Characterization of BoaCRTISO Reveals Its Role in Carotenoid Biosynthesis in Chinese Kale.
Ontology highlight
ABSTRACT:
SUBMITTER: Jiang M
PROVIDER: S-EPMC8160315 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:Chinese kale (Brassica oleracea var. alboglabra) is rich in carotenoids, and neoxanthin is one of the most important carotenoids in Chinese kale. In this study, the function of the neoxanthin synthase gene (BoaNXS) in Chinese kale was investigated. BoaNXS, which had a 699-bp coding sequence, was cloned from the white flower cultivar of Chinese kale and was expressed in all developmental stages and organs of Chinese kale; its expression was highest in young seeds. The subcellular localization indicated that BoaNXS was localized in the chloroplast. BoaNXS-overexpressed plants were obtained via Agrobacterium-mediated transient overexpression methodology, and the gene overexpression efficiencies ranged from 2.10- to 4.24-fold. The color in the leaves of BoaNXS-overexpressed plants changed from green to yellow-green; the content of total and individual carotenoids, such as neoxanthin, violaxanthin, and lutein, was significantly increased, and the expression levels of most carotenoid biosynthetic genes were notably increased. These findings indicated that BoaNXS is of vital importance in carotenoid biosynthesis in Chinese kale and could be used as a candidate gene for enriching the carotenoid accumulation and color of Chinese kale and other Brassica vegetables.
| S-EPMC8393425 | biostudies-literature
Project description:Glucosinolates are Brassicaceae-specific secondary metabolites that act as crop protectants, flavor precursors, and cancer-prevention agents, which shows strong evidences of anticarcinogentic, antioxidant, and antimicrobial activities. MYB28, the R2R3-MYB28 transcription factor, directly activates genes involved in aliphatic glucosinolate biosynthesis. In this study, the MYB28 homology (BoaMYB28) was identified in Chinese kale (Brassica oleracea var. alboglabra Bailey). Analysis of the nucleotide sequence indicated that the cDNA of BoaMYB28 was 1257 bp with an ORF of 1020 bp. The deduced BoaMYB28 protein was a polypeptide of 339 amino acid with a putative molecular mass of 38 kDa and a pI of 6.87. Sequence homology and phylogenetic analysis showed that BoaMYB28 was most closely related to MYB28 homologs from the Brassicaceae family. The expression levels of BoaMYB28 varies across the tissues and developmental stages. BoaMYB28 transcript levels were higher in leaves and stems compared with those in cotyledons, flowers, and siliques. BoaMYB28 was expressed across all developmental leaf stages, with higher transcript accumulation in mature and inflorescence leaves. Over-expression and RNAi studies showed that BoaMYB28 retains the basic MYB28 gene function as a major transcriptional regulator of aliphatic glucosinolate pathway. The results indicated that over-expression and RNAi lines showed no visible difference on plant morphology. The contents of aliphatic glucosinolates and transcript levels of aliphatic glucosinolate biosynthesis genes increased in over-expression lines and decreased in RNAi lines. In over-expression lines, aliphatic glucosinolate contents were 1.5- to 3-fold higher than those in the wild-type, while expression levels of aliphatic glucosinolate biosynthesis genes were 1.5- to 4-fold higher than those in the wild-type. In contrast, the contents of aliphatic glucosinolates and transcript levels of aliphatic glucosinolate biosynthesis genes in RNAi lines were considerably lower than those in the wild-type. The results suggest that BoaMYB28 has the potential to alter the aliphatic glucosinolates contents in Chinese kale at the genetic level.
| S-EPMC5478679 | biostudies-literature
Project description:The carotenoid isomerase gene (BoaCRTISO) of Chinese kale was targeted and edited using the CRISPR/Cas9 system in the present study. The results showed a high mutation rate (81.25%), and 13 crtiso mutants were obtained. Only two types of mutations, insertions and replacements, were found. Both the total and individual carotenoid and chlorophyll concentrations of the biallelic and homozygous mutants were reduced, and the total levels declined by 11.89-36.33%. The color of the biallelic and homozygous mutants changed from green to yellow, likely reflecting a reduction in the color-masking effect of chlorophyll on carotenoids. The expression levels of most carotenoid and chlorophyll biosynthesis-related genes, including CRTISO, were notably lower in the mutants than in the WT plants. In addition, the functional differences between members of this gene family were discussed. In summary, these findings indicate that CRISPR/Cas9 is a promising technique for the quality improvement of Chinese kale and other Brassica vegetables.
