Β-carotene Accumulation Research Articles

Impacts of long-term enhanced UV-B radiation on bryophytes in two sub-Arctic heathland sites of contrasting water availability.

Anthropogenic depletion of stratospheric ozone in Arctic latitudes has resulted in an increase of ultraviolet-B radiation (UV-B) reaching the biosphere. UV-B exposure is known to reduce above-ground biomass and plant height, to increase DNA damage and cause accumulation of UV-absorbing compounds in polar plants. However, many studies on Arctic mosses tended to be inconclusive. The importance of different water availability in influencing UV-B impacts on lower plants in the Arctic has been poorly explored and might partially explain the observed wide variation of responses, given the importance of water in controlling bryophyte physiology. This study aimed to assess the long-term responses of three common sub-Arctic bryophytes to enhanced UV-B radiation (+UV-B) and to elucidate the influence of water supply on those responses. Responses of three sub-Arctic bryophytes (the mosses Hylocomium splendens and Polytrichum commune and the liverwort Barbilophozia lycopodioides) to +UV-B for 15 and 13 years were studied in two field experiments using lamps for UV-B enhancement with identical design and located in neighbouring areas with contrasting water availability (naturally mesic and drier sites). Responses evaluated included bryophyte abundance, growth, sporophyte production and sclerophylly; cellular protection by accumulation of UV-absorbing compounds, β-carotene, xanthophylls and development of non-photochemical quenching (NPQ); and impacts on photosynthesis performance by maximum quantum yield (F(v) /F(m)) and electron transport rate (ETR) through photosystem II (PSII) and chlorophyll concentrations. Responses were species specific: H. splendens responded most to +UV-B, with reduction in both annual growth (-22 %) and sporophyte production (-44 %), together with increased β-carotene, violaxanthin, total chlorophyll and NPQ, and decreased zeaxanthin and de-epoxidation of the xanthophyll cycle pool (DES). Barbilophozia lycopodioides responded less to +UV-B, showing increased β-carotene and sclerophylly and decreased UV-absorbing compounds. Polytrichum commune only showed small morphogenetic changes. No effect of UV-B on bryophyte cover was observed. Water availability had profound effects on bryophyte ecophysiology, and plants showed, in general, lower growth and ETR, together with a higher photoprotection in the drier site. Water availability also influenced bryophyte responses to +UV-B and, in particular, responses were less detectable in the drier site. Impacts of UV-B exposure on Arctic bryophytes were significant, in contrast to modest or absent UV-B effects measured in previous studies. The impacts were more easily detectable in species with high plasticity such as H. splendens and less obvious, or more subtle, under drier conditions. Species biology and water supply greatly influences the impact of UV-B on at least some Arctic bryophytes and could contribute to the wide variation of responses observed previously.

Induced point mutations in the phytoene synthase 1 gene cause differences in carotenoid content during tomato fruit ripening

In tomato, carotenoids are important with regard to major breeding traits such as fruit colour and human health. The enzyme phytoene synthase (PSY1) directs metabolic flux towards carotenoid synthesis. Through TILLING (Targeting Induced Local Lesions IN Genomes), we have identified two point mutations in the Psy1 gene. The first mutation is a knockout allele (W180*) and the second mutation leads to an amino acid substitution (P192L). Plants carrying the Psy1 knockout allele show fruit with a yellow flesh colour similar to the r, r mutant, with no further change in colour during ripening. In the line with P192L substitution, fruit remain yellow until 3 days post-breaker and eventually turn red. Metabolite profiling verified the absence of carotenoids in the W180* line and thereby confirms that PSY1 is the only enzyme introducing substrate into the carotenoid pathway in ripening fruit. More subtle effects on carotenoid accumulation were observed in the P192L line with a delay in lycopene and β-carotene accumulation clearly linked to a very slow synthesis of phytoene. The observation of lutein degradation with ripening in both lines showed that lutein and its precursors are still synthesised in ripening fruit. Gene expression analysis of key genes involved in carotenoid biosynthesis revealed that expression levels of genes in the pathway are not feedback-regulated by low levels or absence of carotenoid compounds. Furthermore, protein secondary structure modelling indicated that the P192L mutation affects PSY1 activity through misfolding, leading to the low phytoene accumulation.

