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1.
Anticancer Drugs ; 34(10): 1146-1150, 2023 11 01.
Article in English | MEDLINE | ID: mdl-36728908

ABSTRACT

The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) dramatically improve the clinical outcomes of non-small cell lung cancer (NSCLC) patients harboring EGFR -sensitive mutations. Despite the remarkable efficacy of first-and second-generation EGFR TKIs, disease relapse is inevitable. EGFR T790M mutation is a primary contributor to the acquired resistance to first- and second-generation EGFR TKIs. Osimertinib, which is an irreversible third-generation EGFR TKI, was designed for EGFR -activating mutations as well as the EGFR T790M mutation in patients with advanced NSCLC and has demonstrated a convincing efficacy. However, acquired resistance to osimertinib after treatment inevitably occurs. The acquired resistance mechanisms to osimertinib are highly complicated and not fully understood, encompassing EGFR -dependent as well as EGFR -independent mechanisms. Treatment approaches for patients progressing from osimertinib have not been established. We present a case of a stage IV lung adenocarcinoma patient harboring EGFR L858R, acquired T790M after treatment with first-line gefitinib. She then acquired a new EML4-ALK gene fusion after treatment with osimertinib. A combination targeted therapy of osimertinib plus alectinib was initiated, with a progression-free survival of 5 months without any serious adverse reaction. After disease progression, EGFR C797S in cis was detected with a loss of the EML4-ALK fusion by targeted next-generation sequencing. Then therapy was changed to pemetrexed combined with bevacizumab plus camrelizumab, but no obvious effect was observed. The patient had achieved an overall survival of 31 months. As far as we know, this was the first reported case that an EGFR -mutant NSCLC patient-acquired ALK fusion mediating resistance to osimertinib, and sequential EGFR C797S mutation mediating resistance to combined targeted therapy with osimertinib and alectinib. Our case shows that EML4-ALK fusion is a rare but critical resistance mechanism to osimertinib, and C797S mutation in cis may be an underlying mechanism of acquired resistance mutation in double TKIs therapy. Furthermore, molecular detection and rebiopsy play important roles in the selection of therapeutic strategies when the disease progresses.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Female , Humans , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/pathology , ErbB Receptors/genetics , Mutation , Protein Kinase Inhibitors/therapeutic use , Aniline Compounds/therapeutic use , Aniline Compounds/pharmacology , Oncogene Proteins, Fusion
2.
Zhongguo Gu Shang ; 34(4): 315-20, 2021 Apr 25.
Article in Chinese | MEDLINE | ID: mdl-33896128

ABSTRACT

OBJECTIVE: To explore the dynamic changes of lumbosacral sagittal parameters after real-time three-dimensional navigation assisted minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) and traditional open TLIF for treatment of lumbar degenerative disease. METHODS: The clinical data of 61 patients with lumbar degenerative disease underwent single-segment surgery from September 2017 to September 2019 were retrospectively analyzed. Among them, 31 cases underwent MIS-TLIF with 3D navigation techniques (MIS-TLIF group) and another 30 cases underwent conventional open TLIF (traditional open TLIF group). The basic information, operative time and intraoperative blood loss were collected. The sagittal radiologic parameters were measured before surgery and 3 months after surgery, including lumbar lordosis (LL), segmental lordosis (SL), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), anterior disc height (ADH), posterior disc height(PDH).And the average disc height(DH) and pelvic incidence to lumbar lordosis mismatch (PI-LL) were calculated. RESULTS: Operative time and intraoperative blood loss in MIS-TLIF group were significantly less than in traditional open TLIF group(P<0.05). In MIS-TLIF group, LL, SL, PI-LL, and DH were significantly improved at 3 months after surgery (P<0.05), while PI, PT, and SS were not statistically different from those before surgery (P>0.05). LL, PI-LL, and DH of patients in the traditional open TLIF group were significantly improved at 3 months after surgery (P<0.05), while the PI, PT, SS, and SL were not statistically different from those before surgery (P>0.05). LL change showed a significant correlation with SL change (r= 0.433, P<0.001). Change in SL closely correlated to change in ADH (r=0.621, P<0.05) and PDH(r=0.527, P<0.05). CONCLUSION: Real-time navigation-assisted MIS-TLIF and traditional open TLIF can recover DH in a short term for lumbar degenerative diseases, improve LL and PI-LL, and make the arrangement of the sagittal plane of the lumbosacral region more coordinated after surgery. But only the navigation assisted MIS -TLIF can significantly improve SL. Compared with traditional open TLIF, real-time navigation assisted MIS-TLIF in the treatment of degenerative lumbar diseases has the advantages of short operation time and less intraoperative bleeding.


