The Renin Angiotensin System (RAS) plays a vital role in the regulation of blood pressure and fluid homeostasis. RAS is regulated via the hormone Angiotensin II through an association with the Na+/H+ exchanger NHE6. Here, NHE6 was found to be activated by Angiotensin II through the Angiotensin II AT1 receptor. Furthermore, it was shown that NHE6 requires phosphorylation for activation and this phosphorylation signaling mechanism does not involve phospholipase C. The elucidation of the signaling pathway associated with NHE6 and AT1 allows for the greater understanding of function and regulation of the NHE6 protein. The Angiotensin receptor AT2 is a G-coupled protein receptor (GPCR) that is highly expressed in infant neural tissue. The S1 subunit of the pertussis toxin can inhibit GPCR signaling via ADP-ribosylation of the cognate Gi protein, suggesting that the S1 subunit may interfere with AT2 signaling. In order to observe whether S1 associates with AT2, Chinese hamster ovary cells were transfected with plasmids expressing AT2 or mutants of AT2. The lysates of these cells were incubated with His-tagged S1 subunit and it was observed that only the wild-type AT2 co-immunoprecipitated with S1. These results imply that there is a direct interaction between the S1 subunit and AT2. Municipal wastewater can be considered as an effective growth medium for the cultivation of microorganisms due to organic material found in the water. Oleaginous microorganisms produce large amounts of triacylglycerols (TAGs) when cultivated on medium containing high sugar content and low nitrogen. These TAGs can then be converted into biodiesel. To determine if the oleaginous yeast Rhodotorula glutinis could survive and synthesize lipids using wastewater as a cultivation medium, R. glutinis was inoculated into primary effluent wastewater supplemented with glucose. Results indicated that R. glutinis was able to survive and synthesize lipids in the wastewater which is suggestive that R. glutinis can successfully compete with indigenous microorganisms in the wastewater.

No description available.

Foi realizado um estudo para verificar a ocorrência de leveduras do gênero Rhodotorula, em hemocultura por um período de 8 anos. Os pacientes identificados foram descritos clinicamente segundo variáveis de interesse incluindo dados sobre terapêutica e desfecho. Determinou-se também as concentrações inibitórias mínimas de 20 cepas frente a diferentes antifúngicos de acordo com NCCLS e EUCAST. Realizou-se tipagem molecular através da cariotipagem eletroforética em campo pulsátil / A study was conducted to verify the frequency of occurrence of Rhodotorula spp. from blood cultures over an 8-year period, clinically and microbiologically characterizing patients affected, including data regarding antifungal treatment and outcome. The minimal inhibitory concentrations of antifungal agents were determined against 20 isolates. Molecular typing of the strains were performed using pulsed field gel electrophoresis method

Foi realizado um estudo para verificar a ocorrência de leveduras do gênero Rhodotorula, em hemocultura por um período de 8 anos. Os pacientes identificados foram descritos clinicamente segundo variáveis de interesse incluindo dados sobre terapêutica e desfecho. Determinou-se também as concentrações inibitórias mínimas de 20 cepas frente a diferentes antifúngicos de acordo com NCCLS e EUCAST. Realizou-se tipagem molecular através da cariotipagem eletroforética em campo pulsátil / A study was conducted to verify the frequency of occurrence of Rhodotorula spp. from blood cultures over an 8-year period, clinically and microbiologically characterizing patients affected, including data regarding antifungal treatment and outcome. The minimal inhibitory concentrations of antifungal agents were determined against 20 isolates. Molecular typing of the strains were performed using pulsed field gel electrophoresis method

Rhodotorula glutinis (Rhodosporidium toruloides) est une levure oléagineuse dont les fortes capacités d’accumulation lipidique à partir de glucose comme source carbonée ont fait de la souche un modèle d’étude. La capacité de cette levure à utiliser le glycérol ou le xylose en simple ou co-substrat avec le glucose est toutefois encore peu explorée. De l’analyse des travaux antérieurs il a été possible de dégager les verrous scientifiques qui nécessitent une amélioration des connaissances du comportement physiologique de cette levure pour la conversion des substrats cités. Des stratégies expérimentales adaptées à la quantification rationnelle des dynamiques de Rhodotorula glutinis en conditions de croissance et accumulation lipidique à partir de xylose et de glycérol en simple ou co-substrats avec le glucose ont été développées. Des résultats originaux ont été obtenus dont :- la mise en évidence des potentialités de co-consommation des substrats xylose et glucose ou glycérol et glucose sans accumulation de substrat ni production de métabolites en conditions contrôlées des flux de substrats. Il a été possible de déterminer la vitesse spécifique maximale de consommation du carbone de la souche qui diminue lorsque la part de xylose ou glycérol augmente dans l’apport de carbone total.