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UV/vis spectra and characteristic ions of carotenoids from six maturation stages of loquat fruits, obtained by HPLC-PDA-MS
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Background
Carotenoids are the main colouring substances found in orange-fleshed loquat fruits. The aim of this study was to unravel the carotenoid biosynthetic pathway of loquat fruit (cv. ‘Obusa’) in peel and flesh tissue during distinct on-tree developmental stages through a targeted analytical and molecular approach.
Results
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Contexts in source publication
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... amplification cycle was followed by a melting curve run, carrying out 61 cycles with 0.5 °C increment from 65 to 95 °C. The annealing temperature of previ- ously published primers for loquat carotenoid biosynthetic genes (58 to 65 °C) is shown in Additional file 1: Table S1. Loquat's actin gene was used as a housekeeping reference gene (EjACT). ...
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... in the peel and in the flesh were identified and quantified (Table 1, Additional file 1: Figure S1). Thirty-two carotenoids were detected by HPLC-DAD and LC-MS techniques. ...
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... identification was based on their relative retention time values, their UV-Vis spectra, their mass spectra, information from the literature and comparison with authentic standards when possible. Table 1 summarizes the identification data for each ca- rotenoid, including chromatographic and spectroscopic values. ...
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... carotenoid profiling of loquat flesh was found to be quite different from the peel. The most abundant carotenoid in mature fruits was trans-β-cryptoxanthin, followed by trans-β-carotene, compounds 18 and 31, and 5,8-epoxy-β-carotene (peak 30) ( Table 1, Fig. 3, Additional file 1: Table S4). An increment in the concentration of all carotenoids during on-tree development except for trans- neoxanthin, trans-neochrome and trans-lutein was found (Fig. 3, Additional file 1: Table S4). ...
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... trans-lutein in the peel registered the highest contents during the initial develop- mental stages and went descending thereafter, meanwhile it was found in detectable amounts in the flesh only at stages 1 and 2, yet substantially lower compared to the peel. Citranaxanthin and phytoene have also been identi- fied, although they were not quantified (Table 1). ...
Citations
... The color of the fruit distinguishes red and white-fleshed loquat varieties, with red-fleshed varieties exhibiting a reddish hue due to their high carotenoid content, while white-fleshed loquats have a lower carotenoid content [4,5]. The ripening of loquat fruit is marked by carotenoid biosynthesis-induced color transformation and changes in fruit firmness [6,7]. Carotenoids exhibit diverse functions across animals, plants, and microorganisms. ...
The yellow-fleshed loquat is abundant in carotenoids, which determine the fruit’s color, provide vitamin A, and offer anti-inflammatory and anti-cancer health benefits. In this research, the impact of abscisic acid (ABA), a plant hormone, on carotenoid metabolism and flesh pigmentation in ripening loquat fruits was determined. Results revealed that ABA treatment enhanced the overall content of carotenoids in loquat fruit, including major components like β-cryptoxanthin, lutein, and β-carotene, linked to the upregulation of most genes in the carotenoid biosynthesis pathway. Furthermore, a transcription factor, EjWRKY6, whose expression was induced by ABA, was identified and was thought to play a role in ABA-induced carotenoid acceleration. Transient overexpression of EjWRKY6 in Nicotiana benthamiana and stable genetic transformation in Nicotiana tabacum with EjWRKY6 indicated that both carotenoid production and genes related to carotenoid biosynthesis could be upregulated in transgenic plants. A dual-luciferase assay proposed a probable transcriptional control between EjWRKY6 and promoters of genes associated with carotenoid production. To sum up, pre-harvest ABA application could lead to carotenoid biosynthesis in loquat fruit through the EjWRKY6-induced carotenoid biosynthesis pathway.
... The function of LCYB is to cyclase the lycopene [29]. There is a direct link between the synthesis and accumulation of β-cryptoxanthin and the expression of BCH [30]. D27 β-carotene isomerase is highly specific to the C9-C10 double bond and catalyzes the conversion of all-trans into 9-cis-β-carotene [31]. ...
