan (Brgy. Balayhangin, Cells solitary, elongated to cylindrical, 365.0–520.0 Javier Eco Farm), E. DLR. Arguelles s.n. Photomicrograph μm in length and 40.0–47.0 μm in width. Semicells are prepared from the mounted specimen.

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References: Coesel and Meesters 2007, Desmids of the mucilaginous sheath is colorless and not lamellated. Lowlands: Mesotaeniaceae and Desmidiaceae of the European Lowlands, 112p., Pl. 61, Fig. 13. Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph Class: Zygnematophyceae prepared from the mounted specimen. Order: Desmidiales Family: Closteriaceae References: Komárek 2013, Czech Polar Reports 3(2): 130, Fig. 7; Martinez 1984, A Checklist of Blue-Green Algae of Genus: Closterium Corda ex Ralfs the Philippines, 31p.; Desikachary 1959, Cyanophyta, p. 1. Closterium acerosum Ehrenberg ex Ralfs 104–105, Pl. 24, G.4 and Pl. 26, G. 4 & 15. Pl. I, Fig. 8 Cells are usually solitary, straight and greenish in color, Class Cyanophyceae central part of the cell is cylindrical and narrowly fusiform Order: Synechococcales towards the apex; middle part of the cell without curvature; Family: Pseudanabaenaceae chloroplast are divided into two parts and are constricted at Genus: Pseudanabaena Lauterborn the middle portion of the cell; cells 9–16 times longer than 1. Pseudanabaena catenata Lauterborn Pl. II, Fig. 2 broad; 225.0–400.0 μm in length and 16.0–28.0 μm in width. Cells are blue-green in color with homogenous protoplasm, Specimen: LUZON, Laguna, Calauan (Brgy. Balayhangin, usually 1.5–2.0 times longer than wide, 2.5–3.0 μm Javier Eco Farm), E.DLR. Arguelles s.n. Photomicrograph in length and 0.9–1.5 μm in width; anterior as well as prepared from the mounted specimen. posterior end cell are rounded. Trichomes usually are solitary, isopolar, straight, and cylindrical with observable Reference: Shakhmatov et al. 2018, Folia Cryptogamica constrictions at the cross walls of the filament. Estonica, 8p., Fig. 2-1. Specimen: LUZON, Laguna, Calauan (Brgy. Balayhangin, Phylum: CYANOBACTERIA Javier Eco Farm), E. DLR. Arguelles s.n. Photomicrograph Class Cyanophyceae prepared from the mounted specimen. Order: Chroococcales References: Arguelles 2019d, Journal of Microbiology, Family: Chroococcaceae Biotechnology and Food Science 9(1): 3, Fig. 6; Park Genus: Chroococcus Nägeli 2012, Algal Flora of Korea: Cyanophyta: Cyanophyceae: 1. Chroococcus subnudus (Hansgirg) G. Cronberg & Chroococcales, Oscillatoriales, 5(1): 48, Fig. 16G, 17A,B; J. Komárek Pl. I, Fig. 9 McGregor 2007, Freshwater Cyanoprokaryota of North- Basionym: Chroococcus turgidus var. subnudus Hansgirg Eastern Australia 1: Oscillatoriales, 36p., Fig. 2f. Cells are oval to spherical and sometimes subspherical, Class Cyanophyceae dark green or pale olive-green in color, 25.0–32.0 μm Order: Oscillatoriales in diameter; cells normally in clusters of 2–4 cells but Family: Oscillatoriaceae at times occurring as solitary cells and rarely up to 16 Genus: Phormidium Kützing ex Gomont cells. The cells are covered with a thin, hyaline (not 1. Phormidium breve (Kützing ex Gomont) Anagnostidis lamellated) mucilage copying the cell margin sometimes & Komárek Pl. II, Fig. 3 indistinguishable and would require staining. Basionym: Oscillatoria brevis Kützing ex Gomont Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit Cells are cylindrical and usually shorter than wide, Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph 1.0–3.5 μm in length and 4.5–7.0 μm in width, smooth, prepared from the mounted specimen. homogenous protoplast with cross-walls that are not granulated. Apical cells with obtuse-conical end and Reference: Park 2012, Algal Flora of Korea (Cyanophyta: without calyptra. Filaments are blue-green in color without Cyanophyceae: Chroococcales, Oscillatoriales) 20, Figs. sheath, forming bundles of entangled mats. Trichomes are 8A and B. slightly curved or straight, isopolar, no constriction at the 2. Chroococcus minutus (Kützing) Nägeli Pl. II, Fig. 1 cross-walls, and slightly attenuated at the ends. Basionym: Protococcus minutus Kützing Specimen: LUZON, Laguna, Calauan (Brgy. Balayhangin, Javier Eco Farm), E. DLR. Arguelles s.n. Photomicrograph Cells are spherical or oval, bluish-green in color and prepared from the mounted specimen. are enclosed in a mucilage, occurring as solitary cells or in groups (2–4); cells 3.0–5.0 μm in diameter without References: Park 2012, Algal Flora of Korea: Cyanophyta: mucilage and 6.0–8.0 μm with mucilaginous sheath; Cyanophyceae: Chroococcales, Oscillatoriales 5(1): 62, smooth and homogenous protoplast without granules; Fig. 31A–C; McGregor 2007, Freshwater Cyanoprokaryota

