Chlorella Strains Research Articles

Development of novel microsatellite markers for strain-specific identification of Chlorella vulgaris.

A strain-specific identification method is required to secure Chlorella strains with useful genetic traits, such as a fast growth rate or high lipid productivity, for application in biofuels, functional foods, and pharmaceuticals. Microsatellite markers based on simple sequence repeats can be a useful tool for this purpose. Therefore, this study developed five novel microsatellite markers (mChl-001, mChl-002, mChl-005, mChl-011, and mChl-012) using specific loci along the chloroplast genome of Chlorella vulgaris. The microsatellite markers were characterized based on their allelic diversities among nine strains of C. vulgaris with the same 18S rRNA sequence similarity. Each microsatellite marker exhibited 2~5 polymorphic allele types, and their combinations allowed discrimination between seven of the C. vulgaris strains. The two remaining strains were distinguished using one specific interspace region between the mChl-001 and mChl-005 loci, which was composed of about 27 single nucleotide polymorphisms, 13~15 specific sequence sites, and (T)n repeat sites. Thus, the polymorphic combination of the five microsatellite markers and one specific locus facilitated a clear distinction of C. vulgaris at the strain level, suggesting that the proposed microsatellite marker system can be useful for the accurate identification and classification of C. vulgaris.

Screening and characterization of oleaginous Chlorella strains and exploration of photoautotrophic Chlorella protothecoides for oil production

The growth and oil production of nine Chlorella strains were comparatively assessed and Chlorellaprotothecoides CS-41 demonstrated the greatest lipid production potential. The effects of different nitrogen forms and concentrations, phosphorus concentrations and light intensities on growth and oil production were studied in laboratory columns. C. protothecoides CS-41 accumulated lipids up to 55% of dry weight, with triacylglycerol and oleic acid being 71% of total lipids and 59% of total fatty acids, respectively. High biomass and lipid productivities were achieved in outdoor panel PBRs, up to 1.25 and 0.59gL−1day−1, or 44. 1 and 16.1gm−2day−1, respectively. A two-stage cultivation strategy was proposed to enhance the algal biomass and lipid production. This is the first comprehensive investigation of both indoor and outdoor photoautotrophic C. protothecoides cultures for oil production, and C. protothecoides CS-41 represents a promising biofuel feedstock worthy of further exploration.

Growth relationships of a lipid-producing Chlorella-alga with common microalgae in laboratory co-cultures

Co-existence growth relationships were studied in communities consisting of a lipid-producing alga Chlorella sp. HQ, another green alga and one cyanobacterium: I Scenedesmus obliquus and Microcystis aeruginosa; II Chlamydomonas reinhardtii and Anabaena flos-aquae; III Selenastrum capricornutum and Microcystis wesenbergii. The cyanobacteria and green algae except for Chlorella sp. HQ were commonly detected in Chinese reservoir and wastewater. The rate of increase of apparent cell number difference with other algae (k app), inhibition/stimulation ratio (ISR) and the parameters of logistic model and co-existence model were determined for Chlorella. Chlorella strains were the most competitive in Combination I, and were stimulated during 75% of the cultivation time for all three combinations. Anabaena growth exceeded those of Chlorella and Chlamydomonas on the 5th cultivation day under 1 : 1 : 1 inoculum ratio. Scenedesmus colonies consisted of fewer cells, whose average length significantly shortened after the 5th cultivation day under 1 : 1 : 1 inoculum ratio. The developed co-existence model can identify the concrete growth inhibitor or stimulator among three species compared with the single method of cell number monitoring. Good correlation was found between transformed and non-transformed co-existence model through a mn and b mn values. Allelopathy and nutrient competition are both possible mechanisms in the above growth relationships.

Chlorella for protein and biofuels: from strain selection to outdoor cultivation in a Green Wall Panel photobioreactor.

