F-Box Family Genes, LTSF1 and LTSF2, Regulate Low-Temperature Stress Tolerance in Pepper (Capsicum chinense)
Abstract
:1. Introduction
2. Results
2.1. LTSF1 and LTSF2 Sequence Analysis
2.2. VIGS Analysis
2.3. LTSF1 and LTSF2 Interaction with SKP1
2.4. The Transgenic Expression of LTSF1 Promotes Low-Temperature Stress Tolerance in N. benthamiana Plants
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Phylogenetic Analysis and Identification of Conserved Motifs
4.3. VIGS
4.4. Subcellular Localization of the LTSF1 and LTSF2 Proteins
4.5. Y2H Assays
4.6. Bimolecular Fluorescence Complementation (BiFC) Analysis
4.7. Genomic DNA Extraction
4.8. Agrobacterium-Mediated Transformation of N. benthamiana
4.9. Low-Temperature Stress Treatments
4.10. Total RNA Extraction and Reverse-Transcription Quantitative PCR (RT-qPCR)
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Primer Name | Sequence | Purpose |
---|---|---|
Y2H: LTSF1/2: NdeF1 | GAGACATATGATGCCTGTCAAAGTAGCA | Y2H vector cloning |
Y2H: LTSF1/2: BamR1 | GAGAGGATCCTTAGATAACTAATTTTGGAGA | Y2H vector cloning |
Y2H: LTSF1/2: NdeF2 | GAGACATATGATAGATTATGATCAGCAGGCAAT | Y2H vector cloning |
Y2H: LTSF1/2: BamR2 | GAGAGGATCCCTATACATCCTCAAAAGTGGATA | Y2H vector cloning |
Y2H: LTSF1/2: NdeF3 | TATACATATGGTACAACAACTCGATCCCCCT | Y2H vector cloning |
SKP1-F-EcoRI | TCGCGAATTCATGTCTGCCCCAAAGAAAAT | Y2H vector cloning |
SKP1-R-BamHI | TATAGGATCCTCACTCAAAGGCCCAAGCAT | Y2H vector cloning |
SKP1:F-BamHI | CAATGGATCCATGTCTGCCCCAAAGAAAAT | BiFC vector cloning |
SKP1:R-XhoI | ATATCTCGAGCTCAAAGGCCCAAGCA | BiFC vector cloning |
LTSF1/2-F-BamHI | ACAGGGATCCATGCCTGTCAAAGTAGCA | BiFC vector cloning |
LTSF1/2-R-XhoI | ACAGCTCGAGGATAACTAATTTTGGAGAA | BiFC vector cloning |
VIGS-LTSF1UT-1F | CGACGACAAGACCCTAGGTGTTATTTTTGTCCTTTCC | TRV2 vector cloning |
VIGS-LTSF1UT-1R | GAGGAGAAGAGCCCTCTGCAAACTCACTGTACAATTTAG | TRV2 vector cloning |
VIGS-LTSF2UT-1F | CGACGACAAGACCCTGAAAAATGATTTTATGATATC | TRV2 vector cloning |
VIGS-LTSF2UT-1R | GAGGAGAAGAGCCCTCAATGTATAATTTAGTCCACTGAAAAT | TRV2 vector cloning |
VIGS-LTSF1/2-1F | CGACGACAAGACCCTTCCAAAACTTGGGAG | TRV2 vector cloning |
VIGS-LTSF1/2-1R | GAGGAGAAGAGCCCTGCGGTTTCTCTTGTTAAGGGGT | TRV2 vector cloning |
TRV2:LIC-F | TGTTACTCAAGGAAGCACGATGAGCT | LIC cloning confirmation |
TRV2:LIC-R | CAGGCACGGATCTACTTAAAGAACGTAG | LIC cloning confirmation |
LTSF1/2.Spe–F | GCTCACTAGTATGCCTGTCAAAGTAGCA | pMDC83 vector cloning/Transgene confirmation |
LTSF1.Asc–R | AATATTGGCGCGCCAGATAACTAATTTTGGAGAA | |
LTSF2.Asc–R | AATATTGGCGCGCCAAATAACTAATTTTGGAGAA | |
Hpt-F | CCTGAACTCACCGCGACG | |
Hpt-R | AAGACCAATGCGGAGCATAT |
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Venkatesh, J.; Kang, M.-Y.; Liu, L.; Kwon, J.-K.; Kang, B.-C. F-Box Family Genes, LTSF1 and LTSF2, Regulate Low-Temperature Stress Tolerance in Pepper (Capsicum chinense). Plants 2020, 9, 1186. https://doi.org/10.3390/plants9091186
Venkatesh J, Kang M-Y, Liu L, Kwon J-K, Kang B-C. F-Box Family Genes, LTSF1 and LTSF2, Regulate Low-Temperature Stress Tolerance in Pepper (Capsicum chinense). Plants. 2020; 9(9):1186. https://doi.org/10.3390/plants9091186
Chicago/Turabian StyleVenkatesh, Jelli, Min-Young Kang, Li Liu, Jin-Kyung Kwon, and Byoung-Cheorl Kang. 2020. "F-Box Family Genes, LTSF1 and LTSF2, Regulate Low-Temperature Stress Tolerance in Pepper (Capsicum chinense)" Plants 9, no. 9: 1186. https://doi.org/10.3390/plants9091186