Comparison of carotenoid,chlorophyll concentrations and their biosynthetic transcript levels in different coloured cauliflower

Carotenoids and chlorophylls are among the most widely distributed pigments in nature that play essential roles in the photosynthetic apparatus and confer diverse colours in plants.Among all vegetables,cauliflower(Brassica oleracea L.ssp.var.botrytis)is rich in phytochemicals and is an important crop grown all over the world.This study investigates carotenoid and chlorophyll concentrations in differently pigmented cultivars and elucidates the role of transcriptional regulation of carotenoid accumulation including lutein andβ-carotene.Here,we characterised changes in pigments by UHPLC-DAD-ToF-MS and changes in transcript levels of carotenoid metabolic genes by qRT-PCR in florets and leaves of orange(‘Jaffa'and‘Sunset'),purple(‘Di Sicilia Violetto'and‘Graffiti'),green(‘Trevi')and white(‘Clapton')cultivars.Transcript levels of all carotenoid metabolic genes showed different transcript level patterns in the leaves and florets.Compared to the other cultivars,the orange cultivars had the highest levels ofβ-carotene in the florets and lutein in the leaves resulting in changes lutein/β-carotene ratios.In the green cultivar,higher transcript levels were also found,especially for phytoene synthase and phytoene desaturase genes of the core biosynthesis pathway.However,no increased carotenoid concentrations were observed,possibly due to a higher carotenoid turnover induced by the carotenoid cleavage dioxygenase 4 in the green cultivar.In the white(‘Clapton')and purple(‘Di Sicilia Violetto'and‘Graffiti')cultivars the phytoene desaturase transcript levels as well as carotenoid concentrations were low.Chlorophyll concentrations changed in trend comparable to the carotenoid concentrations and were only significantly lower in the leaves of the orange cultivar‘Jaffa'.Also,the chlorophyll a/b ratio changed in‘Jaffa'.In florets the highest chlorophylls concentrations were observed for the green cultivar(‘Trevi')and the purple cultivar(‘Di Sicilia Violetto').Taken together,the study demonstrates the complex source-sink relationship of carotenoid accumulation in different coloured cauliflower.

Epigenetic changes in the regulation of carotenoid metabolism during honeysuckle flower development

Flower development is one of the most vital pathways in plant development, during which the epigenetic regulation of gene expression is essential. DNA methylation, the most conserved epigenetic modification, participates in gene expression regulation and transposable element silencing. Honeysuckle(Lonicera japonica) is an important medicinal plant renowned for its colorful and fragrant flowers. Honeysuckle flowers change color from white to gold as a result of carotenoid accumulation during development. However, the role of DNA methylation in flower color changes is not well understood in L. japonica. Here, we performed whole-genome bisulfite sequencing and transcriptome sequencing during flowering development in honeysuckle. The results showed that a decrease in the levels of genome-wide average DNA methylation during flower development and changes in DNA methylation were associated with the expression of demethylase genes. Moreover, many genes involved in carotenoid biosynthesis and degradation, such as Lj PSY1, LjPDS1, LjLCYE, and LjCCD4, have altered expression levels because of hypomethylation, indicating that DNA methylation plays an important role in flower color changes in honeysuckle. Taken together, our data provide epigenetic insights into flower development and color change in honeysuckles.

Transcriptomes of Litopenaeus vannamei reveal modulation of antioxidant system induced by dietary archaeal carotenoids

Oxidative stress induced by factors such as ammonia nitrogen has become a major issue in shrimp farming.The effects of carotenoids on the growth and antioxidant capability of Litopenaeus vannamei juveniles were investigated in this study using dietary archaeal carotenoids supplementation.For four weeks,shrimp were given diets containing 0 mg/kg(Ctrl)and 55.98 mg/kg(Car)archaeal carotenoids.Dietary archaeal carotenoids significantly enhanced the astaxanthin content in shrimp muscles and carapaces,as well as the superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)activity(P<0.05).The malonaldehyde(MDA)content in Car group significantly decreased(P<0.05).The transcriptome analysis was conducted to determine the molecular processes in response to archaeal carotenoids supplementation.A total of 1536 differentially expressed genes(DEGs)were detected,including 538 upregulated DEGs and 998 downregulated DEGs.GO functional enrichment analysis between Ctrl and Car indicated that 26 GO terms including extracellular region,metabolic process,and proteolysis were enriched.The KEGG pathway enrichment analysis revealed that the amino sugar and nucleotide sugar metabolism,cysteine and methionine metabolism,glycine serine and threonine metabolism,and amino acid biosynthesis were enriched.Archaeal carotenoids influenced the expression of several important genes involved in reactive oxygen species(ROS)generation,Nrf2 signaling,and antioxidant enzymes.Seven DEGs were chosen to confirm the RNA-Seq data using qRT-PCR.The genes and pathways discovered in this work assist to elucidate the molecular processes through which archaeal carotenoid enhances L.vannamei antioxidative system.