BackgroundChlorella is one of the few microalgae employed for human consumption. It typically has a high protein content, but it can also accumulate high amounts of lipids or carbohydrates under stress conditions and, for this reason, it is of interest in the production of biofuels. High production costs and energy consumption are associated with its cultivation. This work describes a strategy to reduce costs and environmental impact of Chlorella biomass production for food, biofuels and other applications.ResultsThe growth of four Chlorella strains, selected after a laboratory screening, was investigated outdoors in a low-cost 0.25 m2 GWP-II photobioreactor. The capacity of the selected strains to grow at high temperature was tested. On the basis of these results, in the nitrogen starvation trials the culture was cooled only when the temperature exceeded 40°C to allow for significant energy savings, and performed in a seawater-based medium to reduce the freshwater footprint. Under nutrient sufficiency, strain CH2 was the most productive. In all the strains, nitrogen starvation strongly reduced productivity, depressed protein and induced accumulation of carbohydrate (about 50%) in strains F&M-M49 and IAM C-212, and lipid (40 - 45%) in strains PROD1 and CH2. Starved cultures achieved high storage product productivities: 0.12 g L−1 d−1 of lipids for CH2 and 0.19 g L−1 d−1 of carbohydrates for F&M-M49. When extrapolated to large-scale in central Italy, CH2 showed a potential productivity of 41 t ha−1 y−1 for biomass, 16 t ha−1 y−1 for protein and 11 t ha−1 y−1 for lipid under nutrient sufficiency, and 8 t ha−1 y−1 for lipid under nitrogen starvation.ConclusionsThe environmental and economic sustainability of Chlorella production was enhanced by growing the organisms in a seawater-based medium, so as not to compete with crops for freshwater, and at high temperatures, so as to reduce energy consumption for cooling. All the four selected strains are good candidates for food or biofuels production in lands unsuitable for conventional agriculture. Chlorella strain CH2 has the potential for more than 80 tonnes of biomass, 32 tonnes of protein and 22 tonnes of lipid per year under favourable climates.

Comparative Analyses of Three Chlorella Species in Response to Light and Sugar Reveal Distinctive Lipid Accumulation Patterns in the Microalga C. sorokiniana

While photosynthetic microalgae, such as Chlorella, serve as feedstocks for nutritional oils and biofuels, heterotrophic cultivation can augment growth rates, support high cell densities, and increase triacylglycerol (TAG) lipid content. However, these species differ significantly in their photoautotrophic and heterotrophic characteristics. In this study, the phylogeny of thirty Chlorella strains was determined in order to inform bioprospecting efforts and detailed physiological assessment of three species. The growth kinetics and lipid biochemistry of C. protothecoides UTEX 411, C. vulgaris UTEX 265, and C. sorokiniana UTEX 1230 were quantified during photoautotrophy in Bold's basal medium (BBM) and heterotrophy in BBM supplemented with glucose (10 g L−1). Heterotrophic growth rates of UTEX 411, 265, and 1230 were found to be 1.5-, 3.7-, and 5-fold higher than their respective autotrophic rates. With a rapid nine-hour heterotrophic doubling time, Chlorella sorokiniana UTEX 1230 maximally accumulated 39% total lipids by dry weight during heterotrophy compared to 18% autotrophically. Furthermore, the discrete fatty acid composition of each strain was examined in order to elucidate lipid accumulation patterns under the two trophic conditions. In both modes of growth, UTEX 411 and 265 produced 18∶1 as the principal fatty acid while UTEX 1230 exhibited a 2.5-fold enrichment in 18∶2 relative to 18∶1. Although the total lipid content was highest in UTEX 411 during heterotrophy, UTEX 1230 demonstrated a two-fold increase in its heterotrophic TAG fraction at a rate of 28.9 mg L−1 d−1 to reach 22% of the biomass, corresponding to as much as 90% of its total lipids. Interestingly, UTEX 1230 growth was restricted during mixotrophy and its TAG production rate was suppressed to 18.2 mg L−1 d−1. This constraint on carbon flow raises intriguing questions about the impact of sugar and light on the metabolic regulation of microalgal lipid biosynthesis.

Planktochlorella nurekis gen. et sp. nov. (Trebouxiophyceae, Chlorophyta), a novel coccoid green alga carrying significant biotechnological potential.

