Flavonoid Metabolism in Tetrastigma hemsleyanum Diels et Gilg Based on Metabolome Analysis and Transcriptome Sequencing
Abstract
:1. Introduction
2. Results
2.1. Flavonoid Components Differ in Leaves and Root Tubers of T. hemsleyanum
2.2. Transcriptome Sequencing of Leaves and Root Tubers of T. hemsleyanum
2.3. Differentially Expressed Genes of Leaves and Root Tubers of T. hemsleyanum
2.4. Analysis Correlation between Flavonoid Biosynthetic Gene Expression and Flavonoid Concentration
2.5. Validation of the DEGs
2.6. Key Gene Expression and Total Flavonoid Content Trends for T. hemsleyanum from Different Locations
3. Discussion
3.1. Key Role of DEGs in Photosynthesis-Antenna Proteins in Flavonoid Metabolism in Leaves and Root Tubers
3.2. CHI and UFGT Are Critical Genes for Flavonoid Biosynthesis in T. hemsleyanum
4. Materials and Methods
4.1. Plant Materials
4.2. Determination of Flavonoid Metabolomes
4.3. RNA Extraction and Transcriptome Sequencing
4.4. De Novo Assembly and Sequence Clustering
4.5. FPKM Calculation and DEGs Screening
4.6. Functional Annotation and Classification of Genes
4.7. Quantitative Real-Time PCR
4.8. Total Flavonoid Determination
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
T. hemsleyanum | Tetrastigma hemsleyanum Diels et Gilg |
PAL | Phenylalanine ammonia lyase |
C4H | Cinnamate 4-hydroxylase |
4CL | 4-coumarate-coenzyme A |
CHI | Chalcone isomerase |
CHS | Chalcone synthase |
F3H | Flavanone-3-hydroxylase |
F3′H | Flavanone-3′-hydroxylase |
F3′5′H | Flavonoid-3′,5′-hydroxylase |
FLS | Flavonol synthase |
FNS | flavone synthase |
DFR | Dihydroflavonol 4-reductase |
LDOX | Leucoanthocyanidin dioxygenase |
3-Glct | Flavonoid-3-O-glucosyltransferase |
7-Glct | Flavonoid-3-O-glucosyltransferase |
1,2-Rhat | 1,2-rhamnosyltransferase |
1,6-Rhat | 1,6-rhamnosyltransferase |
UGTs | UDP-Glycosyltransferases |
OMT | O-methyltransferase |
UFGT | UDP-glycose flavonoid glycosyltransferase |
MAPK | Mitogen-activated protein kinase. |
KOG | Clusters of orthologous groups for eukaryotic complete genomes |
NR | Non-Redundant Protein Sequence Database |
GO | Gene Ontology |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
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Term | All > 300 bp | ≥500 bp | ≥1000 bp | N50 | Total Length | Max Length | Min Length | Average Length |
---|---|---|---|---|---|---|---|---|
Y-Unigene | 53,039 | 29,973 | 14,690 | 1275 | 47,471,677 | 15,654 | 301 | 895.03 |
G-Unigene | 60,953 | 33,607 | 16,023 | 1236 | 53,338,272 | 15,654 | 301 | 875.07 |
Compound | Regression Equation | r | Linear Range |
---|---|---|---|
Vitexin | Y = 8,542,235.9 X + 18,454,664.6 | 0.9987 | 20.000–2000.000 |
Orientin | Y = 9,071,337.1 X + 33,093.6 | 0.9996 | 1.000–100.000 |
Afzelin | Y = 9,658,807.6 X + 92,259.2 | 0.9993 | 0.001–0.050 |
Rutin | Y = 4,950,860.8 X + 231,551.2 | 0.9994 | 0.010–0.500 |
Isoquercitrin | Y = 3,356,305.1 X + 593,847.8 | 0.9989 | 0.020–1.000 |
Luteolin | Y = 9,109,434.3 X + 35,359.9 | 0.9998 | 0.020–1.000 |
