A Gain-of-Function Mutant of IAA7 Inhibits Stem Elongation by Transcriptional Repression of EXPA5 Genes in Brassica napus
Abstract
:1. Introduction
2. Results
2.1. Morphology and Agronomic Traits of ndf-2
2.2. Aux/IAA-Mediated Auxin Signaling Is Involved in Dwarfism Formation of ndf-2
2.3. The Transcription of BnaEXPA5 Is Regulated by Aux/IAA Mediated Signaling Pathway
2.4. BnaARF6/BnaARF8 Protein Interacts with BnaA03.IAA7 Protein and Binds to AuxRE on the Promoter of BnaEXPA5 Gene
2.5. Overexpression of BnaEXPA5 Resulted in Longer Stems and Larger Leaves in the Transformed Plants
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Growth Conditions
4.2. Genetic Analysis
4.3. Mapping and Identification of ndf-2 Gene
- the SNPs and InDels are heterozygous in wild-type parent and WT-pool;
- the SNPs and InDels are inconsistent between wild-type parent and WT-pool;
- the SNPs and Indels are the same between Df-pool and WT-pool;
- the SNPs and Indels are heterozygous in mutant parent.
4.4. Transcriptome Analysis
4.5. Fluorescent Quantitative Analysis
4.6. Yeast Two-Hybrid Screening Assay
4.7. Hormone Treatment
4.8. Y1H Assays
4.9. Development of AS-PCR Markers
4.10. Scanning Electron Microscopy
4.11. Protoplast Transformation and Transient Expression Assays
4.12. Transgenic Arabidopsis Thaliana
4.13. Data Statistics and Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ARF | Auxin response factor |
Aux/IAA | Auxin/Indole-3-Acetic Acid |
BC | Back cross |
BR | Brassinosteroid |
BSA-seq | Bulked Segregant Analysis-sequencing |
DEGs | differential expression genes |
DES | Diethyl sulfate |
DH | Double haploid |
FPKM | Fragments Per Kilobase of exon model per Million mapped fragments |
GA | Gibberellin |
IAA | Indole-3-Acetic |
InDel | Insertion-deletion |
NPA | 1-naphthylphthalamic acid |
SNP | Single nucleotide polymorphism |
EXP | EXPANSIN |
qRT-PCR | Quantitative reverse transcription PCR |
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Line | Height (cm) | PPEB (cm) | LMI (cm) | NPB | NSB | NPPP | NSPP | WS (g) | YPP (g) |
---|---|---|---|---|---|---|---|---|---|
ndf-1 | 71.4 ± 3.3 | 5.8 ± 2.7 | 8.2 ± 3.2 | 6.6 ± 2.4 | 5.5 ± 2.2 | 237.6 ± 55.7 | 11.8 ± 3.7 | 3.6 ± 0.2 | 10.3 ± 2.6 |
ndf-2 | 86.8 ± 4.9 | 7.1 ± 2.3 | 11.0 ± 2.2 | 7.1 ± 1.8 | 5.8 ± 1.7 | 256.1 ± 41.4 | 10.6 ± 4.1 | 3.5 ± 0.3 | 12.9 ± 3.5 |
3529 (ck) | 198.7 ± 15.9 | 22.8 ± 5.2 | 67.8 ± 8.4 | 9.1 ± 2.3 | 14.7 ± 1.3 | 388.0 ± 35.8 | 17.5 ± 2.9 | 3.4 ± 0.2 | 23.4 ± 4.3 |
Population | Number of Plants (Height Range) | Segregation Ratio | Expected Ratio | χ2 | p | ||
---|---|---|---|---|---|---|---|
Tall | Semi-Dwarf | Dwarf | |||||
3529 | 60 (172–213 cm) | 60 | 0 | 0 | 1:0:0 | - | - |
ndf-2 | 60 (73–95 cm) | 0 | 0 | 60 | 0:0:1 | - | - |
F1 | 56 (121–167 cm) | 0 | 56 | 0 | 0:1:0 | - | - |
F2 | 576 (47–219 cm) | 139 (≥170 cm) | 268 (110–170 cm) | 159 (≤110 cm) | 1:2:1 | 3.125 | >0.05 |
B11 (F1 × 3529) | 164 (115–215 cm) | 73 (≥170 cm) | 91 (110–170 cm) | 0 (≤110 cm) | 1:1:0 | 1.97 | >0.05 |
B12 (F1 × ndf-2) | 234 (56–164 cm) | 0 (≥170 cm) | 127 (110–170 cm) | 107 (≤110 cm) | 0:1:1 | 1.71 | >0.05 |
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Wei, T.; Zhang, L.; Zhu, R.; Jiang, X.; Yue, C.; Su, Y.; Ren, H.; Wang, M. A Gain-of-Function Mutant of IAA7 Inhibits Stem Elongation by Transcriptional Repression of EXPA5 Genes in Brassica napus. Int. J. Mol. Sci. 2021, 22, 9018. https://doi.org/10.3390/ijms22169018
Wei T, Zhang L, Zhu R, Jiang X, Yue C, Su Y, Ren H, Wang M. A Gain-of-Function Mutant of IAA7 Inhibits Stem Elongation by Transcriptional Repression of EXPA5 Genes in Brassica napus. International Journal of Molecular Sciences. 2021; 22(16):9018. https://doi.org/10.3390/ijms22169018
Chicago/Turabian StyleWei, Tao, Li Zhang, Ruijia Zhu, Xuefei Jiang, Chu Yue, Ying Su, Hongpei Ren, and Maolin Wang. 2021. "A Gain-of-Function Mutant of IAA7 Inhibits Stem Elongation by Transcriptional Repression of EXPA5 Genes in Brassica napus" International Journal of Molecular Sciences 22, no. 16: 9018. https://doi.org/10.3390/ijms22169018