Changhai Wang

Nutrient deprivation enhances lipid content in marine microalgae.

The lipid content of microalgae is important for biodiesel production. In this study, we determined photosynthetic performance, biomass composition and lipid production of marine unicellular microalgae Chaetoceros muelleri (Bacillariophyceae) and Dunaliella salina (Chlorophyceae) cultured under different nutrient deprivation conditions. The results showed the highest lipid content of C. muelleri (46.32±3.53%) and D. salina (54.15±2.71%) could be achieved by nitrogen and complete nutrient deprivation, respectively. Analysis of fatty acid methyl esters (FAMEs) profile under those conditions revealed that the lipid accumulated by the two species was suitable to produce biodiesel. From these studies, we concluded that C. muelleri and D. salina would be highly suitable candidates for biodiesel production.

Breeding of high biomass and lipid producing Desmodesmus sp. by Ethylmethane sulfonate-induced mutation

To improve the biomass yield and lipid productivity, two desert microalgae, Desmodesmus sp. S81 and G41 were induced mutagenesis by Ethylmethane sulfonate (EMS), and obtained two potential mutants, Desmodesmus sp. S5 and G3 from the mutagenic clones for their greatly promoted biomass and lipid production. The results showed that the biomass yield, lipid content and lipid productivity of the mutant strains S5 and G3 were 778.10mg·L(-1), 48.41% and 19.83mg·L(-1)·d(-1), 739.52mg·L(-1), 46.01%, and 17.92mg·L(-1)·d(-1), respectively, which presented the increment of 45.50%, 8.00% and 74.24%, 20.67%, 10.35% and 55.77% than those of S81 and G41. Comparing with the wild strains, the mutants showed reduced PUFAs and glycol lipids, elevated MUFAs and neutral lipids contents, which were appropriate for biodiesel production.

Effect of kelp waste extracts on the growth and lipid accumulation of microalgae.

Kelp waste extracts (KWE) contained massive soluble sugars, amino acids and various mineral elements. To probe the effects of KWE on microalgal physiological and biochemical responses, the cultures were carried out under the different dilutions. The results showed that 8.0% KWE increased the biomass productivities and total lipid contents of Chlorella strains dramatically, which were 1.83-31.86 times and 20.78-25.91% higher than that of the control. Phaeodactylum tricornutum and Spirulina maxima presented a better growth performance in 1.0% and 4.0% treatment respectively, while their lipid accumulation were not enhanced. In Chlorella-Arc, Chlorella sorokiniana and P. tricornutum, the contents of saturated and monounsaturated fatty acids could be increased, and polyunsaturated fatty acids could be decreased under the conditions of high concentration of KWE (6.0-8.0%). Briefly, KWE facilitated to enhance the biomass productivity and lipid content of Chlorella strains, also improved the fatty acid compositions for biodiesel production.

Combining and Comparing Coalescent, Distance and Character-Based Approaches for Barcoding Microalgaes: A Test with Chlorella-Like Species (Chlorophyta)

Several different barcoding methods of distinguishing species have been advanced, but which method is the best is still controversial. Chlorella is becoming particularly promising in the development of second-generation biofuels. However, the taxonomy of Chlorella–like organisms is easily confused. Here we report a comprehensive barcoding analysis of Chlorella-like species from Chlorella, Chloroidium, Dictyosphaerium and Actinastrum based on rbcL, ITS, tufA and 16S sequences to test the efficiency of traditional barcoding, GMYC, ABGD, PTP, P ID and character-based barcoding methods. First of all, the barcoding results gave new insights into the taxonomic assessment of Chlorella-like organisms studied, including the clear species discrimination and resolution of potentially cryptic species complexes in C. sorokiniana, D. ehrenbergianum and C. Vulgaris. The tufA proved to be the most efficient barcoding locus, which thus could be as potential “specific barcode” for Chlorella-like species. The 16S failed in discriminating most closely related species. The resolution of GMYC, PTP, P ID, ABGD and character-based barcoding methods were variable among rbcL, ITS and tufA genes. The best resolution for species differentiation appeared in tufA analysis where GMYC, PTP, ABGD and character-based approaches produced consistent groups while the PTP method over-split the taxa. The character analysis of rbcL, ITS and tufA sequences could clearly distinguish all taxonomic groups respectively, including the potentially cryptic lineages, with many character attributes. Thus, the character-based barcoding provides an attractive complement to coalescent and distance-based barcoding. Our study represents the test that proves the efficiency of multiple DNA barcoding in species discrimination of microalgaes.

Growth and lipid accumulation in three Chlorella strains from different regions in response to diurnal temperature fluctuations.

It was economically feasible to screen strains adaptive to wide temperature fluctuation for outdoor cultivation without temperature control. In this research, three Chlorella strains from arctic glacier, desert soil and temperate native lake were isolated and identified. The growth, biochemical composition, lipid content and fatty acid composition of each strain cultured under the mode of diurnal temperature fluctuations were compared. All the three Chlorella strains showed desirable abilities of accumulating lipid under diurnal temperature fluctuations and their fatty acid profiles were suitable for biodiesel production, although the growth and biochemical composition were seemed to be region-specific. The highest lipid content was at 51.83±2.49% DW, 42.80±2.97% DW and 36.13±2.27% DW under different temperature fluctuation of 11 °C, 25 °C, 7 °C, respectively. The results indicated that the three Chlorella strains could be promising biodiesel feedstock for outdoor cultivation by the cultural mode of diurnal temperature fluctuations.