Phylogenetic position, morphology and ultrastructure were investigated for biotechnologically remarkable strain Chlorella vulgaris IFR C-111, utilized in various commercial applications. Molecular phylogenetic analyses based on the SSU and ITS rDNA data revealed that the strain IFR C-111 forms a distinct lineage within the Parachlorella-clade in Chlorellaceae. We describe this organism as a new genus and species, Planktochlorella nurekis. Vegetative cells of this newly recognized species are spherical, possessing a single pot-shaped chloroplast with starch-covered pyrenoid. Asexually it reproduces by the formation of 2-16(-32) autospores. Cell wall is composed of two layers, the outer layer containing extended microfibrillar material. The fuzzy cell wall structure improves the buoyancy resulting in low sedimentation rate of P. nurekis. To resolve the phylogenetic position of Planktochlorella and its relationship to the closely related genera, nucleotide saturation present in the ITS rDNA data was reduced by four different approaches. The resulting topologies pointed to the poor phylogenetic signal in generally utilized SSU and ITS rDNA data and the need of sequencing other molecular markers.

Starch and lipid accumulation in eight strains of six Chlorella species under comparatively high light intensity and aeration culture conditions

The microalgae family Chlorella species are known to accumulate starch and lipids. Although nitrogen or phosphorous deficiencies promote starch and lipids formation in many microalgae, these deficiencies also limit their growth and productivity. Therefore, the Chlorellaceae strains were attempted to increase starch and lipids productivity under high-light-intensity conditions (600-μmol photons m(-2)s(-1)). The 12:12-h light-dark (LD) cycle conditions elicited more stable growth than the continuous light (LL) conditions, whereas the starch and lipids yields increased in LL conditions. The amount of starch and lipids per cell increased in Chlorella viscosa and Chlorella vulgaris in sulfur-deficient medium, and long-chain fatty acids with 20 or more carbon atoms accumulated in cells grown in sulfur-deficient medium. Accumulation of starch and lipids was investigated in eight strains. The accumulation was strain-dependent, and varied according to the medium and light conditions. Five of the eight Chlorella strains exhibited similar accumulation patterns.

Effect of Increased CO2 And Temperature on Growth, Photosynthesis And Lipid Content of Tropical Algae

Increasing temperatures causally linked with anthropogenically elevated atmospheric carbon dioxide (CO2 ) levels characterise current ‘global warming’. Very little is known about how these factors will affect tropical microalgae. The aim of this study was to investigate the CO2 and temperature sensitivity of the photosynthetic process of two tropical Chlorella strains (Chlorella vulgaris UMACC 001 and Chlorella vulgaris UMACC 014). We grew cultures of the two strains under four conditions: (1) 28°C and 375 ppm CO2 (control), (2) 28°C and 750 ppm CO2 (high CO2 ), (3) 32°C and 375 ppm CO2 (high temperature), and (4) 32°C and 750 ppm CO2 (combined factors). Based on specifi c growth rate (μ), both Chlorella strains grew best under the combined factors of high CO2 level (750 ppm) and high temperature (32°C). Elevated CO2 stimulated the production of lipid in both strains. (Keywords: Chlorella, temperature, CO2 , photosynthesis )

Lipid productivity and fatty acid composition-guided selection of Chlorella strains isolated from Malaysia for biodiesel production

The need to develop biomass-based domestic production of high-energy liquid fuels (biodiesel) for transportation can potentially be addressed by exploring microalgae with high lipid content. Selecting the strains with adequate oil yield and quality is of fundamental importance for a cost-efficient biofuel feedstock production based on microalgae. This work evaluated 29 strains of Chlorella isolated from Malaysia as feedstock for biodiesel based on volumetric lipid productivity and fatty acid profiles. Phylogenetic studies based on 18S rRNA gene revealed that majority of the strains belong to true Chlorella followed by Parachlorella. The strains were similarly separated into two groups based on fatty acid composition. Of the 18 true Chlorella strains, Chlorella UMACC187 had the highest palmitic acid (C16:0) content (71.3 ± 4.2 % total fatty acids, TFA) followed by UMACC84 (70.1 ± 0.7 %TFA), UMACC283 (63.8 ± 0.7 %TFA), and UMACC001 (60.3 ± 4.0 %TFA). Lipid productivity of the strains at exponential phase ranged from 34.53 to 230.38 mg L−1 day−1, with Chlorella UMACC050 attaining the highest lipid productivity. This study demonstrated that Chlorella UMACC050 is a promising candidate for biodiesel feedstock production.

