Expression of cowpea mosaic virus coat protein precursor in transgenic tobacco plants

Nida, Debbie L.; Anjos, J. R. N. dos; Lomonossoff, George P.; Ghabrial, Said A.

doi:10.1099/0022-1317-73-1-157

Cited by 22 publications

(7 citation statements)

References 37 publications

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“…Notable for its failure, tobaccos transformed with the 60-kDa CP precursor of cowpea mosaic comovirus were not protected (even locally) against cowpea mosaic comovirus inoculation, the precursor was not cleaved, and no virus-like capsids were seen (67).…”

Section: On Cp-mediated Protection and Proposed Mechanismsmentioning

confidence: 99%

Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms.

Wilson¹

1993

Proc. Natl. Acad. Sci. U.S.A.

230

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Since 1986, the ability to confer resistance against an otherwise devastating virus by introducing a single pathogen-derived or virus-targeted sequence into the DNA of a potential host plant has had a marked influence on much of the research effort, focus, and shortterm objectives of plant virologists throughout the world. The vast literature on coat protein-mediated protection, for example, attests to our fascination for unraveling fundamental molecular mechanism(s), our (vain) Despite extensive and sometimes elegant experimentation, the molecular mechanism(s) of this viral "crossprotection" have remained elusive and controversial. In some cases, the coat protein (CP) of the protectant virus was thought to be primarily responsible, either by preventing particle disassembly or by re-encapsidating the incoming genome of the more severe challenge virus. However, viroids [240-to 380-nt-long, naked circular single-stranded (ss)RNA pathogens] and mutant viruses making assembly-defective or no detectable CP could also cross-protect against their more severe relatives. Such observations prompted models based on inhibitory interactions between sense and antisense RNAs or between the replicational machineries ofthe two competing pathogens (6). Controversy arose largely because available molecular technology could not resolve which regulatory or coding sequence(s) or polypeptide product(s) from the actively replicating genome of the primary (protectant) pathogen were responsible for interfering with the many replicative processes essential to establish infection by the secondary, related, and more severe virus. In contrast, multiple infections by unrelated viruses sharing a common host are very prevalent in nature. Many aspects of this older story have their parallels in current hypotheses and lack of a unified model for pathogenderived resistance in transgenic plants.The advent of improved cell and tissue culture techniques, efficient protocols for Agrobacterium tumefaciens-mediated transformation and plantlet regeneration in dicotyledonous species (7) [and more recent methods for monocot crops (8-10)], has permitted, among other applications (11,12), the theory of pathogenderived virus resistance to be tested in practice.Collaboration between researchers at Monsanto and Washington University (St. Louis) led to the first report of CPmediated protection (CPMP) against tobacco mosaic virus (TMV) in tobacco in 1986 (4). Since then, CPMP has been reported for over 20 viruses in at least 10 different taxonomic groups, in a wide 3134 variety of dicot plant species, and the list is increasing rapidly.

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Section: On Cp-mediated Protection and Proposed Mechanismsmentioning

confidence: 99%

Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms.

Wilson¹

1993

Proc. Natl. Acad. Sci. U.S.A.

230

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“…Recently, Nida et al [25] made transgenic tobacco plants expressing the 60 k coat proteins-precursor (CP-P) of CPMV. Because CPMV neither induces symptoms in tobacco nor moves systemically, they evaluated the reaction of the transgenic plants of CPMV inoculation based on virus accumulation in the inoculated leaves.…”

Section: Expectations From the Expression Of Sqmv Coat Protein Genesmentioning

confidence: 99%

The coat protein genes of squash mosaic virus: cloning, sequence analysis, and expression in tobacco protoplasts

