Transfection of whole plants from wounds inoculated with Agrobacterium tumefaciens containing cDNA of tobacco mosaic virus.
We engineered cDNA of tobacco mosaic tobamovirus (TMV) into Agrobacterium tumefaciens for inoculation of plant cells. The resulting bacterial strains were used to transfect tobacco (Nicotiana tabacum cv. Xanthi and Xanthi/nc) with wild type and a defective virus. Lesion formation on Xanthi/nc tobacco was used to measure the timing and efficiency of transfection. Infections mediated by Agrobacterium produced lesions an average of two days later than infections produced by inoculation with virions. The addition of approximately 80 bp of non-viral sequences to the 5'-end of TMV transcripts abolished transfection. Transcripts with non-viral sequences at the 3'-end initiated infections, while precise transcript termination with a synthetic ribozyme sequence increased transfection frequencies two-fold. Culture conditions reported to induce genes of the vir region of the Agrobacterium Ti plasmid also increased the transfection frequency approximately two-fold. Therefore, in addition to the pararetroviruses and geminiviruses previously described, 'agroinoculation' may be used to infect plants with plus-sense RNA viruses.
Turpen
T H
TH
Biosource Genetics Corporation, Vacaville, CA 95688.
Turpen
A M
AM
Weinzettl
N
N
Kumagai
M H
MH
Dawson
W O
WO
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D010957:Plasmids / Q000235:genetics; D016337:RNA, Catalytic / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000382:microbiology; D014027:Tobacco Mosaic Virus / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D037521:Virulence Factors
D001426:Bacterial Proteins; D016337:RNA, Catalytic; D012367:RNA, Viral; D037521:Virulence Factors