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Plate II. Photomicrographs of (1) Chroococcus minutus (Kützing) Nägeli; (2) Pseudanabaena catenata Lauterborn; (3) Phormidium breve (Kützing ex Gomont) Anagnostidis & Komárek; (4) Oscillatoria subbrevis Schmidle; (5) Arthrospira jenneri Stizenberger ex Gomont; and (6) Phacus monilatus (Stokes) Lemmerman. All scale bars = 10 μm.

of North-Eastern Australia 1: Oscillatoriales, 55, Fig. 9D, Lakes Area, 491p., Pl. 107, G. 23. Pl. 7D. Martinez 1984, A Checklist of Blue-Green Algae of the Philippines, 57p.; Velasquez 1962, Philippine Journal Class Cyanophyceae of Science 91(3): 296, Pl. 2., Fig. 31. Order: Oscillatoriales Family: Microcoleaceae Class Cyanophyceae Genus: Arthrospira Stizenberger ex Gomont Order: Oscillatoriales 1. Arthrospira jenneri Stizenberger ex Gomont Family: Oscillatoriaceae Pl. II, Fig. 5 Genus: Oscillatoria Vaucher ex Gomont Trichomes are cylindrical and spirally coiled with or 1. Oscillatoria subbrevis Schmidle Pl. II, Fig. 4 without a thin sheath. Filaments have rounded apical Trichomes are straight, solitary, and scattered, 6.0–7.0 cells and without a calyptra. Cells within the filaments μm broad; cross walls at the filaments are slightly have cross-walls that are slightly constricted and with constricted; anterior cells are narrow, without calyptra granulations, usually the cells are shorter than wide; and not capitated; cells are bluish-green, 1.5–4.0 μm in 5.0–7.0 μm in width and 2.5–4.5 μm in length. length and 3.5–5.0 μm in width, granulated protoplast, cross-walls without granulation. Specimen: LUZON, Laguna, Calauan (Brgy. Balayhangin, Javier Eco Farm), E. DLR. Arguelles s.n. Photomicrograph Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit prepared from the mounted specimen. Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph prepared from the mounted specimen. Reference: Nowicka-Krawczyk et al. 2019, Scientific Reports (Nature) 9: 694, Fig. 1 and 2A–C. References: Martinez 1984, A Checklist of Blue-Green Algae of the Philippines, 65p.; Pantastico 1977, Taxonomy Phylum: EUGLENOPHYTA of the Freshwater Algae of Laguna de Bay and Vicinity, Class: Euglenophyceae 53, Pl. IV, G. 2; Prescott 1962, Algae of the Western Great Order: Euglenida Family: Phacidae

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Genus: Phacus Dujardin inventory list of phytotelm microalgae. To date, there 1. Phacus monilatus (Stokes) Lemmerman has been only one report in Asia of the occurrence of Pl. II, Fig. 6 microalgae in phytotelmata (bromeliad tanks), which Basionym: Chloropeltis monilata A. Stokes was documented in Thailand (Poniewozik et al. 2020). Comparison of the phytotelm microalgae identified in Cells are oval to round with a small eyespot characterized the Philippines (current study) with those from Thailand by having a short, linear process; 34.5–36.0 μm long and indicated Chroococcus minutus and Kirchneriella lunaris 22.0–24.0 μm wide; several discoid chloroplasts; pellicles as the only two algal taxa in common. The current are longitudinally striated covering the cell with tiny study reported thirteen new taxa as additional records granules; a small papilla is present at the anterior part of microalgae in phytotelm microhabitat of Asia, which of the cell; flagellum is shorter than the size of the cell, includes Phacus monilatus, Monoraphidium lunare, two ring-shaped grains of paramylon bodies are present. Coelastrum proboscideum, Klebsormidium flaccidum, A new record for the Philippines. Pleurotaenium trabecula, Cosmarium granatum, Specimen: LUZON, Laguna, Calauan (Brgy. Imok, Gapit Cosmarium contractum var. rotundatum, Closterium Fruit Farm), E. DLR. Arguelles s.n. Photomicrograph acerosum, Pseudanabaena catenata, Oscillatoria prepared from the mounted specimen. subbrevis, Phormidium breve, Arthrospira jenneri, and Chroococcus