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RNA-dependent RNA polymerase (NIb) of the potyviruses is an avirulence factor for the broad-spectrum resistance gene Pvr4 in Capsicum annuum cv. CM334.

Kim SB, Lee HY, Seo S, Lee JH, Choi D - PLoS ONE (2015)

Bottom Line: To identify the avirulence factor corresponding to Pvr4 in potyviruses, we performed Agrobacterium-mediated transient expressions of potyvirus protein coding regions in potyvirus-resistant (Pvr4) and -susceptible (pvr4) pepper plants.Hypersensitive response (HR) was observed only when a RNA-dependent RNA polymerase (NIb) of PepMoV, PepSMV, or PVY was expressed in Pvr4-bearing pepper leaves in a genotype-specific manner.Our results clearly demonstrate that NIbs of PepMoV, PepSMV, and PVY serve as avirulence factors for Pvr4 in pepper plants.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Science, Plant Genomics and Breeding Institute, Seoul National University, Seoul, Korea.

ABSTRACT
Potyviruses are one of the most destructive viral pathogens of Solanaceae plants. In Capsicum annuum landrace CM334, a broad-spectrum gene, Pvr4 is known to be involved in resistance against multiple potyviruses, including Pepper mottle virus (PepMoV), Pepper severe mosaic virus (PepSMV), and Potato virus Y (PVY). However, a potyvirus avirulence factor against Pvr4 has not been identified. To identify the avirulence factor corresponding to Pvr4 in potyviruses, we performed Agrobacterium-mediated transient expressions of potyvirus protein coding regions in potyvirus-resistant (Pvr4) and -susceptible (pvr4) pepper plants. Hypersensitive response (HR) was observed only when a RNA-dependent RNA polymerase (NIb) of PepMoV, PepSMV, or PVY was expressed in Pvr4-bearing pepper leaves in a genotype-specific manner. In contrast, HR was not observed when the NIb of Tobacco etch virus (TEV), a virulent potyvirus, was expressed in Pvr4-bearing pepper leaves. Our results clearly demonstrate that NIbs of PepMoV, PepSMV, and PVY serve as avirulence factors for Pvr4 in pepper plants.

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Genotypes and genotype-specific accumulation of PepMoV in pepper plants.(A) Identification of genotype in relation to Pvr4 using the CAPS marker (PCAPS15). Pvr4-harboring pepper genotypes have 550- and 270-bp fragments, while pvr4-plants have 470- and 350-bp fragments. RR; a resistant homozygotic F2, SS; a susceptible homozygotic F2. Genotype of each plant is depicted under the cultivar name, and phenotypes of plants are also described under the images. R denotes resistant, and S denotes susceptible. (B) Detection of accumulated PepMoV by ELISA. Resistance against PepMoV was confirmed by ELISA with PepMoV antibody, which presents an accumulation of virus. Genotype of each plant is depicted under the cultivar name. Pepper leaves were sampled at 15 dpi. Error bars represent standard deviations. This result and subsequent figures show a representative experiment of three biological replicates.
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pone.0119639.g001: Genotypes and genotype-specific accumulation of PepMoV in pepper plants.(A) Identification of genotype in relation to Pvr4 using the CAPS marker (PCAPS15). Pvr4-harboring pepper genotypes have 550- and 270-bp fragments, while pvr4-plants have 470- and 350-bp fragments. RR; a resistant homozygotic F2, SS; a susceptible homozygotic F2. Genotype of each plant is depicted under the cultivar name, and phenotypes of plants are also described under the images. R denotes resistant, and S denotes susceptible. (B) Detection of accumulated PepMoV by ELISA. Resistance against PepMoV was confirmed by ELISA with PepMoV antibody, which presents an accumulation of virus. Genotype of each plant is depicted under the cultivar name. Pepper leaves were sampled at 15 dpi. Error bars represent standard deviations. This result and subsequent figures show a representative experiment of three biological replicates.

Mentions: For detection of Pvr4-linked markers, PCR products that were amplified with the marker primer were digested with XhoI. Pvr4-linked CAPS marker (PCAPS15) allows discernment of the Pvr4 allele as Pvr4/Pvr4, Pvr4/pvr4, or pvr4/pvr4 [18]. As shown in Fig. 1A, XhoI digestion of the PCR products generated 550- and 270-bp fragments for Pvr4 and 470- and 350-bp fragments for pvr4.