Subject(s)
Lumbar Vertebrae , Spinal Fusion , Humans , Lumbar Vertebrae/surgery , Lumbosacral Region , Minimally Invasive Surgical Procedures , Retrospective Studies , Treatment Outcome
3.
Plant J ; 106(2): 351-365, 2021 04.
Article in English | MEDLINE | ID: mdl-33486798

ABSTRACT

Lotus plumule, the embryo of the seed of the sacred lotus (Nelumbo nucifera), contains a high accumulation of secondary metabolites including flavonoids and possesses important pharmaceutical value. Flavonoid C-glycosides, which accumulate exclusively in lotus plumule, have attracted considerable attention in recent decades due to their unique chemical structure and special bioactivities. As well as mono-C-glycosides, lotus plumule also accumulates various kinds of di-C-glycosides by mechanisms which are as yet unclear. In this study we identified two C-glycosyltransferase (CGT) genes by mining sacred lotus genome data and provide in vitro and in planta evidence that these two enzymes (NnCGT1 and NnCGT2, also designated as UGT708N1 and UGT708N2, respectively) exhibit CGT activity. Recombinant UGT708N1 and UGT708N2 can C-glycosylate 2-hydroxyflavanones and 2-hydroxynaringenin C-glucoside, forming flavone mono-C-glycosides and di-C-glycosides, respectively, after dehydration. In addition, the above reactions were successfully catalysed by cell-free extracts from tobacco leaves transiently expressing NnCGT1 or NnCGT2. Finally, enzyme assays using cell-free extracts of lotus plumule suggested that flavone di-C-glycosides (vicenin-1, vicenin-3, schaftoside and isoschaftoside) are biosynthesized through sequentially C-glucosylating and C-arabinosylating/C-xylosylating 2-hydroxynaringenin. Taken together, our results provide novel insights into the biosynthesis of flavonoid di-C-glycosides by proposing a new biosynthetic pathway for flavone C-glycosides in N. nucifera and identifying a novel uridine diphosphate-glycosyltransferase (UGT708N2) that specifically catalyses the second glycsosylation, C-arabinosylating and C-xylosylating 2-hydroxynaringenin C-glucoside.


Subject(s)
Flavonoids/metabolism , Glycosides/metabolism , Nelumbo/metabolism , Glycosylation , Glycosyltransferases/genetics , Glycosyltransferases/metabolism , Metabolic Networks and Pathways , Nelumbo/enzymology , Nelumbo/genetics , Phylogeny , Plants, Genetically Modified , Nicotiana
4.
Chem Biodivers ; 18(2): e2000848, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33403807

ABSTRACT

Paeonia cultivars are famous ornamental plants, and some of them are also traditional Chinese medicinal resources. Intersubgeneric hybrids of Paeonia (IHPs) are formed by the hybridization of herbaceous peony (Paeonia lactiflora) and tree peony (Paeonia×suffruticosa or lutea hybrid tree peony). The phenotypic characteristics of IHPs are similar to those of herbaceous peony, and their root systems are large and vigorous. However, their medicinal value has not been reported yet. In this study, the roots of eight IHP samples were analyzed by high performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS/MS). A total of 18 compounds were identified, including phenols, paeonols, monoterpene glycosides, and tannins. The contents of monoterpene glycosides and tannins in IHPs were higher than herbaceous peony and tree peony, exceeding 44.76 mg/g DW and 11.50 mg/g DW, respectively. Three IHPs, 'Prairie Charm', 'Garden Treasure', and 'Yellow Emperor', with more types and a higher content of medicinal compounds, were screened out by cluster analysis. These IHPs have considerable potential for the development of medicinal resources.


