scholarly journals Isolation and comparative proteomic analysis of mitochondria from the pulp of ripening citrus fruit

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Xin Li ◽  
Yingfang Chai ◽  
Hongbin Yang ◽  
Zhen Tian ◽  
Chengyang Li ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Tricarboxylic Acid Cycle ◽  
Gradient Centrifugation ◽  
Citrus Fruit ◽  
Label Free ◽  
High Coverage ◽  
Primary And Secondary Metabolites ◽  
Percoll Density Gradient Centrifugation ◽  
Proteomic Approach ◽  
Mitochondrial Functions

AbstractMitochondria are crucial for the production of primary and secondary metabolites, which largely determine the quality of fruit. However, a method for isolating high-quality mitochondria is currently not available in citrus fruit, preventing high-throughput characterization of mitochondrial functions. Here, based on differential and discontinuous Percoll density gradient centrifugation, we devised a universal protocol for isolating mitochondria from the pulp of four major citrus species, including satsuma mandarin, ponkan mandarin, sweet orange, and pummelo. Western blot analysis and microscopy confirmed the high purity and intactness of the isolated mitochondria. By using this protocol coupled with a label-free proteomic approach, a total of 3353 nonredundant proteins were identified. Comparison of the four mitochondrial proteomes revealed that the proteins commonly detected in all proteomes participate in several typical metabolic pathways (such as tricarboxylic acid cycle, pyruvate metabolism, and oxidative phosphorylation) and pathways closely related to fruit quality (such as γ-aminobutyric acid (GABA) shunt, ascorbate metabolism, and biosynthesis of secondary metabolites). In addition, differentially abundant proteins (DAPs) between different types of species were also identified; these were found to be mainly involved in fatty acid and amino acid metabolism and were further confirmed to be localized to the mitochondria by subcellular localization analysis. In summary, the proposed protocol for the isolation of highly pure mitochondria from different citrus fruits may be used to obtain high-coverage mitochondrial proteomes, which can help to establish the association between mitochondrial metabolism and fruit storability or quality characteristics of different species and lay the foundation for discovering novel functions of mitochondria in plants.

scholarly journals Unraveling Arbuscular Mycorrhiza-Induced Changes in Plant Primary and Secondary Metabolome

Metabolites ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 335 ◽  
Author(s):  
Sukhmanpreet Kaur ◽  
Vidya Suseela
Keyword(s):  
Secondary Metabolites ◽  
Mycorrhizal Fungi ◽  
Tricarboxylic Acid Cycle ◽  
Abiotic Stress Tolerance ◽  
Plant Performance ◽  
Growth And Yield ◽  
Primary Metabolism ◽  
Symbiotic Association ◽  
Phytochemical Content ◽  
Primary And Secondary Metabolites

Arbuscular mycorrhizal fungi (AMF) is among the most ubiquitous plant mutualists that enhance plant growth and yield by facilitating the uptake of phosphorus and water. The countless interactions that occur in the rhizosphere between plants and its AMF symbionts are mediated through the plant and fungal metabolites that ensure partner recognition, colonization, and establishment of the symbiotic association. The colonization and establishment of AMF reprogram the metabolic pathways of plants, resulting in changes in the primary and secondary metabolites, which is the focus of this review. During initial colonization, plant–AMF interaction is facilitated through the regulation of signaling and carotenoid pathways. After the establishment, the AMF symbiotic association influences the primary metabolism of the plant, thus facilitating the sharing of photosynthates with the AMF. The carbon supply to AMF leads to the transport of a significant amount of sugars to the roots, and also alters the tricarboxylic acid cycle. Apart from the nutrient exchange, the AMF imparts abiotic stress tolerance in host plants by increasing the abundance of several primary metabolites. Although AMF initially suppresses the defense response of the host, it later primes the host for better defense against biotic and abiotic stresses by reprogramming the biosynthesis of secondary metabolites. Additionally, the influence of AMF on signaling pathways translates to enhanced phytochemical content through the upregulation of the phenylpropanoid pathway, which improves the quality of the plant products. These phytometabolome changes induced by plant–AMF interaction depends on the identity of both plant and AMF species, which could contribute to the differential outcome of this symbiotic association. A better understanding of the phytochemical landscape shaped by plant–AMF interactions would enable us to harness this symbiotic association to enhance plant performance, particularly under non-optimal growing conditions.

