<Emphasis Type="Italic">Bacillus rhizosphaerae</Emphasis> sp. nov., an novel diazotrophic bacterium isolated from sugarcane rhizosphere soil

A Gram-positive, non-pigmented, rod-shaped, diazotrophic bacterial strain, designated SC-N012^T, was isolated from rhizosphere soil of sugarcane and was subjected to a polyphasic taxonomic study. The strain exhibited phenotypic properties that included chemotaxonomic characteristics consistent with its classification in the genus Bacillus. Sequence analysis of the 16S rRNA gene of SC-N012^T revealed the closest match (98.9% pair wise similarity) with Bacillus clausii DSM 8716^T. However, DNA–DNA hybridization experiments indicated low levels of genomic relatedness (32%) with this strain. The major components of the fatty acid profile are iso-C_15:0, anteiso-C_15:0, iso-C_17:0 and anteiso-C_17:0. The diagnostic cell-wall diamino acid was meso-diaminopimelic acid. The G+C content of the genomic DNA is 43.0 mol%. The lipids present in strain SC-N012^T are diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol and two unknown phospholipids. Their predominant respiratory quinone was MK-7. Studies of DNA-DNA relatedness, morphological, physiological and chemotaxonomic analyses and phylogenetic data based on 16S rRNA gene sequencing allowed strain SC-N012^T to be described as members of novel species of the genus Bacillus, for which the name Bacillus rhizosphaerae sp. nov. is proposed. The type strain is SC-N012^T (=DSM 21911^T = NCCB 100267^T).     

Endophytic bacillus sp. isolated from the interior of balloon flower root

A bacterial strain, designated CY22, was isolated from the interior of balloon flower (Platycodon grandiflorum) root in the Republic of Korea. The isolate coproduced an iturin-like antifungal compound and a surfactin-like potent biosurfactant. Analysis of the 16S-rDNA of strain CY22 showed that the isolate was a member of Bacillus. High similarities were observed between strain CY22 and Bacillus sp. TKSP 24, and between strain CY22 and B. subtilis 168. Phylogenetic analysis based on 16S-rDNA sequences showed that strain CY22 was closely related to Bacillus sp. The main whole-cell fatty acids were anteiso-C15:0 (37%), C17:0 (5.1%), and iso-C15:0 (27.7%). DNA G+C content was 54 mol%. Based on phylogenetic inference, phenotypic and chemotaxonomic characteristics, this endophytic strain Bacillus sp. CY22 was assigned to the genus Bacillus.     

Bacillus gaemokensis sp. nov., isolated from foreshore tidal flat sediment from the Yellow Sea

A Gram-positive, rod-shaped, endospore-forming organism, strain BL3-6^T, was isolated from tidal flat sediments of the Yellow Sea in the region of Tae-An. A 16S rRNA gene sequence analysis demonstrated that this isolate belongs to the Bacillus cereus group, and is closely related to Bacillus mycoides (99.0% similarity), Bacillus thuringiensis (99.0%), Bacillus weihenstephanensis (99.0%), Bacillus cereus (98.9%), Bacillus anthracis (98.8%), and Bacillus pseudomycoides (98.1%). The phylogenetic distance from any validly described Bacillus species outside the Bacillus cereus group was less than 95.6%. The DNA G+C content of the strain was 39.4 mol% and the major respiratory quinone was menaquinone-7. The major cellular fatty acids were iso-C_14:0 (17.8%), iso-C_16:0 (15.8%), and iso-C_12:0 (11.3%). The diagnostic amino acid of the cell wall was meso-diaminopimelic acid and the major cell wall sugar was galactose. The results of DNA-DNA hybridization (<55.6%) and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain BL3-6^T from the published Bacillus species. BL3-6^T therefore represents a new species, for which the name Bacillus gaemokensis sp. nov. is proposed, with the type strain BL3-6^T (=KCTC 13318^T =JCM 15801^T).     

