Comparative chloroplast genomes of Ulva prolifera and U. linza (Ulvophyceae) provide genetic resources for the development of interspecific markers

Journal of Oceanology and Limnology - The green seaweeds Ulva linza and U. prolifera are closely related species. They usually co-occur widely and have important ecological significance as primary...     

Investigation of the decline of Ulva prolifera in the Subei Shoal and Qingdao based on physiological changes

It has been a scientific consensus that Ulva prolifera green tide that break out annually between May and August in the recent decade off eastern coast of Yellow Sea. To investigate the ecological mechanism of the decline of U. prolifera green tide, we compared the physiological characteristics of U. prolifera samples collected in three stages of the green tide in Subei Shoal north Jiangsu, the initiation point, on May 10(early, the initiation), July 10(middle, the outbreak), and in Taipingjiao, Qingdao on August 10(late, the decline). Compared to the early samples, the middle, and late samples showed a lower chlorophyll content and photosynthetic e fficiency at a higher lipid peroxidation level. The middle and later samples had fewer chloroplasts in the cells of U. prolifera than those in the early samples. In addition, the cells of U. prolifera sampled in the late stage showed ruptured and deformed cell walls. Differences in physiological function were reflected in the growth rate. Both the middle and late U. prolifera samples presented negative growth rates. Correlation analysis results show that the rising temperature was mainly responsible for the local decline of U. prolifera green tide. The high light-intensity that could induce the photoinhibition was also an important factor for the decline. Therefore, U. prolifera population that remained in the Subei Shoal and those that northward drifted to Qingdao experienced the same decline processes: rising temperature and continuous high light conditions in the late phase of green tide reduced the photosynthetic capacity and destroyed the antioxidant system of the thallus, which lead to decrease of biomass. The only difference is that the temperature in the Subei Shoal rose earlier than that in Qingdao; thus, U. prolifera remaining in the Subei Shoal went disappeared first.     

Sequences and phylogeny analysis of <Emphasis Type="Italic">rbcL</Emphasis> gene in marine chlorophyta

The rbcL gene of Ulva pertusa,Enteromorpha prolifera and Monostroma grevillei was amplified,sequenced and analyzed.By comparing the rbcL sequences with seven other Ulvales species retrieved from GenBank,the sequence divergences and the phyletic evolution were analyzed and the phylogenetic tree was constructed.From the phylogenetic tree,it can be found that U.pertusa,E.prolifera and U.californica group in one branch,while E.compressa,U.rigida and U.fenestrata cluster in another clade.Obviously,unlike the Enteomorpha species,the Ulva species do not gather in one branch.Therefore Ulva and Enteomorpha might be affiliates of one genus.E.compressa and E.intestinalis gathered together,which coincided with the morphological characters.However,the thallus of U.pertusa is thick and with many holes,which is different from E.prolifera in morphology.They cluster together in the phylogenetic tree with a genetic distance of 0.005.The results indicate that Ulva and Enteromorpha are not distinguished strictly.     

A New Molecular Label Applied to the Study of the Yellow Sea Green Tide

From 2007 to 2017, large-scale green tides occurred every year in the Yellow Sea of China, and Ulva prolifera was the main species leading to the green tide. In this study, we used the Polymerase chain reaction and 3' Rapid-amplification of c DNA ends technique to amplify the nr DNA-LSU and IGS sequences in U. prolifera and one species of Blidingia. These techniques showed 3259 bp of nrD NA-LSU and 3388 bp of IGS in U. prolifera and 3282 bp nr DNA-LSU and 3059 bp IGS in Blidingia sp. At the same time, tandem repeats, short dyads, palindromic and multiple simple repeat sequences in the IGS sequence were found by analyzing the structure of the IGS sequence in U. prolifera and Blidingia sp. G + C contents of the IGS sequence in U. prolifera and Blidingia sp. were 52.42% and 53.09%, respectively. We divided the U. prolifera into two types according to the morphological characteristics. Although the specimens of U. prolifera from the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea have different morphologies, their ITS and IGS sequences are almost identical. Therefore, the main species of the green tides in the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea are the same and have the same origin.     

Analysis of expressed sequence tags from the Ulva prolifera (Chlorophyta)

In 2008, a green tide broke out before the sailing competition of the 29th Olympic Games in Qingdao. The causative species was determined to be Enteromorpha prolifera (Ulva prolifera O. F. Müller), a familiar green macroalga along the coastline of China. Rapid accumulation of a large biomass of floating U. prolifera prompted research on different aspects of this species. In this study, we constructed a nonnormalized cDNA library from the thalli of U. prolifera and acquired 10 072 high-quality expressed sequence tags (ESTs). These ESTs were assembled into 3 519 nonredundant gene groups, including 1 446 clusters and 2 073 singletons. After annotation with the nr database, a large number of genes were found to be related with chloroplast and ribosomal protein, GO functional classification showed 1 418 ESTs participated in photosynthesis and 1 359 ESTs were responsible for the generation of precursor metabolites and energy. In addition, rather comprehensive carbon fixation pathways were found in U. prolifera using KEGG. Some stress-related and signal transduction-related genes were also found in this study. All the evidences displayed that U. prolifera had substance and energy foundation for the intense photosynthesis and the rapid proliferation. Phylogenetic analysis of cytochrome c oxidase subunit I revealed that this green-tide causative species is most closely affiliated to Pseudendoclonium akinetum (Ulvophyceae).     