subnudus. Also, this survey documented Reference: Wolowski et al. 2013, Polish Botanical Journal the occurrence of two rare microalgae – namely, Phacus 58: 659–685, Fig. 67a,b, 101. monilatus (Stokes) Lemmerman and Monoraphidium lunare Nygaard, J. Komárek, J. Kristiansen, & O.M. Skulberg – for the first time in the Philippines. In general, microalgal species observed in pineapple phytotelmata DISCUSSION found in the Philippines are somewhat different as Phytotelmata are a unique habitat for the existence of compared to those observed from other plant species new and rare species of microalgae. However, only a with at least six reported species common in other few studies were documented on microalgal diversity countries. Such observation proves that this overlooked in these microhabitats (Sophia 1999). Globally, a aquatic microhabitat contain rare and endemic species total of 459 taxa of microalgae have been reported in of microalgae that can be further studied to deepen our phytotelmata (Ramos and Moura 2019; Poniewozik et understanding on the diversity of these microorganisms. al. 2020). Among these taxonomic studies, 118 taxa The number of phytotelm algal species described and were reported in phytotelmata from bromeliads of identified in this survey (15 species) is lower as compared Thailand and 215 algal species were documented in with other taxonomic survey done in other countries tropical plants of Brazil. This corresponds to around like that of Poniewozik et al. (2020) and Gebühr et al. 72.5% of the overall number of reported species for this (2006), who identified 118 and 78 taxa, respectively microhabitat proving the significant contribution of these as well as Hernandez-Rodriguez et al. (2014), who countries in the taxonomy and ecology of phytotelm identified 16 species. The small diversity of algal flora algae. In the Philippines, there have been no documented described in this study may be due to a limited amount reports of the occurrence of microalgae associated with of microcosm observed in the bromeliad phytotelmata. phytotelmata. This survey presents the first taxonomic The microcosm in these plants can be considered as an study of phytotelm algae from bromeliad tanks of extreme environment that can favor and select the growth pineapple [Ananas comosus (L.) Merr] from Calauan, of a specific microalgal group in the phytotelmata. The Laguna. A total of 15 microalgal taxa were taxonomically presence of high organic matter (detritus) can favor the identified and described from the collected samples, growth of pigmented and colorless euglenophytes that of which six species (five genera) are classified as are tolerant to these conditions (Sachertt Mendes et al. members of Cyanophyceae, four species (three genera) 2019). Therefore, euglenoids are usually profuse in belong to Zygnematophyceae, three species (three aquatic environments that are eutrophic, grazing on both genera) to Chlorophyceae, and one species (one genus) organic solutes and other bacteria. Also, high species each for Euglenophyceae and Klebsormidiophyceae. diversity of phytotelm algae in pineapple from Calauan, Among these taxa, nine species (Phacus monilatus, Laguna can be attributed to its plant architecture. Monoraphidium lunare, Coelastrum proboscideum, This bromeliad is capable of producing several (over Klebsormidium flaccidum, Cosmarium granatum, 20–30) broad leaves that form several water cavities Cosmarium contractum var. rotundatum, Phormidium (greater complexity) where the accumulation of water breve, Arthrospira jenneri, and Chroococcus subnudus) occurs. These leaves are positioned in open angles that are new additional records of microalgae in the global allow good illumination that supports the growth and