Fiber-specific increase of carotenoid content promotes cotton fiber elongation by increasing abscisic acid and ethylene biosynthesis

Cotton fiber is a raw material for the global textile industry and fiber quality is essential to its industrial application.Carotenoids are plant secondary metabolites that may serve as dietary components,regulate light harvesting,and scavenge reactive oxygen species.Although carotenoids accumulate predominantly in rapidly elongating cotton fibers,their roles in cotton fiber development remain poorly understood.In this study,a fiber-specific promoter proSCFP was applied to drive the expression of GhOR1Del,a positive regulator of carotenoid accumulation,to upregulate the carotenoid level in cotton fiber in planta.Fiber length,strength,and fineness were increased in proSCFP:GhOR1Del transgenic cotton and abscisic acid(ABA)and ethylene contents were increased in elongating fibers.The ABA downstream regulator GhbZIP27a stimulated the expression of the ethylene synthase gene GhACO3 by binding to its promoter,suggesting that ABA promoted fiber elongation by increasing ethylene production.These findings suggest the involvement of carotenoids and ABA signaling in promoting cotton fiber elongation and provide a strategy for improving cotton fiber quality.

Molecular mechanism of lycopene cyclases regulating carotenoids ratio in different branches during tea leaf and flower development

Carotenoids are essential components in tea quality, contributing to leaf color and aroma. However, little information about carotenoids in different tea cultivars and their biosynthesis regulation mechanism during leaf development is known. Here we analyzed carotenoids by HPLC in the buds and leaves of 113 tea cultivars harvested on the same day. By profile clustering, carotenoids were divided into five groups. Same group cultivars displayed divergence in the total content of carotenoids but a similar molar ratio. To figure out the molecular mechanisms of this phenomenon, we further characterized all functional lycopene cyclases, which are the branch point of the carotenoid biosynthesis pathway. Two β-lycopene cyclases(CsLCYB1 and CsLCYB2) and one ε-lycopene cyclase(CsLCYE1) were cloned. Subcellular localization analysis showed that all cloned CsLCYs were localized in plastids. Enzyme activity assays in E. coli indicated both CsLCYBs catalyzed lycopene into β-carotene, and CsLCYE1 produced δ-carotene and ε-carotene. We found CsLCYB1 and CsLCYE1 predominantly expressed in leaf, while CsLCYB2was mainly expressed during flowering stages. Suppression by antisense oligonucleotides reduced CsLCYB1 and CsLCYE1 transcripts and led to reduction of both β,β-branch and β,ε-branch carotenoids in leaf. The expression levels of CsLCYB1 showed a significant positive correlation withβ,β-branch carotenoids in leaf. Our study provides carotenoid profiles of different tea cultivars, which can assist tea producers in selecting cultivars of interest. Meanwhile, we proposed the molecular mechanism of carotenoids reflecting the tenderness of tea plant leaf from a metabolic flux perspective, and suggested lycopene cyclase that could be applied to the breeding of tea varieties with different branch carotenoids.