DNA barcoding assessment of green macroalgae in coastal zone around Qingdao,China

An assessment with assistance of DNA barcoding was conducted on green macroalgae in coastal zone around Qingdao, China, during the period of April–December, 2011. Three markers were applied in molecular discrimination, including the plastid elongation factor tufA gene, the internal transcribed spacer (ITS) region of the ribosomal cistron and rubisco large subunit gene 3′ regions (rbcL-3P). DNA barcoding discriminated 8 species, excluding species of genus Cladophora and Bryopsis due to failures in amplification. We ascertained and corrected 4 species identified by morphological methods for effectively assisting the classification. The gene tufA presented more advantages as an appropriate DNA marker with the strongest amplification success rate and species discrimination power than the other two genes. The poorest sequencing success largely handicapped the application of ITS. Samples identified by tufA and rbcL as Ulva flexuosa were clustered into the clade of U. prolifera by ITS in the neighbor-joining tree. Confusion with discrimination of the complex of U. linza, U. procera and U. prolifera (as the LPP complex) still existed for the three DNA markers. Based on our results, rbcL is recommended as a preferred marker for assisting tufA to discriminate green macroalgae. In distinguishing green-tide-forming Ulva species, the free-floating sample collected from the green tide in 2011 was proved to be identical with U. prolifera in Yellow Sea for ITS and rbcL genes. This study presents a preliminary survey of green macroalgae distributed in the coastal area around Qingdao, and proves that DNA barcoding is a powerful tool for taxonomy of green macroalgae.     

First record of Ulva californica in the mainland of China: a single alien parthenogenetic population in discontinuous distribution

Journal of Oceanology and Limnology - Molecular investigations have raised concerns about the ecological risks of green tides caused by alien Ulva species in new habitats. The green tide-forming...     

Phylogenetic analyses of four species of Ulva and Monostroma grevillei using ITS, rbc L and 18S rDNA sequence data

Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer (ITS) regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-1 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulva prolifera (or U. pertusa) gene sequences from Qingdao and Nan’ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U. flexuosa had the least evolutionary distance from U. californica in both the ITS regions (0.009) and the 18S rDNA (0.002). These data verified that Ulva and Enteromorpha are not separate genera.     

Dynamic Diurnal Changes in Green Algae Biomass in the Southern Yellow Sea Based on GOCI Images

Macroalgae blooms of Ulva prolifera have occurred every summer in the southern Yellow Sea since 2007, inducing severe ecological problems and huge economic losses. Genesis and secular movement of green algae blooms have been well monitored by using remote sensing and other methods. In this study, green algae were detected and traced by using Geostationary Ocean Color Imager(GOCI), and a novel biomass estimation model was developed from the relationship between biomass measurements and previously published satellite-derived biomass indexes. The results show that the green algae biomass can be determined most accurately with the biomass index of green algae for GOCI(BIGAG), which is calculated from the Rsurf data that had been atmospherically corrected by ENVI/QUAC method. For the first time, dynamic changes in green algae biomass were studied over an hourly scale. Short-term biomass changes were highly influenced by Photosynthetically Available Radiation(PAR) and tidal phases, but less by sea surface temperature variations on a daily timescale. A new parameter of biomass changes(PBC), calculated by the ratio of the biomass growth rate to movement velocity, could provide an effective way to assess and forecast green tide in the southern Yellow Sea and similar areas.     

The distribution of green algal micro-propagules and macroalgae at the early stage of green tide in the coastal area of South Jiangsu Province in 2014

An integrated investigation was carried out during late April to late May of 2014 in the coastal area of South Jiangsu Province to understand the early development of green tide in the Yellow Sea and discover the temporal and spatial distribution of green algal micro-propagules and macroalgae at the early stage of green tide. The results showed that green algal micro-propagules distributed in all stations from late April to late May, and micro-propagules of high density( 300 ind. L~(-1)) concentrated in the adjacent Porphyra aquaculture area. Floating macroalgae were initially observed in the northern survey areas, and increased gradually. The dominant species was U. prolifera in the floating macroalgae, ranging between 90% and 100%. The seawater surface temperature and salinity in the above mentioned time period were suitable for the growth of macroalgae. This work can help government to strengthen management to reduce the blooms of macroalgae in the coastal area of South Jiangsu Province and also facilitate the decision-making for managers at the early stage of green tide.     

