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SUMMARY: The color of purple carrot taproots mainly depends on the anthocyanins sequestered in the vacuoles. Glutathione S‐transferases (GSTs) are key enzymes involved in anthocyanin transport. However, the precise mechanism of anthocyanin transport from the cytosolic surface of the endoplasmic reticulum (ER) to the vacuoles in carrots remains unclear. In this study, we conducted a comprehensive analysis of the carrot genome, leading to the identification of a total of 41 DcGST genes. Among these, DcGST1 emerged as a prominent candidate, displaying a strong positive correlation with anthocyanin pigmentation in carrot taproots. It was highly expressed in the purple taproot tissues of purple carrot cultivars, while it was virtually inactive in the non‐purple taproot tissues of purple and non‐purple carrot cultivars. DcGST1, a homolog of Arabidopsis thaliana TRANSPARENT TESTA 19 (TT19), belongs to the GSTF clade and plays a crucial role in anthocyanin transport. Using the CRISPR/Cas9 system, we successfully knocked out DcGST1 in the solid purple carrot cultivar 'Deep Purple' ('DPP'), resulting in carrots with orange taproots. Additionally, DcMYB7, an anthocyanin activator, binds to the DcGST1 promoter, activating its expression. Compared with the expression DcMYB7 alone, co‐expression of DcGST1 and DcMYB7 significantly increased anthocyanin accumulation in carrot calli. However, overexpression of DcGST1 in the two purple carrot cultivars did not change the anthocyanin accumulation pattern or significantly increase the anthocyanin content. These findings improve our understanding of anthocyanin transport mechanisms in plants, providing a molecular foundation for improving and enhancing carrot germplasm. Significance Statement: These findings improve our understanding of anthocyanin transport mechanisms in plants, providing a molecular foundation for improving and enhancing carrot germplasm. [ABSTRACT FROM AUTHOR]

The first domesticated carrots were thought to be purple carrots rich in anthocyanins. The anthocyanins biosynthesis in solid purple carrot taproot was regulated by DcMYB7 within P3 region containing a gene cluster of six DcMYBs. Here, we described a MYB gene within the same region, DcMYB11c, which was highly expressed in the purple pigmented petioles. Overexpression of DcMYB11c in 'Kurodagosun' (KRDG, orange taproot carrot with green petioles) and 'Qitouhuang' (QTHG, yellow taproot carrot with green petioles) resulted in deep purple phenotype in the whole carrot plants indicating anthocyanins accumulation. Knockout of DcMYB11c in 'Deep Purple' (DPPP, purple taproot carrot with purple petioles) through CRISPR/Cas9‐based genome editing resulted in pale purple phenotype due to the dramatic decrease of anthocyanins content. DcMYB11c could induce the expression of DcbHLH3 and anthocyanins biosynthesis genes to jointly promote anthocyanins biosynthesis. Yeast one‐hybrid assay (Y1H) and dual‐luciferase reporter assay (LUC) revealed that DcMYB11c bound to the promoters of DcUCGXT1 and DcSAT1 and directly activated the expression of DcUCGXT1 and DcSAT1 responsible for anthocyanins glycosylation and acylation, respectively. Three transposons were present in the carrot cultivars with purple petioles but not in the carrot cultivars with green petioles. We revealed the core factor, DcMYB11c, involved in anthocyanins pigmentation in carrot purple petioles. This study provides new insights into precise regulation mechanism underlying anthocyanins biosynthesis in carrot. The orchestrated regulation mechanism in carrot might be conserved across the plant kingdom and useful for other researchers working on anthocyanins accumulation in different tissues. Summary statement: Here, we described a MYB gene, DcMYB11c, which was highly expressed in the purple pigmented petioles. The orchestrated regulation mechanism in carrot might be conserved across the plant kingdom and useful for other researchers working on anthocyanins accumulation in different tissues. [ABSTRACT FROM AUTHOR]

The germination of carrot seeds is a complex regulatory process and the relationship between phytohormone and carrot seed germination is unclear. We performed morphological observation, transcriptome sequencing, phytohormone content determination, and gene expression profiles analysis. The results showed that the white hypocotyls of carrot seeds grew rapidly on the 3 d, and cotyledons unfold on the 8 d. A total of 745 differentially expressed genes (DEGs) related to plant hormone signal transduction were identified at different germination stages (3, 5, 8 d), and most of the genes were up-regulated obviously in the later stage. The content of GA3 reached its peak on the first day of germination, with a content of 7.12 ng/g FW, and showed a trend of first decreasing, then increasing and decreasing. And the trend of GA4 content change was similar to GA3 content from 2 d to 8 d. The contents of indole-3-acetic acid (IAA) and methyl jasmonate (MeJA) showed a gradual increasing trend, while the content of abscisic acid (ABA) gradually decreased. The results of RT-qPCR showed that DcGA3ox1 and DcAOC expressed decreased, while DcGA20ox2 and DcYUCCA increased in the whole process. These results provide potential new insights for the efficient production of carrots.. [ABSTRACT FROM AUTHOR]