| S-EPMC7527958 | biostudies-literature
Project description:Ultraviolet-A (UVA) (315-400 nm) is an essential environmental signal that regulates plant development and affects phytochemicals biosynthesis, including glucosinolate biosynthesis. The effects of different UVA (380 ± 10 nm, 40 μmol/m2/s) exposure durations, including 0 h/d (UV0), 6 h/d (UV6) and 12 h/d (UV12), on the growth and phytochemicals of Chinese kale (Brassica alboglabra) under white 250 μmol/m2/s LEDs were investigated. UVA exposure of different durations influenced the growth and phytochemicals biosynthesis of Chinese kale. Prolonging UVA irradiation throughout the growth cycle positively affected the growth and the development of Chinese kale, with evident increases in the dry weights of shoots and roots, plant height, stem diameter, specific leaf weight and flower budding rate. The application of UVA increased the soluble sugar content, whereas higher flavonoid content and antioxidant capacity (FRAP) and lower nitrate content were only observed in Chinese kale exposed to UV6 treatment. Besides, the qPCR assay showed that supplemental UVA-radiation exposure up-regulated the gene expressions of UVR8, transcription factors genes and genes related to the glucosinolate biosynthesis pathway, thereby promoting the accumulation of glucosinolates. Therefore, supplemental UVA-radiation exposure for 12 h/d was more conducive to plant growth, while supplemental UVA-radiation exposure for 6 h/d was better for phytochemical biosynthesis in Chinese kale in an artificial-light plant factory.
| S-EPMC9320135 | biostudies-literature
Project description:Chinese kale is a native vegetable in Southern China and the flowering stalk is the most commonly used edible part due to its high glucosinolate content and other nutritional qualities. The GTR protein played important roles in the glucosinolate transport process. In this study, three BocGTR1 genes were cloned from Chinese kale for the first time. Their gene structure, physicochemical properties, signal peptides, transmembrane structures, functional domains, second and third-order protein structures, and phylogenetic relationships were predicted. The expression levels of BocGTR1a and BocGTR1c were much higher than those of BocGTR1b in various tissues, especially in leaves and buds. In addition, the expression patterns of three genes were examined under various abiotic stresses or hormone treatment, including those induced by wounding, heat stress, methyl jasmonate, salicylic acid, salt, and MgCl₂ treatment. BocGTR1a and BocGTR1c were strongly induced by wounding and heat stress. The expression of BocGTR1a and BocGTR1c was significantly silenced in plants transformed by RNAi technology. Total glucosinolate content was significantly decreased in mature leaves and increased in roots of RNAi-transformed plants compared to wild-type plants. In addition, we found that BocGTR1a and BocGTR1c may participate in glucosinolate accumulation in different tissues with a selection for specific glucosinolates. These results indicated that BocGTR1a and BocGTR1c may be the key genes involved in the glucosinolate accumulation in different tissues of Chinese kale.