Antioxidant system responses in two co-occurring green-tide algae under stress conditions

Green tides have occurred every year from 2007 to 2014 in the Yellow Sea. Ulva prolifera(Müller) J. Agardh has been identified as the bloom-forming alga,co-occurring with U. intestinalis. We observed distinct strategies for both algal species during green tides. U. prolifera exhibited a high abundance initially and then decreased dramatically,while U. intestinalis persisted throughout. The antioxidant system responses of these two macroalgae were compared in the late phase of a green tide(in-situ) and after laboratory acclimation. Lipid peroxidation and antioxidant system responses differed significantly between the two. Malondialdehyde and hydrogen peroxide contents increased significantly in-situ in U. prolifera,but not in U. intestinalis. In U. prolifera,we observed a significant decrease in total antioxidant ability(T-AOC),antioxidant enzymes(SOD and Apx),and non-enzyme antioxidants(GSH and As A) in-situ. U. intestinalis showed the same pattern of T-AOC and SOD,but its Gpx,Apx,and GSH responses did not differ significantly. The results suggest that U. prolifera was more susceptible than U. intestinalis to the harsh environmental changes during the late phase of a Yellow Sea green tide. The boom and bust strategy exhibited by U. prolifera and the persistence of U. intestinalis can be explained by differences in enzyme activity and antioxidant systems.     

The possibility analysis of habitats, origin and reappearance of bloom green alga （Enteromorpha prolifera） on inshore of western Yellow Sea

Combining some information from field investigation of algae along the coastal areas in China and a few pictures materialized from the western Yellow Sea in 2008,authors analyze the necessary conditions and possible water area in China producing a large biomass,some reasons for firestorm,and the possibility of the reappearance of marine bloom green alga Enteromorpha prolifera.The change of habitats and the increase of nutritional levels related to the water area could be considered as direct reasons.It was ...     

Karyological observations of Ulva linza chromosomes

Journal of Oceanology and Limnology - Ulva linza is one of the species that causes green tides in the Yellow Sea, China. Due to the difficulties in chromosomal preparation, the large numbers of...     

The appearance of Ulva laetevirens (Ulvophyceae,Chlorophyta) in the northeast coast of the United States of America

Introduced species may outcompete or hybridize with native species, resulting in the loss of native biodiversity or even alteration of ecosystem processes. In this study, we reported an alien distromatic Ulva species, which was found in an embayment (Holly Pond) connected with Long Island Sound, USA. The morphological and anatomical observations in combination with molecular data were used for its identification to species. Anatomy of collected specimens showed that the cell shape in rhizoidal and basal regions was round and the marginal teeth along the basal and median region were not found. These characteristics were primarily identical to the diagnostic characteristics of Ulva laetevirens Areschoug (Chlorophyta). The plastid-encoding tufA and nucleusencoding ITS1 were used for its molecular identification. Phylogenetic analysis for the tufA gene placed the specimens from Holly Pond in a well-supported clade along with published sequences of U. laetevirens identified early without any sequence divergence. In ITS tree, the sample also formed well-supported clades with the sequences of U. laetevirens with an estimated sequence divergence among the taxa in these clades as low as 1%. These findings confirmed the morpho-anatomical conclusion. Native to Australia, this species was reported in several countries along the Mediterranean coast after the late of 1990s. This is the first time that U. laetevirens is found in the northeast coast of United States and the second record for Atlantic North America.     

Long interspersed elements in three species of Gracilaria （Gracilariaceae, Rhodophyta）

In order to find out whether long interspersed elements (LINEs) existed in macro-algae ge- nomes or not, we tested the LINE homologues in representative families (species): Gracilaria (G. eucheumoides Harv., G. tenuistipitata Chang et Xia, and G. textorii (Sur) De-Toni), Laminaria (L. longis- sima Miyabe and L. japonica Aresch.), and Ulva (U. lactuca L. and U. pertusa Kjellm.) during 2004 to 2005. Polymerase Chain Reaction (PCR) was carried out with degenerate oligonucleotide primers de- signed from LINEs of rice homologues and Cin4 of maize. Cloning and nucleotide sequencing of the PCR products revealed that 4 clones that derived from 3 species of Gracilaria have LINE homologues. The nucleotide sequences of the 4 LINE homologues diverged greatly, but the amino acid sequences deduced from them were relatively conserved. The endonuclease regions of the LINE homologues greatly di- verged from that of other plants, but they had closer phylogenetic relationship to Zepp elements in Chlor- ella sp., which indicated that sequence divergence by vertical transmission has been a major influence on the evolution of algal LINEs.     