Optimization of nutritional compositions of growth medium for Chlorella sp. FJ3 growth kinetics in batch and continuous-flow photoreactors

This study investigates improvement to culture medium for specific growth rate of Chlorella sp. FJ3 using a fractional factorial design for 32 experiments with six variable components. Six tested components were NaNO3 (0.5 or 3.0 g/l), K2HPO4 (0.01 or 0.06 g/l), MgSO4·7H2O (0.05 or 1.0 g/l), CaCl2·2H2O (0.01 or 0.06 g/l), ferric ammonium citrate (0.002 or 0.02 g/l) and NaCl (0.5 or 5.0 g/l). Magnesium sulphate and interaction between magnesium sulphate and ferric ammonium citrate were found to be critical for the cultivation of Chlorella sp. FJ3. The optimal concentrations of MgSO4·7H2O and ferric ammonium citrate were found to be 2.0 and 0.35 g/l, respectively. The concentration of carbonate () in effluent confirmed that the optimized culture medium was associated with a high carbonate utilization rate and specific growth rate during a transient period in batch and continuous-flow tests. The extent of growth of strain FJ3 in the optimized medium was 1.61 times greater than that in a non-optimized medium in the batch test. In the continuous-flow test, the maximum growth of Chlorella strain FJ3 in the optimized medium was 1.77 times higher than that in a non-optimized medium. The rate of fixation in the non-optimized and the optimized media was 339 mg/l-day and 887 mg/l-day, respectively, in the steady state. These experimental and modelling results indicated that optimization of concentration in nutritional compositions in the culture medium enhanced the capacity of Chlorella sp. FJ3 for inorganic carbon fixation in batch and continuous-flow modes of photoreactors.

Rapid Detection and Quantification of Triacylglycerol by HPLC–ELSD inChlamydomonas reinhardtiiandChlorellaStrains

Triacylglycerol (TAG) analysis and quantification are commonly performed by first obtaining a purified TAG fraction from a total neutral lipid extract using thin-layer chromatography (TLC), and then analyzing the fatty acid composition of the purified TAG fraction by gas chromatography (GC). This process is time-consuming, labor intensive and is not suitable for analysis of small sample sizes or large numbers. A rapid and efficient method for monitoring oil accumulation in algae using high performance liquid chromatography for separation of all lipid classes combined with detection by evaporative light scattering (HPLC–ELSD) was developed and compared to the conventional TLC/GC method. TAG accumulation in two Chlamydomonas reinhardtii (21 gr and CC503) and three Chlorella strains (UTEX 1230, CS01 and UTEX 2229) grown under conditions of nitrogen depletion was measured. The TAG levels were found to be 3–6 % DW (Chlamydomonas strains) and 7–12 % DW (Chlorella strains) respectively by both HPLC–ELSD and TLC/GC methods. HPLC–ELSD resolved the major lipid classes such as carotenoids, TAG, diacylglycerol (DAG), free fatty acids, phospholipids, and galactolipids in a 15-min run. Quantitation of TAG content was based on comparison to calibration curves of trihexadecanoin (16:0 TAG) and trioctadecadienoin (18:2 TAG) and showed linearity from 0.2 to 10 μg. Algal TAG levels >0.5 μg/g DW were detectable by this method. Furthermore TAG content in Chlorella kessleri UTEX 2229 could be detected. TAG as well as DAG and TAG content were estimated at 1.6 % DW by HPLC–ELSD, while it was undetectable by TLC/GC method.

The Effect of Light on the Growth and Product Accumulation of <i>Chlorella </i>

Microalgae are microscopic heterotrophicautotrophic photosynthesizing organisms with enormous potential as a source of biofuel. In the masses of biomasses, microalgae has the advantages of high lipid, large biomass, short growth cycle and easy cultivation and with microalgae as the raw material. Light condition is crucial to cultivation of microalgae.In this paper, the microalgae strain of Chlorella sp. was supplied in BG11 medium, and Cultivation optimization were experimented. The results were showed that growth of chlorella was associated with a saturated light intensity, light intensity which was suitable for the growth of chlorella was 2000Lux. Growth of chlorella required an appropriate light-dark alternation to facilitate matching of photoreaction of photosynthetic process with dark reaction of photosynthetic, appropriate light-dark photoperiod of illumination time which was suitable for the growth of chlorella was 12:12.