| S-EPMC6471314 | biostudies-literature
Project description:The car gene cluster of the ascomycete Fusarium fujikuroi encodes two enzymes responsible for torulene biosynthesis (CarRA and CarB), an opsin-like protein (CarO), and a putative carotenoid cleaving enzyme (CarX). It was presumed that CarX catalyzes the formation of the major carotenoid in F. fujikuroi, neurosporaxanthin, a cleavage product of torulene. However, targeted deletion of carX did not impede neurosporaxanthin biosynthesis. On the contrary, DeltacarX mutants showed a significant increase in the total carotenoid content, indicating an involvement of CarX in the regulation of the pathway. In this work, we investigated the enzymatic activity of CarX. The expression of the enzyme in beta-carotene-accumulating Escherichia coli cells led to the formation of the opsin chromophore retinal. The identity of the product was proven by high-performance liquid chromatography and gas chromatography-mass spectrometry. Subsequent in vitro assays with heterologously expressed and purified CarX confirmed its beta-carotene-cleaving activity and revealed its capability to produce retinal also from other substrates, such as gamma-carotene, torulene, and beta-apo-8'-carotenal. Our data indicate that the occurrence of at least one beta-ionone ring in the substrate is required for the cleavage reaction and that the cleavage site is determined by the distance to the beta-ionone ring. CarX represents the first retinal-synthesizing enzyme reported in the fungal kingdom so far. It seems likely that the formed retinal is involved in the regulation of the carotenoid biosynthetic pathway via a negative feedback mechanism.
| S-EPMC1865656 | biostudies-literature
Project description:Watercress (Nasturtium officinale R. Br.) is an important aquatic herb species belonging to the Brassicaceae family. It has various medicinal properties and has been utilized for the treatment of cancer and other diseases; however, currently available genomic information regarding this species is limited. Here, we performed the first comprehensive analysis of the carotenoid biosynthesis pathway (CBP) genes of N. officinale, which were identified from next-generation sequencing data. We identified and characterized 11 putative carotenoid pathway genes; among these, nine full and two partial open reading frames were determined. These genes were closely related to CBP genes of the other higher plants in the phylogenetic tree. Three-dimensional structure analysis and multiple alignments revealed several distinct conserved motifs, including aspartate or glutamate residues, carotene-binding motifs, and dinucleotide-binding motifs. Quantitative reverse transcription-polymerase chain reaction results showed that the CBP was expressed in a tissue-specific manner: expression levels of NoPSY, NoPDS, NoZDS-p, NoCrtISO, NoLCYE, NoCHXE-p, and NoCCD were highest in the flower, whereas NoLCYB, NoCHXB, NoZEP, and NoNCED were highest in the leaves. Stems, roots, and seeds did not show a significant change in the expression compared to the leaves and flowers. High-performance liquid chromatography analysis of the same organs showed the presence of seven distinct carotenoid compounds. The total carotenoid content was highest in the leaves followed by flowers, seeds, stems, and roots. Among the seven individual carotenoids, the levels of six carotenoids (i.e., 13-Z-β-carotene, 9-Z-β-carotene, E-β-carotene, lutein, violaxanthin, and β-cryptoxanthin) were highest in the leaves. The highest content was observed for lutein, followed by E-β-carotene, and 9-Z-β-carotene; these carotenoids were much higher in the leaves compared to the other organs. The results will be useful references for further molecular genetics and functional studies involving this species and other closely related species.
| S-EPMC8756599 | biostudies-literature
Project description:Vitamin D deficiency poses a major global health challenge, shaped by environmental and genetic determinants. In this genetic context, the recently discovered rs11542462 nonsense variant in the short-chain dehydrogenase/reductase 42E1 (SDR42E1) gene emerges as a critical regulator, though its biological implications remain largely unexplored. Our comprehensive bioinformatic assessments revealed pronounced expression of SDR42E1 in skin keratinocytes and the analogous HaCat human keratinocyte cell lines, prompting us to select the latter as an experimental model. Employing CRISPR/Cas9 technology and an integrated multi-omics approach, combining transcriptomics and proteomics, we explored SDR42E1's functions, focusing on its localization, role in lipid and steroid biosynthesis, and the consequent effects on vitamin D biosynthesis. We highlighted the significant cytoplasmic presence of the SDR42E1 protein, essential for lipid and steroid production. Our whole transcriptomics analysis of the SDR42E1 depleted model showed a notable disruption in the steroid biosynthesis pathway by 1.6-fold (P = 0.03). This disruption was accompanied by considerable alterations in genes involved in vitamin D synthesis, a finding supported by our proteomics data. Notably, SERPINB2 (P = 2.17E−103), EBP (P = 2.46E−13), and DHCR7 (P = 8.03E−09) showed significant elevations by ~2-3 fold, while ALPP (P < 2.2E−308), SLC7A5 (P = 1.96E−215), and CYP26A1 (P = 1.06E−08) decreased by ~1.5-3 fold. These alterations led to an accumulation of 7-dehydrocholesterol, preventing its conversion to vitamin D3, as evidenced by our drug enrichment analysis (P = 4.39E−06). This investigation unveils SDR42E1's pivotal role in vitamin D homeostasis and its broader impact on metabolic pathways. Our findings provide a foundation for future research and highlight the potential of precision medicine in addressing vitamin D deficiency, offering significant insights into the genetic regulation of vitamin D biosynthesis.