Camellia nitidissima is famous for its golden flowers. Its flowers are rich in secondary metabolites, and they have ornamental, medicinal, and edible value. Pigment composition and regulation has been studied in the golden petals, but there has been little research on pigment composition or the molecular mechanism underlying yellow stamens in C. nitidissima. To explore the molecular mechanism of yellow stamen formation, three developmental stages (S0, S1, and S2) were used for transcriptome and pigment analyses. Pigment analysis showed that the flavonoid content increased sharply from the S0 to S1 stage and decreased from the S1 to S2 stage, and the carotenoid content increased sharply during yellow stamen formation (from the S1 to S2 stage). RNA-seq analysis showed that a total of 20,483 differentially expressed genes (DEGs) were identified. KEGG and heatmap analyses showed that flavonoid and carotenoid biosynthesis pathways were enriched, and we identified 14 structural genes involved in flavonoid biosynthesis and 13 genes involved in carotenoid biosynthesis and degradation. In addition, the expression of carotenoid- and flavonoid-related genes was consistent with carotenoid and flavonoid content. In addition, correlation network analysis indicated that the WARYK, MYB, bHLH, and AP2/ERF transcription factor families were screened for involvement in the biosynthesis of flavonoids and carotenoids. In this study, we describe the pathway associated with color formation in the stamens of C. nitidissima.
... Moreover, the levels of α-carotene, β-carotene, and γ-carotene in chili peppers were correlated with the expression of LCYB and LCYE [12]. Research has highlighted a close correlation between the expression levels of the LCYB and LCYE genes and the content of trans-β-carotene in loquat fruit [30]. Zhou et al. analyzed the transcriptome and metabonomics and found that PSY, NCED1 and CCD4 were the key genes that informed the significant differences in carotenoid content in the apricot fruits of two cultivars [28]. ...
Carotenoids are important pigments in pepper fruits. The colors of each pepper are mainly determined by the composition and content of carotenoid. The ‘ZY’ variety, which has yellow fruit, is a natural mutant derived from a branch mutant of ‘ZR’ with different colors. ZY and ZR exhibit obvious differences in fruit color, but no other obvious differences in other traits. To investigate the main reasons for the formation of different colored pepper fruits, transcriptome and metabolome analyses were performed in three developmental stages (S1–S3) in two cultivars. The results revealed that these structural genes (PSY1, CRTISO, CCD1, CYP97C1, VDE1, CCS, NCED1 and NCED2) related to carotenoid biosynthesis were expressed differentially in the two cultivars. Capsanthin and capsorubin mainly accumulated in ZR and were almost non-existent in ZY. S2 is the fruit color-changing stage; this may be a critical period for the development of different color formation of ZY and ZR. A combination of transcriptome and metabolome analyses indicated that CCS, NCED2, AAO4, VDE1 and CYP97C1 genes were key to the differences in the total carotenoid content. These new insights into pepper fruit coloration may help to improve fruit breeding strategies.
... Sadana (1949) first revealed that b-carotene is the predominant pigment positively associated with fruit color of cultivated loquats. Then, 23 (Zhou et al., 2007), 25 (De Faria et al., 2009) and 30 (Hadjipieri et al., 2017) carotenoid compounds were identified via HPLC and HPLC-PDA-MS/MS. With violaxanthin palmitate, rubixanthin laurate, bcryptoxanthin laurate, b-cryptoxanthin, rubixanthin palmitate, bcryptoxanthin palmitate, lutein dilaurate, b-cryptoxanthin oleate, violaxanthin-myristate-caprate, b-cryptoxanthin myristate and lutein dipalmitate newly identified here, we identified the most carotenoid constituents (38 molecules) from loquat fruit via UPLC-MS/MS (Figures 2A, B, Table S1). ...
... As a key enzyme in the carotenoid biosynthesis pathway, ZDS can catalyze zcarotene to form lycopene. EjZDS was also up-regulated during fruit pigmentation of the orange-colored 'Obusa' loquat (Hadjipieri et al., 2017). Overexpression of apple MdZDS notably improved both carotenoid biosynthesis and saline-alkali stress tolerance in transgenic plants . ...
