Post-transcriptional gene silencing in plum pox virus resistant transgenic European plum containing the plum pox potyvirus coat protein gene.
Transgenic plums containing the plum pox potyvirus coat protein (PPV-CP) gene were inoculated with PPV. Infection was monitored by evaluating symptoms, ELISA, and IC-RT-PCR. Transgenic clone C5 was highly resistant to PPV during four years of testing and displayed characteristics typical of post-transcriptional gene silencing (PTGS), including a high level of transgene transcription in the nucleus, low levels of transgene mRNA in the cytoplasm, a complex multicopy transgene insertion with aberrant copies, and methylation of the silenced PPV-CP transgene. The PPV-CP transgene was also methylated in seedlings of C5 and these seedlings were resistant to PPV. Our results show, for the first time, that PTGS functions as a mechanism for virus resistance in a woody perennial species.
Scorza
R
R
United States Department of Agriculture, Agricultural Research Service, Kearnevsville, WV 25430, USA. rscorza@afrs.ars.usda.gov
Callahan
A
A
Levy
L
L
Damsteegt
V
V
Webb
K
K
Ravelonandro
M
M
D015139:Blotting, Southern; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D004797:Enzyme-Linked Immunosorbent Assay; D005638:Fruit / Q000235:genetics* / Q000378:metabolism / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D020868:Gene Silencing; D005814:Genes, Viral / Q000235:genetics; D020022:Genetic Predisposition to Disease; D010935:Plant Diseases / Q000235:genetics / Q000821:virology; D030821:Plants, Genetically Modified; D017801:Plum Pox Virus / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*
D012333:RNA, Messenger