Blue light induces leaf color change by modulating carotenoid metabolites in orange-head Chinese cabbage(Brassica rapa L.ssp. pekinensis)

Carotenoids are involved in the formation of plant leaf color as well as photosystem photoprotection. This study showed that blue light significantly induced up-regulation of the total carotenoid content in the inner leaves of orange-head Chinese cabbage(OHCC). Furthermore, the transcriptomic analysis revealed that blue light treatment induced upregulation of genes in photosynthesis(BrHY5-2, BrCOP1 and BrDET1) and the methylerythritol 4-phosphate pathways(BrGGPS, BrDXS and BrHDR) upstream of the carotenoid metabolic pathway. Carotenoid metabolomic analysis revealed that the accumulation of several orange and red carotenoids(lycopene, zeaxanthin, β-carotene, lutein, and β-cryptoxanthin) after blue light treatment contributed to the deepening of the leaf coloration, suggesting that short-term blue light treatment could be used to boost nutritional quality. The light signal gene BrHY5-2 participated in the blue light-induced transcriptional regulation of carotenoid biosynthesis in OHCC. Overexpression of BrHY5-2 in Arabidopsis significantly increased the total carotenoid content and the sensitivity to blue light. The above findings revealed new insights about blue-light-induced carotenoid synthesis and accumulation in OHCC lines. They suggested a new engineering approach to increase the nutritional value of vegetables.

Natural variations and geographical distributions of seed carotenoids and chlorophylls in 1167 Chinese soybean accessions

Understanding the composition and contents of carotenoids in various soybean seed accessions is important for their nutritional assessment.This study investigated the variability in the concentrations of carotenoids and chlorophylls and revealed their associations with other nutritional quality traits in a genetically diverse set of Chinese soybean accessions comprised of cultivars and landraces.Genotype,planting year,accession type,seed cotyledon color,and ecoregion of origin significantly influenced the accumulation of carotenoids and chlorophylls.The mean total carotenoid content was in the range of 8.15–14.72μg g–1 across the ecoregions.The total carotenoid content was 1.2-fold higher in the landraces than in the cultivars.Soybeans with green cotyledons had higher contents of carotenoids and chlorophylls than those with yellow cotyledons.Remarkably,lutein was the most abundant carotenoid in all the germplasms,ranging from 1.35–37.44μg g–1.Carotenoids and chlorophylls showed significant correlations with other quality traits,which will help to set breeding strategies for enhancing soybean carotenoids without affecting the other components.Collectively,our results demonstrate that carotenoids are adequately accumulated in soybean seeds,however,they are strongly influenced by genetic factors,accession type,and germplasm origin.We identified novel germplasms with the highest total carotenoid contents across the various ecoregions of China that could serve as the genetic materials for soybean carotenoid breeding programs,and thereby as the raw materials for food sectors,pharmaceuticals,and the cosmetic industry.

Retrieval of Winter Wheat Canopy Carotenoid Content with Ground-and Airborne-Based Hyperspectral Data

Accurate assessment of canopy carotenoid content(CC_(x+c)C)in crops is central to monitor physiological conditions in plants and vegetation stress,and consequently supporting agronomic decisions.However,due to the overlap of absorption peaks of carotenoid(C_(x+c))and chlorophyll(C_(a+b)),accurate estimation of carotenoid using reflectance where carotenoid absorb is challenging.The objective of present study was to assess CC_(x+c)C in winter wheat(Triticum aestivum L.)with ground-and aircraft-based hyperspectral measurements in the visible and near-infrared spectrum.In-situ hyperspectral reflectance were measured and airborne hyperspectral data were acquired during major growth stages of winter wheat in five consecutive field experiments.At the canopy level,a remarkable linear relationship(R^(2)=0.95,p<0.001)existed between C_(x+c) and Ca+b,and correlation between CC_(x+c)C and wavelengths within 400 to 1000 nm range indicated that CC_(x+c)C could be estimated using reflectance ranging from visible to near-infrared wavebands.Results of Cx+c assessment based on chlorophyll and carotenoid indices showed that red edge chlorophyll index(CI red edge)performed with the highest accuracy(R^(2)=0.77,RMSE=22.27μg/cm^(2),MAE=4.97μg/cm^(2)).Applying partial least square regression(PLSR)in CC_(x+c)C retrieval emphasized the significance of reflectance within 700 to 750 nm range in CC_(x+c)C assessment.Based on CI red edge index,use of airborne hyperspectral imagery achieved satisfactory results in mapping the spatial distribution of CC_(x+c)C.This study demonstrates that it is feasible to accurately assess CC_(x+c)C in winter wheat with red edge chlorophyll index provided that C_(x+c) correlated well with C_(a+b) at the canopy scale.it is therefore a promising method for CC_(x+c)C retrieval at regional scale from aerial hyperspectral imagery.

