Carotenoid Metabolic Pathway Research Articles

Biochemical Characterization of Orange-Colored Rice Calli Induced by Target Mutagenesis of OsOr Gene.

We generated an orange-colored (OC) rice callus line by targeted mutagenesis of the orange gene (OsOr) using the CRISPR-Cas9 system. The OC line accumulated more lutein, β-carotene, and two β-carotene isomers compared to the WT callus line. We also analyzed the expression levels of carotenoid biosynthesis genes by qRT-PCR. Among the genes encoding carotenoid metabolic pathway enzymes, the number of transcripts of the PSY2, PSY3, PDS, ZDS and β-LCY genes were higher in the OC line than in the WT line. In contrast, transcription of the ε-LCY gene was downregulated in the OC line compared to the WT line. In addition, we detected increases in the transcript levels of two genes involved in carotenoid oxidation in the OC lines. The developed OC lines also showed increased tolerance to salt stress. Collectively, these findings indicate that targeted mutagenesis of the OsOr gene via CRISPR/Cas9-mediated genome editing results in β-carotene accumulation in rice calli. Accordingly, we believe that this type of genome-editing technology could represent an effective alternative approach for enhancing the β-carotene content of plants.

Temperature-induced zeaxanthin overproduction in Synechococcus elongatus PCC 7942.

The exogenous crtZ gene from Brevundimonas sp. SD212, coding for a 3,3' β-car hydroxylase, was expressed in Synechococcus elongatus PCC 7942 under the control of a temperature-inducible promoter in an attempt to engineer the carotenoid metabolic pathway, to increase the content of zeaxanthin and its further hydroxylated derivatives caloxanthin and nostoxanthin. These molecules are of particular interest due to their renowned antioxidant properties. Cultivation of the engineered strain S7942Z-Ti at 35°C, a temperature which is well tolerated by the wild-type strain and at which the inducible expression system is activated, led to a significant redistribution of the relative carotenoid content. β-Carotene decreased to about 10% of the pool that is an excess of a threefold decrease with respect to the control, and concomitantly, zeaxanthin became the dominant carotenoid accounting for about half of the pool. As a consequence, zeaxanthin and its derivatives caloxanthin and nostoxanthin collectively accounted for about 90% of the accumulated carotenoids. Yet, upon induction of CrtZ expression at 35°C the S7942Z-Ti strain displayed a substantial growth impairment accompanied, initially, by a relative loss of carotenoids and successively by the appearance of chlorophyll degradation products which can be interpreted as markers of cellular stress. These observations suggest a limit to the exploitation of Synechococcus elongatus PCC 7942 for biotechnological purposes aimed at increasing the production of hydroxylated carotenoids.

Integrated Metabolomic and Transcriptomic Analyses Reveal the Basis for Carotenoid Biosynthesis in Sweet Potato (Ipomoea batatas (L.) Lam.) Storage Roots.

Carotenoids are important compounds of quality and coloration within sweet potato storage roots, but the mechanisms that govern the accumulation of these carotenoids remain poorly understood. In this study, metabolomic and transcriptomic analyses of carotenoids were performed using young storage roots (S2) and old storage roots (S4) from white-fleshed (variety S19) and yellow-fleshed (variety BS) sweet potato types. S19 storage roots exhibited significantly lower total carotenoid levels relative to BS storage roots, and different numbers of carotenoid types were detected in the BS-S2, BS-S4, S19-S2, and S19-S4 samples. β-cryptoxanthin was identified as a potential key driver of differences in root coloration between the S19 and BS types. Combined transcriptomic and metabolomic analyses revealed significant co-annotation of the carotenoid and abscisic acid (ABA) metabolic pathways, PSY (phytoene synthase), CHYB (β-carotene 3-hydroxylase), ZEP (zeaxanthin epoxidase), NCED3 (9-cis-epoxycarotenoid dioxygenase 3), ABA2 (xanthoxin dehydrogenase), and CYP707A (abscisic acid 8'-hydroxylase) genes were found to be closely associated with carotenoid and ABA content in these sweet potato storage roots. The expression patterns of the transcription factors OFP and FAR1 were associated with the ABA content in these two sweet potato types. Together, these results provide a valuable foundation for understanding the mechanisms governing carotenoid biosynthesis in storage roots, and offer a theoretical basis for sweet potato breeding and management.