Optimization of β-carotene production by a mutant of the lactose-positive yeast Rhodotorula acheniorum from whey ultrafiltrate

The present work investigated on carotenogenesis with high β-carotene content by a new isolated high-activity strain-producer Rhodotorula acheniorum mutant MRN in cheese whey ultrafiltrate. After a serial of UV, ethymethanesurfonate (EMS), and nitrosoguanidine (NTG) mutagenesis, a mutant named MRN of the red lactose-positive yeast strain R. acheniorum was obtained. Then, the effects of different growth medium factors on carotenoid production by this mutant at batch-scale level were identified and optimized by means of response surface methodology (RSM) in order to achieve high-level production of β-carotene. The optimum conditions required to achieve the highest level of β-carotene (262.12±1.01 mg/L) were determined as follows: whey ultrafiltrate (WU) lactose concentration 55 g/L, pH 5.85, ammonium sulfate concentration 3.5 g/L, temperature 23°C, and aeration rate 1.56 vvm. The medium optimization resulted in a 6.45-fold increase in volumetric production (262.12±1.01 mg/L) and a 4.62-fold increase in the cellular accumulation (10.69±0.19 mg/g) of β-carotene.

Evaluation of the genotype, environment and their interaction on carotenoid and ascorbic acid accumulation in tomato germplasm

Tomatoes are an important source of antioxidants (carotenoids, vitamin C, etc.) owing to their high level of consumption. There is great interest in developing cultivars with increased levels of lycopene, β-carotene or L-ascorbic acid. There is necessary to survey new sources of variation. In this study, the potential of improvement for each character in tomato breeding programmes, in a single or joint approach, and the nature of genotype (G), environment (E) and G × E interaction effects in the expression of these characters were investigated. The content of lycopene, β-carotene and ascorbic acid determined was very high in some phenotypes (up to 281, 35 and 346 mg kg(-1) respectively). The important differences in the three environments studied (with some stressing conditions in several situations) had a remarkable influence in the phenotypic expression of the functional characters evaluated. Nevertheless, the major contribution came from the genotypic effect along with a considerable G × E interaction. The joint accumulation of lycopene and β-carotene has a high genetic component. It is possible to select elite genotypes with high content of both carotenoids in tomato breeding programmes but multi-environment trials are recommended. The improvement of ascorbic acid content is more difficult because the interference of uncontrolled factors mask the real genetic potential. Among the accessions evaluated, there are four accessions with an amazing genetic potential for functional properties that can be used as donor parents in tomato breeding programmes or for direct consumption in quality markets.

Mayonnaise Contributes to Increasing Postprandial Serum β-Carotene Concentration through the Emulsifying Property of Egg Yolk in Rats and Humans

We performed in vitro, animal, and human studies to clarify the effect of mayonnaise on β-carotene intake and its mechanism. In an artificial gastric juice model, we examined the transfer of β-carotene from grated carrot to mayonnaise or vegetable oil. Mayonnaise was more easily dispersed in artificial gastric juice than vegetable oil. The β-carotene concentration was greater in mayonnaise than vegetable oil. In rats, the postprandial serum β-carotene concentration in the mayonnaise group (β-carotene with mayonnaise) was higher than that in the control (β-carotene only) and vegetable oil (β-carotene with vegetable oil) groups. Continuous feeding of dietary β-carotene (14 d), employing mayonnaise or egg yolk, resulted in an increased accumulation of β-carotene in the liver. In a human study, diets were provided in the form of (1) carrot as a control (CON), (2) carrot juice (JU), (3) carrot with oil (OIL) and (4) carrot with mayonnaise (MS). Following collection of fasting blood samples, nine adult males consumed one of the four diets in random order. Fasting and postprandial changes in serum β-carotene were assessed at 2, 3, 4, 6 and 8 h following ingestion of each diet. The incremental areas under the curves of serum β-carotene concentration were higher following MS than following both CON and JU. In conclusion, we suggest that mayonnaise contributes to raising the serum β-carotene concentration when consumed with carrots rich in β-carotene, and that its mechanism is related to the emulsifying property of the egg yolk contained in mayonnaise.