Subject(s)
Paeonia/chemistry , Plant Roots/chemistry , Acetophenones/analysis , Glycosides/analysis , Monoterpenes/analysis , Phenols/analysis , Tannins/analysis
5.
Phytochemistry ; 174: 112331, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32146385

ABSTRACT

Paeonia Radix Rubra (PRR) is a very common traditional Chinese medicine (TCM). The roots of Paeonia lactiflora and Paeonia anomala subsp. veitchii are used for the production of PRR. However, other species of section Paeonia in China are also used to produce PRR. The roots of section Paeonia from 20 populations of seven species and two subspecies in China were analyzed by high performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF- MS). A total of 21 metabolites were identified, including nine monoterpene glycosides, seven tannins, three phenols, one paeonol and one flavonoid. There were significant differences in the composition and content of metabolites among different populations. The relative contents of monoterpene glycosides and tannins were generally higher in most samples. Cluster analysis showed that 20 populations could be divided into four groups. Among them, the populations of P. lactiflora and Paeonia mairei were clustered together in one group. The content of paeoniflorin in P. lactiflora was high (>22.20 mg g-1, dry weight, the same below), and the content of lactiflorin in P. mairei (>17.50 mg g-1) was significantly higher than in other species (<2.30 mg g-1). The monoterpene glycoside content in P. mairei (>51.60 mg g-1) was significantly higher than in other species (<43.40 mg g-1), suggesting that it could be useful medicinal germplasm for the development of monoterpene glycosides. In addition, some populations of Paeonia emodi, Paeonia sterniana and P. mairei may also be used as potential germplasm resources for use in PRR. Genetic and environmental factors resulted in differences in the composition and content of metabolites in different populations of the same species. Therefore, it is necessary to carefully consider the origin of Paeonia germplasm when selecting medicinal materials to be used as resources for the production of PRR.


Subject(s)
Drugs, Chinese Herbal , Paeonia , China , Monoterpenes , Phytochemicals , Plant Roots
6.
J Sep Sci ; 42(18): 2888-2899, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31282097

ABSTRACT

Elm fruits were once an important food source in the years of famine. Research on the functional compounds in elm fruits was almost unavailable. In this study, we established an efficient high-performance liquid chromatography method for the simultaneous separation of eight chlorogenic acids and 28 flavonoids in elm fruits for the first time. Total flavonoid contents ranged from 286 mg/100 g (Ulmus laciniata) to 1228 mg/100 g (U. pumila). High concentrations of rutin, quercetin 3-O-glucoside, and kaempferol derivatives were present in U. laevis, U. castaneifolia, and U. pumila, respectively. Furthermore, the fruit extracts of U. americana, U. castaneifolia, U. davidiana, and U. pumila showed higher antioxidant activity. These results suggest that fruits of these species can be used as bioresources for the extraction of the corresponding functional compounds. This work provides informative data and can be an important reference for future research on elm fruits as a renewed food resource.


Subject(s)
Antioxidants/analysis , Chlorogenic Acid/analysis , Flavonoids/analysis , Fruit/chemistry , Ulmus/chemistry , Antioxidants/pharmacology , Benzothiazoles/antagonists & inhibitors , Biphenyl Compounds/antagonists & inhibitors , Chlorogenic Acid/pharmacology , Flavonoids/pharmacology , Picrates/antagonists & inhibitors , Sulfonic Acids/antagonists & inhibitors
7.
J Pharm Biomed Anal ; 161: 455-463, 2018 Nov 30.
Article in English | MEDLINE | ID: mdl-30216794

ABSTRACT

Gentianopsis barbata, Halenia corniculata, and Gentianella acuta were widely distributed throughout China and commonly used in traditional Chinese medicine. However, owing to similar living environments and morphological features, locals often had trouble distinguishing between these three species. In this present study, chromatograms at 350 nm were obtained and the composition and content of their chemical compounds determined using HPLC-DAD/ESI-MS2. In total, 35 chemical compounds were detected, 32 of which were identified, 25 of which were xanthones, 6 flavonoids, and 1 chlorogenic acid. The 350 nm chromatograms of these three species displayed evident differences. The individual compounds and their occurrence and content in different parts of the plant within different species were included in our results. This basic data will be useful for future pharmacological study. The total compositions of flavonoids and xanthones were approximately comparable in G. barbata and H. corniculata. Meanwhile, xanthones were predominant in G. acuta. From the perspective of chemical compound compositions, the leaf is recommended as the most valuable medicinal section for each of these three species.