Evaluation of selenium and carotenoid concentrations of 200 foxtail millet accessions from China and their correlations with agronomic performance

As selenium and carotenoids are essential micronutrients, the determination of their concentrations in different varieties is important in the breeding of foxtail milet （Setaria italicaL. P.Beauv.）. To identify selenium- and carotenoid-enriched foxtail milet varieties and to analyze correlations between trace elements and agronomic traits, we measured the selenium and carotenoid concentrations of 200 Chinese accessions by high-performance liquid chromatography and atomic lfuorescence spectrometry. Our analysis revealed that lutein concentration in 200 foxtail milet accessions folowed normal distribution and average was 3.1 μg g–1. The mean value of zeaxanthin concentration in 200 accessions was 8.6 μg g–1. Lutein and zeaxanthin concentrations were higher in the foxtail milet from Liaoning than in varieties from other locations, with averages of 10.0 and 3.5 μg g–1, respectively. The average measured selenium concentration was 100.3 μg kg–1. The highest average selenium concentration, 110.3 μg kg–1, was found in varieties from Shanxi. Varieties from Inner Mongolia had the lowest average selenium concentration, 84.7 μg kg–1, which was signiifcantly lower （P〈0.05） than that of Shanxi. Selenium con-centrations of 23 varieties were higher than 117.9 μg kg–1, accounting for 11.5% of the total, thereby were considered to be enriched in selenium. In addition, we identiifed 29 lutein-enriched varieties （〉4.27 μg g–1） and 30 zeaxanthin-enriched ones （〉12.63 μg g–1）, which corresponded to 14.5 and 15% of tested accessions, respectively. Correlation analysis revealed that selenium concentration was signiifcantly positively correlated with spikelet length （P〈0.01）, while zeaxanthin concentration was signiifcantly correlated with grass weight （P〈0.05） and spikelet length （P〈0.01）. No correlation was found between lutein concentration and agronomic characters, selenium content or zeaxanthin content. Our results should contribute substantialy to the selection of suitable varieties for the development of plants with desired levels of these nutritionaly important elements. These results wil signiifcantly contribute towards selection of the most suitable varieties for obtaining plants with desired levels of these nutritionaly important elements.

Carotenoid Pigment Accumulation in Horticultural Plants

Carotenoids are a group of widely distributed natural pigments.They give many horticultural plants the bright red,orange,and yellow colors,as well as the aroma and flavor.Carotenoids enhance the health value and represent an essential quality trait of horticultural products.Significant efforts have been made to correlate specific carotenoid production with pathway gene expression.Some transcription factors that directly regulate transcription of the pathway genes have been identified.Horticultural crops have evolved with complicated and multifaceted regulatory mechanisms to generate the enormous diversity in carotenoid content and composition.However,the diverse and complex control of carotenoid accumulation is still not well understood.In this review,we depict carotenoid accumulation pathways and highlight the recent progress in the regulatory control of carotenoid accumulation in horticultural plants.Because of the critical roles of chromoplasts for carotenoid hyperproduction,we evaluate chromoplast ultrastructures and carotenoid sequestrations.A perspective on carotenoid research in horticultural crops is provided.

Enhanced Carotenoid Production by a Mutant of the Marine Yeast Rhodotorula sp. hidai

After a serial of UV, EMS and NTG mutagenesis, a mutant named MM of the red marine yeast strain Rhodotorula sp. hidai was obtained. The mutant MM could produce 603.93 μg g-1 of carotenoid within 5 days in the medium containing 4.0 g sucrose, 1.5 g yeast extract, 0.1 g MgSO4, and 100 mL of sea water, with pH 6.0 and at 30 ℃, while only 213.18 μg g-1 of carotenoid was pro-duced by the wild type under the same conditions.