- la quantification de la dynamique de croissance sur xylose et glycérol pur en terme de taux de croissance et de rendement : sur xylose µmax= 0.034h-1 et RS/X= 0.28 Cmolx.Cmolxylose-1; sur glycérol µmax=0.04h-1 RS/X=0.31Cmolx.Cmolglycérol-1.- la quantification des vitesses spécifiques et rendements de production de lipides à partir de xylose ou de glycérol en simple ou co-substrat avec du glucose : 20%xylose-80%glucose : qp=0.065CmolTAG.Cmolbiomasse.h-1, RS/P=0.3CmoleTAG.Cmolesubstrat-1 100%xylose : qp=0.035065CmolTAG.Cmolbiomasse.h-1, RS/P=0.31CmoleTAG.Cmolesubstrat-1, 25% glycérol-75%glucose : qp=0.07065CmolTAG.Cmolbiomasse.h-1, RS/P=0.25CmoleTAG.Cmolesubstrat-1 , 100% glycérol : qp=0.03065CmolTAG.Cmolbiomasse.h-1, RS/P= 0.29CmoleTAG.Cmolesubstrat-1.- L’impact de la nature des substrats sur le profil lipidique de Rhodotorula glutinis demeure léger : il apparait que le xylose entraîne une surproduction de C16:0 et C18:3et le glycérol favorise l’accumulation de C18:0 / Rhodotorula glutinis (Rhodosporidium toruloides) is an oleaginous yeast. The micro-organism has demonstrated high lipid accumulation when utilizing glucose as a substrate, and has become a model for oil production. Glycerol and xylose are interesting as substrates for production of oil from renewable resources, but the capacity of R. glutinis to utilize glycerol and xylose as substrates has not been characterized well. Fermentation strategies were designed to quantify growth and lipid accumulation dynamics of R. glutinis when utilizing glycerol and xylose - either as pure substrates, or as co-substrates with glucose. Several original results have been found, including: - Co-consumption of xylose or glycerol along with glucose was observed, without carbon substrate accumulation or byproduct formation, when the carbon feed rate was carefully controlled. The specific carbon consumption rate decreases when the proportion of the second substrate (glycerol or xylose) increases in the feed, relative to glucose. - Growth capacities were characterized on pure xylose and pure glycerol in terms of growth rate and carbon yields: on xylose μmax= 0.034h-1 and RS/X= 0.28 Cmolx.Cmolxylose-1; on glycerol μmax=0.04h-1 RS/X=0.31Cmolx.Cmolglycerol-1. - specific production rate of lipid production and substrate to product carbon conversion yields from xylose or glycerol as single or cosubstrate with glucose were determinated: 20%xylose-80%glucose : qp=0.065CmolTAG.Cmolbiomasse.h-1, RS/P=0.3CmoleTAG.Cmolesubstrat-1 100%xylose : qp=0.035065CmolTAG.Cmolbiomasse.h-1, RS/P=0.31CmoleTAG.Cmolesubstrat-1, 25% glycerol-75%glucose : qp=0.07065CmolTAG.Cmolbiomasse.h-1, RS/P=0.25CmoleTAG.Cmolesubstrat-1 , 100% glycerol : qp=0.03065CmolTAG.Cmolbiomasse.h-1, RS/P= 0.29CmoleTAG.Cmolesubstrat-1. - Substrate diversification slightly impacts Rhodotorula glutinis´s lipid profile: xylose leads to an overproduction of C16:0 and C18:3 and glycerol increases C18:0 accumulation

Over recent years microbial oil has attracted much attention due to its potential to replace traditional oil sources in the production of biofuels and nutraceuticals. Its advantages arise from its independence of the food supply chain and its ease of production compared to conventional plant oils. Also, as concerns for the environment grow, microbially-synthesized oil emerges as potential competitor for the sustainable production of biodiesel. However, the high cost of its production currently hinders its large scale application. The bottlenecks to industrial microbial oil production are the cost of substrate and cultivation. Current research is focusing on process improvements to make microbial oil more competitive and worthwhile to produce. Several types of microorganisms have been explored so far and waste substrates have been utilised as cheap feedstocks. The overall cost is affected by the fermentation stage, therefore it is imperative to design cultivations with little operating requirements and high yields. Consequently, the present thesis aims to contribute to the field by developing and investigating a simple process for oleaginous yeast