The cauliflower Orange gene enhances petiole elongation by suppressing expression of eukaryotic release factor 1

The cauliflower (Brassica oleracea var. botrytis) Orange (Or) gene affects plant growth and development in addition to conferring β-carotene accumulation. This study was undertaken to investigate the molecular basis for the effects of the Or gene mutation in on plant growth. The OR protein was found to interact with cauliflower and Arabidopsis eukaryotic release factor 1-2 (eRF1-2), a member of the eRF1 family, by yeast two-hybrid analysis and by bimolecular fluorescence complementation (BiFC) assay. Concomitantly, the Or mutant showed reduced expression of the BoeRF1 family genes. Transgenic cauliflower plants with suppressed expression of BoeRF1-2 and BoeRF1-3 were generated by RNA interference. Like the Or mutant, the BoeRF1 RNAi lines showed increased elongation of the leaf petiole. This long-petiole phenotype was largely caused by enhanced cell elongation, which resulted from increased cell length and elevated expression of genes involved in cell-wall loosening. These findings demonstrate that the cauliflower Or gene controls petiole elongation by suppressing the expression of eRF1 genes, and provide new insights into the molecular mechanism of leaf petiole regulation.

Carotenoid content and expression of phytoene synthase and phytoene desaturase genes in bitter melon (Momordica charantia)

Momordica charantia, a tropical plant, produces a fruit that has a β-carotene concentration five times higher than that of carrot. To elucidate the molecular basis of β-carotene accumulation in M. charantia, the gene expression levels of phytoene synthase (McPSY) and phytoene desaturase (McPDS) were determined. These levels were particularly high in the flowers of M. charantia. During fruit maturation, the expression levels of McPSY and McPDS decreased during the mid-stages but increased in the fully mature fruit. In addition, carotenoids accumulated as the peel changed from green to orange. Thus, McPSY and McPDS expression correlated with carotenoid accumulation during fruit maturation. Principal component analysis (PCA) also was used to evaluate the differences among the profiles of seven carotenoids identified in the fruit at several maturation stages. Riper fruits had higher carotenoid concentrations than less ripe fruits.

Expression of lycopene cyclase genes and their regulation on downstream carotenoids during fruit maturation of Guoqing No. 1 Satsuma mandarin and Cara Cara navel orange

Lycopene β-cyclase ( LCYb) and lycopene ɛ-cyclase ( LCYe) are believed to be crucial genes to lycopene cyclization and downstream carotenoid accumulation. To illuminate the gene expression profiles and their regulation on downstream carotenoids, expression profiles of these two genes and contents of several corresponding carotenoids were determined by real-time PCR and HPLC, respectively, in relative shorter intervals during breaker and maturation stage of Guoqing No. 1 satsuma mandarin ( Citrus unshiu Marcow) and Cara Cara navel orange ( Citrus sinensis Osbeck). Data showed that the expression profiles of these two cyclase genes and their relative expression levels between the peels and the pulps at the same stage all varied with cultivars. But in general, accumulation of lutein and β-carotene experienced a similar tendency in both cultivars and were up-regulated by expression of LCYb and LCYe in spite of poor correlation in the pulps. Meanwhile, lycopene abundance was negatively correlated with the expression of LCYe as expected, but hardly correlated with LCYb in the pulps of Cara Cara navel orange, indicating that LCYe might exert a more prominent influence on lycopene cyclization and downstream carotenoids formation. Results of carotenoids accumulation suggest that the biosynthetic process of carotenoids would shift from upstream β, ɛ- or β, β-carotenoids such as lutein and β-carotene to downstream β, β-carotenoids during fruit development.

Characterization of pigment apparatus in winter-green and summer-green leaves of a shade-tolerant plant Ajuga reptans

Comparative study was performed to assess the content and proportions of photosynthetic pigments and the violaxanthin cycle (VXC) activity in winter-green and summer-green leaves of bugleweed (Ajuga reptans L.) plants grown in shaded (photosynthetically active radiation, PAR 150 μmol/(m2 s)) and sunny (PAR 1200 μmol/(m2 s)) habitats in the Botanic Garden of Jagiellonian University (Krakow, Poland). In overwintered and newly formed leaves of shade plants, the content of green and yellow pigments was two times higher than in leaves of sun plants. The shade plants were distinguished by accumulation of β-carotene, while lutein was predominant in leaves of sun plants. Under the action of strong light (2000 μmol/(m2s)), the level of violaxanthin deepoxidation in winter-green leaves of shade and sun plants increased five- to sixfold, whereas it changed insignificantly in summer-green leaves of shade plants. It is concluded that, in a shadetolerant species A. reptans, the photosynthetic apparatus of winter-green leaves in sun and shade plants and of summer-green leaves in sun plants is protected against excess insolation by high activity of VXC. The carotenoids of summer-green leaves in shade plants are supposed to function mainly as light-harvesting pigments.

