Transcriptome profiling of sulfate deprivation responses in two agarophytes Gracilaria changii and Gracilaria salicornia (Rhodophyta)

Seaweeds survive in marine waters with high sulfate concentration compared to those living at freshwater habitats. The cell wall polymer of Gracilaria spp. which supplies more than 50% of the world agar is heavily sulfated. Since sulfation reduces the agar quality, it is interesting to investigate the effects of sulfate deprivation on the sulfate contents of seaweed and agar, as well as the metabolic pathways of these seaweeds. In this study, two agarophytes G. changii and G. salicornia were treated under sulfate deprivation for 5 days. The sulfate contents in the seaweed/agar were generally lower in sulfate-deprivated samples compared to those in the controls, but the differences were only statistically significant for seaweed sample of G. changii and agar sample of G. salicornia. RNA sequencing (RNA-Seq) of sulfate-deprivated and untreated seaweed samples revealed 1,292 and 3,439 differentially expressed genes (DEGs; ≥1.5-fold) in sulfate-deprivated G. changii and G. salicornia, respectively, compared to their respective controls. Among the annotated DEGs were genes involved in putative agar biosynthesis, sulfur metabolism, metabolism of sulfur-containing amino acids, carbon metabolism and oxidative stress. These findings shed light on the sulfate deprivation responses in agarophytes and help to identify candidate genes involved in agar biosynthesis.

[1]  W. Yaphe,et al.  The agar polysaccharides of Gracilaria species , 1971 .

[2]  J. Mclachlan,et al.  Gracilaria (Gigartinales, Rhodophyta) and productivity☆ , 1986 .

[3]  A. Grossman,et al.  Purification and biosynthesis of a derepressible periplasmic arylsulfatase from Chlamydomonas reinhardtii , 1988, The Journal of cell biology.

[4]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[5]  U. Karsten,et al.  The effect of light intensity and daylength on the β‐dimethylsulphoniopropionate (DMSP) content of marine green macroalgae from Antarctica* , 1990 .

[6]  G. Kirst,et al.  The effect of nitrogen deficiency, methionine and inhibitors of methionine metabolism on the DMSP contents of Tetraselmis subcordiformis (Stein) , 1992 .

[7]  [Roles of anions in cells: studies on a Cl(-)-translocating ATPase and sulfate uptake system in Acetabularia acetabulum]. , 1994, Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan.

[8]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[9]  D W Nebert,et al.  The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. , 1997, Pharmacogenetics.

[10]  A. Hanson,et al.  Identification and stereospecificity of the first three enzymes of 3-dimethylsulfoniopropionate biosynthesis in a chlorophyte alga , 1998 .

[11]  T. Leustek,et al.  PATHWAYS AND REGULATION OF SULFUR METABOLISM REVEALED THROUGH MOLECULAR AND GENETIC STUDIES. , 2000, Annual review of plant physiology and plant molecular biology.

[12]  Kazuki Saito Regulation of sulfate transport and synthesis of sulfur-containing amino acids. , 2000 .

[13]  O. Schofield,et al.  Characterization of sulfate assimilation in marine algae focusing on the enzyme 5'-adenylylsulfate reductase. , 2000, Plant physiology.

[14]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[15]  F. Speleman,et al.  Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes , 2002, Genome Biology.

[16]  T. Kuroiwa,et al.  The Second Serine Acetyltransferase, Bacterial-type O-Acetylserine (thiol) Lyase and Eukaryotic-type O-Acetylserine (thiol) Lyase from the Primitive Red Alga Cyanidioschyzon merolae , 2001, Journal of Plant Research.

[17]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[18]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[19]  T. Leustek,et al.  The sac Mutants of Chlamydomonas reinhardtii Reveal Transcriptional and Posttranscriptional Control of Cysteine Biosynthesis1 , 2002, Plant Physiology.

[20]  Holger Hesse,et al.  Transcriptome analysis of sulfur depletion in Arabidopsis thaliana: interlacing of biosynthetic pathways provides response specificity. , 2003, The Plant journal : for cell and molecular biology.

[21]  Akiko Maruyama-Nakashita,et al.  Transcriptome Profiling of Sulfur-Responsive Genes in Arabidopsis Reveals Global Effects of Sulfur Nutrition on Multiple Metabolic Pathways1[w] , 2003, Plant Physiology.