Screening for ω−1-hydroxy fatty acid over-producing mutants for bioconversion of oleic acid by combining general mutagenesis and specific selection

Chemical mutation of a strain producing hydroxy-fatty acid from oleic acid (OA) using NTG (N-methyl-N’-nitro-N-nitrosoguanidine) resulted in a high percentage of improved mutants. A positive screening procedure yielded several high producers, specifically the strain Bacillus pumilus M-F641 (BP M-F641). This strain produced predominantly ω_?1-hydroxy fatty acid and could utilize higher concentrations of OA than the parent strain. In shake flask culture, the best ω_?1-hydroxy fatty acid concentration and yield (the ratio of ω_?1-hydroxy fatty acid accumulation to OA consumption) reached 570 mg L^?1 and 11.5%, respectively. Repeated tests showed that the mutant BP M-F641 was genetically stable.     

A thermoacidophilic bacterium similar to Bacillus acidocaldarius

The thermoacidophilic strain TAC1 was isolated from a sulphatara field. It grows heterotrophically on a synthetic medium, containing yeast extract and carbohydrates at 50 to 70°C and pH 2 to 5. The cells are motile spore-forming rods utilizing, for instance, glucose, lactose, or sucrose as carbon substrate. They also used concentrated whey as nutrient medium. The maximum specific growth rate calculated from batch culture data of the strain TAC1 on glucose is 0.9 h^?1 at 65°C and pH 3. The yield coefficient determined in a chemostat culture of the strain TAC1 on glucose, is 0.15 to 0.31 grams of cells produced per gram of glucose consumed (63 to 70°C, pH 2.2 to 4.0, dilution rate 0.2 to 0.4 h^?1). The lipid fraction extracted from the cells consists of 72 to 93% of ω-cyclohexyl C₁₇ and C₁₉ fatty acids. The composition of lipid fraction varied with the pH value and the dilution rate but not with the temperature. In regard to the morphology and physiology of the isolated strain as well as the high percentage of ω-cyclohexyl fatty acids of the cell material, the strain TAC1 is similar to Bacillus acidocaldarius.     

Specificity of translation of spore polypeptides in bacillus in vitro systems

Cell free extracts prepared from exponentially growing Escherichia coli and Bacillus cereus as well as from Bacillus cereus at the end of exponential growth were optimized for various factors required for amino acid incorporation when programmed with Bacillus ribonucleic acid. All three preparations synthesized glutamine synthetase antigen when ribonucleic acid from a Bacillus subtilis strain that overproduces glutamine synthetase was added. The post exponential Bacillus cereus extract, however, was most active for the synthesis of Bacillus cereus spore coat antigen when supplemented with the appropriate ribonucleic acid. There appears to be some specificity in the translation of at least this sporulation messenger RNA.Non-Standard Abbreviations PMSF phenyl methyl sulfonylfluoride - GS glutamine synthetase - UDS 8 M urea, 1% (W/V) sodium dodecyl sulfate, 50 mM dithioerythritol, 2 mM PMSF, 5 mM cyclohexylaminoethane sulfonic acid, pH 9.6     

The Polyunsaturated Fatty Acids of Marine and Freshwater Cryptomonads1

SYNOPSIS Fatty acids were examined of photosynthetic and non-photosynthetic marine and freshwater cryptomonads cultured as photoauxotrophs, photoheterotrophs and heterotrophs at various incubation temperatures and constant light intensity. Photo-synthetic marine and freshwater forms contained octadecatrienoic, octadecatetraenoic, eicosapentaenoic and docosahexaenoic (all-cis, ω3 acids) as the major polyunsaturates, and a freshwater heterotroph contained mostly the octadecatrienoic acid. The polar lipids of a marine, photosynthetic form, Cryptomonas sp., included the usual thylakoid membrane lipids of the chloroplasts of eukaryotic, photosynthetic cells: galactosyl diglycerides, phosphatidyl glycerol and a sulfolipid. Also present were 2 choline-containing phospholipids: phosphatidyl choline and an unknown. Ninhydrin-positive and inositol-containing lipids were not detected. Octadecatetraenoic acid comprised 75% of the total fatty acids of the monogalactosyl diglyceride fraction. The phosphatidyl glycerol was acylated mostly by ω13 trans-hexadecaenoic acid and the eicosapentaenoic acid. Evolutionary relationships of cryptomonads as mirrored in lipid composition are discussed.     