POSSIBILITIES OF PRACTICAL USE OF JASMONATES IN PRODUCTION OF FRUITS AND VEGETABLES

Jasmonates, mostly jasmonic acid (JA) and methyl jasmonate (JA-Me), are important signal transducers and greatly stimulate the biosynthesis of a wide range of secondary metabolites in plant cell cultures and in different intact plants. In this review we focused on the role of jasmonates in regulation of some physiological processes in growth and development of fruits and vegetables. For example, preharvest application of JA-Me improved fruit quality of blackberry and strawberry by stimulation of the biosynthesis of flavonoids and anthocyanins, and increased antioxidant capacities. In the case of sweet basil, JA-Me enhanced accumulation of alkaloids, terpenoids and phenolics. Moreover, JA-Me has been demonstrated to increase anthocyanin content in apple and strawberry fruits. It is well known that flavonoids, mostly anthocyanins, show preventive and therapeutic activities in many illness, for example extracts of blackberry fruits showed anticancer activity against human lung cells and leukemia cells. Besides, it has been found that exogenous JA-Me inhibits lycopene in tomatoes and stimulates β-carotene accumulation in tomatoes, apples, banana, guava, mango, and greatly stimulates ethylene production in tomatoes at different stages of ripening, as well as in preclimacteric apples, Japanese plum, and mangoes. JA-Me appears to be effective in reducing chilling injury in avocados, grapefruits, mangoes, peppers, cucumbers and others. JA-Me inhibits postharvest sprouting and improves storage quality of radishes. It is interesting also that JA-Me extends shelf life and reduces microbial contamination of fresh-cut celery, peppers, pineapple and tomatoes. Cooperative cross-talk among jasmonates and other phytohormones, especially ethylene occurs in regulation of growth and development and in defence responses against a wide of abiotic and biotic stressors. Moreover, it should be mentioned that jasmonates have medical importance due to inducing suppression of proliferation and death in several human cancer cells (leukemia, prostate, breast, lung and melanoma cell lines). These recently published findings suggest that the natural jasmonates may potentially be a novel class of anticancer drugs.

Transcriptional-Metabolic Networks in β-Carotene-Enriched Potato Tubers: The Long and Winding Road to the Golden Phenotype

Vitamin A deficiency is a public health problem in a large number of countries. Biofortification of major staple crops (wheat [Triticum aestivum], rice [Oryza sativa], maize [Zea mays], and potato [Solanum tuberosum]) with β-carotene has the potential to alleviate this nutritional problem. Previously, we engineered transgenic "Golden" potato tubers overexpressing three bacterial genes for β-carotene synthesis (CrtB, CrtI, and CrtY, encoding phytoene synthase, phytoene desaturase, and lycopene β-cyclase, respectively) and accumulating the highest amount of β-carotene in the four aforementioned crops. Here, we report the systematic quantitation of carotenoid metabolites and transcripts in 24 lines carrying six different transgene combinations under the control of the 35S and Patatin (Pat) promoters. Low levels of B-I expression are sufficient for interfering with leaf carotenogenesis, but not for β-carotene accumulation in tubers and calli, which requires high expression levels of all three genes under the control of the Pat promoter. Tubers expressing the B-I transgenes show large perturbations in the transcription of endogenous carotenoid genes, with only minor changes in carotenoid content, while the opposite phenotype (low levels of transcriptional perturbation and high carotenoid levels) is observed in Golden (Y-B-I) tubers. We used hierarchical clustering and pairwise correlation analysis, together with a new method for network correlation analysis, developed for this purpose, to assess the perturbations in transcript and metabolite levels in transgenic leaves and tubers. Through a "guilt-by-profiling" approach, we identified several endogenous genes for carotenoid biosynthesis likely to play a key regulatory role in Golden tubers, which are candidates for manipulations aimed at the further optimization of tuber carotenoid content.