[22]  John Quackenbush,et al.  TIGR Gene Indices clustering tools (TGICL): a software system for fast clustering of large EST datasets , 2003, Bioinform..

[23]  E. Murano Chemical structure and quality of agars from Gracilaria , 1995, Journal of Applied Phycology.

[24]  I. Kohane,et al.  Gene regulation and DNA damage in the ageing human brain , 2004, Nature.

[25]  S. Phang,et al.  Studies on Gracilaria changii (Gracilariales, Rhodophyta) from Malaysian mangroves , 1996, Hydrobiologia.

[26]  C. Ho,et al.  Optimisation of RNA extraction from Gracilaria changii (Gracilariales, Rhodophyta) , 2004, Journal of Applied Phycology.

[27]  Jeff Shrager,et al.  Insights into the Survival of Chlamydomonas reinhardtii during Sulfur Starvation Based on Microarray Analysis of Gene Expression , 2004, Eukaryotic Cell.

[28]  K. Saito Sulfur Assimilatory Metabolism. The Long and Smelling Road1 , 2004, Plant Physiology.

[29]  M. Hirai,et al.  Integration of transcriptomics and metabolomics for understanding of global responses to nutritional stresses in Arabidopsis thaliana. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[30]  F. Kauffman Sulfonation in Pharmacology and Toxicology , 2004, Drug metabolism reviews.

[31]  Juan Miguel García-Gómez,et al.  BIOINFORMATICS APPLICATIONS NOTE Sequence analysis Manipulation of FASTQ data with Galaxy , 2005 .

[32]  Shu-Dong Zhang,et al.  Bioinformatics Original Paper Effect of Pooling Samples on the Efficiency of Comparative Studies Using Microarrays , 2022 .

[33]  Lin Fang,et al.  WEGO: a web tool for plotting GO annotations , 2006, Nucleic Acids Res..

[34]  V. Irihimovitch,et al.  The sulfur acclimation SAC3 kinase is required for chloroplast transcriptional repression under sulfur limitation in Chlamydomonas reinhardtii , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[35]  T. Kajiwara,et al.  Physical and Chemical Characterization of Agar Polysaccharides Extracted from the Thai and Japanese Species of Gracilaria , 2006 .

[36]  Yuki Moriya,et al.  KAAS: an automatic genome annotation and pathway reconstruction server , 2007, Nucleic Acids Res..

[37]  J. Léger,et al.  Response of the transcriptome of the intertidal red seaweed Chondrus crispus to controlled and natural stresses. , 2007, The New phytologist.

[38]  A. J. Smit,et al.  Dimethylsulfoniopropionate (DMSP) accumulation in abalone Haliotis midae (Mollusca: Prosobranchia) after consumption of various diets, and consequences for aquaculture , 2007 .

[39]  M. Stephens,et al.  RNA-seq: an assessment of technical reproducibility and comparison with gene expression arrays. , 2008, Genome research.

[40]  C. Ho,et al.  Profiling the Transcriptome of Gracilaria changii (Rhodophyta) in Response to Light Deprivation , 2009, Marine Biotechnology.

[41]  J. Rupprecht,et al.  Transcriptome for Photobiological Hydrogen Production Induced by Sulfur Deprivation in the Green Alga Chlamydomonas reinhardtii , 2008, Eukaryotic Cell.

[42]  A. Khodursky,et al.  Gene expression patterns of sulfur starvation in Synechocystis sp. PCC 6803 , 2008, BMC Genomics.

[43]  B. Williams,et al.  Mapping and quantifying mammalian transcriptomes by RNA-Seq , 2008, Nature Methods.

[44]  A. Grossman,et al.  The Central Role of a SNRK2 Kinase in Sulfur Deprivation Responses1[W][OA] , 2008, Plant Physiology.

[45]  Daniel R. Zerbino,et al.  Pebble and Rock Band: Heuristic Resolution of Repeats and Scaffolding in the Velvet Short-Read de Novo Assembler , 2009, PloS one.

[46]  B. Yan,et al.  Cloning and analysis of the galactose-1-phosphate uridylyltransferase (galt) gene of Gracilariopsis lemaneiformis (Rhodophyta) and correlation between gene expression and agar synthesis , 2010, Journal of Applied Phycology.

[47]  M. Gerstein,et al.  RNA-Seq: a revolutionary tool for transcriptomics , 2009, Nature Reviews Genetics.