Eriobotrya is an evergreen fruit tree native to South-West China and adjacent countries. There are more than 26 loquat species known in this genus, while E. japonica is the only species yet domesticated to produce fresh fruits from late spring to early summer. Fruits of cultivated loquat are usually orange colored, in contrast to the red color of fruits of wild E. henryi (EH). However, the mechanisms of fruit pigment formation during loquat evolution are yet to be elucidated. To understand these, targeted carotenoid and anthocyanin metabolomics as well as transcriptomics analyses were carried out in this study. The results showed that β-carotene, violaxanthin palmitate and rubixanthin laurate, totally accounted for over 60% of the colored carotenoids, were the major carotenoids in peel of the orange colored ‘Jiefangzhong’ (JFZ) fruits. Total carotenoids content in JFZ is about 10 times to that of EH, and the expression levels of PSY, ZDS and ZEP in JFZ were 10.69 to 23.26 folds to that in EH at ripen stage. Cyanidin-3-O-galactoside and pelargonidin-3-O-galactoside were the predominant anthocyanins enriched in EH peel. On the contrary, both of them were almost undetectable in JFZ, and the transcript levels of F3H, F3’H, ANS, CHS and CHI in EH were 4.39 to 73.12 folds higher than that in JFZ during fruit pigmentation. In summary, abundant carotenoid deposition in JFZ peel is well correlated with the strong expression of PSY, ZDS and ZEP, while the accumulation of anthocyanin metabolites in EH peel is tightly associated with the notably upregulated expressions of F3H, F3’H, ANS, CHS and CHI. This study was the first to demonstrate the metabolic background of how fruit pigmentations evolved from wild to cultivated loquat species, and provided gene targets for further breeding of more colorful loquat fruits via manipulation of carotenoids and anthocyanin biosynthesis.
... This response indicates that red wild-type tomato is deficient in the CrtL-e gene (Ronen et al., 1999). Gene regulation of carotenoid biosynthesis has been studied intensively in many fruits, such as apricot , citrus (Lu et al., 2021), loquat (Hadjipieri et al., 2017), mango (Ma et al., 2018), papaya (Zhou et al., 2019), peach (Cao et al., 2017), persimmon (Qi et al., 2019), and tomato (Lu et al., 2015). There are only a few studies, however, into gene regulation of carotenoid biosynthesis in durian fruit. ...
Durian (Durio zibethinus Murray) is an important economic crop in Southeast Asian countries. Analysis of the composition of durian pulp indicates that it has high nutraceutical value that is related to the presence of bioactive antioxidant compounds, with carotenoids being one of the important constituents. The major carotenoids are β‐ and α‐carotene, with the minor carotenoids being lutein and zeaxanthin. Carotenoid biosynthesis in durian pulp involves the coordinated expression of many genes with at least nine genes encoding enzymes having been identified. Carotenoid accumulation increases as fruit maturity advances and is highest in ripe fruit. The concentration of carotenoids in ripe durian pulp varies among cultivars. Ripening‐induced carotenoid accumulation is regulated by endogenous ethylene that controls the expression of key genes.
... In contrast, a* showed an increase as the fruit ripened. It is known that color is related to ripening due to the accumulation of pigmentation and the variation in sugar and acid content in the fruit; (Samaniego et al., 2020); as mentioned, lightness varied between 40.09 and 61.3, similar to that reported by Hadjipieri et al. (2017) for loquat (52.12-74.32). The tendency to decrease brightness, according to Samaniego et al. (2020), expresses the intensity of fruit color as the fruit ripens. ...
... The tendency to decrease brightness, according to Samaniego et al. (2020), expresses the intensity of fruit color as the fruit ripens. The chromatic coordinate a* ranged from 1.3 to 13.2, and b* ranged from 9.2 to 29.1 with a certain tendency to decrease in the three states (Table 1), which are values approximating those obtained in loquat in the study of Hadjipieri et al. (2017). These coordinates indicate the variation of red or yellow color in the fruit, related to the accumulation of anthocyanins (Samaniego et al., 2020). ...