scholarly journals Integrative Analyses of Widely Targeted Metabolic Profiling and Transcriptome Data Reveals Molecular Insight into Metabolomic Variations during Apple (Malus domestica) Fruit Development and Ripening

2020 ◽  
Vol 21 (13) ◽  
pp. 4797 ◽  
Author(s):  
Jidi Xu ◽  
Jinjiao Yan ◽  
Wenjie Li ◽  
Qianying Wang ◽  
Caixia Wang ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Fruit Quality ◽  
Fruit Development ◽  
Large Scale ◽  
Tricarboxylic Acid Cycle ◽  
Flavonoid Pathway ◽  
Primary And Secondary Metabolites ◽  
Development Stages ◽  
Fruit Development And Ripening ◽  
Fruit Stage

The apple is a favorite fruit for human diet and is one of the most important commercial fruit crops around the world. Investigating metabolic variations during fruit development can provide a better understanding on the formation of fruit quality. The present study applied a widely targeted LC-MS-based metabolomics approach with large-scale detection, identification and quantification to investigate the widespread metabolic changes during “Pinova” apple development and ripening. A total of 462 primary and secondary metabolites were simultaneously detected, and their changes along with the four fruit-development stages were further investigated. The results indicated that most of the sugars presented increasing accumulation levels while organic acid, including Tricarboxylic acid cycle (TCA) intermediates, showed a distinct decreasing trend across the four fruit-development stages. A total of 207 secondary metabolites consisted of 104 flavonoids and 103 other secondary metabolites. Many flavonoids maintained relatively high levels in the early fruit stage and then rapidly decreased their levels at the following developmental stages. Further correlation analyses of each metabolite–metabolite pair highlighted the cross talk between the primary and secondary metabolisms across fruit development and ripening, indicating the significant negative correlations between sugars and secondary metabolites. Moreover, transcriptome analysis provided the molecular basis for metabolic variations during fruit development. The results showed that most differentially expressed genes (DEGs) involved in the TCA cycle were upregulated from the early fruit stage to the preripening stage. The extensive downregulation of controlling genes involved in the flavonoid pathway is probably responsible for the rapid decrease of flavonoid content at the early fruit stage. These data provide a global view of the apple metabolome and a comprehensive analysis on metabolomic variations during fruit development, providing a broader and better understanding on the molecular and metabolic basis of important fruit quality traits in commercial apples.

scholarly journals Biosynthesis of Natural Products

2021 ◽  
Author(s):  
Stella O. Bruce ◽  
Felix A. Onyegbule
Keyword(s):  
Natural Products ◽  
Secondary Metabolites ◽  
Cultural History ◽  
Shikimic Acid ◽  
Tricarboxylic Acid Cycle ◽  
Primary And Secondary Metabolites ◽  
Methylerythritol Phosphate ◽  
Acid Pathway ◽  
Living Organisms ◽  
Nitrogen Containing Compounds

Natural products are in the form of primary and secondary metabolites and are isolated chemical compounds or substances from living organisms. Terpenes, Phenolic compounds, and Nitrogen-containing compounds are secondary metabolites. The biosyntheses of secondary metabolites are derived from primary metabolism pathways, which consist of a tricarboxylic acid cycle (TCA), methylerythritol phosphate pathway (MEP), mevalonic and shikimic acid pathway. This chapter provides an overview of the diversity of secondary metabolites in plants, their multiple biological functions, and multi-faceted cultural history.