cultivation, focusing mainly on enhancing the yields during the bioreactor stage. Oleaginous yeasts were screened for their ability to grow on glycerol and the most promising strain was selected for further research. Then, the necessary conditions for its growth and oil accumulation were defined. Shake-flask cultivations showed that the specific growth rate and glycerol consumption of Rh. glutinis were higher at lower glycerol concentrations (smaller or equal to40 g/L), while higher C/N elemental ratios enhanced oil content. Experimental data were used to construct an unstructured kinetic model to describe and predict the system's behaviour. The Monod-based model took into account double substrate growth dependence and substrate inhibition. Following that, bioreactor cultivations extended the range of parameters studied, to include the influence of aeration rate and oxygen supply on cellular growth and microbial oil production. Cultivations at different air flow rates were performed in a 2 L bioreactor and showed that a low aeration rate of 0.5 L/min gave the best glycerol and nitrogen uptake rates, resulting in a concentration of biomass of 5.3 g/L with oil content of 33% under simple batch operation. This was improved by 68% to 16.8 g/L (cellular biomass) with similar oil content (34%) by applying a fed-batch strategy. Finally, different glycerol feeding schemes were evaluated in terms of their effect on oil accumulation. The concept of targeting first a cell proliferation stage, limited by the availability of nitrogen, followed by a lipid accumulation stage, fuelled by glycerol was tested. Continual feeding and pulsed feedings, delivering the same total amount of nitrogen (and glycerol), resulted in similar elevated values of both cellular biomass (~25 g/L) and oil content (~40%). Addition of glycerol at higher rates but giving the same total amount of nitrogen led to a further increase in oil content to 53%, resulting in an overall oil yield of more than 16 g/L (the highest achieved throughout the project). With comparable yields to those reported in the literature but achieved with a much poorer medium, there is every reason to be optimistic that microbial oil production from glycerol could be commercially viable in the future.

Das MMS stellt eine lebensbedrohliche Erkrankung großwüchsiger Hunde dar. Die ansteigenden Inzidenzzahlen der letzten Jahre, die ungeklärte Ätiologie sowie die hohe Mortalitätsrate rechtfertigen weitere Untersuchungen zu diesem Krankheitskomplex. Daher wurden in der vorliegenden Arbeit Mageninhalte (30 Tiere mit MMS / 13 Kontrolltiere) und Blutproben (102 Tiere mit MMS / 116 Kontrolltiere) mikrobiologisch, biochemisch (kurz-kettige Fettsäuren, Amylase, Lipase, Laktat, pH) und immunologisch (BotNt, CRP, Gesamt-immunglobuline, IgA-, IgG- und IgM-Spiegel ausgewählter mikrobieller Antigene) untersucht. Die Untersuchung des Mageninhaltes der Tiere mit MMS erbrachte signifikant gesteigerte Nachweishäufigkeiten für Hefen, hier v.a. von Rhodotorula mucilaginosa. Des Weiteren konnten in der Gruppe der Tiere mit MMS signifikant gesteigerte pH-Werte sowie signifikant erhöhte Mengen an Acetat, Butyrat und Gesamtfettsäuren im Vergleich zu den Kontrolltieren festgestellt werden. Die gesteigerte bakterielle Fermentation, die ursächlich mit der Entstehung des MMS zusammenhängen dürfte, ist vermutlich auf ein gesteigertes Vorkommen gasbildender Kokken zurückzuführen. Ein Einfluss von Clostridium perfringens an der Entstehung des MMS lässt sich anhand der ermittelten bakteriologischen Ergebnisse nicht belegen. BotNt haben nach den in der vorliegenden Arbeit gewonnenen Daten keinen Einfluss auf die Entstehung des MMS. Serologisch konnten in der Gruppe der Tiere mit MMS signifikant erhöhte CRP-Spiegel gemessen werden. Das CRP ließ aber keine prognostische Aussage über die Überlebenschancen der Tiere zu. Der erhöhte Gesamtimmunglobulin A-Spiegel, bei signifikant erniedrigtem Gesamtimmunglobulin G-Titer weist auf eine vermehrte Auseinandersetzung mit Antigenen hin, die über die geschädigten Schleimhäute eindringen können. Während der Serumlaktatspiegel eine prognostische Aussage über das Outcome der Tiere erlaubt, erscheint eine Bestimmung der Amylase und Lipase beim MMS nicht sinnvoll. Über einen Fragebogen konnte ein gesteigertes Risiko mit zunehmendem Alter, steigender Futtermenge pro Mahlzeit und niedriger Fütterungsfrequenz ermittelt werden. Eine gesteigerte Freßgeschwindigkeit übt einen tendenziellen Einfluss auf die Ausbildung des MMS aus. Keinen Einfluss dahingegen haben das Geschlecht und der Charakter der Tiere, bestehende gastrointestinale Störungen sowie vorangegangene Antibiotikumgaben.

