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(Cladophorales, Ulvophyceae) –A Bloom Forming Marine Algae from Goa, India

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(Cladophorales, Ulvophyceae) –A Bloom Forming Marine Algae from Goa, India Indian Journal of Geo-Marine Sciences Vol. 44(12), December 2015, pp. 1874-1879 Cladophora goensis sp. nov. (Cladophorales, Ulvophyceae) –a bloom forming marine algae from Goa, India Felix Bast*, Aijaz Ahmad John & Satej Bhushan Centre for Biosciences, Central University of Punjab, Bathinda, Punjab, 151001, India * [Email: [email protected]] Received 22 April 2014; revised 12 June 2014 A new species of green seaweed, Cladophora goensis sp. nov. (Cladophorales, Ulvophyceae), had been discovered from Vasco-da-Gamma, Goa, India. This species formed algal bloom of moderate intensity in the Bay of Mormugao, on the west coast of India. Observations suggest combination of a number of morphological characteristics of this alga distinct from previously described members of this genus, including parietal chloroplast surrounding central hollow and bilenticular pyrenoids. Molecular sequence data at Nuclear ribosomal DNA Internal Transcribed Spacer-1 and 2 (ITS1 and ITS2) regions along with intervening 5.8S rRNA indicated Kimura-2-Parameter (T3P) pair-wise distance of 1.77 x 10-1 between this species and the nearest phylogenetic accession of Cladophora glomerata. In phylogenetic reconstructions using Bayesian Inference and Maximum Likelihood, this species was not part of any monophyletic clades comprising any of the previously described species of this genus at the locus studied, thereby ascertaining conformity with phylogenetic species concept. With this discovery, a new phylogenetically primitive morphological synapomorphy of “pseudo dichotomous profuse branching” has been revealed for cladophoralean algae, and this is the single most important morphological characteristic of this bloom- forming seaweed. [Keywords: Green tide; ITS1; systematics, phylogenetics; seaweed] Introduction Genus Cladophora (Kützing) consists of some adaptation to environments of various salinity of the ubiquitous bloom-forming green algae and wave-exposure regimes, with with cosmopolitan in distribution. This representative species of macroalgal genus is renowned for the genus has been described from marine, the main axis, cell shape and wall thickness, brackish and freshwater habitats. Its massive shape of the apical cell tip-if tapered, ephemeral blooms have been described from apical/intercalary growth, arrangement of marine, as well as freshwater, habitats chlorophytes and so on. Presence of epi- throughout the world1-3 and, therefore, algae of endophyte Ulvella leptochaete on this genus this genus is considered to play a major role in from India has also been reported recently5. marine, estuarine and limnetic ecosystem There are 188 morphospecies of this genus dynamics. currently accepted taxonomically; almost all of them described based solely on microscopic Algal thalli of the genus Cladophora characters and, therefore, species-level consist of moderately branched uniseriate identification remains very cumbersome for filaments with coenocytic cells having a the field phycologists6. Species level typical banded pattern under low identification based on both morphology and magnification. Dominant form of life history molecular phylogeny has lead to discovery of in this genus is isomorphic diplohaplontic many new species which otherwise would alternation and exhibit considerable have gone un-noticed7. A number of recent morphological plasticity in relation with molecular phylogenetic studies have revealed environmental factors4. This genus is known polyphyly of this genus owing to the for lack of clear-cut species delineating unaccountability of morphological morphological characters. Species delineation synapomorphies, with members of this genus within this genus have been traditionally based clustered within at least two orders; upon combinations of various microscopic Cladophorales and Siphonocladales8,9. For characteristics, such as insertion of branches in taxonomically reliable and reproducible BAST et al.: CLADOPHORA GOENSIS SP. NOV. (CLADOPHORALES, ULVOPHYCEAE) 1875 description of cladophoralean species, 10 g wet weight of thalli was used for molecular data and its phylogenetic DNA extraction and sequencing. Unabridged reconstruction have recently been integrated in protocols used for DNA extraction, a European species10. amplification and sequencing used in the present study are as per Bast, 201317. In Previously reported species of genus summary, total genomic DNA was extracted Cladophora in Indian subcontinent include from dried specimens using HiPurATM Algal Cladophora callicoma11, Cladophora 12 13 Genomic Extraction Kit (HiMedia uberrima and Cladophora crispata . At Laboratories, India). A region consisting of least three other