Tobacco Rattle Virus as a Tool for Rapid Reverse-Genetics Screens and Analysis of Gene Function in Cannabis sativa L.
Abstract
:1. Introduction
2. Results and Discussion
2.1. VIGS in Cannabis Using a TRV Vector
2.2. VAGE in Cannabis Using a TRV Vector
3. Materials and Methods
3.1. Plant Material
3.2. Construction of pTRV2 Vectors
3.3. Agro-Inoculation of pTRV Vectors
3.4. RNA Extraction, cDNA Synthesis, and Quantitative Real-Time PCR (qRT)
3.5. Imaging
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer Number | Product | Forward Primer | Reverse Primer |
---|---|---|---|
1 | PDS424 | 5′-AATTCTCGAGCTTCAGCTCCCACCAGAGTC-3’ | 5’-ATTCTAGATCACCGTCATCATCTTTCCA-3’ |
2 | PDS486 | 5’-AATTCTCGAGACTGGAAAGAGATTCCGTATTTCA-3’ | 5’-ATTCTAGAACAAAACCGCACCTTCCAT-3’ |
3 | TRV2-CP | 5′-ACGATTCTTGGGTGGAATCA-3′ | 5′-TCGTAACCGTTGTGTTTGGA-3′ |
4 | TRV2-DsRed | 5′-ACGATTCTTGGGTGGAATCA-3′ | 5′-CCCATGGTCTTCTTCTGCAT-3′ |
5 | PDS | 5′-ACTGTTCCTGATTGCGAACC-3′ | 5′-CTCGGCCAAAATTCTCTGAC-3′ |
6 | UBQ5 | 5′-AAGCTCGCTCTTCTCCAGTTC-3′ | 5′-CACACTTGCCGCAGTAATGTC-3′ |
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Alter, H.; Peer, R.; Dombrovsky, A.; Flaishman, M.; Spitzer-Rimon, B. Tobacco Rattle Virus as a Tool for Rapid Reverse-Genetics Screens and Analysis of Gene Function in Cannabis sativa L. Plants 2022, 11, 327. https://doi.org/10.3390/plants11030327
Alter H, Peer R, Dombrovsky A, Flaishman M, Spitzer-Rimon B. Tobacco Rattle Virus as a Tool for Rapid Reverse-Genetics Screens and Analysis of Gene Function in Cannabis sativa L. Plants. 2022; 11(3):327. https://doi.org/10.3390/plants11030327
Chicago/Turabian StyleAlter, Hanan, Reut Peer, Aviv Dombrovsky, Moshe Flaishman, and Ben Spitzer-Rimon. 2022. "Tobacco Rattle Virus as a Tool for Rapid Reverse-Genetics Screens and Analysis of Gene Function in Cannabis sativa L." Plants 11, no. 3: 327. https://doi.org/10.3390/plants11030327