Subject(s)
Chlorogenic Acid/analysis , Drugs, Chinese Herbal/chemistry , Flavonoids/analysis , Gentianaceae/chemistry , Xanthones/analysis , Chromatography, High Pressure Liquid , Plant Structures/chemistry , Spectrometry, Mass, Electrospray Ionization
8.
Plant Sci ; 274: 212-222, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30080606

ABSTRACT

α-linolenic acid (ALA) deficiency and a skewed ω6: ω3 fatty acid ratio in the diet are thought to be a major cause for the high incidence of cardiovascular, inflammatory, and autoimmune diseases. Recent years, tree peony (Paeonia suffruticosa Andr.) with the high proportion of ALA (more than 45% in seed oil) is widely concerned. However, the underlying accumulation mechanism of the ALA in tree peony seeds remains unknown. In this study, comparative transcriptome analysis was performed between two cultivars ('Saiguifei' and 'Jingshenhuanfa') with different ALA contents. The analysis of the metabolic enzymes associated with ALA biosynthesis and temporal accumulation patterns of unsaturated fatty acids demonstrated the importance of microsomal ω-3 fatty acid desaturase 3 (FAD3). Moreover, PsFAD3 gene was identified from tree peony seeds, which was located in endoplasmic reticulum and the expression levels of PsFAD3 were consistent with ALA accumulation patterns in seeds. Heterologous expression in Saccharomyces cerevisiae and Arabidopsis thaliana confirmed that the isolated PsFAD3 protein could catalyze ALA synthesis. These results indicated that PsFAD3 was involved in the synthesis of ALA in seeds and could be exploited by the genetic breeding of new cultivars with high ALA content in tree peony as well as other potential crops.


Subject(s)
Fatty Acid Desaturases/metabolism , Paeonia/enzymology , Transcriptome , alpha-Linolenic Acid/metabolism , Fatty Acid Desaturases/genetics , Fatty Acids, Unsaturated/metabolism , Paeonia/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Seeds/enzymology , Seeds/genetics
9.
Gene ; 666: 72-82, 2018 Aug 05.
Article in English | MEDLINE | ID: mdl-29738839

ABSTRACT

MicroRNAs (miRNAs) and long noncoding RNAs (lncRNAs) act as important molecular regulators in a wide range of biological processes during plant development and seed formation, including oil production. Tree peony seeds contain >90% unsaturated fatty acids (UFAs) and high proportions of α-linolenic acid (ALA, > 40%). To dissect the non-coding RNAs (ncRNAs) pathway involved in fatty acids synthesis in tree peony seeds, we construct six small RNA libraries and six transcriptome libraries from developing seeds of two cultivars (J and S) containing different content of fatty acid compositions. After deep sequencing the RNA libraries, the ncRNA expression profiles of tree peony seeds in two cultivars were systematically and comparatively analyzed. A total of 318 known and 153 new miRNAs and 22,430 lncRNAs were identified, among which 106 conserved and 9 novel miRNAs and 2785 lncRNAs were differentially expressed between the two cultivars. In addition, potential target genes of the microRNA and lncRNAs were also predicted and annotated. Among them, 9 miRNAs and 39 lncRNAs were predicted to target lipid related genes. Results showed that all of miR414, miR156b, miR2673b, miR7826, novel-m0027-5p, TR24651|c0_g1, TR24544|c0_g15, and TR27305|c0_g1 were up-regulated and expressed at a higher level in high-ALA cultivar J when compared to low-ALA cultivar S, suggesting that these ncRNAs and target genes are possibly involved in different fatty acid synthesis and lipid metabolism through post-transcriptional regulation. These results provide a better understanding of the roles of ncRNAs during fatty acid biosynthesis and metabolism in tree peony seeds.


Subject(s)
MicroRNAs/genetics , Paeonia/genetics , RNA, Long Noncoding/genetics , Seeds/genetics , alpha-Linolenic Acid/biosynthesis , Biosynthetic Pathways , Gene Expression Regulation, Plant , Gene Ontology , High-Throughput Nucleotide Sequencing , MicroRNAs/metabolism , Molecular Sequence Annotation , Paeonia/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , RNA Interference , RNA, Long Noncoding/metabolism , RNA, Plant/genetics , RNA, Plant/metabolism , Seeds/metabolism , Transcriptome
10.
Phytochemistry ; 146: 16-24, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29207319

ABSTRACT

Mu Dan Pi is a traditional Chinese medicine used to treat inflammation, cancer, allergies, diabetes, angiocardiopathy, and neurodegenerative diseases. In this study, the metabolome variation within Mu Dan Pi collected from 372 tree peony cultivars was systematically investigated. In total, 42 metabolites were identified, comprising of 14 monoterpene glucosides, 11 tannins, 8 paeonols, 6 flavonoids, and 3 phenols. All cultivars revealed similar metabolite profiles, however, they were further classified into seven groups on the basis of their varying metabolite contents by hierarchical cluster analysis. Traditional cultivars for Mu Dan Pi were found to have very low metabolite contents, falling into clusters I and II. Cultivars with the highest amounts of metabolites were grouped in clusters VI and VII. Five potential cultivars, namely, 'Bai Yuan Qi Guan', 'Cao Zhou Hong', 'Da Zong Zi', 'Sheng Dan Lu', and 'Cheng Xin', with high contents of monoterpene glycosides, tannins, and paeonols, were further screened. Interestingly, the majority of investigated cultivars had relatively higher metabolite contents compared to the traditional medicinal tree peony cultivars.