Overexpression of PSY1 increases fruit skin and flesh carotenoid content and reveals associated transcription factors in apple (Malus × domestica).

Knowledge of the transcriptional regulation of the carotenoid metabolic pathway is still emerging and here, we have misexpressed a key biosynthetic gene in apple to highlight potential transcriptional regulators of this pathway. We overexpressed phytoene synthase (PSY1), which controls the key rate-limiting biosynthetic step, in apple and analyzed its effects in transgenic fruit skin and flesh using two approaches. Firstly, the effects of PSY overexpression on carotenoid accumulation and gene expression was assessed in fruit at different development stages. Secondly, the effect of light exclusion on PSY1-induced fruit carotenoid accumulation was examined. PSY1 overexpression increased carotenoid content in transgenic fruit skin and flesh, with beta-carotene being the most prevalent carotenoid compound. Light exclusion by fruit bagging reduced carotenoid content overall, but carotenoid content was still higher in bagged PSY fruit than in bagged controls. In tissues overexpressing PSY1, plastids showed accelerated chloroplast to chromoplast transition as well as high fluorescence intensity, consistent with increased number of chromoplasts and carotenoid accumulation. Surprisingly, the expression of other carotenoid pathway genes was elevated in PSY fruit, suggesting a feed-forward regulation of carotenogenesis when this enzyme step is mis-expressed. Transcriptome profiling of fruit flesh identified differentially expressed transcription factors (TFs) that also were co-expressed with carotenoid pathway genes. A comparison of differentially expressed genes from both the developmental series and light exclusion treatment revealed six candidate TFs exhibiting strong correlation with carotenoid accumulation. This combination of physiological, transcriptomic and metabolite data sheds new light on plant carotenogenesis and TFs that may play a role in regulating apple carotenoid biosynthesis.

Overexpression of Sweet Potato Carotenoid Cleavage Dioxygenase 4 (IbCCD4) Decreased Salt Tolerance in Arabidopsis thaliana.

Salt stress has a serious impact on normal plant growth and yield. Carotenoid cleavage dioxygenase (CCD) degrades carotenoids to produce apocarotenoids, which are involved in plant responses to biotic and abiotic stresses. This study shows that the expression of sweet potato IbCCD4 was significantly induced by salt and dehydration stress. The heterologous expression of IbCCD4 in Arabidopsis was induced to confirm its salt tolerance. Under 200 mM NaCl treatment, compared to wild-type plants, the rosette leaves of IbCCD4-overexpressing Arabidopsis showed increased anthocyanins and carotenoid contents, an increased expression of most genes in the carotenoid metabolic pathway, and increased malondialdehyde (MDA) levels. IbCCD4-overexpressing lines also showed a decreased expression of resistance-related genes and a lower activity of three antioxidant enzymes: peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT). These results indicate that IbCCD4 reduced salt tolerance in Arabidopsis, which contributes to the understanding of the role of IbCCD4 in salt stress.

Methylome and transcriptome analyses of three different degrees of albinism in apple seedlings

BackgroundLeaf colour mutations are universally expressed at the seedling stage and are ideal materials for exploring the chlorophyll biosynthesis pathway, carotenoid metabolism and the flavonoid biosynthesis pathway in plants.ResultsIn this research, we analysed the different degrees of albinism in apple (Malus domestica) seedlings, including white-leaf mutants (WM), piebald leaf mutants (PM), light-green leaf mutants (LM) and normal leaves (NL) using bisulfite sequencing (BS-seq) and RNA sequencing (RNA-seq). There were 61,755, 79,824, and 74,899 differentially methylated regions (DMRs) and 7566, 3660, and 3546 differentially expressed genes (DEGs) identified in the WM/NL, PM/NL and LM/NL comparisons, respectively.ConclusionThe analysis of the methylome and transcriptome showed that 9 DMR-associated DEGs were involved in the carotenoid metabolism and flavonoid biosynthesis pathway. The expression of different transcription factors (TFs) may also influence the chlorophyll biosynthesis pathway, carotenoid metabolism and the flavonoid biosynthesis pathway in apple leaf mutants. This study provides a new method for understanding the differences in the formation of apple seedlings with different degrees of albinism.