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proliferation of phytotelm algae. A similar observation CONCLUSION was noted by Ramos et al. (2018b) in Alcantarea nahoumii where high species richness of algae can be The current study is the first floristic survey that attributed to the complex structure of the bromeliad. characterizes the taxonomy and species composition They observed that leaves of A. nahoumii associated of phytotelm algae associated with bromeliad tanks at the lateral tanks of the bromeliad are held at good of pineapple. The study reports an additional of nine open angles, allowing a substantial amount of light species as new records in the global inventory list of penetration. Ecological parameters like pH, dissolved phytotelm microalgae and thirteen new taxa as additional oxygen, light intensity, temperature, and conductivity records of microalgae in phytotelm microhabitat of Asia. can affect the species richness of phytotelm algae in Also, the occurrence of a rare photosynthetic euglenoid bromeliad tanks. High algal species diversity is favored [Phacus monilatus (Stokes) Lemmerman] and a green in bromeliad tanks with high temperatures and slightly microalga [Monoraphidium lunare Nygaard, J. Komárek, acidic water, exposed to high rainfall, high dissolved J. Kristiansen, & O.M. Skulberg] are documented for the oxygen, moderate conductivity, high light intensity, and first time in the Philippines. This study provided a quick high available nutrients (Ramos et al. 2018b; Marino et assessment and identifies some of the dominant and rare al. 2011; Sophia et al. 2004). species of phytotelm algae in bromeliad tanks of pineapple found in the Philippines. To date, several species of bromeliads (ornamental bromeliad species) that form phytotelmata are generally classified as critically endangered and highly vulnerable species of plant (Ramos et al. 2018b). Thus, it is ACKNOWLEDGMENTS important to emphasize the need for conservation and The author is grateful for the assistance of the Philippine protection of these naturally occurring plants against National Collection of Microorganisms, BIOTECH, excessive harvesting (to be used for decorative purposes) University of the Philippines Los Baños (UPLB) in and burning (Ramos and Moura 2019). The conservation this study. Also, the author acknowledges the kind help of these plants is important in preserving and maintaining provided by Mrs. Maria Lynette B. Aquino and the the rare and endemic species of microalgae present in University Library of UPLB for assisting the author to these phytotelmata communities. retrieve some of the important references needed in the The current study is the first floristic survey that study. The constructive suggestions and comments of the highlights the taxonomy of phytotelm algae associated external reviewers are also acknowledged with gratitude. with bromeliad tanks of pineapple. An additional comprehensive survey will, therefore, be needed to deepen our understanding and knowledge of phytotelm algae in these overlooked environments, not only in REFERENCES the Philippines but also to other countries throughout ARGUELLES EDLR. 2020. Species composition of algal the tropical region. The majority of the studies done on epiphyton of Pink Lotus (Nymphaea pubescens Willd) phytotelm algae of bromeliad tanks came from those found in Laguna de Bay (Philippines). Walailak J Sci planted in soil, and no comprehensive studies are done & Tech 17(3): 237–256. focusing on algal flora of phytotelmata in bromeliads that are epiphytic to other plants (observed in tree canopies). ARGUELLES EDLR. 2019a. Morphotaxonomic study of Phytotelmata on these bromeliads may contain a greater algal epiphytes from Ipomoea aquatica Forssk. (Con- diversity of phytotelm algae because of high exposure volvulaceae) found in Laguna de Bay (Philippines). to light (Ramos and Moura 2019). Also, thorough and Pertanika J Trop Agric Sci 42(2): 817–832. in-depth taxonomic and ecological (light intensity, ARGUELLES EDLR. 2019b. Systematic study of some temperature, pH, dissolved oxygen, and conductivity) epiphytic algae (non-diatoms) on the submerged parts studies on other phytotelmata such as plant axils and of water hyacinth [Eichhornia crassipes (Mart.) Solms- modified leaves (such as those found in Pandanaceae Loubach] found in Laguna de Bay, Philippines. Trop and Nepenthaceae, respectively) will provide us basal Life Sci Res 30(1): 1–21. information to better understand the distribution pattern plus biological and ecological significance of these ARGUELLES EDLR. 2019c. Descriptive study of some microorganisms. epiphytic algae growing on Hydrilla verticillata (L.f) Royle (Hydrocharitaceae) found in the shallow freshwater lake, Laguna de Bay (Philippines). Egypt J Aquat Biol Fish 23(2): 15–28.