Isolation and characterization of lipid-degrading Bacillus thermoleovorans IHI-91 from an icelandic hot spring

An efficient lipid-degrading thermophilic aerobic bacterium was isolated from an icelandic hot spring and classified as Bacillus thermoleovorans IHI-91. The aerobic bacterium grows optimally at 65°C and pH 6.0 and secretes a high level of lipase (300 U l⁻¹). The newly isolated strain utilizes several lipids such as palmitic acid, stearic acid, lanolin, olive oil, sunflower seed oil, soya oil, and fish oil as sole carbon and energy source without an additional supply of growth factors. The degradation of about 93% of triolein, which is present in olive oil, was observed after only 7 h of fermentation at a maximal growth rate of 1.0 h⁻¹. During growth at optimal conditions on yeast extract, the doubling time was only 15 min. Based on 16S rDNA studies, DNA–DNA hybridization and morphological and physiological properties, the isolate IHI-91 was identified as Bacillus thermoleovorans IHI-91 sp. nov. Because of its production of high concentrations of thermoactive lipases and esterases and the capability of degrading a wide range of lipids at high temperatures, the isolated strain is an ideal candidate for application in various biotechnological processes such as wastewater treatment. Received: August 25, 2000 / Accepted: September 26, 2000     

Fibrivirga algicola gen. nov., sp. nov., an algicidal bacterium isolated from a freshwater river

An aerobic, gram-stain-negative, pink-colored, non-motile and rod-shaped algicidal bacterium, designated as JA-25^T was isolated from freshwater in Geumgang River, Republic of Korea. Strain JA-25^T grew at 15–30 °C and pH 6–9, and did not require NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain JA-25^T belongs to the family ‘Spirosomaceae’ and is most closely related to Fibrella aestuarina BUZ 2^T (93.6%). Strain JA-25^T showed?<?90% sequence similarity to other members of the family ‘Spirosomaceae’. The average nucleotide identity(ANI), in silico DNA-DNA hybridization and average amino acid identity(AAI) values based on the genomic sequences of JA-25^T and F. aestuarina BUZ 2^T were 74.4, 20.5, and 73.6%, respectively. Strain JA-25^T showed an algicidal effect on the marine flagellate alga Heterocapsa triquetra, but no effect on fresh water cyanobacterium (Nostoc). In genome analysis, RIPP-like peptides were detected and predicted to resemble the indolmycin biosynthetic gene cluster, which possibly influence its algicidal effect. Furthermore, a bacteriorhodopsin gene with photoheterotrophic characteristics was detected. The genomic DNA G?+?C content was 52.5 mol%. The major cellular fatty acids were summed feature 3 (C16:1 ω6c/C16:1 ω7c), C16:1 ω5c, C16:0 (>?10%). The major respiratory quinone was menaquinone 7 and major polar lipids were phosphatidylethanolamine, two unidentified aminolipids, two phospholipids, and five unidentified lipids. Considering the phylogenetic inference, phenotypic, and chemotaxonomic data, strain JA-25^T should be classified as a novel species in the novel genus Fibrivirga, with the proposed name Fibrivirga algicola sp. nov. The type strain is JA-25^T (=?KCCM 43334^T?=?NBRC 114259^T).

     

Polysaccharide and lipid composition of the brown seaweed <Emphasis Type="Italic">Laminaria gurjanovae</Emphasis>

Polysaccharide and lipid composition of the Pacific brown seaweed Laminaria gurjanovae is determined. Alginic acid is shown to be the main polysaccharide of its biomass (about 28%); it consists of mannuronic and guluronic acid residues at a ratio of 3: 1. The yield of water-soluble polymannuronic acid is low and does not exceed 1.1% of dry biomass. High laminaran content (about 22%) is found, whereas the yield of fucoidan is no more than 3.6%. Laminaran consists of two fractions, soluble and insoluble in cold water, their ratio is 2.5: 1. Insoluble laminaran is a practically linear 1,3-β-D-glucan, and the soluble fraction was shown to be 1,3;1,6-β-D-glucan. The oligosaccharide products of desulfation or partial acidic hydrolysis of fucoidan were studied by MALDI TOF MS; they were found to be fuco- and galactooligosaccharides. The fucoidan is suggested to be a highly sulfated partially acetylated galactofucan (Fuc/Gal is ～1: 1). The main lipid components of the dried L. gurjanovae are neutral lipids and glyceroglycolipids, whereas phospholipids are found in minor amounts. The main fatty acid components of lipids are 14:0, 16:0, 16:1 ω-7, 18:1 ω-7 and 18:2 ω-6 acids.     

