An induced hypersensitive-like response limits expression of foreign peptides via a recombinant TMV-based vector in a susceptible tobacco.
BACKGROUND
By using tobacco mosaic virus (TMV)-based vectors, foreign epitopes of the VP1 protein from food-and-month disease virus (FMDV) could be fused near to the C-terminus of the TMV coat protein (CP) and expressed at high levels in susceptible tobacco plants. Previously, we have shown that the recombinant TMV vaccines displaying FMDV VP1 epitopes could generate protection in guinea pigs and swine against the FMDV challenge. Recently, some recombinant TMV, such as TMVFN20 that contains an epitope FN20 from the FMDV VP1, were found to induce local necrotic lesions (LNL) on the inoculated leaves of a susceptible tobacco, Nicotiana tabacum Samsun nn. This hypersensitive-like response (HLR) blocked amplification of recombinant TMVFN20 in tobacco and limited the utility of recombinant TMV vaccines against FMDV.


RESULTS
Here we investigate the molecular mechanism of the HLR in the susceptible Samsun nn. Histochemical staining analyses show that these LNL are similar to those induced in a resistant tobacco Samsun NN inoculated with wild type (wt) TMV. The recombinant CP subunits are specifically related to the HLR. Interestingly, this HLR in Samsun nn (lacking the N/N'-gene) was able to be induced by the recombinant TMV at both 25°C and 33°C, whereas the hypersensitive response (HR) in the resistant tobacco plants induced by wt TMV through the N/N'-gene pathways only at a permissive temperature (below 30°C). Furthermore, we reported for the first time that some of defense response (DR)-related genes in tobacco were transcriptionally upregulated during HLR.


CONCLUSIONS
Unlike HR, HLR is induced in the susceptible tobacco through N/N'-gene independent pathways. Induction of the HLR is associated with the expression of the recombinant CP subunits and upregulation of the DR-related genes.
Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai, China.
Mangmang
M
Li
Ping
P
Li
Rentao
R
Song
Zhengkai
Z
Xu
D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D000939:Epitopes / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D005822:Genetic Vectors / Q000235:genetics; D054884:Host-Pathogen Interactions; D010935:Plant Diseases / Q000235:genetics* / Q000821:virology; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000235:genetics* / Q000502:physiology
D036022:Capsid Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; C442501:VP1 protein, Foot-and-mouth disease virus