Application of two bicistronic systems involving 2A and IRES sequences to the biosynthesis of carotenoids in rice endosperm

Coordination of multiple transgenes is essential for metabolic engineering of biosynthetic pathways. Here, we report the utilization of two bicistronic systems involving the 2A sequence from the foot-and-mouth disease virus and the internal ribosome entry site (IRES) sequence from the crucifer-infecting tobamovirus to the biosynthesis of carotenoids in rice endosperm. Two carotenoid biosynthetic genes, phytoene synthase (Psy) from Capsicum and carotene desaturase (CrtI) from Pantoea, were linked via either the synthetic 2A sequence that was optimized for rice codons or the IRES sequence under control of the rice globulin promoter, generating PAC (Psy-2A-CrtI) and PIC (Psy-IRES-CrtI) constructs, respectively. The transgenic endosperm of PAC rice had a more intense golden color than did PIC rice, demonstrating that 2A was more efficient than IRES in coordinating gene expression. The 2A and IRES constructs were equally effective in driving transgene transcription. However, immunoblot analysis of CRTI, a protein encoded by the downstream open reading frame of the bicistronic constructs, revealed that 2A was ninefold more effective than IRES in driving translation. The PAC endosperms accumulated an average of 1.3 μg/g of total carotenoids, which was ninefold higher than was observed for PIC endosperms. In particular, accumulation of β-carotene was much higher in PAC endosperms than in PIC endosperms. Collectively, these results demonstrate that both 2A and IRES systems can coordinate the expression of two biosynthetic genes, with the 2A system exhibiting greater efficiency. Thus, the 2A expression system described herein is an effective new tool for multigene stacking in crop biotechnology.

Carotenoid Accumulation by Carrot Storage Roots in Relation to Nitrogen Fertilization Level

The accumulation of carotenoid compounds in carrot (Daucus carota L.) roots of five cultivars, in relation to different nitrogen fertilization levels was investigated. The experiment was carried out at the Warsaw University of Life Sciences. Carrot cultivars ‘Karotan’ F1, ‘Trafford’ F1, ‘Krakow’ F1, ‘Komarno’ F1 and ‘HY 7842’ were used in the study. Nitrogen fertilization was applied as urea form, in doses ranging from 0 to 120 kg N ha–1, and on two terms: pre-sowing and in the middle of the growing season. The carrot seeds were sown at the beginning of May and the roots were harvested in mid-October at maturity. Total carotenoids and β-carotene contents in the carrot roots were determined after the harvest by means of the standard spectrophotometric method. CIE L*a*b* colour parameters of the roots and the juice, as well as dry matter in the roots were determined. Results of the experiment showed that carotenoid accumulation in the roots was significantly affected by carrot genotype. ‘HY7842’ carrot showed the highest accumulation of total carotenoids and β-carotene. Nitrogen in the rates applied, did not significantly influence carotenoid content in the roots. Moderate correlation between carotenoid content and colour a* parameter (redness intensity) of the roots and the juice was found.

Hepatic stellate cells are an important cellular site for β-carotene conversion to retinoid

Hepatic stellate cells (HSCs) are responsible for storing 90–95% of the retinoid present in the liver. These cells have been reported in the literature also to accumulate dietary β-carotene, but the ability of HSCs to metabolize β-carotene in situ has not been explored. To gain understanding of this, we investigated whether β-carotene-15,15′-monooxygensase ( Bcmo1) and β-carotene-9′,10′-monooxygenase ( Bcmo2) are expressed in HSCs. Using primary HSCs and hepatocytes purified from wild type and Bcmo1-deficient mice, we establish that Bcmo1 is highly expressed in HSCs; whereas Bcmo2 is expressed primarily in hepatocytes. We also confirmed that HSCs are an important cellular site within the liver for accumulation of dietary β-carotene. Bcmo2 expression was found to be significantly elevated for livers and hepatocytes isolated from Bcmo1-deficient compared to wild type mice. This elevation in Bcmo2 expression was accompanied by a statistically significant increase in hepatic apo-12′-carotenal levels of Bcmo1-deficient mice. Although apo-10′-carotenal, like apo-12′-carotenal, was readily detectable in livers and serum from both wild type and Bcmo1-deficient mice, we were unable to detect either apo-8′- or apo-14′-carotenals in livers or serum from the two strains. We further observed that hepatic triglyceride levels were significantly elevated in livers of Bcmo1-deficient mice fed a β-carotene-containing diet compared to mice receiving no β-carotene. Collectively, our data establish that HSCs are an important cellular site for β-carotene accumulation and metabolism within the liver.