Recently, there has been a growing interest in bioactive compounds metabolized by plants, which are an important nutritional source for the human diet and are found in almost all vegetables and fruits. The objective was to evaluate the changes of bioactive compounds during the ripening of loquat (Eriobotrya japonica) fruit. For this purpose, fruits were collected at three different stages of ripening from three different production sites in the Amazon region, located in northeastern Peru. Color, total phenolic content (Folin-Ciocalteu method), antioxidant activity (DPPH free radical method) and total flavonoids (colorimetric assay) were determined for all samples. Data were subjected to analysis of variance and means were compared by Tukey's test (p ≤ 0.05). Color and bioactive compounds depend on the stage of ripening and, to a lesser extent, on the origin of the fruit. Ripe fruits have a higher content of phenolic compounds and flavonoids (up to five times higher) that can be used in the food and pharmaceutical industry.
... The highest total phenolic contents (TPC) have been reported in 'Mizauto' fruit as compared to other cultivars [4]. Loquat fruit ripening is a complex process of different physiological and metabolic changes, mainly including: ethylene biosynthesis and respiratory modifications [5], colour modifications due to carotenoid biosynthesis [6], sugar and acid metabolism contributing to the variation in fruit sensory attributes [7] and changes in the lignin, polysaccharide, and pectin contents during fruit ripening resulting in cell wall modifications and fruit firmness changes during ripening [8]. These changes are associated with complex transcriptional elucidations and interlinked metabolic changes in loquat fruit. ...
Loquat (Eriobotrya japonica Lindl.) fruit is a rich source of carotenoids, flavonoids, phenolics, sugars, and organic acids. Although it is classified as a non-climacteric fruit, susceptibility to mechanical and physical bruising causes its rapid deterioration by moisture loss and postharvest decay caused by pathogens. Anthracnose, canker, and purple spot are the most prevalent postharvest diseases of loquat fruit. Cold storage has been used for quality management of loquat fruit, but the susceptibility of some cultivars to chilling injury (CI) consequently leads to browning and other disorders. Various techniques, including cold storage, controlled atmosphere storage, hypobaric storage, modified atmosphere packaging, low-temperature conditioning, heat treatment, edible coatings, and postharvest chemical application, have been tested to extend shelf life, mitigate chilling injury, and quality preservation. This review comprehensively focuses on the recent advances in the postharvest physiology and technology of loquat fruit, such as harvest maturity, fruit ripening physiology, postharvest storage techniques, and physiological disorders and diseases.
... In recent years, an increasing number of studies have explained the biosynthesis and regulatory mechanisms of plant metabolites by integrating targeted metabolomes and transcriptomes [38,39]. The discovery of synthetases, transcription factors, and regulatory mechanisms of B vitamin metabolism further enriched our understanding of this field [6]. ...
Areca catechu is well known as a medicinal plant that has high nutritional and medicinal benefits. However, the metabolism and regulatory mechanism of B vitamins during areca nut development remain largely unclear. In this study, we obtained the metabolite profiles of six B vitamins during different areca nut developmental stages by targeted metabolomics. Furthermore, we obtained a panoramic expression profile of genes related to the biosynthetic pathway of B vitamins in areca nuts at different developmental stages using RNA-seq. In total, 88 structural genes related to B vitamin biosynthesis were identified. Furthermore, the integrated analysis of B vitamin metabolism data and RNA-seq data showed the key transcription factors regulating thiamine and riboflavin accumulation in areca nuts, including AcbZIP21, AcMYB84, and AcARF32. These results lay the foundation for understanding metabolite accumulation and the molecular regulatory mechanisms of B vitamins in A. catechu nut.
... We found that the compositions, contents, and proportions of carotenoids differed among the three sweetpotato varieties during tuberous root development (Fig. 3, Additional file 1: Fig. S1). These differences imply that carotenoid metabolism in sweetpotato tuberous roots may be based on the genetics of the varieties, as well as the regulatory mechanisms of tuberous root development and external environmental stimuli, similar to what have been reported in other plant species [35][36][37]. The contents of dry matter and starch increase during sweetpotato tuberous root development. ...
