Transcriptome and Metabolome Integration Reveals the Impact of Fungal Elicitors on Triterpene Accumulation in Sanghuangporus sanghuang
Abstract
:1. Introduction
2. Materials and Methods
2.1. Preparation of Fungal Elicitor
2.2. Strains and Cultivation
2.3. Fungal Elicitor Elicitation Test
2.4. Sample Preparation and Analysis of Metabolism
2.4.1. Specimen Preparation
2.4.2. Liquid Chromatography–Mass Spectrometry Analysis
2.4.3. Metabolomic Data Analysis
2.4.4. RNA Extraction, cDNA Library Construction, and Sequencing
2.4.5. Transcriptome and Metabolomic Assembly and Annotation
2.4.6. Expression of Triterpenoid Biosynthesis-Related Genes in S. sanghuang under Elicitor Treatment
2.5. Determination of Total Triterpene Content
2.6. Detection of NO Content
2.7. Statistical Analysis
3. Results
3.1. Identification of Metabolites
3.2. Variation in DAMs of Triterpenes
3.3. Filtering and Verification of Various Metabolites
3.4. Transcriptomic Analysis of S. sanghuang from ET and WET
3.5. Verification of Differentially Expressed Transcripts (DETs) by qRT-PCR
3.6. Correlation between DAMs and DETs
3.7. Effect of Mevalonate Pathways in ET Group
3.8. Detection of NO Content
3.9. Detection of Total Triterpene Content
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
CC | correlation coefficient |
CE | collision energy |
CMCs | cambial meristematic cells |
COGs | clusters of orthologous groups of proteins |
CUR | curtain gas |
DAMs | differentially accumulated metabolites |
DEGs | differentially expressed genes |
DETs | differentially expressed transcripts |
DP | de-clustering potential |
ESI | electrospray ionization |
ET | elicitor-treated |
GO | gene ontology |
GPI-APs | glycosylphosphatidylinositol-anchored proteins |
GSI | ion source gas I |
GSII | ion source gas I |
HPLC–MS/MS | high-performance liquid chromatography–tandem mass spectrometry |
IS | ion spray voltage |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
KOG | Eukaryotic Orthologous Groups |
LIT | linear ion trap |
MRM | multiple reaction monitoring |
NEB | NEBNext® Ultra™ RNA library prep kit |
NO | nitric oxide |
NOS | nitric oxide synthase |
Nr | non-redundant protein sequences |
OPLS-DA | orthogonal partial least-squares discriminant analysis |
PC1 | principal component 1 |
PC2 | principal component 2 |
PCA | principal component analysis |
QQQ | triple quadrupole |
QRT-PCR | quantitative real-time polymerase chain reaction |
Q TRAP | triple quadrupole linear ion trap mass spectrometer |
RNA-seq | RNA sequencing |
S. sanghuang | Sanghuangporus sanghuang |
SH-CL (ET) | elicitor-treated |
SH-CK (WET) | without elicitor treatment |
TIAs | terpenoid indole alkaloids |
UPLC–MS/MS | ultra-performance liquid chromatography–mass spectrometry |
UV | unit-variance scaling |
VIP | variable importance in projection |
WET | without elicitor treatment |
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Zhou, L.; Fu, Y.; Zhang, X.; Wang, T.; Wang, G.; Zhou, L.; Yu, H.; Tian, X. Transcriptome and Metabolome Integration Reveals the Impact of Fungal Elicitors on Triterpene Accumulation in Sanghuangporus sanghuang. J. Fungi 2023, 9, 604. https://doi.org/10.3390/jof9060604
Zhou L, Fu Y, Zhang X, Wang T, Wang G, Zhou L, Yu H, Tian X. Transcriptome and Metabolome Integration Reveals the Impact of Fungal Elicitors on Triterpene Accumulation in Sanghuangporus sanghuang. Journal of Fungi. 2023; 9(6):604. https://doi.org/10.3390/jof9060604
Chicago/Turabian StyleZhou, Linjiang, Yan Fu, Xinyuan Zhang, Tong Wang, Guangyuan Wang, Liwei Zhou, Hailong Yu, and Xuemei Tian. 2023. "Transcriptome and Metabolome Integration Reveals the Impact of Fungal Elicitors on Triterpene Accumulation in Sanghuangporus sanghuang" Journal of Fungi 9, no. 6: 604. https://doi.org/10.3390/jof9060604