Macroarray Detection of Eleven Potato-Infecting Viruses and <i>Potato spindle tuber viroid</i>

Agindotan, Bright; Perry, Keith L.

doi:10.1094/pdis-92-5-0730

Cited by 42 publications

(19 citation statements)

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“…Perry, unpublished data). The presence of specific viruses can be detected by hybridization with a labeled preparation of randomly primed, reverse-transcribed, and amplified cDNAs derived from infected plant RNAs (Agindotan and Perry, 2008). As expected, labeled cDNA derived from PVX 2.7-infected plants hybridized to PVX sequences present on the macroarray (Fig.…”

Section: Identification Of a Virus Allowing Pvx Infection Of Arabidopsissupporting

confidence: 62%

ARGONAUTE2 Mediates RNA-Silencing Antiviral Defenses againstPotato virus Xin Arabidopsis

et al. 2011

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2R1 (P.M.)RNA-silencing mechanisms control many aspects of gene regulation including the detection and degradation of viral RNA through the action of, among others, Dicer-like and Argonaute (AGO) proteins. However, the extent to which RNA silencing restricts virus host range has been difficult to separate from other factors that can affect virus-plant compatibility. Here we show that Potato virus X (PVX) can infect Arabidopsis (Arabidopsis thaliana), which is normally a nonhost for PVX, if coinfected with a second virus, Pepper ringspot virus. Here we show that the pepper ringspot virus 12K protein functions as a suppressor of silencing that appears to enable PVX to infect Arabidopsis. We also show that PVX is able to infect Arabidopsis Dicer-like mutants, indicating that RNA silencing is responsible for Arabidopsis nonhost resistance to PVX. Furthermore, we find that restriction of PVX on Arabidopsis also depends on AGO2, suggesting that this AGO protein has evolved to specialize in antiviral defenses.

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Section: Identification Of a Virus Allowing Pvx Infection Of Arabidopsissupporting

confidence: 62%

ARGONAUTE2 Mediates RNA-Silencing Antiviral Defenses againstPotato virus Xin Arabidopsis

et al. 2011

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“…Sensitivity is also an important factor. Macroarray assays for plant viruses have recently been developed (1,2,33). RT-PCR has proven to be more sensitive than ELISA (1,12,28).…”

Section: Discussionmentioning

confidence: 99%

“…Commonly used techniques for the detection of plant viruses include ELISA and RT-PCR or PCR (14,23,37,38). Agindotan and Perry (1,2) reported that sensitivity of their macroarray for 11 potato viruses and a viroid was comparable with that of DAS-ELISA but was lower than that for RT-PCR. Macroarray assays for plant viruses have recently been developed (1,2,33).…”

Section: Discussionmentioning

confidence: 99%

“…However, PCR conditions varied by target virus, and no method was available to detect the 12 viruses simultaneously. However, this macroarray was not completely satisfactory; there was cross-hybridization between viruses, making interpretation difficult (2). These viruses were detected by hybridization using synthesized oligonucleotide probes on membranes and chemically labeled targets prepared by the random primed amplified procedure.…”

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confidence: 99%

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Application of cDNA Macroarray for Simultaneous Detection of 12 Potato Viruses

et al. 2010

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Maoka, T., Sugiyama, S., Maruta, Y., and Hataya, T. 2010. Application of cDNA macroarray for simultaneous detection of 12 potato viruses. Plant Dis. 94:1248-1254. A complementary DNA (cDNA) macroarray was developed for simultaneous detection of 12 different potato viruses. A suitable region in the viral genome for each was selected for Alfalfa mosaic virus, Cucumber mosaic virus, Potato aucuba mosaic virus, Potato leafroll virus, Potato mop-top virus, Potato virus A, Potato virus M, Potato virus S, Potato virus X, Potato virus Y, Tomato ringspot virus, and Tomato spotted wilt virus, and their respective cDNAs were cloned into plasmid vectors.Capture probes for each virus ranging from 290 to 577 bp were generated by polymerase chain reaction (PCR) and immobilized on a nylon membrane. Total RNAs were extracted from each of these virus infected-plants, and cDNAs were synthesized from the RNA extracts using a random 9-mer primer. Subsequently, PCR reactions were performed using one primer pair for each of the 12 viruses. During PCR, amplified cDNAs were labeled with biotin and used as a target for hybridization analyses on a macroarray membrane. Hybridization signals between capture probes for the 12 viruses and their respective target cDNAs were observed using chemiluminescent or colorimetric detection. In all viruses, hybridization signals with capture probes were detected only when homologous virus targets were examined, and no hybridization to healthy plant extract was observed, facilitating identification of each virus. The results by colorimetric detection agreed with those obtained using chemiluminescence. The macroarray method developed was 5 × 10 2 to 4 × 10 6 times more sensitive than enzyme-linked immunosorbent assay and 5 to 5 × 10 4 times more sensitive than reverse-transcription PCR, except for Alfalfa mosaic virus. Colorimetric detection and substantial reduction in cross-hybridization signals much improved the method compared with other array-based detection methods for practical use.

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“…Even more recently the use of micro-or macroarrays for potato virus detection and identification has been reported (Boonham et al 2003;Agindotan and Perry 2008;Maoka et al 2010). Still in the developmental stage is a "universal plant virus microarray" that may be able to detect most plants viruses in a single test (Bagewadi et al 2010).…”

Section: Resultsmentioning

confidence: 99%

Standardized RT-PCR Conditions for Detection and Identification of Eleven Viruses of Potato and Potato spindle tuber viroid

Crosslin

Hamlin

2011

Am. J. Pot Res

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Macroarray Detection of Eleven Potato-Infecting Viruses and Potato spindle tuber viroid

Cited by 42 publications

References 34 publications

ARGONAUTE2 Mediates RNA-Silencing Antiviral Defenses againstPotato virus Xin Arabidopsis

ARGONAUTE2 Mediates RNA-Silencing Antiviral Defenses againstPotato virus Xin Arabidopsis

Application of cDNA Macroarray for Simultaneous Detection of 12 Potato Viruses

Standardized RT-PCR Conditions for Detection and Identification of Eleven Viruses of Potato and Potato spindle tuber viroid

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