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The pelagophytes, a morphologically diverse class of marine heterokont algae, have been historically united only by DNA sequences. Recently we described a novel perforated theca (PT) encasing cells from the Pelagophyceae and hypothesized it may be the first morphological feature to define the class. Here we consolidate that observation, describing a PT for the first time in an additional seven pelagophyte genera, including three genera new to science. We established clonal cultures of pelagophytes collected from intertidal pools located around Australia, and established phylogenetic trees based on nuclear 18S rDNA and plastid rbcL, psaA, psaB, psbA and psbC gene sequences that led to the discovery of three new species: Wyeophycus julieharrissiae and Chromopallida australis form a distinct lineage along with Ankylochrysis lutea within the Pelagomonadales, while Pituiglomerulus capricornicus is sister genus to Chrysocystis fragilis in the Chrysocystaceae (Sarcinochrysidales). Using fixation by high-pressure freezing for electron microscope observations, a distinctive PT was observed in the three new genera described in this paper, as well as four genera not previously investigated: Chrysoreinhardia, Sargassococcus, Sungminbooa and Andersenia. The mechanism of PT formation is novel, being fabricated from rafts in Golgi-derived vesicles before being inserted into an established PT. Extracellular wall and/or mucilage layers assemble exterior to the PT in most pelagophytes, the materials likewise secreted by Golgi-derived vesicles, though the mechanism of secretion is novel. Secretory vesicles never fuse with the plasma membrane as in classic secretion and deposition, but rather relocate extracellularly beneath the PT and disintegrate, the contents having to pass through the PT prior to wall and/or mucilage synthesis. This study substantiates the diverse nature of pelagophytes, and provides further evidence that the PT is a sound morphological feature to define the Pelagophyceae, with all 14 of the 20 known genera studied to date by TEM possessing a PT., (© 2022 The Authors. Journal of Phycology published by Wiley Periodicals LLC on behalf of Phycological Society of America.)

Pelagophytes (Heterokonta) are a morphologically diverse class of marine algae historically united only by DNA sequences. We established clonal cultures of sand-dwelling pelagophytes collected from intertidal pools around Australia. Phylogenetic trees based on nuclear 18S rDNA and plastid rbcL, psaA, psaB, psbA, and psbC sequences revealed two new genera, Gazia and Glomerochrysis, related to Aureoumbra in a distinct lineage within the Sarcinochrysidaceae (Pelagophyceae). The three new species (Gazia saundersii, Gazia australica, and Glomerochrysis psammophila), along with an Australian strain of Aureoumbra geitleri, are characterized by dominant benthic stages that differ significantly from one another, while occasionally producing classic heterokont zoospores. The benthic stage of Ga. saundersii has a novel development not observed in any other colonial alga, consisting of large, spherical colonies (up to 140 μm in diameter) containing c. 2,500 cells that eventually differentiate and segregate into a large number of daughter colonies that are subsequently liberated. Alternatively, colonies may differentiate into a mass of zoospores that escape and settle to develop into new colonies. In Gl. psammophila, cubic packets of cells form large sticky clusters that bind sand together, while Ga. australica and A. geitleri are unicellular species. Using fixation by high-pressure freezing, a distinctive perforated theca was observed by TEM in all genera of this lineage, and we hypothesize this unique covering may be the first morphological feature to characterize most, if not all, pelagophytes. This study substantiates the diverse nature of sand-dwelling pelagophytes as well as their mechanisms for thriving in a dynamic habitat., (© 2020 Phycological Society of America.)

Prasinophytes (Chlorophyta) are a diverse, paraphyletic group of planktonic microalgae for which benthic species are largely unknown. Here, we report a sand-dwelling, marine prasinophyte with several novel features observed in clonal cultures established from numerous locations around Australia. The new genus and species, which we name Microrhizoidea pickettheapsiorum (Mamiellophyceae), alternates between a benthic palmelloid colony, where cell division occurs, and a planktonic flagellate. Flagellates are short lived, settle and quickly resorb their flagella, the basal bodies then nucleate novel tubular appendages, termed "microrhizoids", that lack an axoneme and function to anchor benthic cells to the substratum. To our knowledge, microrhizoids have not been observed in any other green alga or protist, are slightly smaller in diameter than flagella, generally contain nine microtubules, are long (3-5 times the length of flagella) and are not encased in scales. Following settlement, cell divisions result in a loose, palmelloid colony, each cell connected to the substratum by two microrhizoids. Flagellates are round to bean-shaped with two long, slightly uneven flagella. Both benthic cells and flagellates, along with their flagella, are encased in thin scales. Phylogenies based on the complete chloroplast genome of Microrhizoidea show that it is clearly a member of the Mamiellophyceae, most closely related to Dolichomastix tenuilepsis. More taxon-rich phylogenetic analyses of the 18S rRNA gene, including metabarcodes from the Tara Oceans and Ocean Sampling Day projects, confidently show the distinctive nature of Microrhizoidea, and that the described biodiversity of the Mamiellophyceae is a fraction of its real biodiversity. The discovery of a largely benthic prasinophyte changes our perspective on this group of algae and, along with the observation of other potential benthic lineages in environmental sequences, illustrates that benthic habitats can be a rich ground for algal biodiscovery., (© 2019 Phycological Society of America.)