Bioenergy application of Dunaliella salina SA 134 grown at various salinity levels for lipid production

The biofuels are receiving considerable attention as a substitute for petro diesel. For microalgae, the cell density or biomass and lipid contents are key components for biodiesel production. This study was conducted to develop favorable culture conditions for Dunaliella salina to maximize its biomass and lipid accumulation. The effect of salinity (0.5 to 2.5 M NaCl) on the cell population, biochemical composition, and lipid output of Dunaliella salina was examined under a controlled environment for 21 days. Maximum growth (6.57 × 107 to 7.17 × 107cells mL−1) potentials were observed at 1.5 to 2 M NaCl. The photosynthetic pigments and carbohydrates also showed trends similar to growth. The maximum carotenoid level (5.16 mg L−1) was recorded at 2 M NaCl. Almost all physicochemical parameters increased with increases in salinity, biomass (1231.66 ± 1.26 mg L−1) and lipid content (248.33 mg L−1), as recorded at 2 M NaCl. Based on fluorescence intensity, the highest values (11.84 × 107cells/ml) of neutral lipids and total lipids (22.28%) were recorded at optimum salinity levels. The present study suggests that a high biomass and lipid accumulation of Dunaliella salina SA 134 could be obtained at the 2 M NaCl level.

Effect of Kelp Waste Extracts on the Growth and Development of Pakchoi (Brassica chinensis L.).

To explore the effects of kelp waste extracts (KWE) on the growth and development of Brassia chinensis L., germination and greenhouse experiments were carried out under different concentrations of KWE. The results showed that a higher germination percentage (95%), associated with high germination index (8.70), germination energy (71.67%) and seedling vigor index (734.67), was obtained under a lower KWE concentration (2%) compared with the control. The radicle length (4.97 cm), fresh weight (0.32 g/10 seedlings) and dry weight (0.015 g/10 seedlings) were significantly increased in the treatment of 2% KWE. KWE also could enhance the root growth, the maximum leaf length × width and the fresh weight of plants, the optimal value of which increased by 8.37 cm, 58.14 cm2 and 7.76 g under the treatment of 10% KWE compared with the control respectively. Meanwhile, the contents of vitamin C and soluble sugars in pakchoi leaf were improved by 19.6 mg/100 g and 1.44 mg/g compared with the control, and the nitrate content was decreased by 212.27 mg/kg. Briefly, KWE could markedly stimulate the pakchoi seeds germination at a lower concentration (2%) and enhance the plant growth and quality at a higher concentration (10%).

Kelp waste extracts combined with acetate enhances the biofuel characteristics of Chlorella sorokiniana

To probe the effect of kelp waste extracts (KWE) combined with acetate on biochemical composition of Chlorella sorokiniana, the cultures were performed under independent/combined treatment of KWE and acetate. The results showed that high cell density and biomass were obtained by KWE combined with acetate treatments, whose biomass productivity increased by 79.69-102.57% and 20.04-35.32% compared with 3.0gL-1 acetate and KWE treatments respectively. The maximal neutral lipid per cell and lipid productivity were gained in KWE combined with 3.0gL-1 acetate treatment, which increased by 16.32% and 129.03% compared with 3.0gL-1 acetate, and 253.35% and 70.74% compared with KWE treatment. Meanwhile, C18:3n3 and C18:2n6c contents were reduced to 4.90% and 11.88%, whereas C16:0 and C18:1n9c were improved to 28.71% and 37.76%. Hence, supplementing appropriate acetate in KWE cultures is supposed to be a great potential method for large-scale cultivation of C. sorokiniana to generate biofuel.

How DNA barcoding can be more effective in microalgae identification: a case of cryptic diversity revelation in Scenedesmus (Chlorophyceae)

Microalgae identification is extremely difficult. The efficiency of DNA barcoding in microalgae identification involves ideal gene markers and approaches employed, which however, is still under the way. Although Scenedesmus has obtained much research in producing lipids its identification is difficult. Here we present a comprehensive coalescent, distance and character-based DNA barcoding for 118 Scenedesmus strains based on rbcL, tufA, ITS and 16S. The four genes, and their combined data rbcL + tufA + ITS + 16S, rbcL + tufA and ITS + 16S were analyzed by all of GMYC, P ID, PTP, ABGD, and character-based barcoding respectively. It was apparent that the three combined gene data showed a higher proportion of resolution success than the single gene. In comparison, the GMYC and PTP analysis produced more taxonomic lineages. The ABGD generated various resolution in discrimination among the single and combined data. The character-based barcoding was proved to be the most effective approach for species discrimination in both single and combined data which produced consistent species identification. All the integrated results recovered 11 species, five out of which were revealed as potential cryptic species. We suggest that the character-based DNA barcoding together with other approaches based on multiple genes and their combined data could be more effective in microalgae diversity revelation.

High-efficiency 18S microalgae barcoding by coalescent, distance and character-based approaches: a test in Chlorella and Scenedesmus

The relatively conserved 18S is often used in the phylogenetic analysis of microalgae. However, whether it can really help in barcoding microalgae needs to be evaluated. In this study the multiple approaches of coalescent, distance and character-based barcoding are first employed in Chlorella and Scenedesmus to test the efficiency of 18S sequences for barcoding green microalgae. We show that most Chlorella and Scenedesmus species, including the cryptic species, can be distinguished by 18S sequences with all coalescent General Mixed Yule-coalescent (GMYC), poisson tree process (PTP), and P ID, distance (ABGD) and character-based approaches. Both GMYC and PTP analyses produce more genetic groups. The P ID and ABGD analyses only cluster some species. All species (apart from a few of lineages) can be separated in character-based barcoding analysis with more than three character attributes. In comparison with previous barcoding results with rbcL, tufA, ITS and 16S, 18S produces good resolution in identifying Chlorella and Scenedesmus. Our results reveal that 18S is highly efficient in identifying taxa of green microalgae at species level, based on a combination of multiple barcoding approaches. Combining 18S with other gene markers may be useful in barcoding microalgae.