Preference of the herbivorous marine teleost Siganus canaliculatus for different macroalgae

The decomposition of a large amount of unexploited macroalgal resource along the coast of China often results in heavy environmental pollution. In order to pave a way of using macroalgae as the dietary ingredient of rabbitfish Siganus canaliculatus, one of a few farmed herbivorous marine teleosts in China, its preference （feeding selectivity） for different macroalgae was determined in this study. Seven seaweed species abundantly inhabiting the coast of east Guangdong Province were exposed simultaneously to rab-bitfish juveniles in laboratory （multiple-choice feeding） with their content and absolute intake assayed. It was found that the most preferred algae were Ulva prolifera, Gracilaria lemaneiformis and Chaetomorpha linum, less preferred algae were U. pertusa and Porphyra haitanensis, and least preferred ones were Sargassum fusiforme and Corallina sessilis. Such an order did not change when one to four relatively preferred seaweeds were removed. The preferred seaweeds were richer in protein and soluble sugar thus higher in energy than the least preferred. In addition, this fish was found to favor filamentous and flat algae rather than calcified ones. Ac-cordingly, the richness of nutrients and morphological characteristics determined the preference of S. canaliculatus for tested macro-algae.     

Genome-Wide Patterns of Codon Usage in the Pacific Oyster Genome

The use of synonymous codons with varying frequencies has been observed in many species, and the magnitude varies among closely related species and genes within the same genome. Mutational processes or natural selective pressures affect this bias.However, a systematic investigation of codon usage pattern for molluscan species and its association with the two factors hasn't been conducted. In this study, the whole genome codon usage patterns in oyster genome is investigated using the relative synonymous codon usage(RSCU) method, and 17 putative optimal codons were identified, wherein most had either a cytosine(C) or guanine(G)residue at the third position. Results showed that codon bias was positively associated with gene expression levels and breadth. Optimal codons had different mutational bias patterns compared with nonoptimal codons. Moreover, the results show that codon bias is negatively associated with nucleotide diversity. In the oyster genome, the fourfold degenerate codons are affected by different selective pressures, which can be regarded as an evidence that natural selection shapes codon usage patterns. This research will help to understand how natural selection and mutation bias affect codon usage in Mollusca genomes.     

The green tide in Yingkou,China in summer 2021 was caused by a subtropical alga—Ulva meridionalis (Ulvophyceae,Chlorophyta)

Journal of Oceanology and Limnology - The large-scale green tide caused by Ulva has occurred successively in the Yellow Sea since 2007, and new events of green tide also continued to appear in...     

Effect of UV-B irradiation on interspecific competition between Ulva pertusa and Grateloupia filicina

We report the effect of UV-B irradiation (9.6 kJ m^?2 day^?1) on interspecific competition between two species of macroalgae, Ulva pertusa (U) and Grateloupia filicina (G), in co-culture. Growth of U. pertusa and G. filicina was inhibited by UV-B irradiation in mono-culture and specific growth rate (μ) declined as a result. Interspecific competition between U. pertusa and G. filicina was closely related to the initial weights when co-cultured. When initial ratios of U. pertusa (U) to G. filicina (G) were U:G=1.2:1 and 1:1, U. pertusa was the dominant algae. When the initial U:G ratio was 1:1.2, G. filicina was competitively dominant in the earlier stage, but U. pertusa grew faster, superseding G. filicina in the later stage. At initial ration U:G = 1:1.4, G. filicina was predominant. Under UV-B irradiation, the competitive ability of G. filicina was weakened and the interspecific competitive balance favored U. pertusa, which suggests that G. filicina was more sensitive to UV-B irradiation. We also probed the potential allelopathic effects between the two species, which led to mutual growth inhibition.     

Pyrolytic characteristics and kinetics of the marine green tide macroalgae, Enteromorpha prolifera

The marine macroalgae Enteromorpha prolifera was one of the main algal genera that occurred in the widespread green tides in Qingdao, China, during the summers of 2007, 2008 and 2010. It is thus a plentiful source of biomass and could be used as a biofuel. In this study, the pyrolytic characteristics and kinetics of E. prolifera were investigated using thermogravimetric analysis (TGA) method. Cornstalk and sawdust were used as comparisons. Pyrolytic characteristics were studied using TG-DTG (thermogravimetry-derivative thermogravimetry) curves. Three stages in the pyrolytic process were determined: dehydration, dramatic weight loss and slow weight loss. E. prolifera was pyrolyzed at a lower initial temperature than the two terrestrial biomass forms. The apparent activation energy values for the three types of biomass were calculated and the mechanism functions were determined using 16 different mechanism functions, frequently used in thermal kinetics analysis. Activation energy values varied with mechanism function and the range of activation energy values for E. prolifera, cornstalk, and sawdust were 25-50 kJ/mol, 60-90 kJ/mol and 120-155 kJ/mol, respectively. This indicates that E. prolifera has low thermal stability for pyrolysis and good combustion characteristics.