Membrane fractions from the outer layers of spores of Bacillus thuringiensis with toxicity to lepidopterous larvae.

Two membrane fractions, F1 and F2, have been purified from the outer layers of spores of Bacillus thuringiensis. Both fractions contain 6-7% cysteine and appear to be similar in composition. Amino acids account for about 75% of the dry weight, carbohydrate for about 2% and lipids for about 25%. The fractions are both toxic to Pieris brassicae and the toxicity is inactivated by antiserum to the toxic crystal of Bacillus thuringiensis. The fractions can be distinguished by examination under the electron microscope; both fractions show similar hexagonal patterns but with different spacings. The same fractions from an acrystaliferous mutant (cr) were prepared. These were identical in density and in appearance under the electron microscope; the amino acid analysis of fraction F2 from both strains was identical. However, the spores and fractions F1 and F2 from this strain lacked toxicity. Fraction F2 from the cr strain was used to prepare antiserum specific to fraction F2. Using this anti-serum and anticrystal serum, crystal and F2 antigens were shown to appear simultaneously in sporulating cultures. Crystal and F2 antigens appeared some time before the maximum rate of uptake of [35s]cysteine. It is concluded that fraction F2 is derived from the exosporium and that fraction F1 probably originates from the spore coat. The exosporium in Bacillus thuringiensis appears to be synthesised during stages II and III of sporulation although uptake of [35S]cysteine occurs much later.     

Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea

Thermal stress is considered one of the main causes of mass scleractinian coral degradation; however, it is still unknown how corals can adapt to future global warming. In this study, 11 strains of coral-associated Flavobacteria were shown to produce zeaxanthin, a carotenoid antioxidant, which may help coral holobionts to alleviate thermal stress. In addition, a novel zeaxanthin-producing Flavobacterium, designated R38^T, was identified using polyphasic taxonomy. Although strain R38^T shared a maximum 16S rRNA gene sequence similarity of 93% with Mesoflavibacter aestuarii KYW614^T, phylogenetic analyses based on whole genome and 16S rRNA gene sequences revealed that strain R38^T forms a distinct branch in a robust cluster composed of strain R38^T and Leptobacterium flavescens KCTC 22160^T under the family Flavobacteriaceae. Strain R38^T exhibited average nucleotide identities of 70.2% and 72.5% for M. aestuarii KYW614^T and L. flavescens KCTC 22160^T, respectively. The only detected respiratory quinone was menaquinone 6 (MK-6). The genomic DNA G?+?C content was 33.2 mol%. The major polar lipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids. The major cellular fatty acids were iso???C_15:?0, iso???C_15:?0 ω6c, C_16:2 DMA, and C_13:1 ω3c. The distinct biochemical, chemotaxonomic, phylogenetic, and phylogenomic differences from validly published taxa suggest that strain R38^T represents a new species of a new genus, for which Prasinibacter corallicola gen. nov., sp. nov. is proposed. The type strain R38^T (=?MCCC 1K03889^T?=?KCTC 72444^T).

     

Isolation and characterization of thermophilic,alkaliphilic, cellulose‐degrading bacillus thermoalcaliphilus sp. nov. from termite (odontotermes obesus) mound soil of a semiarid area

A new heterotrophic, thermophilic, alkaliphilic, facultatively anaerobic, cellulose‐degrading bacterium strain STS1 was isolated from mound soils infested with the higher termite Odontotermes obesus in the semiarid ecosystem of Delhi (India). The gram‐positive, spore‐forming, catalase‐positive Bacillus sp. grew on natural and raw celluloses. The taxonomic position of the organism was investigated. The guanine plus cytosine content of the isolate was found to be 48.6 mol% (melting temperature profile). Addition of peptone or yeast extract stimulated growth. The isolate did not grow on silica gel plates or on agar media in which the agar was the sole source of carbon and energy. The high growth temperature of 70°C and the pH of 9.0 are characteristic of this species. The role of this bacterium in the semiarid ecosystem is discussed. Because of its high optimum temperature and high optimum pH for growth, the name Bacillus thermoalcaliphilus is proposed.     

