Expression of artificial microRNAs in transgenic Arabidopsis thaliana confers virus resistance

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...

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Bibliographic Details
Published in:	Nature biotechnology 2006-11, Vol.24 (11), p.1420-1428
Main Authors:	Chua, Nam-Hai, Niu, Qi-Wen, Lin, Shih-Shun, Reyes, Jose Luis, Chen, Kuan-Chun, Wu, Hui-Wen, Yeh, Shyi-Dong
Format:	Article
Language:	eng
Subjects:	Arabidopsis - genetics Arabidopsis - virology Arabidopsis thaliana Biological and medical sciences Biotechnology Botany Coat protein Disease resistance Fundamental and applied biological sciences. Psychology Gene Expression Profiling Gene sequencing Gene Silencing Genetic engineering Genetic technics Genetics Immunity, Innate - genetics Infections Methods. Procedures. Technologies MicroRNAs MicroRNAs - biosynthesis MicroRNAs - metabolism MicroRNAs - pharmacology miRNA Mosaics mRNA Plant Diseases - virology Plants, Genetically Modified - genetics Plants, Genetically Modified - virology Proteins Ribonucleic acid RNA RNA Interference RNA-mediated interference siRNA Suppressors Temperature Transgenic animals and transgenic plants Transgenic plants Turnip mosaic virus Turnip yellow mosaic virus Turnips Tymovirus - pathogenicity Viruses
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Summary:	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.
ISSN:	1087-0156 1546-1696