The protein-protein interaction of virus and host is essential for virus infection and host defense. The coat protein (CP) of tomato mosaic virus (ToMV) has been proved to be involved in cell-to-cell and long-distance movements of viruses that are presumably related with the protein-protein interactions. However, the host proteins that interact with the ToMV coat protein (ToCP) are largely unknown. In this study, we isolated a cDNA from a tobacco library through yeast two-hybrid system, which encodes a protein designated the ToMV CP-interacting protein-L (IP-L) that interacted with ToCP in vitro and in vivo. Sequencing analysis revealed that the putative coding region of IP-L gene was identical to that of an 'elicitor responsive protein' gene from N. tabacum (Genbank: #AB040409). A homology was also found between the cDNA sequence of IP-L and two senescence-related cDNAs (SENU1: Z75523 and AY479987) isolated from tomato and pepper. Northern blotting analysis showed that the mRNA level of IP-L was elevated after infection of ToMV. Then, we investigated the in vivo function of IP-L using virus-induced gene silencing (VIGS) and virus challenging assay. Semi-quantitative RT-PCR and Northern blotting results showed that the endogenous mRNA of IP-L in N. benthamiana plant was silenced at 10 days post inoculation with the in vitro transcripts of PVX-IP-L that were produced from the potato virus X (PVX)-based gene silencing plasmid pPC2S.IP-L. The IP-L silent plant developed a delayed systemic symptom at 7 days post challenging with ToMV, indicating that a high expression of IP-L was necessary for the interaction with ToCP to assist the viral transportation. Together, our data suggested that IP-L is a novel plant factor that interacts with the coat protein of ToMV and facilitates the long-distance movement of virus, which may provide a valuable clue for us to further investigate the mechanisms of plant virus infection and to control plant virus diseases.
-specific monoclonal antibodies did not recognize PVY-AGA. The ability of these new PVY recombinants to overcome resistance genes in potato producing mild or no symptoms, combined with the lack of serological reactivity towards at least one PVY N -specific antibody may present a significant threat posed by these isolates to seed potato production areas.
Potato virus Y (PVY) causes substantial losses in potato production by decreasing yields and affecting the quality of potato tubers. Management of PVY in potato is dependent primarily on potato seed certification programs to prevent or limit initial levels of virus inoculum. Prior to 1990, the ordinary strain of PVY (PVYO) was the predominant virus in North America. PVYO induces clear foliar symptoms in many potato cultivars, allowing successful management in seed potato through a combination of visual inspections and limited laboratory testing. In recent years, necrotic strains of PVY (PVYN, PVYNTN, and PVYN:O) have begun to spread in the United States, many of which induce mild symptoms in potato, making them more difficult to manage through visual inspections. In addition to reducing yield, necrotic isolates may also cause external and internal damage in tubers of susceptible cultivars, which is known as potato tuber necrotic ringspot disease (PTNRD). Tuber necrotic strains of PVY have been reported across the northern United States (1,2,4), although limited information is available on their incidence and spread in commercial potato production. During June and July of 2007, 38 random samples were collected from three different commercial fields displaying disease problems (cvs. Russet Ranger, Alturas, and Russet Burbank) in the vicinity of Idaho Falls, ID. Plants collected showed various degrees of mosaic and leaf yellowing. By using double-antibody sandwich (DAS)-ELISA and reverse transcription (RT)-PCR, 25 of these plants were identified as PVY positive. The mutiplex RT-PCR assay (3) confirmed that nine plants were infected with PVYNTN and 11 with PVYN:O. No RT-PCR products were amplified from five samples. During September and October of 2007, 25 tuber samples (cv. Russet Burbank) showing various degrees of unusual internal symptoms (e.g., brown spots) were collected near Idaho Falls, ID. Twenty-two tubers were found PVY positive by DAS-ELISA, and multiplex RT-PCR determined 13 of those were PVYNTN, three were PVYO, one was a PVYNTN/N:O mixture, and one was a PVYO/N:O mixture. No RT-PCR products were amplified from four samples. In October 2007, six tubers showing distinct external tuber damage characteristic of PTNRD (cv. Highland Russet) were collected near Twin Falls, ID. All six tubers were determined to be PVY positive by DAS-ELISA, and RT-PCR identified five as infected with PVYNTN and one with PVYN:O. All the mixtures were easily separated by inoculating tobacco plants followed by subsequent testing of individual plants. Asymptomatic tubers from the same lot not showing PTNRD damage were found PVY negative by DAS-ELISA and RT-PCR. All PVYNTN isolates collected during 2007 were inoculated into tobacco plants (Nicotiana tabacum L. cv. Xanthi) and confirmed to induce systemic vein necrosis. Limited sequencing of four of the PVYNTN isolates determined that they contained recombinant junctions 2 and 3, identifying them as being related to the European strain of PVYNTN (3). The data suggest an increase in distribution and incidence of necrotic strains of PVY in commercial, potato-production areas in Idaho during an outbreak in 2007 and the potential for an increase in PTNRD. References: (1) P. M. Baldauf et al. Plant Dis. 90:559, 2006. (2) J. M. Crosslin et al. Plant Dis. 90:1102, 2006. (3) J. H. Lorenzen et al. Plant Dis. 90:935, 2006. (4) L. M. Piche et al. Phytopathology 94:1368, 2004.
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