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ARGUELLES EDLR. 2019d. New records of corticolous GUIRY MD, GUIRY GM. 2020. AlgaeBase. National microalgae and cyanobacteria for Philippine algal flora University of Ireland, Galway. Retrieved on 22 Mar from Mt. Makiling Forest Reserve. J Microb Biotech 2020 from http://www.algaebase.org Food Sci 9(1): 1–8. HERNÁNDEZ‐RODRÍGUEZ B, ESTRADA‐VARGAS ARGUELLES EDLR, MARTINEZ-GOSS MR. 2019. L, NOVELO E. 2014. Las microalgas de Tillandsia Diversity of Philippine photosynthetic euglenophytes multicaulis Steud. (Bromeliaceae) de la Reserva and their potential biotechnological uses: a review. Int Ecológica “La Martinica”, Veracruz. TIP Rev Espec J Emerg Technol 10(4): 24–31. Cienc Quím‐Biol 17(2): 117–125 ARGUELLES EDLR, MONSALUD RG. 2017. Morpho- JOHN DM, TSARENKO M. 2011. Order Chlorococcales. taxonomy and diversity of terrestrial microalgae and In: The Freshwater Algal Flora of the British Isles: An cyanobacteria in biological crusts of soil from paddy Identification Guide to Freshwater and Terrestrial Al- fields of Los Baños, Laguna (Philippines). Philipp J gae. John DM, Whitton BA, Brook AJ eds. Cambridge: System Biol 11(2): 25–36. Cambridge University Press. p. 327–409. ARGUELLES EDLR, MARTINEZ-GOSS MR, SHIN W. KILLICK SA, BLANCHON DJ, LARGE MF. 2014. Algal 2014. Some noteworthy photosynthetic euglenophytes communities in phytotelmata: a comparison of native from Laguna and vicinities. Philipp Sci 51: 1–36. Collospermum and exotic bromeliads (Monocotyle- donae) in New Zealand. Telopea 17: 303–310. https:// BROUARD O, CÉRÉGHINO R, CORBARA B, LE- doi.org/10.7751/telopea20147847 ROY C, PELOZUELO L, DEJEAN A, CARRIAS JF. 2012. Understorey environments influence functional KITCHING RL. 2000. Food webs and container habitats: diversity in tank‐bromeliad ecosystems. Freshw Biol the natural history and ecology of phytotelmata. Cam- 57(4): 815–823. https://doi.org/10.1111/j.1365‐ bridge: Cambridge University Press. 449p. 2427.2012.02749.x KOMÁREK J. 2013. Phenotypic and ecological diversity COESEL PFM, MEESTERS KJ. 2007. Desmids of the of freshwater coccoid cyanobacteria from maritime Lowlands: Mesotaeniaceae and Desmidiaceae of the Antarctica and islands of NW Weddell Sea. I. Syn- European Lowlands. Zeist, the Netherlands: KNNV echococcales. Czech Pol Rep 3(2): 130–143. Publishing. 352p. LYRA LT. 1971. Algumas diatomaceas encontradas COMAS A, KRIENITZ L. 1997. Comparative LM- and em Bromeliaceas. Brasil Mem Inst Oswaldo Cruz SEM- studies on Coelastrum (Chlorophyta, Chlorococ- 69(1): 129–139. https://doi.org/10.1590/S0074- cales) under culture conditions. Algol Stud 87: 87–98. 02761971000100010 DESIKACHARY TV. 1959. Cyanophyta. I.C.A.R. Mono- MARINO NAC, GUARIENTO RB, DIB V, AZEVEDO graph on Algae. New Delhi: ICAR Press. 686p. FD, FARJALLA VF. 2011. Habitat size determine algae biomass in tank-bromeliads. Hydrobiologia DUARTE GSC, ALVES GM, LANSAC‐TÔHA FM, 678: 191–199. VELHO LFM, LANSAC‐TÔHA FA. 2013. Flagel- late protist abundance in phytotelmata of Aechmea MARTINEZ MR. 1984. A checklist of blue-green algae distichantha Lem (Bromeliaceae) in the upper Paraná of the Philippines. National Institute of Molecular river basin. Acta Sci Biol Sci 35(4): 491–498. https:// Biology and Biotechnology (BIOTECH) – University doi.org/10.4025/actascibiolsci.v35i4.17134 of the Philippines Los Baños. 