Association of Dietary Carotenoids Intake with Skeletal Fluorosis in the Coal-burning Fluorosis Area of Guizhou Province

Objective To explore whether the intake of dietary carotenoids could protect against skeletal fluorosis in Guizhou province in which coal-burning fluorosis is endemic. Methods A case-control study of 196 patients with skeletal fluorosis and 196 age and gender-matched controls was conducted in Zhijin, Guizhou Province. Face-to-face interviews were conducted to assess habitual dietary intake using a 75-item food frequency questionnaire and various covariates with structured questionnaires. Urinary fluoride was measured using an ion-selective electrode method. The genotype of superoxide dismutase 2（SOD2） rs11968525 was detected by Taq Man method. Results We observed significant dose-dependent inverse associations of skeletal fluorosis with intake of β-carotene, lutein/zeaxanthin, lycopene, and total carotenoids（P-trend = 0.002 to 0.018）, whereas α-carotene and β-cryptoxanthin intakes were not found to be related to skeletal fluorosis, after adjustment for potential confounders. The adjusted ORs and 95% CI of skeletal fluorosis for the highest versus lowest quartile were 0.30（0.10, 0.86） for β-carotene, 0.23（0.08, 0.66） for lycopene, 0.26（0.10, 0.75） for lutein/zeaxanthin and 0.34（0.14, 0.74） for total carotenoids（all P-trend ＆lt; 0.05）. Stratified analyses showed that the protective effects of lutein/zeaxanthin and total carotenoids on skeletal fluorosis were more evident for individuals with the AG＋AA genotypes of SOD2（rs11968525）. Conclusion Increased intakes of β-carotene, lutein/zeaxanthin, lycopene, and total carotenoids are independently associated with a lower risk of coal-burning skeletal fluorosis. SOD2（rs11968525） polymorphisms might modify the inverse associations between dietary carotenoids and skeletal fluorosis.

Genome-wide Characterization of cis-acting Elements in the Promoters of Key Carotenoid Pathway Genes from the Main Species of Genus Citrus

Carotenoids are indispensable for both human health and plant survival.Citrus,is one of the fruit crops richest in carotenoid compounds,with approximately 115 kinds of carotenoids;tremendous diversity in carotenoids composition and concentration exists among various species,showing different colors from nearly white to crimson.The carotenoid biosynthetic pathway and the key carotenogenic genes have been identified in citrus;however,the underlying regulatory mechanisms remain unclear.In this study,among the main species of genus Citrus(primitive,wild,and cultivated),we detected carotenoids in flavedo using High-Performance Liquid Chromatography,and analyzed variations in cis-acting elements in the promoters of key carotenoid pathway genes.Intriguingly,both carotenoid composition and content were generally increased during the evolution of citrus,and the corresponding variations in the promoters were identified,including the gain or loss of critical environmental stress-responsive elements and hormone-responsive elements,which are closely associated with carotenoid enhancement.In addition,pummelo has the most heat-responsive elements,but the Mangshan mandarin does not have this element in the promoters of PSY,which is highly related to their geographical origin and indicate that temperature is a critical environmental signal influencing carotenoid accumulation.Moreover,the abscisic acid-responsive motif was rich in almost all the seven species,but the ethylene-responsive motif was deficient,which demystified the unique phytohormone regulation mechanism of carotenoid accumulation in citrus.Overall,our study provides new insights into the molecular regulatory mechanism of carotenoid enhancement in the evolution of citrus,which can facilitate breeding and cultivation efforts to improve the nutritional quality and esthetic value in citrus and hopefully other fruit crops.

Expression levels of GSTA2 and APOD genes might be associated with carotenoid coloration in golden pheasant(Chrysolophus pictus) plumage

Carotenoids, which generate yellow, orange, and red colors, are crucial pigments in avian plumage. Investigations into genes associated with carotenoid- based coloration in avian species are important; however, such research is difficult because carotenoids cannot be synthetized in vertebrates as they are only derived from dietary sources. Here, the golden pheasant （Chrysolophus pictus） was used as a model in analysis of candidate gene expression profiles implicated in carotenoid binding and deposition. Using mass and Raman spectrometry to confirm the presence of carotenoids in golden pheasant feathers, we found C40H540 and C40H5602 in feathers with yellow to red colors, and in the rachis of iridescent feathers. The global gene expression profiles in golden pheasant skins were analyzed by RNA-seq and all six carotenoid binding candidate genes sequenced were studied by real- time PCR. STAR4, GSTA2, Scarbl, and APOD in feather follicles showed different expressions in red breast and orange nape feathers compared with that of iridescent mantle feathers. Further comparison of golden pheasant yellow rump and Lady Amherst＇s pheasant （Chrysolophus amherstiae） white nape feathers suggested that GSTA2 and APOD played a potential role in carotenoid-based coloration in golden pheasant.

