The Polyamine Signaling Pathway in Response to Waterlogging Stress of Paeonia lactiflora
Abstract
:1. Introduction
- A preliminary analysis of the polyamine signaling pathway of herbaceous peony in response to waterlogging was carried out, and five key genes of the polyamine signaling pathway were screened: OR552626, OR552627, OR552628, OR552629, and OR552630. Functional validation analyses were performed on the five screened genes at a later stage;
- This study provides a basis for further in-depth research on the mechanism of waterlogging tolerance in herbaceous peony, selection and breeding of excellent waterlogging-tolerant varieties, and promotion of herbaceous peony.
2. Materials and Methods
2.1. Experimental Materials
2.2. Experimental Treatment and Sampling
2.3. Test Methods
2.3.1. Transcriptome Sequencing Analysis
- (1)
- Total RNA extraction from herbaceous peonies
- (2)
- Construction and Sequencing of cDNA Libraries
- (3)
- Fluorescence quantitative PCR expression validation
2.3.2. Determination of Endogenous Polyamine Content
2.3.3. Determination of Growth Hormone and Abscisic Acid Content
2.3.4. Data Processing and Analysis
3. Results and Analysis
3.1. Morphological Changes of Herbaceous Peony under Waterlogging Stress
3.2. Transcriptome Sequencing Analysis
3.2.1. Functional Annotation and Classification of Unigenes
- (1)
- Comparison of homologous functional genes between different species
- (2)
- KOG functional classification
- (3)
- GO functional analysis
- (4)
- Functional analysis of KEGG pathways
3.2.2. Screening and Analysis of Differentially Expressed Genes
3.2.3. GO Pathway Enrichment Analysis of Differentially Expressed Genes in Herbaceous Peony in Response to Waterlogging Stress
3.2.4. KEGG Pathway Enrichment Analysis of Differentially Expressed Genes in Paeonia in Response to Waterlogging Stress
3.2.5. Analysis of Key Differential Genes of the Polyamine Signaling Pathway in Herbaceous Peony
Expression Analysis of Key Differential Genes at Put Synthesis Stage and Quantitative Fluorescence Validation
Expression Analysis of Key Differential Genes in the Synthesis Stage of Spd and Spm and Quantitative Fluorescence Verification
3.2.6. qRT-PCR Validation of Differentially Expressed Genes of the Polyamine Synthesis Pathway
3.2.7. Changes in Endogenous Polyamine Content of Herbaceous Peony Leaves under Waterlogging Stress
3.2.8. Changes in Endogenous Hormone Content of Peony Leaves under Waterlogging Stress
4. Discussion
4.1. Effects of Waterlogging Stress on the Morphology of Herbaceous Peony
4.2. Analysis of Key Genes for Polyamine Signaling in Response to Waterlogging Stress in Herbaceous Peony
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
Appendix A
Synthesis Stage | Protease Name | GeneID | Description | ‘Lihong’ | ‘Qihualushuang’ | |||
---|---|---|---|---|---|---|---|---|
1-CK vs. 1-W | 4-CK vs. 4-W | 12-CK vs. 12-W | 1-CK vs. 1-W | 4-CK vs. 4-W | ||||
Putrescine synthesis | Ornithine decarboxylase | Unigene11573_All | Ornithine decarboxylase | 1.578 | 3.524 | —— | —— | 2.691 |
Arginine decarboxylase | Unigene17303_All | Vitis riparia arginine decarboxylase | —— | —— | —— | —— | 1.001 | |
N-carbamoyl-secantopamine amide hydrolase | CL9124.Contig11_All | Hypothetical predicted protein | —— | —— | 5.858 | —— | —— | |
CL9124.Contig12_All | −1.071 | —— | —— | —— | —— | |||
OR552626 | 2.170 | —— | —— | —— | —— | |||
CL95.Contig1_All | Bromodomain-containing protein 4 | —— | 1.229 | —— | —— | —— | ||
CL95.Contig22_All | —— | —— | 1.689 | —— | —— | |||
CL1627.Contig8_All | Uncharacterized protein | —— | —— | —— | 1.384 | —— | ||
CL280.Contig4_All | —— | —— | —— | −1.194 | —— | |||
CL280.Contig6_All | —— | —— | —— | −1.980 | −3.700 | |||
CL9124.Contig29_All | —— | —— | —— | —— | −1.321 | |||
CL95.Contig18_All | —— | —— | —— | —— | 1.121 | |||
Unigene22574_All | 1.478 | −7.098 | 6.129 | −6.919 | −5.170 | |||
OR552628 | −5.492 | −5.728 | 3.564 | —— | ||||
Unigene22576_All | —— | —— | 5.369 | 2.221 | —— | |||