Bacillus oceani sp. nov., a new slightly halophilic bacterium, isolated from a deep sea sediment environment

A strictly aerobic, Gram-stain positive, slightly halophilic strain, designated SCSIO 04524^T, was isolated from a deep sea sediment sample collected from the northern South China Sea at a depth of 3415 m. The isolate slightly embedded into the medium after 72 h incubation at 30 °C. Growth was found to occur on media with 0–10 % NaCl but extremely weak growth occurred without supplying NaCl. The predominant menaquinone was determined to be MK-7. The major cellular fatty acid identified was iso-C_15:0. The diagnostic polar lipids were determined to be diphosphatidylglycerol, phosphatidyl methylethanolamine, phosphatidylethanolamine and phosphatidylglycerol. The genomic DNA G+C content was determined to be 38 mol%. 16S rRNA gene sequences analysis showed that this strain had the highest similarities with Bacillus carboniphilus JCM 9731^T (94.7 %) and Bacillus endophyticus 2DT^T (94.3 %). Phylogenetic analysis revealed that strain SCSIO 04524^T formed a distinct lineage with Bacillus chungangensis CAU 348^T and B. carboniphilus JCM 9731^T. Physiological characteristics including utilization of sole nitrogen and carbon sources, and chemotaxonomic properties of cellular fatty acids and polar lipids could readily distinguish strain SCSIO 04524^T from its most closely related species. Based on this polyphasic taxonomic data, a new species, Bacillus oceani sp. nov., is proposed, with the type strain SCSIO 04524^T (=DSM 26213^T = KCTC 33077^T).     

Characterization of a Chlorophyta microalga isolated from a microbial mat in Salar de Atacama (northern Chile) as a potential source of compounds for biotechnological applications

Microalgae are an important source of unsaturated fatty acids, phospholipids, glycolipids, and carotenes, which are useful compounds for the food and pharmaceutical industries. The Atacama Desert of northern Chile is one of the driest deserts on Earth and, as such, it is a great natural laboratory in which to study new microorganisms adapted to extreme environments. A microalgal strain, referred to here as CH03, was isolated from a microbial mat in salt flat water in Salar de Atacama. Genetic analysis of the 18S ribosomal RNA gene showed that the strain had homology with other known sequences of the species Chlorella sorokiniana. Our results revealed the adaptability of this microalga to freshwater medium under laboratory conditions, despite coming from an extremely high‐salinity environment. The fatty acid profile of CH03(A) newly isolated in Bold's basal medium differed from that of CH03(B) cultured in vitro in modified F/2 medium and from another five strains of C. sorokiniana and three strains of Chlorella vulgaris in that it had a high stearic acid content and had no polyunsaturated fatty acids. The major biochemical components observed in this strain were proteins (64.3–73.6%) and lipids (26.6–32.6%). This study suggests that the strain CH03 could be a protein source and that this oleaginous microalga is easy to grow in vitro as a biological model for future studies.     

Bacillus manliponensis sp. nov., a new member of the Bacillus cereus group isolated from foreshore tidal flat sediment

A Gram-positive, endospore-forming, new Bacillus species, strain BL4-6^T, was isolated from tidal flat sediment of the Yellow Sea. Strain BL4-6^T is a straight rod, with motility by peritrichate flagella. The cell wall contains meso-diaminopimelic acid, and the major respiratory quinone is menaquinone-7. The major fatty acids are iso-C_15:0 and summed feature 3 (containing C_16:1 ω7c/iso-C_15:0 2OH, and/or iso-C_15:0 2OH/C_16:1 ω7c). Cells are catalase-positive and oxidase-negative. The G+C content of the genomic DNA is 38.0 mol%. Based on a comparative 16S rRNA gene sequence analysis, the isolate belongs to the genus Bacillus, forms a clade with the Bacillus cereus group, and is closely related to Bacillus mycoides (98.5%), Bacillus cereus (98.5%), Bacillus anthracis (98.4%), Bacillus thuringiensis (98.4%), Bacillus weihenstephanensis (98.1%), and Bacillus pseudomycoides (97.5%). The isolate showed less than 85% similarity of the gyrA gene sequence and below 95% similarity of the rpoB gene sequence to the members of this group. DNA-DNA relatedness between strain BL4-6^T and B. cereus group was found to be in a range of 22.8–42.3%, and thus BL4-6^T represents a unique species. On the basis of these studies, strain BL4-6^T (=KCTC 13319^T =JCM 15802^T) is proposed to represent the type strain of a novel species, Bacillus manliponensis sp. nov.     