Comparative metabolomic and transcriptomic analysis reveals a coexpression network of the carotenoid metabolism pathway in the panicle of Setaria italica

BackgroundThe grains of foxtail millet are enriched in carotenoids, which endow this plant with a yellow color and extremely high nutritional value. However, the underlying molecular regulation mechanism and gene coexpression network remain unclear.MethodsThe carotenoid species and content were detected by HPLC for two foxtail millet varieties at three panicle development stages. Based on a homologous sequence BLAST analysis, these genes related to carotenoid metabolism were identified from the foxtail millet genome database. The conserved protein domains, chromosome locations, gene structures and phylogenetic trees were analyzed using bioinformatics tools. RNA-seq was performed for these samples to identify differentially expressed genes (DEGs). A Pearson correlation analysis was performed between the expression of genes related to carotenoid metabolism and the content of carotenoid metabolites. Furthermore, the expression levels of the key DEGs were verified by qRT-PCR. The gene coexpression network was constructed by a weighted gene coexpression network analysis (WGCNA).ResultThe major carotenoid metabolites in the panicles of DHD and JG21 were lutein and β-carotene. These carotenoid metabolite contents sharply decreased during the panicle development stage. The lutein and β-carotene contents were highest at the S1 stage of DHD, with values of 11.474 μg /100 mg and 12.524 μg /100 mg, respectively. Fifty-four genes related to carotenoid metabolism were identified in the foxtail millet genome. Cis-acting element analysis showed that these gene promoters mainly contain ‘plant hormone’, ‘drought stress resistance’, ‘MYB binding site’, ‘endosperm specific’ and ‘seed specific’ cis-acting elements and especially the ‘light-responsive’ and ‘ABA-responsive’ elements. In the carotenoid metabolic pathways, SiHDS, SiHMGS3, SiPDS and SiNCED1 were more highly expressed in the panicle of foxtail millet. The expression of SiCMT, SiAACT3, SiPSY1, SiZEP1/2, and SiCCD8c/8d was significantly correlated with the lutein content. The expression of SiCMT, SiHDR, SiIDI2, SiAACT3, SiPSY1, and SiZEP1/2 was significantly correlated with the content of β-carotene. WGCNA showed that the coral module was highly correlated with lutein and β-carotene, and 13 structural genes from the carotenoid biosynthetic pathway were identified. Network visualization revealed 25 intramodular hub genes that putatively control carotenoid metabolism.ConclusionBased on the integrative analysis of the transcriptomics and carotenoid metabonomics, we found that DEGs related to carotenoid metabolism had a stronger correlation with the key carotenoid metabolite content. The correlation analysis and WGCNA identified and predicted the gene regulation network related to carotenoid metabolism. These results lay the foundation for exploring the key target genes regulating carotenoid metabolism flux in the panicle of foxtail millet. We hope that these target genes could be used to genetically modify millet to enhance the carotenoid content in the future.

The Genetic Components of a Natural Color Palette: A Comprehensive List of Carotenoid Pathway Mutations in Plants.

Carotenoids comprise the most widely distributed natural pigments. In plants, they play indispensable roles in photosynthesis, furnish colors to flowers and fruit and serve as precursor molecules for the synthesis of apocarotenoids, including aroma and scent, phytohormones and other signaling molecules. Dietary carotenoids are vital to human health as a source of provitamin A and antioxidants. Hence, the enormous interest in carotenoids of crop plants. Over the past three decades, the carotenoid biosynthesis pathway has been mainly deciphered due to the characterization of natural and induced mutations that impair this process. Over the year, numerous mutations have been studied in dozens of plant species. Their phenotypes have significantly expanded our understanding of the biochemical and molecular processes underlying carotenoid accumulation in crops. Several of them were employed in the breeding of crops with higher nutritional value. This compendium of all known random and targeted mutants available in the carotenoid metabolic pathway in plants provides a valuable resource for future research on carotenoid biosynthesis in plant species.

