BnSGS3 Has Differential Effects on the Accumulation of CMV, ORMV and TuMV in Oilseed Rape
Abstract
:1. Introduction
2. Results
2.1. Sequence Comparison and Phylogenetic Analysis of SGS3 Genes in Different Hosts
2.2. BnSGS3 Expression in Transgenic and Non-Transgenic Plants of Oilseed Rape
2.3. Dynamics of BnSGS3 Expression in T0 Generation Transgenic and Non-Transgenic Plants of Oilseed Rape after Viral Infection
2.4. Effect of BnSGS3 on the Accumulation of ORMV, TuMV and CMV
2.5. Correlation between BnSGS3 Expression and Virus Accumulation
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Sequence Analysis of BnSGS3 and Vector Construction
Primer | Sequence (5′-3′) | Tm (°C) | Product (bp) |
---|---|---|---|
BnSGS3 cDNA amplification | |||
Bn-F1 | TGAGGTTCTGGACAGGGATC | 55 | 1974 |
Bn-R1 | GCCGTCTTACTGAAAATGGA | ||
Vector construction | |||
Bn-F2 | GTACTCGAGGTTTGCTCTCTGTTTGGTT | 55 | 1848 |
Bn-R2 | CGTCTCGAGGCTAGTAGTCTTCTGTGTC | ||
BnRI-C | TATACTCGAGCAGTGGAAGGGTTTGGGTG | 54 | 404 |
BnRI-KC | GCCCGAATTCTCCCTACTTGACAGTGTTG | ||
BnRI-A | CGAATCTAGACAGTGGAAGGGTTTGGGTG | 54 | 404 |
BnRI-E | GCCGAAGCTTTCCCTACTTGACAGTGTTG | ||
Detection of expression quantity | |||
Bn-F | TGGAAGGGTTTGGGTGAGGAG | 62 | 181 |
Bn-R | GTGGACCATAGGAGTGGCGTG | ||
Actin-F | CTGGAATTGCTGACCGTATGAG | 62 | 145 |
Actin-R | ATCTGTTGGAAAGTGCTGAGGG | ||
Bar-F | TGCCAGAAACCCACGTCAT | 55 | 485 |
Bar-R | CTGCACCATCGTCAACCACTA | ||
Detection of virus accumulation | |||
CMV-F | CCTCACCGGTACTGGTTTATC | 62 | 107 |
CMV-R | CTTTCGCATGTCGCCAATATC | ||
TuMV-F | GGAAGTAAACGCTGGAACCT | 62 | 96 |
TuMV-R | GCCACTCTTTGCTCGTATCT | ||
ORMV-F | CTGTGGCCATTAGGAGTCAA | 62 | 108 |
ORMV-R | GCGCAGTAGTCCAAGGTAATA |
4.3. Production of BnSGS3-Ov and BnSGS3-Si Brassica napus Transformants
4.4. Virus Purification and Inoculation
4.5. Sample Collection and RNA Extraction
4.6. Quantitative RT-PCR
4.7. Correlation Analysis of Virus Accumulation vs. BnSGS3 Expression
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflict of Interests
References
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Chen, Q.; Wang, J.; Hou, M.; Liu, S.; Huang, J.; Cai, L. BnSGS3 Has Differential Effects on the Accumulation of CMV, ORMV and TuMV in Oilseed Rape. Viruses 2015, 7, 4169-4185. https://doi.org/10.3390/v7082815
Chen Q, Wang J, Hou M, Liu S, Huang J, Cai L. BnSGS3 Has Differential Effects on the Accumulation of CMV, ORMV and TuMV in Oilseed Rape. Viruses. 2015; 7(8):4169-4185. https://doi.org/10.3390/v7082815
Chicago/Turabian StyleChen, Quan, Jie Wang, Mingsheng Hou, Shengyi Liu, Junyan Huang, and Li Cai. 2015. "BnSGS3 Has Differential Effects on the Accumulation of CMV, ORMV and TuMV in Oilseed Rape" Viruses 7, no. 8: 4169-4185. https://doi.org/10.3390/v7082815