Comparison Among Five Eucalyptus Species Based on Their Leaf Contents of Some Primary and Secondary Metabolites

2019 ◽  
Vol 20 (7) ◽  
pp. 573-587
Author(s):  
Alyaa Nasr ◽  
Tehmina Saleem Khan ◽  
Shi-Ping Huang ◽  
Bin Wen ◽  
Jian-Wen Shao ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Protocatechuic Acid ◽  
Antioxidant Activities ◽  
Reducing Power ◽  
Renewable Resource ◽  
Total Phenolic ◽  
Eucalyptus Species ◽  
Primary And Secondary Metabolites ◽  
Acetone Water ◽  
Phenolic Constituents

Background: Eucalyptus belongs to the Myrtaceae family. It is the most planted hardwood forest crop worldwide, representing a global renewable resource of fiber, pharmaceuticals and energy. Objective: To compare the five species, E. maidenii, E. robusta, E. citriodora, E. tereticornis and E. camaldulensis, seeking for the richest source of nutrients and pharmaceuticals. Methodology: Eucalyptus samples were subjected to some chemical determinations for both primary and secondary metabolites to verify their nutritional and pharmaceutical importance related to different extracts. GC-MS analysis was applied to detect the presence of some individual phenolic constituents in their leaves. Results: E. robusta recorded the maximum contents of carbohydrates (40.07%) and protein (31.91%). While E. camaldulensis contained the highest contents of total phenolic compounds (46.56 mg/g), tannins (40.01 mg/g) and antioxidant activities assayed by the phosphomolybednum method (57.60 mg/g), followed by E. citridora. However, E. tereticornis exhibited the highest reducing power ability (151.23 mg/g). The GC-MS highlighted 20 phenolic constituents and antioxidants which varied in their abundance in Eucalyptus leaves, 8 individual phenolics (hydroquinone, hesperitin, pyrogallol, resorcinol, protocatechuic acid, naringenin, chlorogenic acid and catechin) were maximally recorded with E. camaldulensis and secondly, with E. citridora in case of at least 5 components. Nevertheless, gallic and quinic acids were more abundant in the leaves of E. tereticornis, which may explain its high corresponding reducing powers. Conclusion: Acetone-water combination has enhanced phenolics extraction from Eucalyptus tissues. This is the first report aiming to compare between the aforementioned Eucalyptus species highlighting either their nutritional or medicinal importance.

Flow cytometric identification and isolation of hypertrophic type II pneumocytes after partial pneumonectomy

1989 ◽  
Vol 257 (3) ◽  
pp. C528-C536 ◽  
Author(s):  
B. D. Uhal ◽  
S. R. Rannels ◽  
D. E. Rannels
Keyword(s):  
Gradient Centrifugation ◽  
Type Ii Cells ◽  
Type Ii ◽  
Acridine Orange Staining ◽  
Flow Cytometric ◽  
Percoll Density Gradient Centrifugation ◽  
Type Ii Pneumocytes ◽  
The Mean ◽  
Cellular Dna ◽  
The Right

Type II pneumocytes were isolated by either Percoll density gradient centrifugation or by immunoglobulin G (IgG) panning from the lungs of normal rats and the right lung of rats subjected to left pneumonectomy. Cells were studied at 7- (pnx-7) and 15- (pnx-15) days postoperative, times during and after, respectively, rapid compensatory growth of the right lung. Acridine orange staining permitted resolution of type II cells from contaminants on the basis of high red fluorescence (greater than 590 nm). Simultaneous measurement of forward-angle light scatter (FALS) suggested a shift of pnx-7 cells toward greater size, which was reversed in pnx-15 cells. By Percoll gradient isolation, approximately 15% of pnx-7 cells analyzed were above the mean FALS of control cells. In contrast, approximately 30% of the pnx-7 cells isolated by IgG panning were above the mean FALS of corresponding control cells. Biochemical analyses of pnx-7 cells separated by cell sorting into "high FALS" and "low FALS" subgroups revealed that high FALS type II cells contained 50% more protein (P less than 0.05) and 140% more RNA (P less than 0.01) than low FALS cells, with no significant change in cellular DNA content. These data are consistent with previous studies of type II cells isolated from the lungs of pneumonectomized animals and confirm the presence of hypertrophic cells in these preparations. They provide a foundation from which to design further flow cytometric studies of the role of hypertrophic type II pneumocytes in compensatory lung growth.

