Occurrence of thraustochytrids: the fungoid protists vis-a-vis marine macroalgae (seaweeds) along the coast of Goa, India

Thraustochytrids are fungoid protists ubiquitous in the marine environment and found to be associated with decaying macroalgae. Not much is known about their association with living macroalgae. Hence in the present study, different macroalgal samples were collected from various beaches of Goa to examine the presence of thraustochytrids during a four-year-long study. Brown, red and green algae were found to be substrata of thraustochytrids. Thraustochytrids were isolated on pine pollen baiting from 17 to 58% of the specimens. Thraustochytrids isolated from various macroalgae belonged to the genera Oblongichytrium, Schizochytrium, Ulkenia, and Thraustochytrium. Labyrinthula sp. was also found once on the green alga Bryopsis hypnoides. These were generally found during dry seasons rather than in monsoons. The seasonal occurrence of thraustochytrids was found to be associated with temporal variation in macroalgal diversity. The statistical analysis supported individual or interactive effects of both factors viz, seasons and macroalgal diversity, on the occurrence of thraustochytrids. Thraustochytrids were also isolated from seawater adjoining macroalgae and from estuarine water at all times of the year. Oblongichytrium sp. was isolated from the green alga Ulva compressa and Anjuna seawater samples at the same time, thus indicating that thraustochytrids from seawater could inhabit the macroalgae.

Ulva compressa from Copper-Polluted Sites Exhibits Intracellular Copper Accumulation, Increased Expression of Metallothioneins and Copper-Containing Nanoparticles in Chloroplasts

In order to analyze the mechanisms involved in copper accumulation in Ulva compressa, algae were collected at control sites of central and northern Chile, and at two copper-polluted sites of northern Chile. The level of intracellular copper, reduced glutathione (GSH), phytochelatins (PCs), PC2 and PC4, and transcripts encoding metallothioneins (MTs) of U. compressa, UcMT1, UcMT2 and UcMT3, were determined. Algae of control sites contained around 20 g of copper g−1 of dry tissue (DT) whereas algae of copper-polluted sites contained 260 and 272 g of copper g−1 of DT. Algae of control sites and copper-polluted sites did not show detectable amounts of GSH, the level of PC2 did not change among sites whereas PC4 was increased in one of the copper-polluted sites. The level of transcripts of UcMT1 and UcMT2 were increased in algae of copper-polluted sites, but the level of UcMT3 did not change. Algae of a control site and a copper-polluted site were visualized by transmission electron microscopy (TEM) and the existence of copper in electrodense particles was analyzed using energy dispersive x-ray spectroscopy (EDXS). Algae of copper-polluted sites showed electrodense nanoparticles containing copper in the chloroplasts, whereas algae of control sites did not. Algae of a control site, Cachagua, were cultivated without copper (control) and with 10 M copper for 5 days and they were analyzed by TEM-EDXS. Algae cultivated with copper showed copper-containing nanoparticles in the chloroplast whereas control algae did not. Thus, U. compressa from copper-polluted sites exhibits intracellular copper accumulation, an increase in the level of PC4 and expression of UcMTs, and the accumulation of copper-containing particles in chloroplasts.

Cadmium Accumulation Involves Synthesis of Glutathione and Phytochelatins, and Activation of CDPK, CaMK, CBLPK, and MAPK Signaling Pathways in Ulva compressa

In order to analyze the effect of cadmium in Ulva compressa (Chlorophyta), the alga was cultivated with 10, 25, and 50 μM of cadmium for 7 days, and the level of intracellular cadmium was determined. Intracellular cadmium showed an increase on day 1, no change until day 5, and an increase on day 7. Then, the alga was cultivated with 10 μM for 7 days, and the level of hydrogen peroxide, superoxide anions, and lipoperoxides; activities of antioxidant enzymes ascorbate peroxidase (AP), dehydroascorbate reductase (DHAR), and glutathione reductase (GR); the level of glutathione (GSH) and ascorbate (ASC); and the level of phytochelatins (PCs) and transcripts encoding metallothioneins (UcMTs) levels were determined. The level of hydrogen peroxide increased at 2 and 12 h, superoxide anions on day 1, and lipoperoxides on days 3 to 5. The activities of AP and GR were increased, but not the DHAR activity. The level of GSH increased on day 1, decreased on day 3, and increased again on day 5, whereas ASC slightly increased on days 3 and 7, and activities of enzymes involved in GSH and ASC synthesis were increased on days 3 to 7. The level of PC2 and PC4 decreased on day 3 but increased again on day 5. The level of transcripts encoding UcMT1 and UcMT2 increased on days 3 to 5, mainly that of UcMT2. Thus, cadmium accumulation induced an oxidative stress condition that was mitigated by the activation of antioxidant enzymes and synthesis of GSH and ASC. Then, the alga cultivated with inhibitors of calcium-dependent protein kinases (CDPKs), calmodulin-dependent protein kinases (CaMKs), calcineurin B-like protein kinases (CBLPKs), and MAPKs and 10 μM of cadmium for 5 days showed a decrease in intracellular cadmium and in the level of GSH and PCs, with the four inhibitors, and in the level of transcripts encoding UcMTs, with two inhibitors. Thus, CDPKs, CaMK, CBLPKS, and MAPKs are involved in cadmium accumulation and GSH and PC synthesis, and GSH and PCs and/or UcMTs may participate in cadmium accumulation.