96p. GEBÜHR C, POHLON E, SCHMIDT AR, KÜSEL K. MARTINEZ-GOSS MR, MANLAPAS JEB, AR- 2006. Development of microalgae communities in GUELLES EDLR. 2019. Cyanobacteria and diatoms the phytotelmata of allochthonous populations of in the cyanobacterial mats in a natural saltwater hot Sarracenia purpurea (Sarraceniaceae). Plant Biology spring in Coron, Palawan, Philippines. Philipp Sci Lett 8: 849–860. https://doi.org/10.1055/s-2006-924474 12(Supplement): 11–32. GERRATH JF. 2003. Conjugating Green Algae and MCGREGOR GB. 2007. Freshwater Cyanoprokaryota of Desmids In: Freshwater Algae of North America: North-Eastern Australia 1: Oscillatoriales. Canberra: Ecology and Classification. Wehr JD, Sheath RG eds. CSIRO Publishing. 124p. Amsterdam: Academic Press. p. 353–379. MIKHAILYUK T, GLASER K, HOLZINGER A, GONZÁLEZ AC, KRIENITZ L. 1997. Taxonomic obser- KARSTEN U. 2015. Biodiversity of Klebsormidium vations on some coccal green algae exsiccatae (Chlo- (Streptophyta) from alpine biological soil crusts (Alps, rococcales, Chlorophyceae). Algol Stud 84: 13–38. Tyrol, Austria, and Italy). J Phycol 51: 750–767.

600 Philippine Journal of Science Arguelles: Microalgae of Pineapple Phytotelmata Vol. 149 No. 3, September 2020

NANDI C, BHOWMICK S, GORAIN CP, PAL R. 2019. mations in Bahia State, Northeastern Brazil. Acta Bot New and Rare Records of Cosmarium (Desmidia- Bras 32(4): 567–577. https://doi.org/10.1590/0102‐ ceae, Zygnematales) from India. Phytomorphology 69 33062018abb0070 (1&2): 41–49. RAMOS GJP, BICUDO CEM, MOURA CWN. 2018c. NOWICKA-KRAWCZYK P, MÜHLSTEINOVÁ R, Some new, rare and interesting desmids from brome- HAUER T. 2019. Detailed characterization of the liad phytotelmata in Brazil. Phytotaxa 346(1): 59–77. Arthrospira type species separating commercially https://doi.org/10.11646/phytotaxa.346.1.3 grown taxa into the new genus Limnospira (Cyano- RAMOS GJP, ALVES-DA-SILVA SM, BICUDO CEM, bacteria). Scientific Reports (Nature) 9: 694. https:// MOURA CWN. 2017a. Euglenophyceae from bro- doi.org/10.1038/s41598-018-36831-0 meliad phytotelmata: new records for Bahia state ORTEGA-CALVO JJ, SANCHEZ-CASTILLO PM, and Brazil. Check List 13(5): 447–454. https://doi. HERNANDEZ- MARINE M, SAIZ-JIMENEZ C. org/10.15560/13.5.447 1993. Isolation and characterization of epilithic Chlo- RAMOS GJP, BICUDO CEM, MOURA CWN. 2017b: rophyta and cyanobacteria from two Spanish cathedrals Cosmarium bahianum, sp. nov. (Desmidiaceae), a (Salamanca and Toledo). Nova Hedwigia 57: 239–253. new desmid species from a phytotelm habitat in the PANTASTICO JB. 1977. Taxonomy of the Fresh-water Brazilian restinga. Phytotaxa 291: 66–72. https://doi. Algae of Laguna de Bay and Vicinity. Taguig City: org/10.11646/phytotaxa.291.1.6 National Research Council of the Philippines. 251p. RAMOS GJP, BICUDO CEM, MOURA CWN. 2016. First PARK JG. 2012. Algal Flora of Korea: Cyanophyta: Cya- record of Parvodinium umbonatum (Stein) Carty (Peri- nophyceae: Chroococcales, Oscillatoriales). Incheon: diniaceae, Dinophyta) for northeast Brazil. Check List NIBR Ministry of Environment. 