Isolation and composition of the carotenoid-containing oil droplets from cone photoreceptors.

A procedure for isolating the carotenoid-containing oil droplets of cone retinal photoreceptors of Gallus domesticus is described. The oil droplets, composed almost entirely of neutral lipids and carotenoids, have been separated into ten chromatographic components. Similar separations have been carried out on the total retinal neutral lipids for comparison. The neutral lipids represented 26.1% of the total retinal lipid. Cholesterol, cholesterol ester, mono-, di- and triacylglycerols represented 92.6% of the total neutral lipid. Each of these and other minor neutral lipid components were also present in the lipids extracted from the isolated oil droplets in correspondingly similar concentrations. However, the concentrations of carotenoids were greatly enriched in the neutral lipids of the oil droplets. Each of the major fatty acyl-containing neutral lipids from the chromatography of oil droplet lipids is greatly enriched in polyunsaturated fatty acids when compared with the corresponding component from the total neutral lipid chromatography. In the acylglycerols and free fatty acid fraction from the oil droplets, linoleic and arachidonic acid together represented 52-83% of the total polyunsaturated fatty acids present. The remainder was generally distributed about equally among six other acids. Except for the diacylglycerol fraction, linoleic acid was usually the most enriched acid in a specific oil droplet fraction when compared with any other polyunsaturated fatty acids. A similar pattern of polyunsaturated fatty acid enrichment observed in the fatty acids of the outer segment phospholipids relative to the corresponding total phospholipid fractions of this cone rich retina (Johnston, D. and Hudson, R.A. (1974) Biochim. Biophys. Acta 369, 269) suggest possible metabolic relationships between the oil droplet neutral lipids and the outer segment membrane phospholipids of the cone photoreceptors. A mechanism for the accumulation of the carotenoids in the oil droplets is also discussed.     

Effects of ω3 Polyunsaturated Fatty Acids on Growth and Tissue Fatty Acid Compositions of Maternal Rats and Their Progeny

The effects of ω3 polyunsaturated fatty acids [PUFA; mainly eicosapentaenoic acid (20: 5, ω3) and docosahexaenoic acid (22:6, ω3)] on the growth, tissue weights and fatty acid compositions of tissue total lipids in female rats and their progeny were investigated. Female rats of the Wistar strain, weighing 77~94g, were fed a 25% casein diet containing 5% of either corn oil (control), sardine oil or PUFA ethyl ester for 8 ~ 9 weeks prior to mating, and during gestation and lactation, and then for a further 2 weeks. The progeny were weaned to the maternal diet and then the latter was administered for a further 2 weeks. Dietary changes in the body weights of the dams were not generally seen, but the body weights at birth and growth of the offspring from the females supplied with the PUFA diet were inferior compared to those of the other groups. The fertility did not differ among the dietary groups. The weights of several tissues in the dams and the progeny increased in proportion to their body weights but not that of the progeny brain, which remained ' almost unchanged by the dietary fats. As to the fatty acid compositions of total lipids in the tissues, on the whole, decreased levels of ωβ fatty acids and increased percentages of ω3 fatty acids were found in the sardine and PUFA groups, the changes being greater in the PUFA group than in the sardine one. Such findings due to the feeding of PUFA were more remarkable in the progeny compared with in the dams. Eicosatrienoic acid (20: 3, ω9) was almost completely undetectable in the tissue total lipids of all the dietary groups.     

Screening of oleaginous algal strains from Chlamydomonas reinhardtii mutant libraries via density gradient centrifugation

Microalgae have a high potential to be utilized as feedstock for biofuels because they have high growth rates and do not compromise food production. Commercialized algae-based biofuel production relies on the development of strains with high lipid content. Based on the relatively low density of lipids compared to other cellular components, density gradient centrifugation was used to isolate high lipid content algal strains from Chlamydomonas reinhardtii mutant libraries. The correlation between cell density and lipid content was confirmed by analysis of Nile red fluorescence intensity, total lipids, and total fatty acid methyl ester content. A strain isolated by this screening method had 50% higher lipid content and 7% lower cell density than the parent wild-type strain. Consequently, we demonstrated that screening of algal strains with low cell density via continuous density gradient centrifugation allows simple, rapid, and inexpensive screening for high lipid content strains.