Unigene22577_All | —— | —— | —— | −2.647 | —— | |||
Unigene22578_All | —— | —— | —— | —— | −5.358 | |||
Unigene22579_All | —— | —— | −1.311 | —— | —— | |||
Unigene22580_All | —— | —— | 2.150 | —— | —— | |||
Unigene29843_All | —— | —— | —— | −8.139 | —— | |||
Unigene3960_All | Synaptonemal complex protein | −1.266 | −1.000 | —— | —— | —— | ||
Unigene4939_All | Hypothetical protein | —— | −6.954 | —— | −7.516 | —— | ||
OR552627 | Transposon protein | −2.273 | —— | —— | —— | —— | ||
Spermidine, spermine synthesis | S-adenosylmethionine decarboxylase | CL2592.Contig2_All | S-adenosylmethionine Decarboxylase proenzyme | 1.513 | ||||
CL2592.Contig3_All | 1.181 | |||||||
Unigene1124_All | 2.329 | |||||||
Spermidine synthase | CL615.Contig2_All | Pistacia vera spermidine synthase 1 | —— | —— | 1.466 | —— | —— | |
CL615.Contig4_All | −4.322 | —— | —— | −4.392 | 5.209 | |||
CL615.Contig6_All | −1.224 | −7.248 | 5.672 | —— | —— | |||
CL615.Contig8_All | —— | 5.585 | —— | −2.939 | —— | |||
CL615.Contig15_All | —— | −1.722 | —— | —— | —— | |||
CL615.Contig16_All | —— | —— | —— | 2.295 | 2.147 | |||
CL615.Contig21_All | —— | —— | −2.778 | —— | —— | |||
CL615.Contig22_All | 4.524 | —— | —— | 2.283 | −4.392 | |||
CL615.Contig23_All | —— | −5.523 | —— | −5.644 | —— | |||
CL615.Contig25_All | −1.295 | 1.924 | —— | —— | —— | |||
CL3232.Contig1_All | Vitis vinifera spermine synthase | —— | −1.150 | —— | −2.296 | —— | ||
CL3232.Contig2_All | —— | —— | —— | −1.683 | —— | |||
CL3232.Contig3_All | Runus pseudocerasus Spermine synthase gene | —— | —— | —— | −2.145 | —— | ||
OR552630 | Vitis riparia spermidine synthase 2 | −1.523 | −1.322 | —— | —— | −1.170 | ||
Unigene3485_All | Ziziphus jujuba spermine synthase | —— | —— | —— | −2.076 | −0.263 | ||
Unigene6488_All | Spermidine synthase 1 | —— | −4.248 | —— | —— | 5.209 | ||
Unigene23250_All | —— | —— | —— | −1.391 | —— | |||
Unigene26129_All | Durio zibethinus Spermidine synthase-like | 1.021 | —— | —— | —— | —— | ||
Unigene32162_All | —— | —— | 5.209 | —— | —— | |||
Polyamine catabolism | Polyamine oxidase | Unigene1094_All | Probable polyamine oxidase 5 | —— | —— | —— | 2.044 | —— |
OR552629 | Polyamine oxidase 5 | —— | —— | —— | 2.770 | —— | ||
CL7378.Contig2_All | Polyamine oxidase 4 | —— | —— | —— | 2.517 | —— | ||
CL6652.Contig6_All | Primary amine oxidase 2 | —— | —— | —— | 2.056 | —— |
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Gene Landing Number | Functional Notes | Primer (5′–3′) |
---|---|---|
Actin | Internal reference gene | F: TTATGCCCTTCCTCACGCTATC |
R: GAGCTGCTTTTGGAAGTCTCCA | ||
OR552630 | Spermidine synthase | F: GTTCAACATGCTTCTACGCCTACC |
R: CTCATCACATTCACAGCCACCTC | ||
OR552629 | Polyamine oxidase | F: GCCTTGCGATTTGATAATGTGTTC |
R: ACCTTCCAGCAGCCATATAGAC | ||
OR552626 | Hypothetical predicted protein | F: CGGACGGAAACGGATTCAAAG |
R: CCATGACATGACACCAATAAGAGG | ||
OR552627 | Transposon protein | F: TGGAGGTTGACATGGGTATCAC |
R: TTCAGATGTAGGCTCGGATGG | ||
OR552628 | Uncharacterized protein | F: TCTTGTGAAACTGGGACCTGATTG |
R: CAACCATACTTCCTACCGCATCTT |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
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Share and Cite
Shi, Y.; Lv, M.; Liu, Z.; Yang, X.; Yang, L.; Dong, L.; Lei, F.; Xie, A.; Zhang, D.; Bao, M.; et al. The Polyamine Signaling Pathway in Response to Waterlogging Stress of Paeonia lactiflora. Horticulturae 2024, 10, 928. https://doi.org/10.3390/horticulturae10090928
Shi Y, Lv M, Liu Z, Yang X, Yang L, Dong L, Lei F, Xie A, Zhang D, Bao M, et al. The Polyamine Signaling Pathway in Response to Waterlogging Stress of Paeonia lactiflora. Horticulturae. 2024; 10(9):928. https://doi.org/10.3390/horticulturae10090928
Chicago/Turabian StyleShi, Yajie, Mengwen Lv, Zemiao Liu, Xiao Yang, Lijin Yang, Lingling Dong, Fuling Lei, Anqi Xie, Dongliang Zhang, Mingyue Bao, and et al. 2024. "The Polyamine Signaling Pathway in Response to Waterlogging Stress of Paeonia lactiflora" Horticulturae 10, no. 9: 928. https://doi.org/10.3390/horticulturae10090928