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Agar and agarose have wide applications in food and pharmaceutical industries. Knowledge on the genome of red seaweeds that produce them is still lacking. To fill the gap in genome analyses of these red algae, we have sequenced the nuclear and organellar genomes of an agarophyte, Gracilaria changii. The partial nuclear genome sequence of G. changii has a total length of 35.8Mb with 10,912 predicted protein coding sequences. Only 39.4% predicted proteins were found to have significant matches to protein sequences in SwissProt. The chloroplast genome of G. changii is 183,855bp with a total of 201 open reading frames (ORFs), 29 tRNAs and 3 rRNAs predicted. Five genes: ssrA, leuC and leuD CP76_p173 (orf139) and pbsA were absent in the chloroplast genome of G. changii. The genome information is valuable in accelerating functional studies of individual genes and resolving evolutionary relationship of red seaweeds., (Copyright © 2017 Elsevier Inc. All rights reserved.)

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.

Many bacterial epiphytes of agar-producing seaweeds secrete agarase that degrade algal cell wall matrix into oligoagars which elicit defense-related responses in the hosts. The molecular defense responses of red seaweeds are largely unknown. In this study, we surveyed the defense-related transcripts of an agarophyte, Gracilaria changii, treated with β-agarase through next generation sequencing (NGS). We also compared the defense responses of seaweed elicited by agarase with those elicited by an agarolytic bacterium isolated from seaweed, by profiling the expression of defense-related genes using quantitative reverse transcription real-time PCR (qRT-PCR). NGS detected a total of 391 differentially expressed genes (DEGs) with a higher abundance (>2-fold change with a p value <0.001) in the agarase-treated transcriptome compared to that of the non-treated G. changii. Among these DEGs were genes related to signaling, bromoperoxidation, heme peroxidation, production of aromatic amino acids, chorismate, and jasmonic acid. On the other hand, the genes encoding a superoxide-generating NADPH oxidase and related to photosynthesis were downregulated. The expression of these DEGs was further corroborated by qRT-PCR results which showed more than 90 % accuracy. A comprehensive analysis of their gene expression profiles between 1 and 24 h post treatments (hpt) revealed that most of the genes analyzed were consistently upregulated or downregulated by both agarase and agarolytic bacterial treatments, indicating that the defense responses induced by both treatments are highly similar except for genes encoding vanadium bromoperoxidase and animal heme peroxidase. Our study has provided the first glimpse of the molecular defense responses of G. changii to agarase and agarolytic bacterial treatments.

Light regulates photosynthesis, growth and reproduction, yield and properties of phycocolloids, and starch contents in seaweeds. Despite its importance as an environmental cue that regulates many developmental, physiological, and biochemical processes, the network of genes involved during light deprivation are obscure. In this study, we profiled the transcriptome of Gracilaria changii at two different irradiance levels using a cDNA microarray containing more than 3,000 cDNA probes. Microarray analysis revealed that 93 and 105 genes were up- and down-regulated more than 3-fold under light deprivation, respectively. However, only 50% of the transcripts have significant matches to the nonredundant peptide sequences in the database. The transcripts that accumulated under light deprivation include vanadium chloroperoxidase, thioredoxin, ferredoxin component, and reduced nicotinamide adenine dinucleotide dehydrogenase. Among the genes that were down-regulated under light deprivation were genes encoding light harvesting protein, light harvesting complex I, phycobilisome 7.8 kDa linker polypeptide, low molecular weight early light-inducible protein, and vanadium bromoperoxidase. Our findings also provided important clues to the functions of many unknown sequences that could not be annotated using sequence comparison.

Galactose-1-phosphate uridylyltransferase (GALT) catalyzes the reversible conversion of glucose-1-phosphate and UDP-galactose to galactose-1-phosphate and UDP-glucose. This enzyme is also responsible for one of the biochemical steps that produce the precursors of agar and agarose. In this study, we report the molecular cloning and sequence analyses of a cDNA encoding GALT, from Gracilaria changii (B. M. Xia et I. A. Abbott) I. A. Abbott, J. Zhang et B. M. Xia, which constitutes a genus of seaweeds that supply more than 60% of the world's agar and agarose. We have subcloned this cDNA into a bacterial expression cloning vector and characterized the enzyme activities of its recombinant proteins in vitro. The GcGALT gene was shown to be up-regulated by salinity stresses. The abundance of transcripts encoding GcGALT was the highest in G. changii, followed by Gracilaria edulis and Gracilaria salicornia in a descending order, corresponding to their respective agar contents. Our findings indicated that GALT could be one of the components that determines the agar yield in Gracilaria species. [ABSTRACT FROM AUTHOR]

Red seaweeds of the genus Gracilaria are agarophytes that produce more than 60% of the world's agar supply. Despite the importance of this genus in agar production, the potential of Gracilaria as a candidate for genomic research has been almost unexplored. In this study, a total of 8,088 expressed sequence tags (ESTs) were generated from Gracilaria changii, and clustered into 4922 tentative unique genes (TUGs), of which approximately 35% showed significant matches (bit scores greater than 50 and E-values less than 10-5) to other genes in the databases. Among the TUGs that have significant similarity to the genes in the databases are ESTs corresponding to diverse functional groups such as metabolism, transcription, signalling, translation, transportation, protein folding, sorting, destination and degradation, cell division, cellular processes, replication and repair, cell structure, and miscellaneous. cDNAs involved in diverse metabolic pathways were identified among the EST collection. The presence and frequency of the transcripts allow us to survey the transcriptomic activities of this tropical agarophyte. [ABSTRACT FROM AUTHOR]

The supply of agar as an important gelling and thickening agent in various industrial applications depends heavily on harvesting of natural seaweed resources and seaweed farming. To facilitate the selection of good seaweed source with higher agar yield and stronger gel strength, accurate and rapid screening method using molecular markers is necessary to replace the tedious, laborious and time-consuming conventional method which involves agar extraction and gel analysis. In this study, we characterized the expression of a number of algal transcripts and proteins from an agar producing seaweed, Gracilaria changii with the aim to identify potential markers for agar yield and gel strength. In total, 15 candidate transcripts that are directly or indirectly related to putative agar biosynthetic pathway were identified based on literature search. The transcript abundance of 4 and 11 of these candidates were found to be significantly (P < 0.05) correlated to the agar yield and gel strength of six G. changii samples, respectively. Among these marker genes, the transcript levels of GcFBPA and GcGALE have the highest linear correlation to both agar yield and gel strength. The protein abundance of GcFBPA and GcGALE was further examined on 13 G. changii samples and was found to have highly significant (P < 0.01) correlation to agar gel strength and agar yield, respectively. GcFBPA and GcGALE may have good potential to be used for molecular screening of yield traits and gel quality of G. changii at both RNA and protein levels. • The abundance of 11 transcripts correlated (P < 0.05) with agar yield and/or gel strength of G. changii. • The protein abundance of GcGALE correlated (P < 0.01) with agar yield. • The protein abundance of GcFBPA correlated (P < 0.01) with gel strength. • GcFBPA and GcGALE have potential to be markers for agar yield or gel strength. [ABSTRACT FROM AUTHOR]