Subject(s)
Drugs, Chinese Herbal/isolation & purification , Paeonia/chemistry , Phytochemicals/isolation & purification , Drugs, Chinese Herbal/chemistry , Medicine, Chinese Traditional , Molecular Structure , Phytochemicals/chemistry , Plant Roots/chemistry
11.
Food Chem ; 219: 490-495, 2017 Mar 15.
Article in English | MEDLINE | ID: mdl-27765256

ABSTRACT

Vaccinium glaucoalbum, a perennial evergreen shrub, is naturally distributed in high-altitude areas. In this study, the composition and content of polyphenolic compounds in the fruit and leaf of V. glaucoalbum were characterized. In total, 24 chemical compounds were detected and identified by HPLC-DAD and HPLC-ESI-MS2. Among all the compounds determined, 15 were anthocyanins and detected in fruit, 5 were flavonols and monitored in leaf, and 4 were chlorogenic acids and found in both fruit and leaf. The total anthocyanin content (TAC) of fruit (682mg/100gFW) was the highest among wild Vaccinium berries in China which have been investigated for now, and the total flavonol content of leaf was 2764mg/100gFW. The antioxidant activity of both fruit and leaf was assessed by DPPH and FRAP assays. Given its high TAC and strong antioxidant activity, the fruit of V. glaucoalbum has great potential in functional food.


Subject(s)
Antioxidants/metabolism , Fruit/metabolism , Plant Extracts/metabolism , Plant Leaves/metabolism , Polyphenols/metabolism , Vaccinium/metabolism , Anthocyanins/metabolism , China , Chromatography, High Pressure Liquid , Flavonols/metabolism , Mass Spectrometry
12.
BMC Genomics ; 17(1): 897, 2016 11 09.
Article in English | MEDLINE | ID: mdl-27829354

ABSTRACT

BACKGROUND: Waterlily (Nymphaea spp.), a perennial herbaceous aquatic plant, is divided into two ecological groups: hardy waterlily and tropical waterlily. Although the hardy waterlily has no attractive blue flower cultivar, its adaptability is stronger than tropical waterlily because it can survive a cold winter. Thus, breeding hardy waterlily with real blue flowers has become an important target for breeders. Molecular breeding may be a useful way. However, molecular studies on waterlily are limited due to the lack of sequence data. RESULTS: In this study, six cDNA libraries generated from the petals of two different coloring stages of blue tropical waterlily cultivar Nymphaea 'King of Siam' were sequenced using the Illumina HiSeq™ 2500 platform. Each library produced no less than 5.65 Gb clean reads. Subsequently, de novo assembly generated 112,485 unigenes, including 26,206 unigenes annotated to seven public protein databases. Then, 127 unigenes could be identified as putative homologues of color-related genes in other species, including 28 up-regulated and 5 down-regulated unigenes. In petals, 16 flavonoids (4 anthocyanins and 12 flavonols) were detected in different contents during the color development due to the different expression levels of color-related genes, and four flavonols were detected in waterlily for the first time. Furthermore, UA3GTs were selected as the most important candidates involved in the flavonoid metabolic pathway, UA3GTs induced blue petal color formation in Nymphaea 'King of Siam'. CONCLUSIONS: This study will improve our understanding of the molecular mechanism of blue flowers in waterlily and provide the basis for molecular breeding of blue hardy waterlily cultivars.