The genus Rhodotorula includes basidiomycetous oleaginous yeast species. Rhodotorula babjevae can produce compounds of biotechnological interest such as lipids, carotenoids, and biosurfactants from low value substrates such as lignocellulose hydrolysate. High-quality genome assemblies are needed to develop genetic tools and to understand fungal evolution and genetics. Here, we combined short- and long-read sequencing to resolve the genomes of two R. babjevae strains, CBS 7808 (type strain) and DBVPG 8058, at chromosomal level. Both genomes are 21 Mbp in size and have a GC content of 68.2%. Allele frequency analysis indicates that both strains are tetraploid. The genomes consist of a maximum of 21 chromosomes with a size of 0.4 to 2.4 Mbp. In both assemblies, the mitochondrial genome was recovered in a single contig, that shared 97% pairwise identity. Pairwise identity between most chromosomes ranges from 82 to 87%. We also found indications for strain-specific extrachromosomal endogenous DNA. A total of 7591 and 7481 protein-coding genes were annotated in CBS 7808 and DBVPG 8058, respectively. CBS 7808 accumulated a higher number of tandem duplications than DBVPG 8058. We identified large translocation events between putative chromosomes. Genome divergence values between the two strains indicate that they may belong to different species.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-24T11:51:36Z No. of bitstreams: 1 Marcelo Rodrigues Figueira de Mello.pdf: 590787 bytes, checksum: 0b21179fd04d627436888523d18d7338 (MD5) / Made available in DSpace on 2017-03-24T11:51:36Z (GMT). No. of bitstreams: 1 Marcelo Rodrigues Figueira de Mello.pdf: 590787 bytes, checksum: 0b21179fd04d627436888523d18d7338 (MD5) Previous issue date: 2009-02-16 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The production of Chinese cabbage (Brassica pekinensis) may be limited by occurrence of diseases, among which the soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc). The effect of inducers and antibiotics on the disease control was studied in laboratory, greenhouse and field. In the first paper acibenzolar-S-metil (ASM) (0.025 g L-1), Ecolife® (2 mL L-1), Agro-Mos® (2 mL L-1) and calcium oxide (0.22 g L-1) were evaluated for in vitro bacterial growth inhibition; epidemiological components, incidence (INC), incubation period (PI), disease final severity (SEVF), diasease index (IDO) and area under disease curve progress (AUPDC); enzymatic activity of peroxidase and polyphenoloxidase and physiological cost of induction. Under greenhouse and field conditions inducers were sprayed seven days after transplant, and seven days after induction strain Pcc120 was inoculated by pricking, except for enzymatic activity and physiological cost experiments. In vitro Pcc120 was not inhibited by any tested product. The INC was not reduced in any experiment. In greenhouse SEVF was reduced by 47.5% by Agro-Mos® and ASM which also reduced IDO by 35.1 and 45.3% respectively and AUDCP. Induced resistance was confirmed by negative antibiosisagainst pathogen and increase of peroxidase and polyphenoloxidase activities in treated plants. In field only ASM confirmed greenhouse results by reducing disease intensity. There was no physiological cost for plants sprayed with ASM or Agro-Mos®. In the second paper it was evaluated the in vitro sensibility of Pcc to bactericides, and the effect of Mycoshield® (oxitetracycline 20%) at 3.0 and 1.5 g L-1, and yeasts (Rh1 and Rh2 - Rhodotorula spp. and Sc1 - Saccharomyces cerevisae at 108 cel mL-1) to control the disease in greenhouse and field. Plants were sprayed with Mycoshield® and yeasts seven days after transplant and inoculated seven days and 12 h after treatment, respectively. In all experiments INC, PI, SEVF, IDO and AUDPC were evaluated. In vitro 40 Pcc isolates were resistant to copper sulfate and sensitive to oxitetracycline, streptomycin, oxitetracycline+ streptomycin and oxitetracycline + copper sulfate all at 0.2 g L-1. Six Pcc isolates were significantly more inhibited