Ascorbate is an antioxidant in plants. Its content is an important index to evaluate the nutritional quality of celery. In higher plant, ascorbate accumulation is effected by CO2 concentration. To study the effects of elevated CO2 (1000 µmol mol−1) on ascorbate accumulation in celery, ascorbate contents and the transcript levels of genes related to ascorbate metabolism in the leaves (leaf blade and petiole) were detected. The results of current study showed that the ascorbate and total ascorbate levels increased during all the treated stages in celery petioles, and they were 1.09–3.91 fold of the control. In leaf blades, the ascorbate contents exhibited a lower level (96.12% and 90.46% of the control) in treatments at 2 and 4 days, and then increased to 1.04 and 1.27 fold of the control in treatments at 6 and 8 days, respectively. Elevated CO2 altered the expression profiles of ascorbate-related genes and the activities of corresponding enzymes. Expression levels of the genes, AgPGI2, AgGMP, AgPMI, AgGalDH, AgGalLDH, GalUR, AgDHAR1, AgAPX1, and AgAO, in the leaf blades corresponded well to the change in ascorbate contents. At 6 days of treatment, the relative expression levels of AgPGI2, AgPMI, AgGGP1, AgGGP2, AgGalUR, AgMDHAR, AgDHAR1, and AgAPX1 in leaf blades increased, and were 4.41, 2.39, 4.65, 3.64, 3.11, 4.53, 4.59, and 3.86 times higher than that of the control. Increased ascorbate accumulation in celery leaves under elevated CO2 treatments might be also related to the change in SOD, POD, and CAT activities, as well as GSH content. These results will enhance our understanding on the effects of elevated CO2 on ascorbate accumulation and potential molecular mechanism regulating ascorbate metabolism in celery. [ABSTRACT FROM AUTHOR]

• Several transgenic A. thaliana and celery lines harboring AgZDS gene were obtained. • The roles of AgZDS gene in carotenoids biosyntheisis were verified. • Overexpression of AgZDS increased the accumulation of lutein and β-carotene. • Antioxidant ability and photosynthetic ability of transgenic plants were improved. ζ-Carotene desaturase (ZDS) is one of the key enzymes regulating carotenoids biosynthesis and accumulation. Celery transgenic efficiency is low and it is difficult to obtain transgenic plants. The study on ZDS was limited in celery. Here, the AgZDS gene was cloned from celery and overexpressed in Arabidopsis thaliana and celery to verify its function. The AgZDS has typical characteristic of ZDS protein and is highly conserved in higher plants. Phylogenetic analysis showed that AgZDS has the closest evolutionary relationship with ZDSs from Solanum lycopersicum , Capsicum annuum and Tagetes erecta. Overexpression of AgZDS gene in A. thaliana and celery resulted in increased accumulations of lutein and β-carotene and up-regulated the expression levels of the genes involved in carotenoids biosynthesis. The contents of lutein and β-carotene in two lines, AtL1 and AgL5, were the highest in transgenic A. thaliana and celery, respectively. The relative expression levels of 5 genes (AtPDS , AtZISO , AtZEP , AtNCED3 , and AtCCD4) were up-regulated compared to the wild type plants. The relative expression levels of most genes in carotenoids biosynthesis pathway, such as AgPDS , AgCRTISO1 , and AgZISO , were up-regulated in transgenic celery plants. The antioxidant capacity of A. thaliana and photosynthetic capacity of celery were also enhanced. This research is the first report on the function of structure gene related to carotenoid biosynthesis in transgenic celery plants. The findings in this study demonstrated the roles of AgZDS in regulating carotenoids metabolism of celery, which laid a potential foundation for quality improvement of celery. [ABSTRACT FROM AUTHOR]

Capsanthin and capsorubin are red κ-xanthophylls exclusively found in a handful of other plant species. Currently, capsanthin and capsorubin are only extracted from red pepper. Here, high purity production of capsanthin and capsorubin has been achieved in carrot taproot by synthetic metabolic engineering strategy. Expression of a capsanthin-capsorubin synthase gene (CaCCS) from pepper resulted in dominant production of capsanthin whereas expression of a LiCCS gene from tiger lily resulted in production of both capsanthin and capsorubin in carrot taproot. The highest content of capsanthin and capsorubin was obtained in LiC-1 carrot taproot hosting the LiCCS gene, 150.09 μg/g DW (dry weight). Co-expression of DcBCH1 with CCS could improve the purity of capsanthin and capsorubin by eliminating the non-target carotenoids (eg. α-carotene and β-carotene). The highest purity of capsanthin and capsorubin was obtained in BLiC-1 carrot taproot hosting DcBCH1+LiCCS genes, 91.10% of total carotenoids. The non-native pigments were esterified partially and stored in the globular chromoplast of carrot taproot. Our results demonstrated the possibility of employing carrot taproot as green factories for high purity production of capsanthin and capsorubin. The capsanthin/capsorubin carrot germplasms were also valuable materials for breeding colorful carrots cultivars., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Many organisms have complex pigmentation patterns. However, how these patterns are formed remains largely unknown. In wild carrot (Daucus carota subsp. carota), which is also known as Queen Anne's lace, one or several purple central flowers occur in white umbels. Here, we investigated the unique central flower pigmentation pattern in wild carrot umbels. Using wild and cultivated carrot (Daucus carota subsp. sativus L.) accessions, transcriptome analysis, protein interaction, stable transformation, and CRISPR/Cas9-mediated knockout, a anthocyanin-activating R2R3-myeloblastosis (MYB) gene, Purple Central Flower (DcPCF), was identified as the causal gene that triggers only central flowers to possess the purple phenotype. The expression of DcPCF was only detected in tiny central flowers. We propose that the transition from purple to nonpurple flowers in the center of the umbel occurred after three separate adverse events: insertion of transposons in the promoter region, premature termination of the coding sequence (caused by a C-T substitution in the open reading frame), and the emergence of unknown anthocyanin suppressors. These three events could have occurred either consecutively or independently. The intriguing purple central flower pattern and its underlying mechanism may provide evidence that it is a remnant of ancient conditions of the species, reflecting the original appearance of Umbelliferae (also called Apiaceae) when a single flower was present., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)