Background
Plant carotenoids are essential for human health, having wide uses in dietary supplements, food colorants, animal feed additives, and cosmetics. With the increasing demand for natural carotenoids, plant carotenoids have gained great interest in both academic and industry research worldwide. Orange-fleshed sweetpotato (Ipomoea batatas) enriched with carotenoids is an ideal feedstock for producing natural carotenoids. However, limited information is available regarding the molecular mechanism responsible for carotenoid metabolism in sweetpotato tuberous roots.
Results
In this study, metabolic profiling of carotenoids and gene expression analysis were conducted at six tuberous root developmental stages of three sweetpotato varieties with different flesh colors. The correlations between the expression of carotenoid metabolic genes and carotenoid levels suggested that the carotenoid cleavage dioxygenase 4 (IbCCD4) and 9-cis-epoxycarotenoid cleavage dioxygenases 3 (IbNCED3) play important roles in the regulation of carotenoid contents in sweetpotato. Transgenic experiments confirmed that the total carotenoid content decreased in the tuberous roots of IbCCD4-overexpressing sweetpotato. In addition, IbCCD4 may be regulated by two stress-related transcription factors, IbWRKY20 and IbCBF2, implying that the carotenoid accumulation in sweeetpotato is possibly fine-tuned in responses to stress signals.
Conclusions
A set of key genes were revealed to be responsible for carotenoid accumulation in sweetpotato, with IbCCD4 acts as a crucial player. Our findings provided new insights into carotenoid metabolism in sweetpotato tuberous roots and insinuated IbCCD4 to be a target gene in the development of new sweetpotato varieties with high carotenoid production.
... Carotenoids synthesis begins with geranylgeranyl diphosphate (GGPP) which is formed by the condensation of isopentenyl pyrophosphate (IPP) and dimethylallyl diphosphate (DMAPP) [20]. GGPP is condensed into colorless phytoene by the activity of phytoene synthase (PSY) [21]. Then, the colorless phytoene forms multiple compounds, such as β-carotene, δ-carotene, zeaxanthin, lutein, violaxanthin and neoxanthin, through a series of reactions [22][23][24]. ...
... In the study of carotenoid metabolites, components of the carotenoids in most species are already well understood, such as in carrot [12], tomato [13][14][15], pepper [16,17] and orange cauliflower and orange heading Chinese cabbage [18,19]. Additionally, Hadjipieri et al. [21] also indicated that trans-lutein and trans-β-carotene were the major carotenoids in the peel of loquat fruit, and trans-β-cryptoxanthin, followed by trans-β-carotene and 5,8-epoxy-β-carotene, to be the most predominant carotenoids in the flesh of loquat fruit. Xu et al. [24] reported significant accumulation of antheraxanthin, zeaxanthin, neoxanthin and β-cryptoxanthin in orange zucchini. ...
Turnip (Brassica rapa ssp. rapa) is considered to be a highly nutritious and health-promoting vegetable crop, whose flesh color can be divided into yellow and white. It is widely accepted that yellow-fleshed turnips have higher nutritional value. However, reports about flesh color formation is lacking. Here, the white-fleshed inbred line, W21, and yellow-fleshed inbred line, W25, were profiled from the swollen root of the turnip at three developmental periods to elucidate the yellow color formation. Transcriptomics integrated with metabolomics analysis showed that the PSY gene was the key gene affecting the carotenoids formation in W25. The coding sequence of BrrPSY-W25 was 1278 bp and that of BrrPSY-W21 was 1275 bp, and BrrPSY was more highly expressed in swollen roots in W25 than in W21. Transient transgenic tobacco leaf over-expressing BrrPSY-W and BrrPSY-Y showed higher transcript levels and carotenoids contents. Results revealed that yellow turnip formation is due to high expression of the PSY gene rather than mutations in the PSY gene, indicating that a post-transcriptional regulatory mechanism may affect carotenoids formation. Results obtained in this study will be helpful for explaining the carotenoids accumulation of turnips.
