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Cloning and Characterization of Three Sugar Metabolism Genes (LBGAE, LBGALA, and LBMS) Regulated in Response to Elevated CO₂ in Goji Berry (Lycium barbarum L.).

ABSTRACT: The composition and content of sugar play a pivotal role in goji berry (Lycium barbarum L.) fruits, determining fruit quality. Long-term exposure of goji berry to elevated CO₂ (eCO₂) was frequently demonstrated to reduce sugar content and secondary metabolites. In order to understand the regulatory mechanisms and improve the quality of fruit in the changing climate, it is essential to characterize sugar metabolism genes that respond to eCO₂. The objectives of this study were to clone full-length cDNA of three sugar metabolism genes-LBGAE (Lycium barbarum UDP-glucuronate 4-epimerase), LBGALA (Lycium barbarum alpha-galactosidase), and LBMS (Lycium barbarum malate synthase)-that were previously identified responding to eCO₂, and to analyze sequence characteristics and expression regulation patterns. Sugar metabolism enzymes regulated by these genes were also estimated along with various carbohydrates from goji berry fruits grown under ambient (400 ?mol mol^-1) and elevated (700 ?mol mol^-1) CO₂ for 90 and 120 days. Homology-based sequence analysis revealed that the protein-contained functional domains are similar to sugar transport regulation and had a high sequence homology with other Solanaceae species. The sucrose metabolism-related enzyme's activity varied significantly from ambient to eCO₂ in 90-day and 120-day samples along with sugars. This study provides fundamental information on sugar metabolism genes to eCO₂ in goji berry to enhance fruit quality to climate change.

SUBMITTER: Ma Y

PROVIDER: S-EPMC7914792 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The traditional Chinese medicinal plants Lycium barbarum L. and L. ruthenicum Murr. are valued for the abundance of bioactive carotenoids and anthocyanins in their fruits, respectively. However, the cellular and molecular mechanisms contributing to their species-specific bioactive profiles remain poorly understood.In this study, the red fruit (RF) of L. barbarum was found to accumulate high levels of carotenoids (primarily zeaxanthin), while they were undetectable in the black fruit (BF) of L. ruthenicum. Cytological and gene transcriptional analyses revealed that the chromoplast differentiation that occurs in the chloroplast during fruit ripening only occurs in RF, indicating that the lack of chromoplast biogenesis in BF leads to no sink for carotenoid storage and the failure to synthesize carotenoids. Similar enzyme activities of phytoene synthase 1 (PSY1), chromoplast-specific lycopene ?-cyclase (CYC-B) and ?-carotene hydroxylase 2 (CRTR-B2) were observed in both L. ruthenicum and L. barbarum, suggesting that the undetectable carotenoid levels in BF were not due to the inactivation of carotenoid biosynthetic enzymes. The transcript levels of the carotenoid biosynthetic genes, particularly PSY1, phytoene desaturase (PDS), ?-carotene desaturase (ZDS), CYC-B and CRTR-B2, were greatly increased during RF ripening, indicating increased zeaxanthin biosynthesis. Additionally, carotenoid cleavage dioxygenase 4 (CCD4) was expressed at much higher levels in BF than in RF, suggesting continuous carotenoid degradation in BF.The failure of the chromoplast development in BF causes low carotenoid biosynthesis levels and continuous carotenoid degradation, which ultimately leads to undetectable carotenoid levels in ripe BF. In contrast, the successful chromoplast biogenesis in RF furnishes the sink necessary for carotenoid storage. Based on this observation, the abundant zeaxanthin accumulation in RF is primarily determined via both the large carotenoid biosynthesis levels and the lack of carotenoid degradation, which are regulated at the transcriptional level.

Project description:Local translocation of small RNAs between cells is proved. Long distance translocation between rootstock and scion is also well documented in the homo-grafting system, but the process in distant-grafting is widely unexplored where rootstock and scion belonging to different genera. Micro RNAs are a class of small, endogenous, noncoding, gene silencing RNAs that regulate target genes of a wide range of important biological pathways in plants. In this study, tomato was grafted onto goji (Lycium chinense Mill.) to reveal the insight of miRNAs regulation and expression patterns within a distant-grafting system. Goji is an important traditional Chinese medicinal plant with enriched phytochemicals. Illumina sequencing technology has identified 68 evolutionary known miRNAs of 37 miRNA families. Moreover, 168 putative novel miRNAs were also identified. Compared with control tomato, 43 (11 known and 32 novels) and 163 (33 known and 130 novels) miRNAs were expressed significantly different in shoot and fruit of grafted tomato, respectively. The fruiting stage was identified as the most responsive in the distant-grafting approach and 123 miRNAs were found as up-regulating in the grafted fruit which is remarkably higher compare to the grafted shoot tip (28). Potential targets of differentially expressed miRNAs were found to be involved in diverse metabolic and regulatory pathways. ADP binding activities, molybdopterin synthase complex and RNA helicase activity were found as enriched terms in GO (Gene Ontology) analysis. Additionally, "metabolic pathways" was revealed as the most significant pathway in KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis. The information of the small RNA transcriptomes that are obtained from this study might be the first miRNAs elucidation for a distant-grafting system, particularly between goji and tomato. The results from this study will provide the insights into the molecular aspects of miRNA-mediated regulation in the medicinal plant goji, and in grafted tomato. Noteworthy, it would provide a basis how miRNA signals could exchange between rootstock and scion, and the relevance to diverse biological processes.

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Cloning and Characterization of Three Sugar Metabolism Genes (LBGAE, LBGALA, and LBMS) Regulated in Response to Elevated CO₂ in Goji Berry (Lycium barbarum L.).

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Cloning and Characterization of Three Sugar Metabolism Genes (LBGAE, LBGALA, and LBMS) Regulated in Response to Elevated CO2 in Goji Berry (Lycium barbarum L.).

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OmicsDI is part of the ELIXIR infrastructure

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Cloning and Characterization of Three Sugar Metabolism Genes (LBGAE, LBGALA, and LBMS) Regulated in Response to Elevated CO₂ in Goji Berry (Lycium barbarum L.).