Guanpin Yang

Silencing UDP-glucose pyrophosphorylase gene in Phaeodactylum tricornutum affects carbon allocation

The effects of the suppression of UDP-glucose pyrophosphorylase (UGPase) on chrysolaminaran biosynthesis and carbon allocation were investigated in Phaeodactylum tricornutum. The 69% decrease in UGPase activity was accompanied by a 4.89 fold reduction in Ugp transcript abundance. Inactivation of UGPase in P. tricornutum led to a significant decrease in chrysolaminaran content and an increase in lipid synthesis. These findings suggest that UGPase is a rate-limiting enzyme and may play an important role in chrysolaminarin biosynthesis and carbon allocation. Our results support a theoretical deduction that Ugp is a good candidate for improving lipid synthesis in diatoms.

Large-scale biodiesel production using flue gas from coal-fired power plants with Nannochloropsis microalgal biomass in open raceway ponds

The potential use of microalgal biomass as a biofuel source has raised broad interest. Highly effective and economically feasible biomass generating techniques are essential to realize such potential. Flue gas from coal-fired power plants may serve as an inexpensive carbon source for microalgal culture, and it may also facilitate improvement of the environment once the gas is fixed in biomass. In this study, three strains of the genus Nannochloropsis (4-38, KA2 and 75B1) survived this type of culture and bloomed using flue gas from coal-fired power plants in 8000-L open raceway ponds. Lower temperatures and solar irradiation reduced the biomass yield and lipid productivities of these strains. Strain 4-38 performed better than the other two as it contained higher amounts of triacylglycerols and fatty acids, which are used for biodiesel production. Further optimization of the application of flue gas to microalgal culture should be undertaken.

CONSTRUCTION AND CHARACTERIZATION OF A TENTATIVE AMPLIFIED FRAGMENT LENGTH POLYMORPHISM–SIMPLE SEQUENCE REPEAT LINKAGE MAP OF LAMINARIA (LAMINARIALES, PHAEOPHYTA)1

A tentative amplified fragment length polymorphism–simple sequence repeat (AFLP–SSR) linkage map of Laminaria was constructed using a haploid population of 40 gametophyte clones isolated from an individual of Dongfang No. 2, the first commercially cultured hybrid of a female gametophyte clone of L. japonica Aresch. [=Saccharina japonica (Aresch.) C. E. Lane, C. Mayes et G. W. Saunders] and a male one of L. longissima Miyabe [=Saccharina longissima (Miyabe) C. E. Lane, C. Mayes et G. W. Saunders]. To the map, 263 markers (255 AFLP, seven SSR, and the gametophyte sex) were assigned. The map consisted of 25 linkage groups (LGs) with ≥ four markers, five triplets, and 15 doublets, which is 1,629.0 centiMorgans (cM) in length, covering 66% of Laminaria genome. The maximum space between loci is 24.63 cM. A putative sex‐determining region was identified in LG2, which was characterized by a dense marker distribution around the gametophyte sex locus. The linkage map itself and the methodology associated with its construction will facilitate the genetic study and further trials of the linkage map construction of Laminaria.

Prediction of the heterosis of Laminaria hybrids with the genetic distance between their parental gametophyte clones

Eighteen microsatellite markers were used to determine the genetic distances between the parental gametophyte clones of 14 Laminaria hybrids, which were then used to establish a linear relationship with the heterosis (hybrid vigor) of economic traits including yield, mean blade try weight, mean blade fresh weight, blade length, blade width and mean blade thickness using regression analysis. Significant regression was found between the genetic distance (x) and the heterosis (y) of yield (y = 115.10x − 77.97, r = 0.8151, p = 0.00038), mean blade dry weight (y = 115.23x  −77.97, r = 0.8154, p = 0.00038), mean blade fresh weight (y = 100.08x − 57.85, r = 0.7306, p = 0.0030) and blade length (y = 204.11x − 46.77, r = 0.6963, p = 0.00566). The prediction of the heterosis of Laminaria hybrids with the genetic distance between parental gametophyte clones will facilitate the selection of elite Laminaria hybrids by avoiding the time-consuming and labor-intensive trait evaluation of a large number of hybridization combinations.