by MycoshieldÒ than by Agri-Micina® (oxitetracycline 1.5% + streptomycin 15%), but there was no inhibition by Kasumin® (kasugamicin 2%). In greenhouse MycoshieldÒ at 3.0 g L-1 reduced SEVF and IDO until 47.4 and 19.0%. The yeast Sc1 reduced SEVF and AUDCP till 27.6 and 39.3% respectively, while Rh1 reduced AUDCP until 33.5%. In field MycoshieldÒ reduced IDO, SEVF and AUDCP by 14.4; 15.5 and 28.9% respectively; while Rh1 reduced IDO by 8.8% and Sc1diminished the AUDCP by 15.7%. The reduction of disease intensity and low physiological cost of induction indicate the potentiality of ASM in an integrate management program of Chinese cabbage soft rot. On the other hand MycoshieldÒ and yeasts showed low efficiency for controlling disease in field. / A produção de couve-chinesa (Brassica pekinensis) pode ser limitada pela ocorrência de doenças, dentre as quais se destaca a podridão-mole causada por Pectobacterium carotovorum subsp. carotovorum (Pcc). O efeito de indutores e antibióticos no controle desta doença foi estudado em laboratório, casa de vegetação e campo. No primeiro trabalho, foram testados acibenzolar-S-metil (ASM) (0,025 g/L), Ecolife® (2 mL/L), Agro-Mos® (2 mL/L) e óxido de cálcio (0,22 g/L). Foram avaliados, a inibição do crescimento bacteriano “in vitro”; os componentes epidemiológicos, incidência (INC), período de incubação (PI), severidade final da doença (SEVF), índice de doença (IDO) e área abaixo da curva de progresso da doença (AACPD); a atividade das enzimas peroxidase e polifenoloxidase e o custo fisiológico da indução. Em casa de vegetação e campo os indutores foram pulverizados sete dias após o transplante e sete dias após a indução, o isolado Pcc120 foi inoculado por picada, exceto nos experimentos de atividade enzimática e custo fisiológico. Pcc120 não foi inibido “in vitro” por nenhum dos produtos testados. Não houve redução da INC em nenhum dos experimentos. Em casa de vegetação, a SEVF da doença foi reduzida em 47,5% pelos tratamentos Agro-Mos® e ASM, os quais também, respectivamente, reduziram o IDO em 35,1% e 45,3% e propiciaram as menores AACPD. A resistência induzida foi evidenciada pela ausência de antibiose dos dois produtos contra Pcc e pelo aumento das atividades da peroxidase e polifenoloxidase. Em campo, apenas o ASM confirmou os resultados de casa de vegetação reduzindo a intensidade da doença. Não houve custo fisiológico para as plantas com aplicação do ASM ou Agro-Mos®. No segundo trabalho avaliou-se a sensibilidade “in vitro” de Pcc a bactericidas, o efeito de Mycoshield® (oxitetraciclina 20%) nas dosagens de 3,0 e 1,5 g/L, e das leveduras (Rh1 e Rh2 - Rhodotorula spp. e Sc1 - Saccharomyces cerevisae a 108 cel/mL) no controle da doença em casa de vegetação e em campo. As plantas foram pulverizadas com Mycoshield® e leveduras sete dias após o transplante, e inoculadas sete dias e 12 h após o tratamento, respectivamente. Foram avaliados INC, PI, SEVF,IDO e AACPD. In vitro, 40 isolados de Pcc testados apresentaram resistência ao sulfato de cobre e sensibilidade a oxitetraciclina, estreptomicina, oxitetraciclina+estreptomicina e oxitetraciclina+sulfato de cobre, todos na concentração de 0,2 g/L. Seis isolados de Pcc foram significativamente mais inibidos por MycoshieldÒ do que por Agri-Micina® (oxitetraciclina 1,5% + estreptomicina 15%), não sendo inibidos por Kasumin® (casugamicina 2%). Em casa de vegetação, o MycoshieldÒ na dosagem 3,0 g/L reduziu a SEVF e o IDO em até 47,4 e 19%; já a levedura Sc1 reduziu a SEVF e a AACPD em até 27,6 e 39,3%, respectivamente, enquanto Rh1 reduziu a AACPD em até 33,5. Em campo, o MycoshieldÒ reduziu o IDO, a SEVF e a AACPD em respectivamente 14,4; 15,5 e 28,9%; enquanto que Rh1 reduziu o IDO em 8,8% e Sc1 reduziu a AACPD em 15,7%. A redução da intensidade da doença e o baixo custo fisiológico da indução indicam a potencialidade do uso do ASM em um programa de manejo integrado da podridão-mole em couve-chinesa. No entanto,o MycoshieldÒ e as leveduras apresentaram baixa eficiência para o controle da doença em campo.