Luteoloside | Y = 3,168,802.6 X + 166,068.3 | 0.9994 | 0.100–5.000 |
Isoorientin | Y = 4,845,698.3 X + 4,222,807.8 | 0.9987 | 1.000–100.000 |
Naringin | Y = 3,972,039.5 X + 64,183.6 | 0.9976 | 0.010–0.500 |
Isorhamnetin-3-O-Glucoside | Y = 8,988,517.2 X + 5,992,231.1 | 0.9982 | 0.020–1.000 |
Isorhamnetin | Y = 1,476,424.2 X + 2,111,521.6 | 0.9986 | 0.001–0.050 |
Astragaline | Y = 2,210,508.5 X + 2,507,321.4 | 0.9986 | 0.010–0.500 |
Eriodictyol | Y = 691,932.1 X + 84659.5 | 0.9991 | 0.010–0.500 |
Taxifolin | Y = 3,575,802.9 X + 284,658.7 | 0.9992 | 0.001–0.050 |
Isovitexin | Y = 8,542,235.9 X + 18,454,664.6 | 0.9987 | 20.000–2000.000 |
Calycosin-7-O-glucoside | Y = 22,042.7 X + 2463.8 | 0.9986 | 0.020–1.000 |
Hispidulin | Y = 207,676.0 X + 105,248.9 | 0.9993 | 0.020–1.000 |
Naringenin | Y = 12,452,147.9 X +74,631.9 | 0.9992 | 0.001–0.050 |
Apigenin | Y = 2,885,631.5 X + 238,287.7 | 0.9989 | 0.001–0.050 |
Quercitrin | Y = 12,206,204.2 X + 1,425,428.8 | 0.9992 | 0.001–0.050 |
Apigenin-7-O-Glycoside | Y = 1,806,515.0 X + 16,658.0 | 0.9996 | 0.010–0.500 |
Gene | Gene ID | Tm (°C) | Primer F (5′-3′) | Primer R (5′-3′) |
---|---|---|---|---|
MDH | \ | 60 | TGTTGCTACGACTGATGT | CCTGAGACTTGTAGATGGAA |
PAL | CL2379.Contig9_All | 56.02 | ACCAACCATGTCCAAAGTGC | CGCCACAAGGTATGTGGAAG |
CL2379.Contig4_All | 56.06 | TGGTGACCTCACCTTCTCAC | GAAGCCCATCCATTCCGAAC | |
CHS | CL1413.Contig5_All | 55.9 | GGAACTGTCCTTCGAACTGC | GTCCGCGGAATGTAACAACA |
CL1413.Contig12_All | 55.99 | TAGCACGTTGAGCGTTTCTG | AGAGTTGGTGGCATAAGGCT | |
CL1413.Contig4_All | 55.71 | CCAACTTGTCTCAGCAGCTC | AATCAAAGTGGGCACAGTGG | |
CL1413.Contig11_All | 55.89 | TGTACCATCAAGGGTGCCAT | GGGCTTGGCCAACTAAAGAG | |
CHI | CL6715.Contig1_All | 56.01 | GTGCAGGGTGTGAAGTTTGT | TTTGGCTTCCTTCCAACAGC |
Unigene19431_All | 56.47 | ACGCCATGGATAGAGAGCAA | CCCATGGTTGAGGATTCGGA | |
CL921.Contig4_All | 55.96 | ACTGAACACCATCCACGACT | GACCAACGAATGCCTCGAAA | |
FNS | Unigene21915_All | 55.99 | AAAGACGAACTCTCCACCGT | GATGTTCGCAACTCCGTTCA |
UFGT | CL11556.Contig3_All | 56.05 | TTGACTTGCCTGAGTGTCCT | AACTCGGATGCTGAGTTGGA |
CL11775.Contig1_All | 55.90 | CAACGGCGGAATGAGCTAAA | TCTTGTGGTCCCTTCGTCAA |
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Bai, Y.; Jiang, L.; Li, Z.; Liu, S.; Hu, X.; Gao, F. Flavonoid Metabolism in Tetrastigma hemsleyanum Diels et Gilg Based on Metabolome Analysis and Transcriptome Sequencing. Molecules 2023, 28, 83. https://doi.org/10.3390/molecules28010083
Bai Y, Jiang L, Li Z, Liu S, Hu X, Gao F. Flavonoid Metabolism in Tetrastigma hemsleyanum Diels et Gilg Based on Metabolome Analysis and Transcriptome Sequencing. Molecules. 2023; 28(1):83. https://doi.org/10.3390/molecules28010083
Chicago/Turabian StyleBai, Yan, Lingtai Jiang, Zhe Li, Shouzan Liu, Xiaotian Hu, and Fei Gao. 2023. "Flavonoid Metabolism in Tetrastigma hemsleyanum Diels et Gilg Based on Metabolome Analysis and Transcriptome Sequencing" Molecules 28, no. 1: 83. https://doi.org/10.3390/molecules28010083