2024-08-22 | GSE262704 | GEO
Project description:Mushroom leaves (MLs) are malformed leaves that develop from the leaf veins in some of Chinese kale genotypes. To study the genetic model and molecular mechanism of ML development in Chinese kale, the F2 segregation population was constructed by two inbred lines, genotype Boc52 with ML and genotype Boc55 with normal leaves (NL). In the present study, we have identified for the first time that the development of mushroom leaves may be affected by the change of adaxial-abaxial polarity of leaves. Examination of the phenotypes of F1 and F2 segregation populations suggested that ML development is controlled by two dominant major genes inherited independently. BSA-seq analysis showed that a major quantitative trait locus (QTL) qML4.1 that controls ML development is located within 7.4 Mb on chromosome kC4. The candidate region was further narrowed to 255 kb by linkage analysis combined with insertion/deletion (InDel) markers, and 37 genes were predicted in this region. According to the expression and annotation analysis, a B3 domain-containing transcription factor NGA1-like gene, BocNGA1, was identified as a key candidate gene for controlling ML development in Chinese kale. Fifteen single nucleotide polymorphisms (SNPs) were found in coding sequences and 21 SNPs and 3 InDels found in the promoter sequences of BocNGA1 from the genotype Boc52 with ML. The expression levels of BocNGA1 in ML genotypes are significantly lower than in the NL genotypes, which suggests that BocNGA1 may act as a negative regulator for ML genesis in Chinese kale. This study provides a new foundation for Chinese kale breeding and for the study of the molecular mechanism of plant leaf differentiation.Supplementary informationThe online version contains supplementary material available at 10.1007/s11032-023-01364-6.
| S-EPMC10248679 | biostudies-literature
Project description:Tomato pericarp tissues at three different stages (green, breaker and ripening) of fruit development from both the above genotypes were collected using a sterile surgical blade and flash frozen in liquid nitrogen. High quality RNA was extracted from the frozen pericarp tissue using TRI Reagent (Ambion, INC. USA) and pooled at least from three independent samples and treated with DNase-I (QIAGEN GmbH, Germany). RNA cleanup was carried out using RNeasy Plant Mini Kit (QIAGEN GmbH, Germany) and 5ug of total RNA from each sample with three biological replications were reverse-transcribed to double stranded cDNA using the GeneChipR One-Cycle cDNA Synthesis Kit. The biotin-labelled cRNA was made using the GeneChipR IVT Labelling Kit (Affymetrix, CA, USA). 15 ug of cRNA samples was fragmented and was hybridized for 16 hours at 45C to the Affymetrix tomato GeneChip Array (Santa Clara, CA, USA). After washing and staining with R-phycoerythrin streptavidin in a Fluidics Station, using the GenechipR Fluidics Station 450, the arrays were scanned by the GenechipR 3000 Scanner. The chip images were scanned and extracted using default settings and the .CEL files were produced by using the Affymetrix GeneChip Operating Software (GCOS 1.2). Samples that passed through various quality checks (data not shown) were normalized with RMA algorithm using GeneSpring GX 10 (Agilent Technologies Inc., Santa Clara, CA, USA). Probes showing twofold up- or down-regulation (in Log2 transformed values) compared to corresponding control condition were taken only into consideration for further analysis
2011-01-04 | E-MEXP-2934 | biostudies-arrayexpress