Screening high oleaginous Chlorella strains from different climate zones

In outdoor cultivation, screening strains adapted to a wide temperature range or suitable strains for different environmental temperatures is of great importance. In this study, triacylglycerol (TAG) content of 23 oil-producing Chlorella strains from different climate zones were analyzed by thin layer chromatography. Four selected Chlorella strains (NJ-18, NJ-7, NMX35N and NMX139N) with rather high TAG content had higher total lipid content compared with Chlorella vulgaris SAG 211-11b. In particular, NJ-18 displayed the highest TAG productivity among the four high oil-producing Chlorella strains. Accumulation of TAGs in strain NMX35N changed a little from 30 to 40°C, showing a desirable characteristic of accumulating TAGs at high temperatures. Our results demonstrated that NJ-18 and NMX35N had suitable fatty acid profiles and good adaption to low and high temperatures respectively. Therefore, cultivation of the two Chlorella strains together might be a good option for economic biodiesel production during the whole seasons of the year.

Effects of Temperature on the Growth and Product Accumulation of <i>Chlorella sp.</i>

In recent years, the use of liquid biofuels in the transport sector has shown rapid global growth, driven mostly by policies focused on achievement of energy security, and mitigation of GHG emissions. Microalgae, as biomass, are a potential source of renewable energy, and they can be converted into energy such as biofuel oil and gas.In this paper, the microalgae strain of Chlorella sp. was supplied in BG11 medium, and Cultivation optimization were experimented. The results were showed that temperature was an important factor which could influence growth and accumulation of secondary metabolite in the cell from accumulation process of biological products, temperature which was suitable for the growth of chlorella was 25°C.

Design, construction and evaluation of solarized airlift tubular photobioreactor

An innovative photobioreactor is developed for growing algae in simulated conditions. The proposed design comprises of a continuous tubular irradiance loop and air induced liquid circulation with gas separation through air lift device. The unique features of air lift system are to ensure the shear free circulation of sensitive algal culture and induce light/dark cycles to the photosynthetic micro-organisms. The design strategy employs to model and construct a 20-liter laboratory scale unit using Boro-silicate glass tubing. The material is selected to ensure maximum photon transmission. All components of the device are designed to have flexibility to be replaced with an alternative design, providing fair chance of modification for future investigators. The principles of fluid mechanics are applied to describe geometrical attributes of the air lift system. Combination of LEDs and Florescent tube lights (Warm white) were used to illuminate the photosynthesis reaction area providing a possibility to control both illumination duration and light intensity. 200 Watt Solar PV system is designed to power up the device which included air pump (100 Watt) and illumination system (100 Watt). Algal strain Chlorella sp was inoculated in photobioreactor which was sparged with air and carbon dioxide. The growth was sustained in the batch mode with daily monitoring of temperature, pH and biomass concentration. The novel photobioreactor recorded a maximum experimental average yield of 0.65 g/l.day (11.3 g/m2.day) as compared to theoretical modeled yield of 0.82 g/l.day (14.26 g/m2.day), suggesting the device can be efficiently and cost-effectively employed in the production of algal biomass for biofuels, concomitantly mitigating CO2.

Biomass and oil content of Chlorella sp., Haematococcus sp., Nannochloris sp. and Scenedesmus sp. under mixotrophic growth conditions in the presence of technical glycerol

The growth of algae strains Chlorella sp., Haematococcus sp., Nannochloris sp. and Scenedesmus sp. under mixotrophic conditions in the presence of different concentrations of technical glycerol was investigated with the aim of increasing biomass growth and algae oil content. The highest concentration of lipid obtained in media with 5 g L−1 glycerol for Chlorella sp., Scenedesmus sp., Nannochloris sp. and Haematococcus sp. was 17.77, 22.34, 27.55 and 34.22 % larger than during the autotrophic growth of these species. Increases in triacylglycerols of up to ten times was observed for Scenedesmus sp. under mixotrophic conditions (using 10 g L−1 glycerol), whereas an increase of 2.28 times was found for Haematococcus sp. The content of saturated fatty acids of Scenedesmus, Chlorella, Haematococcus and Nannochloris was 67.11, 34.63, 23.39 and 24.23 %, and the amount of unsaturated fatty acids was 32.9, 65.06, 79.61 and 75.78 % of total fatty acids, respectively. Growth on technical glycerol of these strains with light produced higher biomass concentrations and lipid content compared with autotrophic growth. The fatty acid content of oils from these species suggests their potential use as biodiesel feedstock.