71p. 12(6): 1–6. https://doi.org/doi: 10.15560/12.6.2024 PONIEWOZIK M, DUANGJAN K, PEKKOH J, SACHERTT MENDES PMS, LANSAC-TÔHA WOŁOWSKI K. 2020. Algae of bromeliad phytotel- FM, MEIRA BR, OLIVEIRA FR, VELHO LFM, mata in the Queen Sirikit Botanical Garden, Chiang LANSAC-TÔHA FA. 2019. Heterotrophic flagellates Mai, Thailand. Phytotaxa 432(1): 17–37. (Amorpha and Diaphoretiches) in phytotelmata bro- meliad (Bromeliaceae). Braz J Biol [Epub]. https:// PRESCOTT GW. 1962. Algae of the Western Great Lakes doi.org/10.1590/1519-6984.218742 Area. Dubuque: Wm. C. Brown Company. 977p. SHAKHMATOV AS, PAVLOVSKIY EV, PAUKOV AG. RAÑOLA MCG, ZAFARALLA MT, VALMONTE RAD. 2018. Desmid algae (Charophyta: Conjugatophyceae) 1990. A preliminary investigation on the epiphyton of of Ekaterinburg, Middle Urals, Russia. Folia Cryptog Eichhornia crassipes (Mart.) Solm. roots in Laguna Estonica 55: 7–15. de Bay. UP Los Baños J 1(1): 53–67. SOPHIA MG, CARMO BP, HUSZAR VL. 2004. Desmids RAMOS GJP, MOURA CWN. 2019. Algae and cyanobac- of phytotelm terrestrial bromeliads from the National teria in phytotelmata: diversity, ecological aspects, and Park of “Restinga de Jurubatiba”, Southeast Brasil. conservation. Biodivers Conserv 28(7): 1667–1697. Algol Stud 114: 99–119. https://doi.org/10.1007/s10531-019-01771-2 SOPHIA MG. 1999. Desmídias de Ambientes Fitoté- RAMOS GJP, LHZ BRANCO, MOURA CWN. 2019. Cy- lmicos Bromelícolas. Rev Bras Biol 59(1): 141–150. anobacteria from bromeliad phytotelmata: new records, morphological diversity, and ecological aspects from STARZOMSKI BM, SUEN D, SRIVASTAVA DS. 2010. northeastern Brazil. Nova Hedwigia 118(1–2): 51–72. Predation and facilitation determine chironomid emer- https://doi.org/10.1127/nova_hedwigia/2018/0499 gence in a bromeliad-insect food web. Ecol Entom 35: 53–60. RAMOS GJP, BICUDO CEM, MOURA CWN. 2018a. Diversity of green algae (Chlorophyta) from bromeliad VELASQUEZ GT. 1962. The blue green algae of the phytotelmata in areas of rocky outcrops and “restinga”, Philippines. Philipp J Sci 91(3): 267–380. Bahia State, Brazil. Rodriguésia 69: 1973–1985. WEHR JD, RG SHEATH. 2003. Freshwater Algae of https://doi.org/10.1590/2175‐7860201869431 North America: Ecology and Classification. Amster- RAMOS GJP, SANTANA LM, MEDINA AM, BICUDO dam: Academic Press. 918p. CEM, BRANCO LHZ, MOURA CWN. 2018b. WHITTON BA. 2002. Phylum Cyanophyta (Cyanobac- Unraveling algae and cyanobacteria biodiversity in teria). In: The Freshwater Algal Flora of the British bromeliad phytotelmata in different vegetation for- Isles: An Identification Guide to Freshwater and Ter-

601 Philippine Journal of Science Arguelles: Microalgae of Pineapple Phytotelmata Vol. 149 No. 3, September 2020

restrial Algae. John DM, Whitton BA, Brook AJ eds. Cambridge: Cambridge University Press. p. 25–122. WOLOWSKI K, PONIEWOZIK M, WALNE PL. 2013. Pigmented Euglenophytes of the genera Euglena, Euglenaria, Lepocinclis, Phacus and Monomorphina from southeastern United States. Polish Botan J 58: 659–685.

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