species had also been reported, ITS1-5.8S-ITS2 was amplified from the viz., Cladophora prolifera14, Cladophora 15 16 extracted DNA using ITS1-forward primer and fasicularis and Cladophora glomerata , ITS4-reverse primer18 and subsequently albeit taxonomy was not the point of focus in subjected to bidirectional Sanger sequencing those reports, and no detailed information on (Applied Biosystems 3730xl Genetic Analyzer, species’ identification was mentioned. Foster City, CA, USA). Sequences were then Cladophora from Indian Subcontinent has assembled and additional sequences of related never been subjected to molecular assessment. taxa were procured from Genbank, with In addition, algal bloom caused by numbers of these accessions listed in Table S1. Cladophora have never been reported from Phylogenetic analyses using distance matrix, India. Maximum Likelihood (ML) and Bayesian Materials and Methods Inference (BI) methods were conducted with Kimura-2-Parameter (K2P) model of In our 2012 field-trip to the west coast nucleotide substitution, gamma distribution of India, a particular Cladophoralean algae and complete deletion of gaps and missing was observed causing bloom of moderate data in dataset, in force. Methodology used for intensity at Mormugao Bay, Vasco-da-Gamma, sequence alignment and Phylogenetic analysis Goa. This bloom was found to be occurring all was as per Bast, 201317 along the bay coastline, from the northern tip of Headland Sada till North-East of Vasco Results and Discussion fishing harbor, with intervening Mormugao Thallus was erect filamentous, light Harbor area. The algae was seen to be green in color with size ranging between 3 cm- profusely growing on mooring lines, buoys, 6 cm (Average 4.34±1cm; n=50) (Fig.1-6). hulls of wooden dinghies and intertidal Under low magnification, irregular branching substrata, including natural rocks and concrete pattern of uniseriate filaments was evident. breakwaters. Cells produced mostly dichotomous branching, A bloom specimen was collected from but third, fourth and fifth order branching were mid-point of the Vasco fishing harbor also occasionally observed. Cells were mostly (15.402639N, 73.814673E), which was cylindrical, with slightly narrowed at posterior observed to be growing attached to the ends. Cell diameter of basal, main axial and mooring line made of coir. Algal specimen apical cells ranged between 65-165 µm, 59- was transported to the laboratory in the ice box 150 µm and 45-141 µm, respectively. Apical (c 4°C) and processed immediately. Samples cells were tapering with obtuse tips. Each cell did not freeze at any point of time and had 100-130 distinct pyrenoids within parietal therefore possibility of the freeze-fracture of network of the chloroplast, most having a chloroplasts can be ruled out. Morphological central hollow. In our understanding, these features were recorded using an upright morphological features are not shared with any microscope (BX53, Olympus, Japan) and of the previously described species of photographs were taken using an attached Cladophora and, therefore, we conclude that digital camera (E450, Olympus, Japan). Public our isolate is a new species. Comparison of domain software ImageJ taxonomic characteristics between this isolate (http://rsbweb.nih.gov/ij/) was used for scale and Cladophora sericea, a closely related calibration and size measurements. bloom-forming species, and Cladophora glomerata, phylogenetically closely related 1876 INDIAN J. MAR. SCI., Vol. 44, NO.12 DECEMBER 2015 Table 1. Distinguishing characters of Cladophora goensis from Cladophora sericea and Cladophora glomerata S. Features Cladophora goensis Cladophora sericea19 Cladophora glomerata20 No Light to grass green, densely Dark green, densely tufted, 1 1 Habit Light green, moderately tufted tufted, upto 20 cm high meter or more high Cell Apical: 45-141 µm, Main axis cells: 59-150 Apical: 19-91 µm, Main axis 2 diameter Apical: 15-70, Matured: 35-55 µm, Basal cells: 65-165 µm cells: up to 150 µm. (µm) Irregular profuse branching of uniseriate Terminal branch systems Pseudodichotomous profuce filaments forming tufts. Branches are inserted acropetal to irregular; main axis branching with terminal bunch Branching 3 apically on the cell without apical meristem pseudodichotomously branched; of arcuate branches. Growth system (pseudodichotomy).Creeping growth mainly by intercalary cell mainly by intercalary cell growth/intercalary cell division absent. division division Apical cells tapering with an obtuse tip; Other Chloroplasts form parietal layer, mostly with Apical cells tapering with an Branches are attached 4 Characters central hollow, and consists of a network of obtuse tip. obliquely, rarely laterally. numerous ovular bilenticular pyrenoids. Common in sheltered marine habitats with Common in intertidal rockpools, Common in freshwater to 5 Ecology stable
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