Evaluation of oil-producing algae as potential biodiesel feedstock

This study attempted to connect the dots between laboratory research and the outdoors. Chlorella sp. NJ-18 was selected among seven oil-producing algae cultivated in this study because it had the highest lipid productivity. The nitrogen and phosphorus concentrations for cultivating this Chlorella strain were optimized indoors. This strain was incubated outdoors in a 70L photobioreactor, containing the favorable nitrogen (8.32mM urea) and phosphorus (0.18mM monopotassium phosphate) concentrations. Semi-continuous cultivation was performed by harvesting 30L biomass and replacing it with fresh medium. The maximum biomass and lipid productivity acquired outdoors were 91.84 and 24.05mgL−1d−1, respectively. Furthermore, biomass productivity could be maintained at a high level throughout the cultivation process when using the semi-continuous mode, whereas it decreased dramatically in batch cultures. More than 95% of the total fatty acids obtained were C16 and C18, which are the main components for biofuel.

Effects of Cultivation Conditions and Media Compositions on Lipid Productivity of Native Microalga <i>Chlorella vulgaris</i> U3-3

A native microalgae strain Chlorella vulgaris U3-3from Baimaxueshan Yunnan province has been isolated and identified as a potential feedstock for biodiesel production. The biomass production, lipid productivity, and fatty acid compositions of U3-3 are investigated and compared under phototrophic, heterotrophic and mixotrophic conditions with various media. U3-3 under phototrophic growth on BG11 medium shows higher cellular lipid content (44.6%) than those under heterotrophic and mixotrophic conditions. And U3-3 display its higher biomass production (5.55g/L) than those under phototrophic and heterotrophic conditions growing on optimized medium with supply 35g/L glucose under mixotrophic condition. However, Chlorella vulgaris U3-3 growing under heterotrophic condition with optimized medium of supply 35g/L glucose exhibited more than 7.26-fold and 1.05-fold increase in lipid productivity compared with that under phototrophic and mixotrophic conditions. Moreover, Chlorella vulgaris U3-3 has suitable fatty acid compositions with high saturated acid contents (28.76%) and favorable degree of unsaturation (92.86) growing under heterotrophic condition with optimized medium of supply 35g/L glucose, suggesting its potential as a biodiesel feedstock on this cultivation condition and medium composition.

Two-stage cultivation of two Chlorella sp. strains by simultaneous treatment of brewery wastewater and maximizing lipid productivity

A cultivation system in the two-stage photoautotrophic–photoheterotrophic/mixotrophic mode was adapted to maximize lipid productivity of two freshwater strains of Chlorella sp. grown in brewery wastewater (BWW). The endogenous Chlorella sp. isolated from BWW had a higher growth rate than wild-type Chlorella vulgaris (UTEX-265) while C. vulgaris (UTEX-265) had a higher maximal biomass and lipid contents than that of endogenous Chlorella sp., resulting in more than 90% of the inorganic nutrients in both total nitrogen (TN) and phosphorus (TP) was removed during the first stage in the two-stage photoautotrophic–photoheterotrophic mode in each Chlorella sp. The maximal biomass and lipid contents of C. vulgaris (UTEX-265) for single stage photoautotrophic cultivation were 1.5g/L and 18%, respectively. Importantly, during two-stage photoautotrophic–photoheterotrophic cultivation for C. vulgaris (UTEX-265), the biomass was increased to 3.5g/L, and the lipid productivity was increased from 31.1 to 108.0mg/Lday.

Symbiosis between hydra and chlorella: Molecular phylogenetic analysis and experimental study provide insight into its origin and evolution

Although many physiological studies have been reported on the symbiosis between hydra and green algae, very little information from a molecular phylogenetic aspect of symbiosis is available. In order to understand the origin and evolution of symbiosis between the two organisms, we compared the phylogenetic relationships among symbiotic green algae with the phylogenetic relationships among host hydra strains. To do so, we reconstructed molecular phylogenetic trees of several strains of symbiotic chlorella harbored in the endodermal epithelial cells of viridissima group hydra strains and investigated their congruence with the molecular phylogenetic trees of the host hydra strains. To examine the species specificity between the host and the symbiont with respect to the genetic distance, we also tried to introduce chlorella strains into two aposymbiotic strains of viridissima group hydra in which symbiotic chlorella had been eliminated in advance. We discussed the origin and history of symbiosis between hydra and green algae based on the analysis.