Improving seedless kelp (Saccharina japonica) during its domestication by hybridizing gametophytes and seedling-raising from sporophytes.

Dongfang no.7 (Saccharina japonica) was bred and maintained by hybridizing gametophytes, self-crossing the best individuals, selecting the best self-crossing line and seedling-raising from yearly reconstructed sporophytes. It increased the air dry yield by 43.2% in average over 2 widely farmed controls. Dongfang no.7 was seedling-raised from bulked sporophytes reconstructed from its representative gametophyte clones. Such strategy ensured it against variety contamination due to possible cross fertilization and occasional mixing and inbred depletion due to self-crossing number-limited sporophytes year after year. It derived from an intraspecific hybrid through 4 rounds of self-crossing and selection and retained a certain degree of genetic heterozygosity, thus being immune to inbred depletion due to purification of unknown detrimental alleles. Most importantly, it can be farmed in currently available system as the seedlings for large scale culture can be raised from reconstructed Dongfang no.7 sporophytes. Breeding and maintaining Dongfang no.7 provided a model that other varieties of kelp (S. japonica) and brown algae may follow during their domestication.

Microsatellite DNA Variation of the Gametophyte Clones Isolated from Introduced Laminaria japonica (Phaeophyta) and L. longissima of China and Varieties Derived from them

The variation of 90 Laminaria gametophyte clones representing the introduced Laminaria japonica (Group 1) and Laminaria longissima (Group 2), the varieties of L. japonica (Group 3) and the varieties derived from interspecific hybrids (Group 4) was determined with 18 microsatellite markers. The allelic diversity and Neis gene diversity of Group 1 were significantly higher than those of Group 2 (2.9 vs. 1.8 and 0.414 vs. 0.161, respectively), demonstrating that the variation of the introduced L. japonica is richer than that of L. longissima. Both allelic diversity and Neis gene diversity of Group 3 were lower than those of Group 1, indicating that only a portion of variation of L. japonica was incorporated into the varieties of L. japonica. Significant genetic differentiation was detected between four groups and between female (Population 1) and male (Population 2) gametophyte clones in each group. The variation among groups accounted for 39.95%, while that among populations accounted for 21.65% of the total. The genetic distance between Group 1 and Group 4 was obviously longer than that between Group 2 and Group 4 (0.686 vs. 0.291), indicating that maternal gametophyte clone contributed more variation to the hybrids than the paternal gametophyte clone did.

Association of triacylglyceride content and transcript abundance of genes involving in lipid synthesis of nitrogen deficient Phaeodactylum tricornutum

Phaeodactylum tricornutum is a diatom that is rich in lipids. Recently, it has received much attention as a feedstock for biodiesel production. Nitrogen deficiency is widely known to increase the content of neutral lipids (mainly triacylglycerides, or TAGs) of microalgae, including P. tricornutum, but the mechanism is unclear. In this study, we deciphered the correlations between TAG content and nine key enzymatic genes involved in lipid synthesis in P. tricornutum. After being cultured under nitrogen-free conditions for 0, 4, 24, 48, 72, 120, and 168 h, the TAG contents of P. tricornutum cells were assayed and the transcript abundances of the target genes were monitored by quantitative real-time PCR. The results show that the abundances of four target gene transcripts (LACS3, G3PDH2, G3PDH3, and G3PDH5) were positively correlated with TAG content, indicating that these genes may be involved in TAG synthesis in P. tricornutum. The findings improve our understanding of the metabolic network and regulation of lipid synthesis and will guide the future genetic improvement of the TAG content of P. tricornutum.