Variability of carotenoid biosynthesis in orange colored Capsicum spp.

Pepper, Capsicum spp., is a worldwide crop valued for heat, nutrition, and rich pigment content. Carotenoids, the largest group of plant pigments, function as antioxidants and as vitamin A precursors. The most abundant carotenoids in ripe pepper fruits are β-carotene, capsanthin, and capsorubin. In this study, the carotenoid composition of orange fruited Capsicum lines was defined along with the allelic variability of the biosynthetic enzymes. The carotenoid chemical profiles present in seven orange pepper varieties were determined using a novel UPLC method. The orange appearance of the fruit was due either to the accumulation of β-carotene, or in two cases, due to only the accumulation of red and yellow carotenoids. Four carotenoid biosynthetic genes, Psy, Lcyb, CrtZ-2, and Ccs were cloned and sequenced from these cultivars. This data tested the hypothesis that different alleles for specific carotenoid biosynthetic enzymes are associated with specific carotenoid profiles in orange peppers. While the coding regions within Psy and CrtZ-2 did not change in any of the lines, the genomic sequence contained introns not previously reported. Lcyb and Ccs contained no introns but did exhibit polymorphisms resulting in amino acid changes; a new Ccs variant was found. When selectively breeding for high provitamin A levels, phenotypic recurrent selection based on fruit color is not sufficient, carotenoid chemical composition should also be conducted. Based on these results, specific alleles are candidate molecular markers for selection of orange pepper lines with high β-carotene and therefore high provitamin A levels.

Modeling Growth of Microalgae <i>Dunaliella Salina</i> under Different Nutritional Conditions

Problem statement: The aim of this study was to find the empirical model that describes the growth kinetics of Dunaliella salina, with low production cost and to estimate parameters of this model. Approach: In this study the strain of D. salina UTEX 200 was cultivated in seawater (0.5 M NaCl) at room temperature with agitation of 150 rpm and luminous intensity of 60 mmols.m-2.s-1. The synthetic medium AS100 (0.2 M NaCl) was used in this study for comparison purposes and in order to determine the optimal growth of the microalgae. Kinetics of growth and β-carotene production was determined in a period of 15 days. Results: After the analysis of the behavior graphic, an unstructured model was used for describing the cell growth (logistic model). It was observed that the model was well adjusted to experimental data for the two conditions of analysis. It was observed that alga produces carotenoids under conditions of stress, in which the cell division are retarded. In the case of cells grown in seawater (higher salt concentration), the cell growth was lower but the concentration of β-carotene was higher. Conclusion: In general, these results suggest that D. salina presents higher potential for β-carotene accumulation and that high salinity decreases cellular concentration (measured by the parameter Xm of the kinetic model proposed); however there is an increase in β-carotene production.

Expression of carotenogenic genes during the development and ripening of watermelon fruit

Five watermelon varieties with different flesh colors: “ZAOHUA” (red); “96B41” (pink); “307CHAOFENG” (yellow); “UNKNOWN” (orange); and “SANBAI” (white) were used to investigate the relationship between carotenoid profiles and expression of carotenoid-related genes during fruit development and ripening. In mature fruit, “ZAOHUA” and “96B41” accumulated high level of lycopene, “SANBAI” contained trace amounts of carotenoids. Through HPLC and dot blot hybridization, carotenoid content was analyzed and compared to the expression of carotenoid-related genes including Psy, Pds, Zds, CrtIso, Lcyb, Chyb, Nced1, Nced2, Nced3 during fruit development. All genes examined were expressed in all five watermelon varieties. In red and pink color varieties, the decrease of gene expression for Lcyb and Chyb at stage C (30 d after pollination) may be consistent with the high accumulation of lycopene and β-carotene. In the yellow variety, expression of the detected genes increased markedly with the onset of maturation, but little carotenoid accumulation was detected, which may be due to high level of carotenoid cleavage dioxygenases. Expression of all detected genes decreased rapidly in the white color variety upon fruit ripening, which may be attributed to a reduction in metabolic flux in the carotenoid pathway. These results imply the complex regulation of the carotenoid biosynthetic pathway in watermelon.