Pang

Nagpala

et al. 1993

Archives of Virology

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Complementary DNA of the middle-component RNA of the melon strain of squash mosaic comovirus (SqMV) was cloned. Clones containing the coat protein genes were identified by hybridization with a degenerate oligonucleotide synthesized according to the amino acid sequence of a purified peptide fragment of the SqMV large coat protein. A clone containing of 2.5 kbp cDNA insert of SqMV M-RNA was sequenced. The total insert sequence of 2510 bp included a 2373 bp open reading frame (ORF) (encoding 791 amino acids), a 123 bp 3'-untranslated region, and a poly(A) region. This ORF is capable of encoding both the 42 and 22 k SqMV coat proteins. Direct N-terminal sequence analysis of the 22 k coat protein revealed its presence at the 3' end of this ORF and the position of the proteolytic cleavage site (Q/S) used to separate the large and small coat proteins from each other. A putative location of the N-terminal proteolytic cleavage site of the 42 k coat protein (Q/N) was predicted by comparisons with the corresponding coat proteins of cowpea mosaic virus, red clover mottle virus, and bean-pod mottle virus. Although the available nucleotide sequences of these viruses revealed little similarity, their encoded coat proteins shared about 47% identity. The identity of the encoded 42 k and 22 k peptides was confirmed by engineering the respective gene regions for expression followed by transfer into tobacco protoplasts using the polyethylene glycol method. Both SqMV coat proteins were expressed in vivo as determined by their reactivity to SqMV coat protein specific antibodies.

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“…Viral coat proteins (CPs) have been used widely to enhance the tolerance levels of different plant species to viral diseases; these plants include tobacco (Nida et al . 1992), tomato (Zrachya et al .…”

Section: Introductionmentioning

confidence: 99%

“…Viral coat proteins (CPs) have been used widely to enhance the tolerance levels of different plant species to viral diseases; these plants include tobacco (Nida et al 1992), tomato (Zrachya et al 2007), orange (Iwanami and Shimizu 2004), papaya (Krubphachaya et al 2007), cantaloupe (Clough and Hamm 1995), melon (Fuchs et al 1998), grapevine (Maghuly et al 2006), sweet orange (Zanek et al 2008), soybean (Tougou et al 2006), squash (Klas et al 2006), sugarcane (Jin et al 2007) and watermelon rootstock (Park et al 2005). Among these genetically modified (GM) plants, strains tolerant to papaya ring and watermelon mosaic virus squash have been successfully commercialized.…”

Section: Introductionmentioning

confidence: 99%

Assessment of the Cucumber Mosaic Virus Coat Protein by Expression Evaluation in a Genetically Modified Pepper and Escherichia Coli Bl21

Bang¹,

Jung²,

Eom

et al. 2011

Journal of Food Biochemistry

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A genetically modified (GM) pepper containing the gene for the cucumber mosaic virus (CMV) coat protein (CP) was investigated to assess the proteolytic behavior and simulated digestion in the stomach and small intestine. CMV‐CP expressed in GM pepper and in Escherichia coli exhibited the molecular mass of about 25 kDa on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. The three internal peptides of CMV‐CP in E. coli BL21 (VLSQQLSR, LLLPDSVTEYDKK and DDALETDELVLHVDIEHQR) were detected by electrospray ionization mass spectrometry/mass spectrometry, and seven peptides (VLSQQLSR, PTINHPTFVGSER, LLLPDSVTEYDKK, FDSTVWVTVRK, LLYDLSAMRA, RKYAVLVYSK and VLHVDIEHQR) of CMV‐CP in GM pepper were verified by the analysis. CMV‐CP expressed in GM pepper was completely degraded in simulated gastric and intestinal fluids, and the immunoreaction of the CMV‐CP was rapidly eliminated by the Western blot assays. PRACTICAL APPLICATIONS A variety of cucumber mosaic virus (CMV‐P0)‐resistant peppers was developed through a breeding program, and commercial cultivars are available. However, a new pathotype virus (CMV‐P1), which can infect CMV‐P0‐resistant peppers, is appeared recently, and this virus causes a heavy loss of productivity. Recently developed new transgenic pepper line is highly tolerable to CMV‐P0 and CMV‐P1. Sequence analysis of the CMV‐coat protein (CMV‐CP) expressed in the new transgenic pepper line and in an Escherichia coli expression system will provide structural information on expressed protein. In addition, in vitro digestibility of the expressed CMV‐CP from transgenic pepper with simulated gastric fluid and simulated intestinal fluid make it possible to understand its proteolytic behavior with simulated digestion fluids.

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Expression of cowpea mosaic virus coat protein precursor in transgenic tobacco plants

Cited by 22 publications

References 37 publications

Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms.

Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms.

The coat protein genes of squash mosaic virus: cloning, sequence analysis, and expression in tobacco protoplasts

Assessment of the Cucumber Mosaic Virus Coat Protein by Expression Evaluation in a Genetically Modified Pepper and Escherichia Coli Bl21

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