Isolation and characterization of a long-chain acyl-coenzyme A synthetase encoding gene from the marine microalga Nannochloropsis oculata

Acyl-coenzyme A synthetases (ACSs) are associated with the anabolism and catabolism of fatty acids and play fundamental roles in various metabolic pathways. The cDNA of long-chain acyl-coenzyme A synthetase (LACS), one of the ACSs, was isolated from Nannochloropsis oculata and named as NOLACS. The predicted amino acid sequence was highly similar to LACSs of other species. NOLACS encodes a long-chain acyl-coenzyme A synthetase; it recovered the function of LACS in Saccharomyces cerevisiae YB525 (a LACS-deficient yeast strain). The substrate specificity of the enzyme was also assayed in yeast. It was found that NOLACS can activate saturated fatty acids (C12:0, C14:0, C16:0, and C18:0) and some unsaturated fatty acids (C18:2Δ9, 12 and C20:2Δ11, 14) with a preference for long-chain fatty acids. Our findings will provide a deep understanding of CoA-dependent fatty acid activation and also make some contribution to understanding the metabolic pathways of lipids in Nannochloropsis. These findings will also facilitate studies on the regulation of gene expression and genetic modification of fatty acid synthesis and storage of N. oculata.

Biomass and nutrient productivities of Tetraselmis chuii under mixotrophic culture conditions with various C:N ratios

Mass microalgal culture plays an irreplaceable role in aquaculture, but microalgal productivity is restricted by traditional autotrophic culture conditions. In the present study, a Tetraselmis chuii strain belonging to the phylum Chlorophyta was isolated from south Yellow Sea. The growth rate and biomass productivity of this strain was higher under mixotrophic conditions with different carbon:nitrogen (C:N) ratios than those under autotrophic conditions. When the C:N ratio was 16, the optical density and biomass productivity were 3.7- and 5-fold higher than their corresponding values under autotrophic culture conditions, respectively. Moreover, T. chuii synthesized more polysaccharides and total lipids under mixotrophic conditions. In addition, T. chuii cultured under mixotrophic conditions synthesized more types of fatty acids than autotrophic culture conditions. At a C:N ratio of 16, the percentage of C16:0 and C18:1 reached 30.08% and 24.65% of the total fatty acid (TFA) content, respectively. These findings may provide a basis for large-scale mixotrophic culture of T. chuii, as a potential bait-microalga.

Tissue-overlapping response of half-smooth tongue sole (Cynoglossus semilaevis) to thermostressing based on transcriptome profiles

Thermal stress, encountered frequently in aquaculture, affects diverse physiological processes and behavior of fish; however, the mechanism underlying these effects may vary among species. Half-smooth tongue sole (Cynoglossus semilaevis) widely inhabits Asian coastal waters and has been intensively cultured in China. In this study, the transcriptomes of three tissues (gill, liver and muscle) of half-smooth tongue sole stressed at temperatures up to 35°C were profiled and compared with those of the fish living at a normal temperature of 25°C. Of 105,228 transcripts, 23,213 were annotated in GO terms, and 718, 1236 and 561 were found to express differentially in gill, liver and muscle from control, respectively. Of the differentially expressed genes, 119 were shared by all three tissues. Tissue-overlapping transcripts and the pathways and functions they defined may underline the primary response of fish to thermostressing. High temperature may cause directly protein mis-folding and hypoxia. Cells suffering thermostress either survive hypoxia or are eliminated by immune system. High temperature may have interacted indirectly through HSPs with HIF-1 and JAKs/STATs signaling pathways. The former up-regulates the expression of hypoxia inducible genes while the later up-regulates the expression of genes associating with insulin and NOD-like receptors signaling pathways. Insulin pathway functions to sustain homeostasis of sugar and lipids, aiding to survive cells, while NOD-like receptors signaling pathway strengthens immune, apoptotic and inflammatory responses, helping to survive cells. These understandings may help improve our culturing practice, the culture performance of half-smooth tongue sole and its breeding.