A marine, sand-dwelling, golden-brown alga is described from clonal cultures established from a high intertidal pool in southeastern Australia. This tiny, unicellular species, which we call the "golden paradox" (Chrysoparadoxa australica gen. et sp. nov.), is benthic, surrounded by a multilayered cell wall and attached to the substratum by a complex adhesive plug. Each vegetative cell gives rise to a single, naked zoospore with heterokont flagella that settles and may become briefly amoeboid prior to dividing. Daughter cells are initially amoeboid, then either permanently attach and return to the benthic stage or become motile again prior to final settlement. Two deeply lobed chloroplasts occupy opposite ends of the cell and are surrounded by only two membranes. The outer chloroplast membrane is continuous between the two chloroplasts via the outer membrane of the nuclear envelope. Only two membranes occupy the chloroplast-nucleus interface, the inner membrane of the nuclear envelope and the inner chloroplast membrane. A small pyrenoid is found in each chloroplast and closely abuts the nucleus or protrudes into it. It contains an unusual, membrane-bound inclusion that stains with SYBR green but is unlikely to be a nucleomorph. Phylogenies inferred from a 10-gene concatenated alignment show an early-branching position within the PX clade. The unusual morphological features and phylogenetic position indicate C. australica should be classified as a new class, Chrysoparadoxophyceae. Despite an atypical plastid, exploration of the C. australica transcriptome revealed typical heterokont protein targeting to the plastid., (© 2018 Phycological Society of America.)

Diatoms are a major component of the slime layers that form on artificial surfaces in marine environments. In this article, the role played by diatoms during the pioneering stages of colonization of three marine antifouling (AF) coatings, viz Intersmooth 360, Super Yacht 800 and a fouling-release (FR) coating Intersleek 700, was investigated. The study was conducted over three distinct seasons in two very different marine environments in Australia, ie temperate Williamstown, Victoria and tropical Cairns, Queensland. Diatom fouling occurred more rapidly on the FR coating Intersleek 700, compared to both biocidal AF paints. However, colonization by diatoms on all three coatings was generally slow during the 16-day study. Benthic diatoms do not subsist by floating around in the water column, rather they only gain the opportunity to colonize new surfaces when they either voluntarily release or are displaced from their benthic habitat, thereafter entering the water column where the opportunity to adhere to a new surface presents itself. However, once settled, fouling diatoms grow exponentially from the site of attachment, spreading out until they populate large areas of the surface. This mode of surface colonization correlates more with an 'infection' type, epidemiology model, a mechanism that accounts for the colonization of significant regions of the coating surface from a single fouling diatom cell, forming 'clonal patches'. This is in comparison to the bacterial colonization of the surface, which exhibits far more rapid recruitment and growth of cells on the substratum surface. Therefore, it is hypothesized that fouling diatoms may be characterized more by their ability to adhere and grow on surfaces already modified by bacterial biofilms, rather than on their strength of adhesion. Cell morphology and the ability to avoid shear may also be an important factor.

The role played by bacteria during the pioneering stages of colonisation on marine coatings was investigated over three distinct seasons in both tropical and temperate environments. Novel methods were developed to facilitate the study of the adhered bacterial population on the test coatings in their native, hydrated state. The approach eliminated destructive sample preparation techniques, including sample dehydration and/or removal from the substratum surface prior to analysis. Bacterial colonisation during initial biofilm formation was evaluated on two antifouling paints, Intersmooth 360 and Super Yacht 800, and a fouling release coating, Intersleek 700. Bacterial colonisation was quantified on all three coating surfaces. Intersleek 700 displayed the quickest colonisation by bacteria, resulting in major modification of the substratum surface within 2-4 days following immersion in the ocean. Whilst fouling accumulated more quickly on Intersleek 700, by 16 days all three coatings were fouled significantly. Bacterial fouling was correlated to both location and season, with fouling occurring at a more rapid rate at the Cairns location, as well as during the summer months, when higher water temperatures were recorded. Successful colonisation of all coatings by bacteria soon after immersion modifies the characteristics of the surfaces at the hull/water interface, and subsequent settlement by higher biofouling organisms must be moderated by these modified surfaces.