Changes of the Main Carotenoid Pigment Contents During the Drying Processes of the Different Harvest Stage Fruits of Lycium barbarum L.

The test analyzed the regularity of biosynthesis and degradation of the main functional components, such as zeaxanthin, β-carotene, and esterified carotenoids in the fruit of Lycium barbarum L. in order to provide theoretical basis for improvement of processing condition, appearance quality, and preservation of carotenoids. RP-HPLC was adapted to assay the changes of the main carotenoids of the different harvested stage fruit during the drying processing. Quantification was realized using external standard with gradient elution. The results showed that zeaxanthin and β-carotene contents in fruits increased dramatically, 2-22 times that of fresh fruits at the beginning of the drying period. In the middle of drying period, degradation occurred to a some extent, and the fall fruit degraded to a large extent. At the end of drying period, zeaxanthin and β-carotene contents increased to a little extent until a balanced state is obtained. Zeaxanthin dipalmitate content had a total degradation to more than 40% at the beginning of the drying period, and increased a little at the middle period, then reached a balanced state finally. The total carotenoid content analysis showed that the summer fruit had higher carotenoid content than the fall fruit. The experiments demonstrated zeaxanthin and β-carotene contents in fruits increased and zeaxanthin dipalmitate decreased during the drying process, which had an effect on the production appearance.

The Mechanism of Carotenoid Degradation in Flue-Cured Tobacco and Changes in the Related Enzyme Activities at the Leaf-Drying Stage During the Bulk Curing Process

The mechanism of carotenoid degradation and the changes in the activities of related enzymes in flue-cured tobacco at the leaf-drying stage during the bulk-curing process were studied in order to provide theoretical basis for optimization of curing technology. The effect of different rising speeds of temperature on the carotenoid degradation and the related enzymes activities at the color-fixing stage during the bulk curing process was studied by using the electric-heated fluecuring barn designed by Henan Agricultural University, China, based on curing technology with yellowing at low temperature and moderate humidity and leaf drying at moderate humidity. The results showed that the carotenoid degradation components （β-carotene, lutein, neoxanthin, and violaxthin） decreased gradually at the color-fixing stage during the bulk curing process. The carotenoid degradation components viz.,β-carotene, lutein, neoxanthin, and violaxthin at the slow heating curing （T1） were relatively higher than the rapid heating curing （T2） accounting for 10, 2, 32 and 32% respectively, but there were no differences among treatments （P〉 0.05）. The effect of different conditions of curing on the activities of enzymes related to carotenoids degradation were significant. The lipoxygenase, phenylalanine ammonialyase, peroxidase, and polyphenol oxidase enzymes had a bidirectional effect on the quality of tobacco leaves and it was beneficial to form more premise matter of aroma based on the higher enzyme activities at the early leaf-drying stage. The slow heating could regulate the change in various enzymes＇ activities reasonably, making cell redox reaction to reach the dynamic balance and make the degradation of carotenoids adequately. Meanwhile, it could avoid the occurrence of browning reaction and provide foundation for improving the quality of tobacco and optimization of technology for bulk curing and further enhancing aroma.

Transcriptome analysis of carotenoid biosynthesis in Dunaliella salina under red and blue light

The quality of light is an important abiotic factor that affects the growth and development of photosynthetic organisms.In this study,we exposed the unicellular green alga Dunaliella salina to red(660 nm)and blue(450 nm)light and analyzed the cell growth,total carotenoid content,and transcriptomes.The growth of D.salina was enhanced by illumination with red light,whereas blue light was not able to promote the algal growth.In contrast,the total carotenoid content increased under both red and blue light.The RNA of D.salina was sequenced and the transcriptomic response of algal cells to red and blue light was investigated.Six transcripts encoding for the blue light receptor cryptochrome were identified,and transcripts involved in the carotenoid metabolism were up-regulated under both red and blue light.Transcripts encoding for photoprotective enzymes related to the scavenging of reactive oxygen species were up-regulated under blue light.The present transcriptomic study provides a more comprehensive understanding of carotenoid biosynthesis in D.salina under different wavelengths of light.