Genetic manipulation for carotenoid production in microalgae an overview

• Carotenoids are natural pigments with significant biological and commercial values. • Microalgae are promising source for carotenoids. • Advanced genetic manipulation tools (Overexpression and gene knockout) are helping to overproduce the carotenoid content. • Manipulating the genes of key enzymes can enhance the carotenoid content in microalgae. Carotenoids are the most diverse pigments found in plants and animals. Carotenoids of various hues such as orange, red, and yellow are abundant in fruits and flowers. Also, they are responsible for the colors seen in some birds and fish. These pigments also offer significant health advantages, such as antioxidant and anti-inflammatory properties, and they have a wide range of applications in the food, cosmetics, and pharmaceutical industries. Carotenoids are key phytocomponents in plants and microalgae for light harvesting, energy transfer during photosynthesis, and protection from photo-oxidative damage. Microalgae are a diverse group of photosynthetic organisms that provide the foundation of the food chain in aquatic habitats. Dunaleilla salina, Haematococcus pluvialis, Chlorella vulgaris, Chlorella pyrenoidosa, and Chlorella zofingiensis are the richest sources for β-carotene, astaxanthin, lycopene, lutein, and zeaxanthin. Microalgae provide renewable supplies for increasing carotenoids because of their perfect host environment, simplicity of culture, and rapid growth rate. Genetic manipulation tools such as overexpression and gene knockout techniques are emerging approaches to study the carotenoid metabolic pathway and reveal information about the critical enzymes in the pathway. In this review, we discussed recent developments in genetic manipulation to overproduce carotenoids. Recent genetic manipulation tools are potential options for increasing productivity in microalgae. These approaches will enhance the production of carotenoids in commercial microalgal species for industrial applications.

Integrated metabolome and transcriptome analysis revealed carotenoid metabolism difference in pepper (<i>Capsicum annuum</i> L.) yellowing mutants under different light quality

Light is an important environmental factor for plant growth and development, different light qualities have different regulatory effects on plants. To investigate the effect of light quality on plants, we determined the physiological characteristics, transcriptome and metabolome analysis of pepper yellowing mutants <italic>yl1</italic> treated with blue, red, green, and purple light. Results showed that the leaf of <italic>yl1</italic> was obviously yellowing, and the contents of chlorophyll, carotenoid and net photosynthetic rate in <italic>yl1</italic> were significantly decreased under purple light. A total of 31,853 genes were quantified under blue, red, green and purple light. The genes related to carotenoid metabolism pathway such as <italic>PSY, LUT5</italic> and<italic> VDE</italic> were significantly increased, while the expression levels of chlorophyll synthesis related genes <italic>POR</italic> and <italic>CAO</italic> were significantly decreased under purple lught. At the same time, twenty-one carotenoid pathway metabolites were detected under the four light, and ten metabolites were more abundant in pepper leaves. <italic>α</italic>-carotene, <italic>β</italic>-carotene, lutein, neoxanthin, <italic>α</italic>-cryptoxanthin and <italic>β</italic>-cryptoxanthin were significantly accumulated under blue, red and green light. However, zeaxanthin and antheraxanthin were accumulated in large quantities under purple light. After silencing <italic>CaVDE</italic> gene under purple light, leaf etiolation degree was significantly weakened, chlorophyll, carotenoids and net photosynthetic rate were significantly increased, and the accumulation of zeaxanthin and antheraxanthin was significantly decreased. These results provide a reference for analyzing the changes of carotenoid components induced by <italic>VDE</italic> in purple light and provide new insights into the mechanism of leaf color change in plants.

Transcriptome sequencing and global analysis of blue light-responsive genes provide clues for high carotenoid yields in Blakeslea trispora.

Blakeslea trispora has great potential uses in industrial production because of the excellent capability of producing a large quantity of carotenoids. However, the mechanisms of light-induced carotenoid biosynthesis even the structural and regulatory genes in pathways remain unclear. In this paper, we reported the first transcriptome study in B. trispora in which we have carried out global survey of expression changes of genes participated in blue light response. We verified that the yield of β-carotene increased 3-fold when transferred from darkness to blue light for 24 h and the enhancement of transcription levels of carRA and carB presented a positive correlation with the increase in carotenoid production. RNA-seq analysis revealed that 1124 genes were upregulated and 740 genes were downregulated respectively after blue light exposure. Annotation through GO, KEGG, Swissprot, and COG databases showed 11119 unigenes compared well with known gene sequences, 5514 unigenes were classified into Gene Ontology, and 4675 unigenes were involved in distinct pathways. Among the blue light-responsive genes, 4 genes (carG1, carG3, carRA and carB) identified to function in carotenoid metabolic pathways were dominantly upregulated. We also discovered that 142 TF genes belonging to 45 different superfamilies showed significant differential expression (p≤ 0.05), 62 of which were obviously repressed by blue light. The detailed profile of transcription data will not only allow us to conduct further functional genomics study in B. trispora, but also enhance our understanding of potential metabolic pathway and regulatory network involved in light-regulated carotenoid synthesis.