β-Carotene accumulation in 3T3-L1 adipocytes inhibits the elevation of reactive oxygen species and the suppression of genes related to insulin sensitivity induced by tumor necrosis factor-α

Objective β-Carotene is an abundant carotenoid with potent antioxidative activities and accumulates in adipose tissue. However, its physiologic functions are poorly understood. In this study, we examined whether accumulation of β-carotene for 4 d in insulin-resistant 3T3-L1 adipocytes alters the expression of genes related to insulin sensitivity. Methods The 3T3-L1 adipocytes were treated with/without 10 or 20 μM β-carotene during differentiation for 4 d. The cells treated with 10 μM β-carotene for 4 d were subsequently incubated with/without 5 ng/mL of tumor necrosis factor-α for 48 h in the medium without β-carotene. The mRNA levels of genes in the cells and adiponectin protein levels in the medium were determined by real-time reverse transcription–polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Reactive oxygen species levels in the cells were assessed by oxidation of 2′,7′-dichlorodihydrofluorescein diacetate. Result β-Carotene treatment at a concentration of 20 μM, but not 10 μM, in 3T3-L1 adipocytes during differentiation for 4 d enhanced the expression of genes related to insulin sensitivity, including adiponectin, adipocyte lipid-binding protein, glucose transporter-4, peroxisome proliferator-activated receptor-γ2, and adiponectin protein in the medium. Tumor necrosis factor-α treatment repressed the expression of these genes and adiponectin protein in the medium and induced reactive oxygen species levels. In contrast, cells that accumulated β-carotene at a concentration of 10 μM did not show these alterations. Conclusion The accumulation of the β-carotene in 3T3-L1 adipocytes restores the expression of genes related to insulin sensitivity and reactive oxygen species levels in insulin-resistant adipocytes.

Effect of pH on growth and biochemical responses of Dunaliella bardawil and Chlorella ellipsoidea

The goal of the present investigation was to study the effect of pH on growth and biochemical responses of Dunaliella bardawil and Chlorella ellipsoidea when exposed to different pH values. The two tested microalgae could grow in a wide range of pH (4-9 for D. bardawil and 4-10 for C. ellipsoidea). The dry weight gain and the biochemical components of D. bardawil were greatly enhanced at pH 7.5. In contrast, dry weight and carbohydrate content of C. ellipsoidea attained their maximum values at the alkaline pH. On the other hand, the protein content of C. ellipsoidea recorded its highest value at pH 4, while the pigment content of the same alga was highest at pH 4, 6, and 7.5 and decreased at alkaline pH. Both pH 6 and pH 9 stimulated the accumulation of β-carotene, vitamin E and vitamin C in D. bardawil, with the highest values of the three compounds recorded at pH 9. In the case of C. ellipsoidea, β-carotene content increased at pH 6 and pH 10 as compared with the control, but the amount of β-carotene was much higher at pH 6 than at pH 10. Vitamin E content was higher in C. ellipsoidea cells at pH 10 than at pH 6. Both pH 6 and pH 10 caused a significant decline in vitamin C content of C. ellipsoidea.

Rice caryopsis structure in relation to distribution of micronutrients (iron, zinc, β-carotene) of rice cultivars including transgenic indica rice

The transgenic indica rice lines of IR68144 and BR29, developed using endosperm-specific promoters were analyzed for their iron, zinc and β-carotene content in the endosperm. Biochemical analysis clearly revealed the presence of higher accumulation of iron, zinc and β-carotene in transgenic rice grains in comparison with control. Prussian blue staining reaction evidenced the presence of iron in the endosperm cells of transgenic rice grains in comparison with control where iron is restricted only to aleurone and embryo. The rice grain structure of IR64, IR72, IR68144, Swarna, BRRI Dhan 29 (BR29), BR28, Taipai 309 (T309) and New Plant Type-3 (NPT3) indicated that the number of aleurone layers, size of the embryo and size of the caryopsis determines the quantity of important micronutrients (iron, zinc) in the grains. Biochemical analysis revealed that iron and zinc content drastically varies in polished and unpolished rice and among the varieties examined. During the polishing process almost entire aleurone and most part of the embryo is removed which are the main storehouse for major micronutrients. It is estimated that more than 70% of micronutrients are lost during polishing process.