scholarly journals Profiles of Secondary Metabolites (Phenolic Acids, Carotenoids, Anthocyanins, and Galantamine) and Primary Metabolites (Carbohydrates, Amino Acids, and Organic Acids) during Flower Development in Lycoris radiata

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 248
Author(s):  
Chang Ha Park ◽  
Hyeon Ji Yeo ◽  
Ye Jin Kim ◽  
Bao Van Nguyen ◽  
Ye Eun Park ◽  
...  
Keyword(s):  
Amino Acids ◽  
Secondary Metabolites ◽  
Organic Acids ◽  
Phenolic Acids ◽  
Flower Development ◽  
Developmental Stages ◽  
Tca Cycle ◽  
Primary And Secondary Metabolites ◽  
Hydrophilic Compounds ◽  
Tca Cycle Intermediates

This study aimed to elucidate the variations in primary and secondary metabolites during Lycorisradiata flower development using high performance liquid chromatography (HPLC) and gas chromatography time-of-flight mass spectrometry (GC-TOFMS). The result showed that seven carotenoids, seven phenolic acids, three anthocyanins, and galantamine were identified in the L. radiata flowers. Most secondary metabolite levels gradually decreased according to the flower developmental stages. A total of 51 metabolites, including amines, sugars, sugar intermediates, sugar alcohols, amino acids, organic acids, phenolic acids, and tricarboxylic acid (TCA) cycle intermediates, were identified and quantified using GC-TOFMS. Among the hydrophilic compounds, most amino acids increased during flower development; in contrast, TCA cycle intermediates and sugars decreased. In particular, glutamine, asparagine, glutamic acid, and aspartic acid, which represent the main inter- and intracellular nitrogen carriers, were positively correlated with the other amino acids and were negatively correlated with the TCA cycle intermediates. Furthermore, quantitation data of the 51 hydrophilic compounds were subjected to partial least-squares discriminant analyses (PLS-DA) to assess significant differences in the metabolites of L. radiata flowers from stages 1 to 4. Therefore, this study will serve as the foundation for a biochemical approach to understand both primary and secondary metabolism in L. radiata flower development.

scholarly journals Metabolomic analysis reveals the interaction of primary and secondary metabolism in white, pale green, and green pak choi (Brassica rapa subsp. chinensis)

2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Hyeon Ji Yeo ◽  
Seung-A Baek ◽  
Ramaraj Sathasivam ◽  
Jae Kwang Kim ◽  
Sang Un Park
Keyword(s):  
Secondary Metabolites ◽  
Brassica Rapa ◽  
High Performance ◽  
Reducing Power ◽  
Total Phenolic ◽  
Pak Choi ◽  
Primary And Secondary Metabolites ◽  
Chlorophyll Contents ◽  
Significant Difference ◽  
Pale Green

AbstractThis study aimed to comprehensively analyze primary and secondary metabolites of three different-colored (white, pale green, and green) pak choi cultivars (Brassica rapa subsp. chinensis) using gas chromatography attached with time-of-flight mass spectrometry (GC-TOFMS) and high-performance liquid chromatography (HPLC). In total, 53 primary metabolites were identified and subjected to partial least-squares discriminant analysis. The result revealed a significant difference in the primary and secondary metabolites between the three pak choi cultivars. In addition, 49 hydrophilic metabolites were detected in different cultivars. Total phenolic and glucosinolate contents were highest in the pale green and green cultivars, respectively, whereas total carotenoid and chlorophyll contents were highest in the white cultivar. Superoxide dismutase activity, 2,2-diphenyl-1-picrylhydraz scavenging, and reducing power were slightly increased in the white, pale green, and green cultivars, respectively. In addition, a negative correlation between pigments and phenylpropanoids was discovered by metabolite correlation analysis. This approach will provide useful information for the development of strategies to enhance the biosynthesis of phenolics, glucosinolates, carotenoids, and chlorophyll, and to improve antioxidant activity in pak choi cultivars. In addition, this study supports the use of HPLC and GC-TOFMS-based metabolite profiling to explore differences in pak choi cultivars.

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