Subject(s)
Flowers/genetics , Flowers/metabolism , Metabolome , Nymphaea/genetics , Nymphaea/metabolism , Transcriptome , Computational Biology/methods , Gene Expression Profiling , Metabolomics , Phenotype
13.
Sci Rep ; 6: 19245, 2016 Jan 12.
Article in English | MEDLINE | ID: mdl-26754912

ABSTRACT

Flavones are important secondary metabolites found in many plants. In Lonicera species, flavones contribute both physiological and pharmaceutical properties. However, flavone synthase (FNS), the key enzyme responsible for flavone biosynthesis, has not yet been characterized in Lonicera species. In this study, FNSII genes were identified from Lonicera japonica Thunb. and L. macranthoides Hand.-Mazz. In the presence of NADPH, the recombinant cytochrome P450 proteins encoded by LjFNSII-1.1, LjFNSII-2.1, and LmFNSII-1.1 converted eriodictyol, naringenin, and liquiritigenin to the corresponding flavones directly. The different catalytic properties between LjFNSII-2.1 and LjFNSII-1.1 were caused by a single amino acid substitution at position 242 (glutamic acid to lysine). A methionine at position 206 and a leucine at position 381 contributed considerably to the high catalytic activity of LjFNSII-1.1. In addition, LjFNSII-1.1&2.1 and LmFNSII-1.1 also biosynthesize flavones that were further modified by O-glycosylation in transgenic tobacco. The expression levels of the FNSII genes were consistent with flavone accumulation patterns in flower buds. Our findings suggested that the weak catalytic activity of LmFNSII-1.1 and the relatively low expression of LmFNSII-1.1 in flowers might be responsible for the low levels of flavone accumulation in flower buds of L. macranthoides.


Subject(s)
Flavones/metabolism , Lonicera/metabolism , Mixed Function Oxygenases/metabolism , Amino Acid Sequence , Biosynthetic Pathways , Catalysis , Enzyme Activation , Gene Expression , Lonicera/enzymology , Lonicera/genetics , Mixed Function Oxygenases/chemistry , Mixed Function Oxygenases/genetics , Models, Molecular , Molecular Sequence Data , Mutation , Phylogeny , Protein Conformation , Protein Transport , Recombinant Proteins
14.
J Exp Bot ; 66(21): 6563-77, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26208646

ABSTRACT

Anthocyanins are major pigments in plants. Methylation plays a role in the diversity and stability of anthocyanins. However, the contribution of anthocyanin methylation to flower coloration is still unclear. We identified two homologous anthocyanin O-methyltransferase (AOMT) genes from purple-flowered (PsAOMT) and red-flowered (PtAOMT) Paeonia plants, and we performed functional analyses of the two genes in vitro and in vivo. The critical amino acids for AOMT catalytic activity were studied by site-directed mutagenesis. We showed that the recombinant proteins, PsAOMT and PtAOMT, had identical substrate preferences towards anthocyanins. The methylation activity of PsAOMT was 60 times higher than that of PtAOMT in vitro. Interestingly, this vast difference in catalytic activity appeared to result from a single amino acid residue substitution at position 87 (arginine to leucine). There were significant differences between the 35S::PsAOMT transgenic tobacco and control flowers in relation to their chromatic parameters, which further confirmed the function of PsAOMT in vivo. The expression levels of the two homologous AOMT genes were consistent with anthocyanin accumulation in petals. We conclude that AOMTs are responsible for the methylation of cyanidin glycosides in Paeonia plants and play an important role in purple coloration in Paeonia spp.


Subject(s)
Methyltransferases/genetics , Paeonia/genetics , Plant Proteins/genetics , Amino Acid Sequence , Anthocyanins/genetics , Anthocyanins/metabolism , Color , Flowers/genetics , Flowers/metabolism , Methylation , Methyltransferases/chemistry , Methyltransferases/metabolism , Molecular Sequence Data , Mutagenesis, Site-Directed , Paeonia/metabolism , Phylogeny , Pigmentation , Plant Proteins/chemistry , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Sequence Alignment , Nicotiana/genetics , Nicotiana/metabolism
15.
Food Chem ; 187: 20-8, 2015 Nov 15.
Article in English | MEDLINE | ID: mdl-25976993

ABSTRACT

Water lily, a member of the Nymphaeaceae family, can be made into tea on the basis of outstanding fragrance characteristics and health care functions. In this study, 16 flavonoids were identified and quantified in tea infusions prepared from the petals of 33 water lily cultivars using HPLC-DAD and HPLC-ESI-MS/MS. The infusions were analyzed with HS-SPME coupled with GC-MS; 29 volatile compounds were detected, of which nine were found to be scent components. The cultivars were clustered into three clusters characterized according to scent components. The 'Conqueror' and 'Virginia' cultivars had the highest antioxidant activities. The concentrations of polyphenols and flavonoids showed significant positive correlations with antioxidant activity as measured by DPPH, ABTS(+), and FRAP assays. This study is valuable for a fuller understanding of this important tea and can also be used for the development of water lily.