Biochemical and transcriptomic analyses reveal that critical genes involved in pigment biosynthesis influence leaf color changes in a new sweet osmanthus cultivar 'Qiannan Guifei'.

BackgroundOsmanthus fragrans (Oleaceae) is one of the most important ornamental plant species in China. Many cultivars with different leaf color phenotypes and good ornamental value have recently been developed. For example, a new cultivar ‘Qiannan Guifei’, presents a rich variety of leaf colors, which change from red to yellow-green and ultimately to green as leaves develop, making this cultivar valuable for landscaping. However, the biochemical characteristics and molecular mechanisms underlying leaf color changes of these phenotypes have not been elucidated. It has been hypothesized that the biosynthesis of different pigments in O. fragrans might change during leaf coloration. Here, we analyzed transcriptional changes in genes involved in chlorophyll (Chl), flavonoid, and carotenoid metabolic pathways and identified candidate genes responsible for leaf coloration in the new cultivar ‘Qiannan Guifei’.MethodsLeaf samples were collected from ‘Qiannan Guifei’ plants at the red (R), yellow-green (YG) and green (G) leaf stages. We compared the different-colored leaves via leaf pigment concentrations, chloroplast ultrastructure, and transcriptomic data. We further analyzed differentially expressed genes (DEGs) involved in the Chl, flavonoid, and carotenoid metabolic pathways. In addition, we used qRT-PCR to validate expression patterns of the DEGs at the three stages.ResultsWe found that, compared with those at the G stage, chloroplasts at the R and YG stages were less abundant and presented abnormal morphologies. Pigment analyses revealed that the leaves had higher flavonoid and anthocyanin levels at the R stage but lower Chl and carotenoid concentrations. Similarly, Chl and carotenoid concentrations were lower at the YG stage than at the G stage. By using transcriptomic sequencing, we further identified 61 DEGs involved in the three pigment metabolic pathways. Among these DEGs, seven structural genes (OfCHS, OfCHI, OfF3H, OfDFR, OfANS, OfUGT andOf3AT) involved in the flavonoid biosynthesis pathway were expressed at the highest level at the R stage, thereby increasing the biosynthesis of flavonoids, especially anthocyanins. Six putativeOfMYB genes, including three flavonoid-related activators and three repressors, were also highly expressed at the R stage, suggesting that they might coordinately regulate the accumulation of flavonoids, including anthocyanins. Additionally, expressions of the Chl biosynthesis-related genes OfHEMA, OfCHLG and OfCAO and the carotenoid biosynthesis-related genes OfHYB and OfZEP were upregulated from the R stage to the G stage, which increased the accumulation of Chl and carotenoids throughout leaf development. In summary, we screened the candidate genes responsible for the leaf color changes of ‘Qiannan Guifei’, improved current understanding of the regulatory mechanisms underlying leaf coloration and provided potential targets for future leaf color improvement in O. fragrans.

Comparative metabolomic and transcriptomic analyses revealed the differential accumulation of secondary metabolites during the ripening process of acerola cherry (Malpighia emarginata) fruit.

Acerola cherry is a famous functional fruit containing plentiful antioxidants and other nutrients. However, studies on the variations among nutrients during the ripening process of acerola fruit are scare. Comparative metabolomic and transcriptomic analyses were performed and identified 31 331 unigenes and 1896 annotated metabolite features in acerola cherry fruit. K Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that several antioxidant and nutrient-related metabolic pathways, such as the flavonoids, vitamins, carotenoids, amino acids, and fatty acids metabolic pathways, were significantly changed during the ripening process. The metabolites related to the vitamin, carotenoid, and fatty acid metabolic pathways were downregulated during the ripening process. Several flavonoid biosynthesis-related genes (including dihydroflavonol 4-reductase, chalcone synthase, flavanone 3-hydroxylase, and anthocyanidin synthase), were significantly upregulated, suggesting their essential functions in the accumulation of flavonoids in mature fruit. Most of the vitamin and carotenoid metabolism-related metabolites significantly accumulated in immature fruit, suggesting that immature acerola fruit is a good material for the extraction of vitamins and carotenoids. For macronutrients, most of the amino acids accumulated in mature fruit and most of the fatty acids greatly accumulated in immature fruit. Our data revealed the differential accumulation of antioxidants and nutrients during the ripening process of acerola cherry fruit. © 2021 Society of Chemical Industry.

