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Published on: May 11, 2020
Expression of artificial microRNAs in transgenic Arabidopsis thaliana confers virus resistance
Qi-Wen Niu1, Shih-Shun Lin, Jose Luis Reyes
1Laboratory of Plant Molecular Biology, Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Abstract:
Plant microRNAs (miRNAs) regulate the abundance of target mRNAs by guiding their cleavage at the sequence complementary region. We have modified an Arabidopsis thaliana miR159 precursor to express artificial miRNAs (amiRNAs) targeting viral mRNA sequences encoding two gene silencing suppressors, P69 of turnip yellow mosaic virus (TYMV) and HC-Pro of turnip mosaic virus (TuMV). Production of these amiRNAs requires A. thaliana DICER-like protein 1. Transgenic A. thaliana plants expressing amiR-P69(159) and amiR-HC-Pro(159) are specifically resistant to TYMV and TuMV, respectively. Expression of amiR-TuCP(159) targeting TuMV coat protein sequences also confers specific TuMV resistance. However, transgenic plants that express both amiR-P69(159) and amiR-HC-Pro(159) from a dimeric pre-amiR-P69(159)/amiR-HC-Pro(159) transgene are resistant to both viruses. The virus resistance trait is displayed at the cell level and is hereditable. More important, the resistance trait is maintained at 15 degrees C, a temperature that compromises small interfering RNA-mediated gene silencing. The amiRNA-mediated approach should have broad applicability for engineering multiple virus resistance in crop plants.
Insights
Researchers engineered artificial microRNAs (amiRNAs) in Arabidopsis plants to confer resistance against specific plant viruses like turnip yellow mosaic virus (TYMV) and turnip mosaic virus (TuMV). This novel amiRNA approach offers broad applicability for developing virus-resistant crop plants.
Area of Science:
- Plant Molecular Biology
- Virology
- Biotechnology
Background:
- Plant microRNAs (miRNAs) are key regulators of gene expression, primarily by degrading target messenger RNAs (mRNAs).
- Viral gene silencing suppressors (e.g., P69, HC-Pro) are crucial for virus replication and spread in plants.
- Engineering plants with specific resistance mechanisms is vital for crop protection.
Purpose of the Study:
- To develop artificial miRNAs (amiRNAs) targeting viral suppressors and coat proteins for plant virus resistance.
- To assess the efficacy and heritability of amiRNA-mediated resistance in Arabidopsis thaliana.
- To investigate the stability of amiRNA-mediated resistance under varying temperature conditions.
Main Methods:
- Modification of an Arabidopsis thaliana miR159 precursor to generate amiRNAs targeting viral sequences.
- Generation of transgenic Arabidopsis thaliana expressing amiRNAs targeting TYMV (P69) and TuMV (HC-Pro, coat protein).
- Virus challenge assays and phenotypic evaluation of resistance in transgenic plants.
Main Results:
- Transgenic plants expressing amiR-P69(159) and amiR-HC-Pro(159) showed specific resistance to TYMV and TuMV, respectively.
- Plants expressing amiR-TuCP(159) were resistant to TuMV, and dual amiRNA expression conferred resistance to both viruses.
- Virus resistance was cell-level, heritable, and maintained at 15°C, a temperature inhibitory to RNA interference.
Conclusions:
- Artificial miRNAs can be effectively engineered to target and suppress viral gene expression in plants.
- The amiRNA-mediated approach provides specific and durable resistance against multiple plant viruses.
- This strategy holds significant potential for developing broad-spectrum virus resistance in economically important crop species.