RNA-dependent RNA polymerase (NIb) of the potyviruses is an avirulence factor for the broad-spectrum resistance gene Pvr4 in Capsicum annuum cv. CM334.

Kim SB, Lee HY, Seo S, Lee JH, Choi D - PLoS ONE (2015)

Genotypes and genotype-specific accumulation of PepMoV in pepper plants.(A) Identification of genotype in relation to Pvr4 using the CAPS marker (PCAPS15). Pvr4-harboring pepper genotypes have 550- and 270-bp fragments, while pvr4-plants have 470- and 350-bp fragments. RR; a resistant homozygotic F2, SS; a susceptible homozygotic F2. Genotype of each plant is depicted under the cultivar name, and phenotypes of plants are also described under the images. R denotes resistant, and S denotes susceptible. (B) Detection of accumulated PepMoV by ELISA. Resistance against PepMoV was confirmed by ELISA with PepMoV antibody, which presents an accumulation of virus. Genotype of each plant is depicted under the cultivar name. Pepper leaves were sampled at 15 dpi. Error bars represent standard deviations. This result and subsequent figures show a representative experiment of three biological replicates.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356556&req=5

pone.0119639.g001: Genotypes and genotype-specific accumulation of PepMoV in pepper plants.(A) Identification of genotype in relation to Pvr4 using the CAPS marker (PCAPS15). Pvr4-harboring pepper genotypes have 550- and 270-bp fragments, while pvr4-plants have 470- and 350-bp fragments. RR; a resistant homozygotic F2, SS; a susceptible homozygotic F2. Genotype of each plant is depicted under the cultivar name, and phenotypes of plants are also described under the images. R denotes resistant, and S denotes susceptible. (B) Detection of accumulated PepMoV by ELISA. Resistance against PepMoV was confirmed by ELISA with PepMoV antibody, which presents an accumulation of virus. Genotype of each plant is depicted under the cultivar name. Pepper leaves were sampled at 15 dpi. Error bars represent standard deviations. This result and subsequent figures show a representative experiment of three biological replicates.
Mentions: For detection of Pvr4-linked markers, PCR products that were amplified with the marker primer were digested with XhoI. Pvr4-linked CAPS marker (PCAPS15) allows discernment of the Pvr4 allele as Pvr4/Pvr4, Pvr4/pvr4, or pvr4/pvr4 [18]. As shown in Fig. 1A, XhoI digestion of the PCR products generated 550- and 270-bp fragments for Pvr4 and 470- and 350-bp fragments for pvr4.

Bottom Line: To identify the avirulence factor corresponding to Pvr4 in potyviruses, we performed Agrobacterium-mediated transient expressions of potyvirus protein coding regions in potyvirus-resistant (Pvr4) and -susceptible (pvr4) pepper plants.Hypersensitive response (HR) was observed only when a RNA-dependent RNA polymerase (NIb) of PepMoV, PepSMV, or PVY was expressed in Pvr4-bearing pepper leaves in a genotype-specific manner.Our results clearly demonstrate that NIbs of PepMoV, PepSMV, and PVY serve as avirulence factors for Pvr4 in pepper plants.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Science, Plant Genomics and Breeding Institute, Seoul National University, Seoul, Korea.

ABSTRACT
Potyviruses are one of the most destructive viral pathogens of Solanaceae plants. In Capsicum annuum landrace CM334, a broad-spectrum gene, Pvr4 is known to be involved in resistance against multiple potyviruses, including Pepper mottle virus (PepMoV), Pepper severe mosaic virus (PepSMV), and Potato virus Y (PVY). However, a potyvirus avirulence factor against Pvr4 has not been identified. To identify the avirulence factor corresponding to Pvr4 in potyviruses, we performed Agrobacterium-mediated transient expressions of potyvirus protein coding regions in potyvirus-resistant (Pvr4) and -susceptible (pvr4) pepper plants. Hypersensitive response (HR) was observed only when a RNA-dependent RNA polymerase (NIb) of PepMoV, PepSMV, or PVY was expressed in Pvr4-bearing pepper leaves in a genotype-specific manner. In contrast, HR was not observed when the NIb of Tobacco etch virus (TEV), a virulent potyvirus, was expressed in Pvr4-bearing pepper leaves. Our results clearly demonstrate that NIbs of PepMoV, PepSMV, and PVY serve as avirulence factors for Pvr4 in pepper plants.

Show MeSH
Related in: MedlinePlus