Evaluation of Carotenoids Accumulation and Biosynthesis in Two Genotypes of Pomelo (Citrus maxima) during Early Fruit Development.

Pomelo is rich in bioactive compounds (carotenoids, phenolics and essential oil) in the early stage of fruit development, but it is often wasted in the cultivation and management process. To gain an insight into the carotenoid metabolism pathway in pomelo, the carotenoid profiles and the expression patterns of carotenogenic genes were investigated in two genotypes of pomelo during early fruit development. The results showed that a higher carotenoid content was observed in honey pomelo as compared with golden pomelo, which may be related to different gene regulation mechanisms. Lutein, α-carotene, and β-carotene were the main carotenoids in pomelo young fruit, and lutein was the highest one. The accumulation of carotenoids during fruit early development in honey pomelo is related to the transcriptional regulation of ZISO and LUT5. In golden pomelo, the rate-limiting gene for carotenoids is PDS and ZDS. In addition, the expression of seven genes except CRTISO in honey pomelo was higher than that in golden pomelo. The results are helpful to further clarify the regulatory mechanism of carotenoid accumulation during early fruit development and provide a direction for the high-value utilization of young fruits in pomelo.

Identification of Key Genes Controlling Carotenoid Metabolism during Apricot Fruit Development by Integrating Metabolic Phenotypes and Gene Expression Profiles.

To explore the metabolic basis of carotenoid accumulation in different developmental periods of apricot fruits, targeted metabonomic and transcriptomic analyses were conducted in four developmental periods (S1-S4) in two cultivars (Prunus armeniaca cv. "Kuchebaixing" with white flesh and P. armeniaca cv. "Shushangganxing" with orange flesh) with different carotenoid contents. 14 types of carotenes and 27 types of carotene lipids were identified in apricot flesh in different developmental periods. In S3 and S4, the carotenoid contents of the two cultivars were significantly different, and β-carotene and (E/Z)-phytoene were the key metabolites that caused the difference in the total carotenoid content between the examined cultivars. Twenty-five structural genes (including genes in the methylerythritol 4-phosphate and carotenoid biosynthesis pathways) related to carotenoid biosynthesis were identified among the differentially expressed genes in different developmental periods of the two cultivars, and a carotenoid metabolic pathway map of apricot fruits was drawn according to the KEGG pathway map. The combined analysis of carotenoid metabolism data and transcriptome data showed that PSY, NCED1, and CCD4 were the key genes leading to the great differences in the total carotenoid content. The results provide a new approach to study the synthesis and accumulation of carotenoids in apricot fruits.

Mapping and predicting a candidate gene for flesh color in watermelon

The color of watermelon flesh is an important trait determined by a series of carotenoids. Herein, we used Cream of Saskatchewan (pale yellow flesh) and PI 186490 (white flesh) as parental materials for an F2 segregation and initial mapping using the bulked segregant analysis sequencing (BSA-seq) strategy. The BSA results revealed a flesh color-related QTL that spans approximately 2.45 Mb on chromosome 6. This region was preliminarily positioned in a 382-kb segment, and then narrowed down into a 66.8-kb segment with 1260 F2 individuals. A total of nine candidate genes were in the fine mapping interval, but only Cla007528 (encoding chlorophyllase) had non-synonymous mutations and was significantly expressed between the parental materials throughout flesh development. We also checked the expression patterns of the carotenoid metabolic pathway genes based on RNA-seq data and qRT-PCR validation. Three genes in the xanthophyll cycle (CICHYB, CINCED-1 and CINCED-7) exhibited differential expression patterns between the two parental lines at different flesh color formation stages. ClPSY1, CIPDS, CIZDS, CICHXE, CICRTISO and CILCYB also exhibited clearly different expression patterns accompanied by carotenoid accumulation.

Red light-induced kumquat fruit coloration is attributable to increased carotenoid metabolism regulated by FcrNAC22.

