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 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.
Niu
Qi-Wen
QW
Laboratory of Plant Molecular Biology, Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Lin
Shih-Shun
SS
Reyes
Jose Luis
JL
Chen
Kuan-Chun
KC
Wu
Hui-Wen
HW
Yeh
Shyi-Dong
SD
Chua
Nam-Hai
NH
D017360:Arabidopsis / Q000235:genetics* / Q000821:virology*; D020869:Gene Expression Profiling; D020868:Gene Silencing; D007113:Immunity, Innate / Q000235:genetics; D035683:MicroRNAs / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology; D010935:Plant Diseases / Q000821:virology; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000821:virology; D034622:RNA Interference; D013696:Temperature; D017806:Tymovirus / Q000472:pathogenicity*
D035683:MicroRNAs