Evaluation of antigenotoxic effects of carotenoids from green algae Chlorococcum humicola using human lymphocytes

Objective:To identify the available phytochemicals and carotenoids in the selected green algae and evaluate the potential genotoxic/antigenotoxic effect using lymphocytes.Methods:Organic,solvent extracts of Chlorococcum humicola(C.humicola)were used for the phytochemical analysis.The available carotenoids were assessed by HPLC,and LC-MS analysis.The genotoxicity was induced by the benzo(a)pyrene in the lymphocyte culture,the genotoxic and antigenotoxic effects of algal carotenoids with and without genotoxic inducer were evaluated by chromosomal aberration(CA),sister chromatid exchange(SCE)and micronucleus assay(MN).Results:The results of the analysis showed that the algae were rich in carotenoids and fatty acids.In the total carotenoids lutein,β-carotene andα-carotene were found to be present in higher concentration.The frequency of CA and SCE increased by benzo(a)pyrene were significantly decreased by the carotenoids(P<0.05 for CA,P<0.001 for SCE).The MN frequencies of the cells were significantly decreased by the treatment with carotenoids when compared with the positive controls(P<0.05).Conclusions:The findings of the present study demonstrate that,the green algae C.humicola is a rich source of bioactive compounds especially carotenoids which effectively fight against environmental genotoxic agents,the carotenoids itself is not a genotoxic substance and should be further considered for its beneficial effects.

Cloning and Functional Characterisation of Carotenoid Cleavage Dioxygenase 4 from Wolfberry

Carotenoid cleavage dioxygenases(CCDs) are a class of enzymes in plants involved in the biosynthesis of apocarotenoids,such as phytohormones,flavour compounds,and other compounds with yet unknown functions.To date,several CCDs have been functionally characterised in plants,but little is known about the CCD4 members.A carotenoid cleavage dioxygenase 4 gene(LcCCD4) was isolated from the leaves of wolfberry(Lycium chinense) to gain insight into its biological function.Multiple sequence alignment and phylogenetic analyses showed that the deduced amino acid sequence of LcCCD4 shares high homology with that of CCD4 proteins from other plants.Expression analysis using semi-quantitative polymerase chain reaction revealed that LcCCD4 was strongly expressed in leaves and flowers and that the expression level was in accordance with β-carotene concentration.LcCCD4 transcripts in fruits tended to decrease as carotenoids accumulated.Recombinant expression of LcCCD4 cleaved β-carotene to produce P-ionone in in vivo assays.These results show that LcCCD4 is a CCD gene that may be involved in producing aromatic apocarotenoids in leaves and flowers,whereas it may be involved in controlling carotenoid accumulation in fruits.

CPTA treatment reveals potential transcription factors associated with carotenoid metabolism in flowers of Osmanthus fragrans

Osmanthus fragrans is one of the top ten traditional flowers in China.It is divided into three different groups according to its color.α-Carotene and β-carotene are the main determinants to distinguish the color differences between three groups.However,the dominant genes and transcription factors involved in carotenoid metabolism remain unclear.CPTA treatment(0.7mmol·L−1)remarkably promoted lycopene,α-carotene and β-carotene contents in flowers.Transcriptome sequencing analysis revealed that CPTA treatment could trigger chain reactions in carotenoid metabolism pathway genes.Four up-regulated and 10 down-regulated transcription factors which have close association with carotenoid variation were significantly induced by CPTA treatment.The up-regulated TFs such as MYB43,MYB123,HSF,were further subjected to transcript expression determination in different cultivars with drastic colors.Among them,transcript expression of four up-regulated TFs coincided with the carotenoid accumulation in different cultivars.We selected up-regulated OfMYB43 to verify its function,which is related to stress tolerance and transcriptional regulation.Transient overexpression of OfMYB43 in O.fragrans flowers showed that it could remarkably promote the expression of PDS,ZISO,LCYE and CCD4,leading to increased accumulation of β-branch carotenoids.OfMYB43 was a potential positive regulator of carotenoid biosynthesis in O.fragrans flowers.This study provides insight into the molecular mechanism of carotenoid metabolism in O.fragrans.