Subject(s)
Antioxidants/analysis , Flavonoids/analysis , Nymphaea/chemistry , Teas, Herbal/analysis , Volatile Organic Compounds/analysis , Chromatography, High Pressure Liquid , Flowers/chemistry , Gas Chromatography-Mass Spectrometry , Polyphenols/analysis , Tandem Mass Spectrometry
16.
Nat Prod Commun ; 10(3): 495-8, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25924537

ABSTRACT

Chrysanthemum is an important traditional Chinese medicine and is drunk daily as a herbal tea. Chlorogenic acids and flavonoids are generally considered as the bioactive compounds. In this work, six kinds of Juhua Tea were analyzed and their active compounds and antioxidant activities were compared. In total, 32 phenolic compounds were profiled and identified using HPLC-DAD/ESI-MSn, composed of chlorogenic acids (10), flavones (8), chalcones (8), flavanones (4) and flavonols (2). Chalcones were the main flavonoids in Kunlun Xueju (Coreopsis tinctoria) extract, while flavones and chlorogenic acids were dominant in the five Chrysanthemum teas. Total chlorogenic acids content (TCA) was highest in Tai Ju (Chrysanthemum morifolium cv. 'Tai Ju') (8.59 ± 0.87 mg/g DW), and total flavonoids content (TF) was highest in Kunlun Xueju (87.2 ± 7.0 mg/g DW), which were both lowest in Ganye Ju (Chrysanthemum eticuspe) (TCA 0.86 ± 0.26 mg/g DW, TF 1.43 ± 0.41 mg/g DW). Huangin Ju (Anthemis tinctoria) possessed the most flavones (19.7 ± 0.6 mg/g DW). Antioxidant capacity of each drink, assessed by Folin-Ciocalteu, DPPH, ABTS and FRAP assays, consistently showed that Kunlun Xueju extract possessed stronger antioxidant activity than the other five, suggesting that the flavonoids content accounted for the free radical scavenging. The present work provides a method for the characterization and quality control of Juhua Tea. Moreover, it is a guideline for consuming choice, due to the different biological functions resulting from chalcones, chlorogenic acids, and flavones.


Subject(s)
Antioxidants/chemistry , Beverages/analysis , Chrysanthemum/chemistry , Animals , Biphenyl Compounds/chemistry , Picrates/chemistry
17.
BMC Genomics ; 16: 208, 2015 Mar 18.
Article in English | MEDLINE | ID: mdl-25887415

ABSTRACT

BACKGROUND: Tree peony (Paeonia section Moutan DC.) is known for its excellent ornamental and medicinal values. In 2011, seeds from P. ostii have been identified as novel resource of α-linolenic acid (ALA) for seed oil production and development in China. However, the molecular mechanism on biosynthesis of unsaturated fatty acids in tree peony seeds remains unknown. Therefore, transcriptome data is needed to better understand the underlying mechanisms. RESULTS: In this study, lipid accumulation contents were measured using GC-MS methods across developing tree peony seeds, which exhibited an extraordinary ALA content (49.3%) in P. ostii mature seeds. Transcriptome analysis was performed using Illumina sequencing platform. A total of 144 million 100-bp paired-end reads were generated from six libraries, which identified 175,874 contigs. In the KEGG Orthology enrichment of differentially expressed genes, lipid metabolism pathways were highly represented categories. Using this data we identified 388 unigenes that may be involved in de novo fatty acid and triacylglycerol biosynthesis. In particular, three unigenes (SAD, FAD2 and FAD8) encoding fatty acid desaturase with high expression levels in the fast oil accumulation stage compared with the initial stage of seed development were identified. CONCLUSIONS: This study provides the first comprehensive genomic resources characterizing tree peony seeds gene expression at the transcriptional level. These data lay the foundation for further understanding of molecular mechanism responsible for lipid biosynthesis and the high unsaturated fatty acids (especially ALA) accumulation. Meanwhile, it provides theoretical base for potential oilseed application in the respect of n-6 to n-3 ratio for human diets and future regulation of target healthy components of oils.