Chloroplast Genomes of the Green-Tide Forming Alga Ulva compressa: Comparative Chloroplast Genomics in the Genus Ulva (Ulvophyceae, Chlorophyta)

To understand the evolution of Ulva chloroplast genomes at intraspecific and interspecific levels, in this study, three complete chloroplast genomes of Ulva compressa Linnaeus were sequenced and compared with the available Ulva cpDNA data. Our comparative analyses unveiled many noticeable findings. First, genome size variations of Ulva cpDNAs at intraspecific and interspecific levels were mainly caused by differences in gain or loss of group I/II introns, integration of foreign DNA fragments, and content of non-coding intergenic spacer regions. Second, chloroplast genomes of U. compressa shared the same 100 conserved genes as other Ulva cpDNA, whereas Ulva flexuosa appears to be the only Ulva species with the minD gene retained in its cpDNA. Third, five types of group I introns, most of which carry a LAGLIDADG or GIY-YIG homing endonuclease, and three of group II introns, usually encoding a reverse transcriptase/maturase, were detected at 26 insertion sites of 14 host genes in the 23 Ulva chloroplast genomes, and many intron insertion-sites have been found for the first time in Chlorophyta. Fourth, one degenerate group II intron previously ignored has been detected in the infA genes of all Ulva species, but not in the closest neighbor, Pseudoneochloris marina, and the other chlorophycean taxa, indicating that it should be the result of an independent invasion event that occurred in a common ancestor of Ulva species. Finally, the seven U. compressa cpDNAs represented a novel gene order which was different from that of other Ulva cpDNAs. The structure of Ulva chloroplast genomes is not conserved, but remarkably plastic, due to multiple rearrangement events.

Comparing Complete Mitochondrion Genome of Bloom-forming Macroalgae from the Southern Yellow Sea, China

The green tide in the Southern Yellow Sea which has been erupting continuously for 14 years. Dominant species of the free-floating Ulva in the early stage of macroalgae bloom were Ulva compressa, Ulva flexuosa, Ulva prolifera, and Ulva linza along the coast of Jiangsu Province. In the present study, we carried out comparative studies on complete mitochondrion genomes of four kinds of bloom-forming green algae, and provided standard morphological characteristic pictures of these Ulva species. The maximum likelihood phylogenetic analysis showed that U. linza is the closest sister species of U. prolifera. This study will be helpful in studying the genetic diversity and identification of Ulva species.

Comparative Transcriptome Analysis Provides Insights into The Response of Ulva Compressa to The Fluctuating Salinity Conditions

Background: Ulva compressa, known as the green tide forming species, was reported that can adapt to hypo-salinity conditions such as estuaries and brackish lakes. To understand the underlying molecular mechanisms of hypo-salinity stress tolerance, a genome-wide gene expression profiles in U. compressa was performed using digital gene expression profile (DGE). Results: The RNA-seq data were analyzed based on the comparison of differently expressed genes involved in specific pathways under hypo-salinity and recovery conditions. Under the long-term hypo-salinity stress, the recovery of photosynthesis and energy metabolism could provide sufficient energy for the tolerance under long-term hypo-saline stress. Multiple strategies were performed to maintain the osmotic homeostasis. Additionally, several long non-coding RNA were detected as differently expressed genes during the stress, which could play important roles in the osmotolerance. Conclusions: Our work will serve as an essential foundation for the understanding of the tolerance mechanism of U. compressa under the fluctuating salinity conditions.