[48]  C. Ho,et al.  TRANSCRIPTOMIC ANALYSIS OF GRACILARIA CHANGII (RHODOPHYTA) IN RESPONSE TO HYPER‐ AND HYPOOSMOTIC STRESSES 1 , 2009, Journal of phycology.

[49]  B. Kloareg,et al.  The Cyclization of the 3,6-Anhydro-Galactose Ring of ι-Carrageenan Is Catalyzed by Two d-Galactose-2,6-Sulfurylases in the Red Alga Chondrus crispus1 , 2009, Plant Physiology.

[50]  Matteo Pellegrini,et al.  RNA-Seq Analysis of Sulfur-Deprived Chlamydomonas Cells Reveals Aspects of Acclimation Critical for Cell Survival[W] , 2010, Plant Cell.

[51]  Harris J. Bixler,et al.  A decade of change in the seaweed hydrocolloids industry , 2011, Journal of Applied Phycology.

[52]  Hideki Takahashi,et al.  Sulfur assimilation in photosynthetic organisms: molecular functions and regulations of transporters and assimilatory enzymes. , 2011, Annual review of plant biology.

[53]  Y. Qi,et al.  Phaeocystis globosa Scherffel, a harmful microalga, and its production of dimethylsulfoniopropionate , 2011 .

[54]  A. Usov,et al.  Polysaccharides of the red algae. , 2011, Advances in carbohydrate chemistry and biochemistry.

[55]  Zhongming Zhao,et al.  RNA-Seq analysis implicates dysregulation of the immune system in schizophrenia , 2012, BMC Genomics.

[56]  C. Ho,et al.  MOLECULAR CLONING AND BIOCHEMICAL CHARACTERIZATION OF GALACTOSE‐1‐PHOSPHATE URIDYLYLTRANSFERASE FROM GRACILARIA CHANGII (RHODOPHYTA) 1 , 2012, Journal of phycology.

[57]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[58]  Wei-Kang Lee,et al.  Effects of sulfate starvation on agar polysaccharides of Gracilaria species (Gracilariaceae, Rhodophyta) from Morib, Malaysia , 2013, Journal of Applied Phycology.

[59]  Susana M. Coelho,et al.  Genome structure and metabolic features in the red seaweed Chondrus crispus shed light on evolution of the Archaeplastida , 2013, Proceedings of the National Academy of Sciences.

[60]  C. Ho,et al.  Molecular cloning and characterization of GDP-mannose-3′,5′-epimerase from Gracilaria changii , 2013, Journal of Applied Phycology.

[61]  Graham J. Etherington,et al.  Transcriptome analysis of the sulfate deficiency response in the marine microalga Emiliania huxleyi. , 2013, The New phytologist.

[62]  W. Scheible,et al.  Transcriptome and metabolome analysis of plant sulfate starvation and resupply provides novel information on transcriptional regulation of metabolism associated with sulfur, nitrogen and phosphorus nutritional responses in Arabidopsis , 2015, Front. Plant Sci..

[63]  Han Liu,et al.  Whole transcriptome expression profiling of mouse limb tendon development by using RNA‐seq , 2015, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[64]  S. Kiel Did shifting seawater sulfate concentrations drive the evolution of deep-sea methane-seep ecosystems? , 2015, Proceedings of the Royal Society B: Biological Sciences.

[65]  S. Benjakul,et al.  Physico-chemical and gel properties of agar from Gracilaria tenuistipitata from the lake of Songkhla, Thailand , 2015 .

[66]  G. Michel,et al.  Sweet and sour sugars from the sea: the biosynthesis and remodeling of sulfated cell wall polysaccharides from marine macroalgae , 2015 .

[67]  Wei-Kang Lee,et al.  Agar properties of Gracilaria species (Gracilariaceae, Rhodophyta) collected from different natural habitats in Malaysia , 2016 .

[68]  M. Tatar,et al.  Nutritional Geometric Profiles of Insulin/IGF Expression in Drosophila melanogaster , 2016, PloS one.

[69]  Martin Krzywinski,et al.  Study Design for Sequencing Studies , 2016, Statistical Genomics.

[70]  C. Ho,et al.  De novo transcriptome analyses of host-fungal interactions in oil palm (Elaeis guineensis Jacq.) , 2016, BMC Genomics.

[71]  Wei-Kang Lee,et al.  Biosynthesis of agar in red seaweeds: A review. , 2017, Carbohydrate Polymers.