Lipids are stored in a cell for providing energy. The main advantages of storing lipids over carbohydrates like glycogen is that, lipids yield more energy after oxidation because they represent the highly reduced form of carbon, needs less space and water for storage. Conservation of chemical energy in the form of biologically inert form is by storing molecules like triacylglycerol (TAG) and Steryl esters (SE). Triacylglycerol is the major storage form of energy in all eukaryotic cells. During the periods of nutritional excess and nutritional stress, all organisms like bacteria, yeast, animals, and plants can able to do the critical function of synthesizing the triacylglycerol. TAG is an energy store and a repository of essential and non-essential fatty acids and precursors for phospholipid biosynthesis. TAG synthesis mainly takes place in endoplasmic reticulum in mammals and in plants, it takes place in plastid and mitochondria. Triacylglycerol synthesis discovered by Kennedy starts with glycerol 3- phosphate. Glycerol 3-phosphate gets acylate to form lysophosphatic acid (LPA), which in turn acylate to form phosphatic acid (PA) and the reactions are catalyzed by glycerol 3-phosphate acyltransferase (GPAT) and LPA acyltransferase (LPAT) respectively. PA undergoes phosphorylation by PA phosphatase enzyme to give diacylglycerol (DAG). Further acylation of DAG gives rise to TAG and the reaction is catalyzed by diacylglycerol acyltransferase (DGAT). There are several DGAT classes were identified they are DGAT1, DGAT2, PDAT and bifunctional TAG/wax ester synthase. However all the enzymes involved in Kennedy TAG biosynthetic pathway as well as the enzymes of all different DGAT classes are membrane bound enzymes. Through our studies an another DGAT class that is soluble and cytosolic DGAT was first identified in peanut and also in yeast, Rhodotorula glutinis in which a soluble cytosolic complex itself has been identified. The biosynthesis of triacylglycerol (TAG) occurs in the microsomal membranes of eukaryotes. Here, we report the identification and functional characterization of diacylglycerol acyltransferase (DGAT), a member of the 10 S cytosolic TAG biosynthetic complex (TBC) in R. glutinis. Both a full-length and an N-terminally truncated cDNA clone of a single gene were isolated from R. glutinis. The DGAT activity of the protein encoded by RgDGAT was confirmed in vivo by the heterologous expression of cDNA in a Saccharomyces cerevisiae quadruple mutant (H1246) that is defective in TAG synthesis. RgDGAT overexpression in yeast was found to be capable of acylating diacylglycerol (DAG) in an acyl-CoA-dependent manner. Quadruple mutant yeast cells exhibit growth defects in the presence of oleic acid, but wild-type yeast cells do not. In an in vivo fatty acid supplementation experiment, RgDGAT expression rescued quadruple mutant growth in an oleate-containing medium. We describe a soluble acyl-CoA-dependent DAG acyltransferase from R. glutinis that belongs to the DGAT3 class of enzymes. The study highlights the importance of alternate TAG biosynthetic pathway in oleaginous yeasts. A key step in the triacylglycerol (TAG) biosynthetic pathway is the final acylation of diacylglycerol (DAG) by DAG acyltransferase. In silico analysis has revealed that the DCR (defective in cuticular ridges) (At5g23940) gene has a typical HX4D acyltransferase motif at the N-terminal end and a lipid binding motif VX2GF at the middle of the sequence. To understand the biochemical function, the gene was overexpressed in Escherichia coli, and the purified recombinant protein was found to acylate DAG specifically in an acyl-CoA-dependent manner. Overexpression of At5g23940 in a Saccharomyces cerevisiae quadruple mutant deficient in DAG acyltransferases resulted in TAG accumulation. At5g23940 rescued the growth of this quadruple mutant in the oleate-containing medium, whereas empty vector control did not. Lipid particles were localized in the cytosol of At5g23940-transformed quadruple mutant cells, as observed by oil red O staining. There was an incorporation of 16-hydroxyhexadecanoic acid into TAG in At5g23940-transformed cells of quadruple mutant. Here we report a soluble acyl-CoA-dependent DAG acyltransferase from Arabidopsis thaliana. Taken together, these data suggest that a broad specific DAG acyltransferase may be involved in the cutin as well as in the TAG biosynthesis by supplying hydroxy fatty acid.