Nucleotide Sequence of the Genomic RNA of Pepper Mild Mottle Virus, a Resistance-breaking Tobamovirus in Pepper

AbstractThe PCR amplification of majority of the ssRNA of both genomic and non-genomic mRNA is accomplished by RT-PCR. The mRNA is subjected to cDNA synthesis using reverse transcriptase and either Oligo(dT)18, or random or gene specific reverse primers based priming strategies. The choice of primer largely depends on the nature of 3 prime terminus of mRNA and length of cDNA synthesized. In general, oligo(dT)18 is the preferred choice for mRNAs having poly(A) tail at 3 prime terminus. In general, tobamoviruses lack any poly(A) tail at their 3 prime untranslated region (UTR) which forms a tRNA like structure and upstream pseudoknot domain except tow viruses viz., Hibiscus latent Fort Pierce virus (HLFPV) and Hibiscus latent Singapore virus (HLSV) which accommodate internal poly(A) sequences of 46 and 87 nucleotides long, respectively in their 3 prime UTR. However, determination of full nucleotide sequence of Pepper mild mottle virus (PMMoV) using an oligo(dT)18 primed cDNA as template indicated the libertinism of oligo(dT)18 in priming cDNA synthesis of RNA template which are known to lack poly(A) tail. at the end or internally at its 3 prime end. Moreover, coat protein (CP) gene of PMMoV and bean common mosaic virus (BCMV) (Potyvirus with a poly(A) tract at its 3 prime end) was amplified using cDNA primed with random primer as well as oligo(dT)18 was successfully amplified but with significant variation in the intensity of the amplification band in case of PMMoV but not in BCMV. This clearly indicated the presence of PMMoV mRNA with polyadenylated 3 prime tail in total RNA isolated from PMMoV infected capsicum leaves with abundance of non-polyadenylated PMMoV genomic RNA (gRNA). Hence, we hypothesize that the generation of polyadenylated RNA population during the infection cycle of PMMoV in pepper may be possible reason for libertinism of oligo(dT)18 in priming cDNA synthesis of RNA template isolated from PMMoV infected leaves followed by amplification of entire PMMoV genome through RT-PCR. This is first study indicating the presence of polyadenylated or polyadenylated rich regions in PMMoV gRNA acquired during the infection cycle in pepper.

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A single amino acid substitution in 126-kDa protein of Pepper mild mottle virus associates with symptom attenuation in pepper; the complete nucleotide sequence of an attenuated strain, C-1421

Archives of Virology ◽

10.1007/s007050200030 ◽

2002 ◽

Vol 147 (4) ◽

pp. 833-840 ◽

Cited By ~ 30

Author(s):

K. Hagiwara ◽

T. U. Ichiki ◽

Y. Ogawa ◽

T. Omura ◽

S. Tsuda

Keyword(s):

Amino Acid ◽

Nucleotide Sequence ◽

Amino Acid Substitution ◽

Complete Nucleotide Sequence ◽

Single Amino Acid Substitution ◽

Mottle Virus ◽

Single Amino Acid ◽

Pepper Mild Mottle Virus

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The complete nucleotide sequence and development of a diferential detection assay for a pepper mild mottle virus (PMMoV) isolate that overcomes L3 resistance in pepper

Journal of Virological Methods ◽

10.1016/s0166-0934(02)00144-1 ◽

2002 ◽

Vol 106 (1) ◽

pp. 135-140 ◽

Cited By ~ 25

Author(s):

Leonardo Velasco ◽

Dirk Janssen ◽

Leticia Ruiz-Garcia ◽

Eduardo Segundo ◽

Isabel M Cuadrado

Keyword(s):

Nucleotide Sequence ◽

Complete Nucleotide Sequence ◽

Mottle Virus ◽

Pepper Mild Mottle Virus ◽

Detection Assay

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Variability and evolution of the plant RNA virus pepper mild mottle virus.

Journal of Virology ◽

10.1128/jvi.63.5.2198-2203.1989 ◽

1989 ◽

Vol 63 (5) ◽

pp. 2198-2203 ◽

Cited By ~ 41

Author(s):

E Rodríguez-Cerezo ◽

A Moya ◽

F García-Arenal

Keyword(s):

Rna Virus ◽

Mottle Virus ◽

Pepper Mild Mottle Virus

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Nucleotide sequence and phylogenetic analysis of the medium (M) genomic RNA segments of three hantaviruses isolated in China

Virus Research ◽

10.1016/s0168-1702(98)00065-3 ◽

1998 ◽

Vol 56 (1) ◽

pp. 69-76 ◽

Cited By ~ 14

Author(s):

Xiaohong Shi ◽

Mifang Liang ◽

Changshou Hang ◽

Gan Song ◽

Conall McCaughey ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Nucleotide Sequence ◽

Genomic Rna

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The coat protein of tobamovirus acts as elicitor of both L 2 and L 4 gene-mediated resistance in Capsicum

Journal of General Virology ◽

10.1099/vir.0.80017-0 ◽

2004 ◽

Vol 85 (7) ◽

pp. 2077-2085 ◽

Cited By ~ 36

Author(s):

P. Gilardi ◽

I. García-Luque ◽

M. T. Serra

Keyword(s):

Hypersensitive Response ◽

Host Response ◽

Expression System ◽

Potato Virus X ◽

Mottle Virus ◽

Coat Proteins ◽

Pepper Mild Mottle Virus ◽

Defence Genes ◽

Glucuronidase Reporter ◽

Capsicum Chacoense

In Capsicum, the resistance conferred by the L 2 gene is effective against all of the pepper-infecting tobamoviruses except Pepper mild mottle virus (PMMoV), whereas that conferred by the L 4 gene is effective against them all. These resistances are expressed by a hypersensitive response, manifested through the formation of necrotic local lesions (NLLs) at the primary site of infection. The Capsicum L 2 gene confers resistance to Paprika mild mottle virus (PaMMV), while the L 4 gene is effective against both PaMMV and PMMoV. The PaMMV and PMMoV coat proteins (CPs) were expressed in Capsicum frutescens (L 2 L 2) and Capsicum chacoense (L 4 L 4) plants using the heterologous Potato virus X (PVX)-based expression system. In C. frutescens (L 2 L 2) plants, the chimeric PVX virus containing the PaMMV CP was localized in the inoculated leaves and produced NLLs, whereas the chimeric PVX containing the PMMoV CP infected the plants systemically. Thus, the data indicated that the PaMMV CP is the only tobamovirus factor required for the induction of the host response mediated by the Capsicum L 2 resistance gene. In C. chacoense (L 4 L 4) plants, both chimeric viruses were localized to the inoculated leaves and produced NLLs, indicating that either PaMMV or PMMoV CPs are required to elicit the L 4 gene-mediated host response. In addition, transient expression of PaMMV CP into C. frutescens (L 2 L 2) leaves and PMMoV CP into C. chacoense (L 4 L 4) leaves by biolistic co-bombardment with a β-glucuronidase reporter gene led to the induction of cell death and the expression of host defence genes in both hosts. Thus, the tobamovirus CP is the elicitor of the Capsicum L 2 and L 4 gene-mediated hypersensitive response.

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