Subject(s)
Fatty Acids/metabolism , Paeonia/genetics , Paeonia/metabolism , Seeds/genetics , Seeds/metabolism , Transcriptome , Biosynthetic Pathways , Computational Biology , Gene Expression Profiling , Gene Expression Regulation, Plant , High-Throughput Nucleotide Sequencing , Lipid Metabolism , Molecular Sequence Annotation , Phenotype
18.
Food Chem ; 173: 133-40, 2015 Apr 15.
Article in English | MEDLINE | ID: mdl-25466004

ABSTRACT

Seeds from Paeonia ostii and Paeoniarockii have been recently identified as novel resources of α-linolenic acid (ALA) in China. To assess whether tree peony cultivars can be used as oil resource, fatty acids (FAs) in 60 cultivars were monitored and evaluated in this study. The results indicated that the composition and content of FAs varied dramatically among different cultivars, in which ALA, linoleic acid, oleic acid, palmitic acid, and stearic acid were the dominant. The 60 cultivars were classified into six clusters by hierarchical cluster analysis, and they were quite distinct from each other. Finally, six cultivars with high yield and high quality were screened out, comprising of 'Liuliguanzhu', 'Hongguanyupei', 'LSS-2', 'LSS-1', 'Jingshenhuanfa' and 'LSS-11'. These cultivars were appropriately applied in practical oil production. Overall, tree peony oil with abundant unsaturated fatty acids especially ALA was proved to be a top-grade source for edible oil and nutritional supplements.


Subject(s)
Fatty Acids/chemistry , Paeonia/chemistry , Plant Oils/chemistry , China , Gas Chromatography-Mass Spectrometry/methods , Paeonia/classification , Seeds/chemistry
19.
PLoS One ; 9(10): e108860, 2014.
Article in English | MEDLINE | ID: mdl-25279809

ABSTRACT

Flavonoids in nine tissues of Nelumbo nucifera Gaertner were identified and quantified by high-performance liquid chromatography with diode array detector (HPLC-DAD) and HPLC-electrospray ionization-mass spectrometry (HPLC-ESI-MSn). Thirty-eight flavonoids were identified; eleven C-glycosides and five O-glycosides were discovered for the first time in N. nucifera. Most importantly, the C-glycosyl apigenin or luteolin detected in lotus plumules proved valuable for deep elucidation of flavonoid composition in lotus tissues and for further utilization as functional tea and medicine materials. Lotus leaves possessed the significantly highest amount of flavonoids (2.06E3±0.08 mg 100 g(-1) FW) and separating and purifying the bioactive compound, quercetin 3-O-glucuronide, from leaves showed great potential. In contrast, flavonoids in flower stalks, seed coats and kernels were extremely low. Simultaneously, the optimal picking time was confirmed by comparing the compound contents in five developmental phases. Finally, we proposed the putative flavonoid biosynthesis pathway in N. nucifera.


Subject(s)
Flavones/metabolism , Flavonoids/metabolism , Glycosides/metabolism , Nelumbo/metabolism , Chromatography, High Pressure Liquid , Flavones/analysis , Flavonoids/analysis , Flowers/chemistry , Glycosides/analysis , Plant Leaves/chemistry , Spectrometry, Mass, Electrospray Ionization
20.
Food Chem ; 160: 357-64, 2014 Oct 01.
Article in English | MEDLINE | ID: mdl-24799249

ABSTRACT

Variation of anthocyanin and flavonol in Vaccinium uliginosum berry from seven locations in the Lesser Khingan Mountains was examined for the first time. A total of 14 anthocyanins and 10 flavonols were identified using HPLC-DAD and HPLC-ESI-MS(2) analysis. The composition of anthocyanins and flavonols in berries were the same in all of the locations investigated. Malvidin 3-O-glucoside was the most abundant anthocyanin, and quercetin 3-O-galactoside was the main flavonol compound. The berries from Wuying region had the highest concentrations of both anthocyanins and flavonols. It was found that the concentrations of both anthocyanins and flavonols increased with rising altitude. Furthermore, the concentrations of polyphenols, and especially anthocyanins, in V. uliginosum berry showed significant positive correlation with antioxidant activity as measured by DPPH, ABTS(+) and FRAP assays. This study is valuable for further basic research and applied development of V. uliginosum in the Lesser Khingan Mountains.


Subject(s)
Anthocyanins/chemistry , Antioxidants/chemistry , Flavonols/chemistry , Fruit/chemistry , Plant Extracts/chemistry , Vaccinium/chemistry , China , Chromatography, High Pressure Liquid , Mass Spectrometry , Molecular Structure
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