Carotenoids play vital roles in the coloration of plant tissues and organs, particularly fruits; however, the regulation of carotenoid metabolism in fruits during ripening is largely unknown. Here, we show that red light promotes fruit coloration by inducing accelerated degreening and carotenoid accumulation in kumquat fruits. Transcriptome profiling revealed that a NAC (NAM/ATAF/CUC2) family transcription factor, FcrNAC22, is specifically induced in red light-irradiated fruits. FcrNAC22 localizes to the nucleus, and its gene expression is up-regulated as fruits change color. Results from dual luciferase, yeast one-hybrid assays and electrophoretic mobility shift assays indicate that FcrNAC22 directly binds to, and activates the promoters of three genes encoding key enzymes in the carotenoid metabolic pathway. Moreover, FcrNAC22 overexpression in citrus and tomato fruits as well as in citrus callus enhances expression of most carotenoid biosynthetic genes, accelerates plastid conversion into chromoplasts, and promotes color change. Knock down of FcrNAC22 expression in transiently transformed citrus fruits attenuates fruit coloration induced by red light. Taken together, our results demonstrate that FcrNAC22 is an important transcription factor that mediates red light-induced fruit coloration via up-regulation of carotenoid metabolism.

Comprehensive transcriptome-based characterization of differentially expressed genes involved in carotenoid biosynthesis of different ripening stages of Capsicum

Pepper (Capsicum annuum L.) is one of the widely cultivated vegetable crops in the world. The color formation of pepper fruit is closely related to the carotenoid composition and content. In order to understand the regulatory mechanism of red and orange pepper formation, comprehensive transcriptome and carotenoid content determination were carried out at 25, 40 and 55 days after flower. At the mature stage (55 DAF), the contents of zeaxanthin, β-cryptoxanthin, β-carotene and capsanthin in red chili pepper(R) were significantly higher than those in orange pepper (O), while the content of lutein in orange pepper(O) was higher than that in red pepper(R). Moreover, it was found that many differentially expressed genes (DEGs) were involved in the biosynthesis of carotenoids, phenylpropanoids, flavonoids and metabolism of phenylalanine. A total of 23 DEGs were found to be involved in carotenoid biosynthesis, among which β-carotene hydroxylase 1(CHYB1) and capsanthin-capsorubin synthase(CCS)were significantly different during fruit development. Further analysis showed that there were complete CCS genes in both red and orange pepper fruits. However, the expression level in orange pepper fruits was maintained at a very low level, on the contrary, the red fruits exhibited a remarkable upward trend and extremely high transcript level. Through weighted gene co-expression network analysis (WGCNA), transcription factors F-box protein SKIP23, GATA transcription factor 26, U-box domain-containing protein 52, zinc finger family FYVE/PHD-type, RING/FYVE/PHD-type and CONSTANS-LIKE 9 might be involved in the regulation of CCS gene. This study will lay a base to further analyze the regulation mechanism of carotenoid metabolism pathway in the process of pepper fruit color formation.

Carotenoid Accumulation and the Expression of Carotenoid Metabolic Genes in Mango during Fruit Development and Ripening

Carotenoids are considered to be important components in mango fruits. However, there is a lack of understanding about the regulation of carotenoids in mango. To gain an insight into the carotenoid metabolism pathway, carotenoid content and the expression of carotenoid metabolic genes were investigated in the peel and pulp of mango during fruit development and ripening in three cultivars, ‘Kaituk’, ‘Nam Dok Mai No.4′, and ‘Nam Dok Mai Sithong’, which are different in color. The highest carotenoid content was observed in ‘Kaituk’, followed by ‘Nam Dok Mai No.4′ and ‘Nam Dok Mai Sithong’, with the major carotenoid being β-carotene. The gene expression analysis found that carotenoid metabolism in mango fruit was primarily regulated at the transcriptional level. The changing patterns of carotenoid biosynthetic gene expression (MiPSY, MiPDS, MiZDS, MiCRTISO, MiLCYb, MiLCYe, MiHYb, and MiZEP) were similar to carotenoid accumulation, and ‘Kaituk’ exhibited a higher expression level than the other two cultivars. In addition, the differential regulation of carotenoid catabolic genes was found to be a mechanism responsible for variability in carotenoid content among the three mango cultivars. The expression of carotenoid catabolic genes (MiCCD1, MiNCED2, and MiNCED3) more rapidly decreased in ‘Kaituk’, resulting in a larger amount of carotenoids in ‘Kaituk’ than the other two cultivars.

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 (0.7 mmol · 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.