Combined BSA-Seq and RNA-Seq Reveal Genes Associated with the Visual Stay-Green of Maize (Zea mays L.)
Abstract
:1. Introduction
2. Results
2.1. Analysis of the Visual Stay-Green
2.2. QTL Mapping Using BSA
2.3. Transcriptome Sequencing Data Analysis
2.4. WGCNA of DEGs in Visual Stay-Green
2.5. Candidate Gene Analysis
3. Discussion
3.1. Excavation of Key Genes in the Stay-Green Mutant Contributes to the Promotion of Maize
3.2. Integrating BSA Data and RNA-seq Data for Candidate Gene Prediction
3.3. Chromosome 9 Is Crucial in Controlling Stay-Green-Associated Parameters
3.4. Candidate Gene Analysis
4. Materials and Methods
4.1. Plant Materials
4.2. Statistics of Visual Stay-Green
4.3. DNA Library Construction and BSA-Seq Analysis
4.4. RNA-Seq Analysis
4.5. WGCNA
4.6. qRT-PCR Validation of Genes
4.7. Combined Analysis of BSA-Seq and RNA-Seq
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Material | Number of Samples | Min (%) | Max (%) | Mean (%) | CV (%) | Kurt | Skew |
---|---|---|---|---|---|---|---|
F2 | 786 | 0.00 | 100.00 | 51.57 | 48.92 | −0.454 | −0.374 |
H-pool | 50 | 80.00 | 100.00 | 90.23 | 8.05 | 0.200 | −1.324 |
L-pool | 50 | 0.00 | 10.00 | 1.18 | - | 2.418 | 4.017 |
Associated Analysis Method | Chromosome | Start of Associated Regions (bp) | End of Associated Regions (bp) | Gene Number in the Associated Regions |
---|---|---|---|---|
Results of ED correlation | Chr9 | 21,439,311 | 97,423,834 | 841 |
Results of Ridit correlation | Chr9 | 20,699,310 | 98,235,998 | 895 |
Results of SmoothG correlation | Chr9 | 22,466,899 | 94,938,048 | 778 |
Results of SNP correlation | Chr9 | 20,964,571 | 98,003,843 | 878 |
The intersection of the four association results | Chr9 | 22,466,899 | 94,938,048 | 778 |
Gene | Annotation |
---|---|
Zm00001eb378880 * | superoxide dismutase |
Zm00001eb380180 | autophagy 2 |
Zm00001eb380210 | ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit |
Zm00001eb380300 | senescence-specific cysteine protease SAG12 |
Zm00001eb382070 | cryptochrome-2 |
Zm00001eb383680 * | phosphoenolpyruvate carboxylase |
Zm00001eb384100 * | cytochrome P450 monooxygenase |
Module | Gene | Annotation |
---|---|---|
green | Zm00001eb240100 | putative GATA transcription factor 22 isoform X1 |
brown | Zm00001eb384100 * | cytochrome P450 monooxygenase |
Zm00001eb405590 | NAC transcription factor 29 | |
Zm00001eb116960 | early light inducible protein1 | |
Zm00001eb118120 | agamous-like MADS-box protein AGL8 | |
pink | Zm00001eb383680 * | phosphoenolpyruvate carboxylase |
Zm00001eb378880 * | superoxide dismutase | |
salmon | Zm00001eb040160 | MYB transcription factor |
Zm00001eb411260 | diacylglycerol kinase 2 | |
Zm00001eb414910 | ZIM-transcription factor 40 | |
Zm00001eb332340 | MYB-RELATED transcription factor |
ID | Sense Primer | Anti-Sense Primer |
---|---|---|
Zm00001eb022220 | GGCCTCAATTCCGTCACTCA | CTTTTTAGCACTGCCGCGTT |
Zm00001eb031990 | AACGGTGTTCTACCCGATGC | TCCATTGGGCTCTTCCTTGG |
Zm00001eb051610 | AGGGATAGTGATGGGCGTCT | TGAGATTTCCTGACATAGCTTTAGA |
Zm00001eb074950 | CGCAGGGGTCTTGAGAGAAG | CTAGCTAGAGCTATCGCGGC |
Zm00001eb269130 | CCGAGAGGGACAGTCCAGTA | GTCGACAAGCTCACGCTTCA |
Zm00001eb377440 | GCGCGCGGGAAATTATAGTG | TGGCCTTCCTCACCTAGCTT |
Zm00001eb377990 | AACCTCGAGAAAGTCTGCGG | CTCAAAGCGATCCCAGGTGT |
Zm00001eb378520 | CGTCAACTGCAACGGGTG | AGCGACGACATGTTCAGAGC |
Zm00001eb380880 | GGCTATGGAAGATGACGCGA | CGACGTCATAAAGTACATAGCGA |
Zm00001eb381410 | TGGTGATGGAGGCTATGGGA | ACAAGGGAGAAGGGTCAACG |
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Zheng, R.; Deng, M.; Lv, D.; Tong, B.; Liu, Y.; Luo, H. Combined BSA-Seq and RNA-Seq Reveal Genes Associated with the Visual Stay-Green of Maize (Zea mays L.). Int. J. Mol. Sci. 2023, 24, 17617. https://doi.org/10.3390/ijms242417617
Zheng R, Deng M, Lv D, Tong B, Liu Y, Luo H. Combined BSA-Seq and RNA-Seq Reveal Genes Associated with the Visual Stay-Green of Maize (Zea mays L.). International Journal of Molecular Sciences. 2023; 24(24):17617. https://doi.org/10.3390/ijms242417617
Chicago/Turabian StyleZheng, Ran, Min Deng, Dan Lv, Bo Tong, Yuqing Liu, and Hongbing Luo. 2023. "Combined BSA-Seq and RNA-Seq Reveal Genes Associated with the Visual Stay-Green of Maize (Zea mays L.)" International Journal of Molecular Sciences 24, no. 24: 17617. https://doi.org/10.3390/ijms242417617