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Possible involvement of maize Rop1 in the defence responses of plants to viral infection

Identifieur interne : 001C34 ( Istex/Corpus ); précédent : 001C33; suivant : 001C35

Possible involvement of maize Rop1 in the defence responses of plants to viral infection

Auteurs : Yanyong Cao ; Yan Shi ; Yongqiang Li ; Yuqin Cheng ; Tao Zhou ; Zaifeng Fan

Source :

RBID : ISTEX:8838450D5AA79FB15B9FC3454F6F9B31F1922616

English descriptors

Abstract

The expression of host genes can be altered during the process of viral infection. To investigate the viral infection‐induced up‐regulated gene expression changes of maize at different time intervals post‐inoculation with Sugarcane mosaic virus (SCMV), a suppression subtractive hybridization cDNA library was constructed. A total of 454 cDNA clones were identified to be viral infection‐induced up‐regulated genes. The influence of Rop1 on the infection of maize by SCMV was investigated. The results showed that transient silencing of the ZmRop1 gene through virus‐induced gene silencing enhanced the accumulation and systemic infection of SCMV and another potyvirus (Pennisetum mosaic virus) in maize plants, whereas transient over‐expression of ZmRop1 in maize protoplasts reduced SCMV accumulation. Furthermore, it was demonstrated that the heterologous expression of ZmRop1 impaired Potato virus X infection in Nicotiana benthamiana plants. These data suggest that ZmRop1 may play a role in plant defence responses to viral infection.

Url:
DOI: 10.1111/j.1364-3703.2011.00782.x

Links to Exploration step

ISTEX:8838450D5AA79FB15B9FC3454F6F9B31F1922616

Le document en format XML

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<json:string>Ding et al., 2006</json:string>
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<json:string>Raghupathy et al., 2006</json:string>
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<json:string>Smith et al., 2004</json:string>
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<json:string>Chen et al., 2008</json:string>
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<p>
<b>Fig. S1</b>
Schematic diagram of pBMV
<sub>A/G</sub>
/Rop1 and pGFP‐ZmRop1s construction. (A) Schematic diagram of the C‐BMV
<sub>A/G</sub>
/Rop1 vector constructs, in which pF3‐5/13′
<sub>A/G</sub>
carries a specific 145‐bp PCR fragment of the
<i>ZmRop1</i>
5′‐terminal untranslated region (UTR) (Rop1
<sub>145</sub>
). Black rectangles represent the viral sequence and grey rectangles represent Rop1
<sub>145</sub>
. The curly lines at both termini of each construct represent bacterial plasmid sequences. The clover‐like symbol represents the tRNA‐like structure at the 3′‐terminus of the viral RNA sequence.
<i>Psh</i>
AI and
<i>Spe</i>
I are restriction sites for plasmid linearization prior to
<i>in vitro</i>
transcription, and
<i>Hin</i>
dIII in pF3‐5/13′
<sub>A/G</sub>
is used for Rop1
<sub>145</sub>
insertion. T3 indicates the T3 promoter sequence (open rectangle) for
<i>in vitro</i>
transcription of viral sequences. (B) Schematic representation of four types of GFP‐ZmRop1 fusion construct. G, green fluorescent protein (GFP); CA, constitutively active; DN, dominant negative; 35 S, 35S promoter derived from
<i>Cauliflower mosaic virus</i>
; T, nopaline synthase (NOS) terminator derived from
<i>Agrobacterium tumefaciens</i>
.</p>
<p>
<b>Fig. S2</b>
Quantitative reverse transcription‐polymerase chain reaction (QRT‐PCR) analysis of the effects of
<i>Sugarcane mosaic virus</i>
(SCMV) infection on the relative mRNA levels of
<i>ZmRop4</i>
(A),
<i>ZmRop9</i>
(B) and
<i>ZmRop8</i>
(C) in maize leaves. Each value is shown as the average of three independent experiments ± standard deviation (SD). IL, inoculated leaves; SL, systemic leaves. Different letters on the columns indicate a significant difference by Student's
<i>t</i>
‐test (
<i>P</i>
< 0.05).</p>
<p>
<b>Fig. S3</b>
The subcellular localization of ZmRop1. (A) Fluorescence micrographs of maize protoplasts transformed with four types of GFP‐ZmRop1 fusion construct using laser scanning confocal microscopy. Scale bar, 10 µm. pGRop1, pGFP‐ZmRop1; pGRop1
<sup>CA</sup>
, pGFP‐ZmRop1
<sup>CA</sup>
; pGRop1
<sup>DN</sup>
, pGFP‐ZmRop1
<sup>DN</sup>
; pGRop1mS
<sup>198</sup>
, pGFP‐ZmRop1mS
<sup>198</sup>
. CA, constitutively active, DN, dominant negative; C, cytoplasm; V, vacuole. (B) The expression levels of fusion proteins were detected by Western blotting using polyclonal antiserum against green fluorescent protein (GFP), as indicated. The Coomassie brilliant blue‐stained 8% sodium dodecylsulphate‐polyacrylamide gel (bottom) for different samples was used as a loading control.</p>
<p>
<b>Table S1</b>
List of up‐regulated gene transcripts post‐
<i>Sugarcane mosaic virus</i>
(SCMV) infection with BLAST hits against the National Center for Biotechnology Information (NCBI) GenBank nonredundant (nr) database.</p>
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<p>The expression of host genes can be altered during the process of viral infection. To investigate the viral infection‐induced up‐regulated gene expression changes of maize at different time intervals post‐inoculation with
<i>Sugarcane mosaic virus</i>
(SCMV), a suppression subtractive hybridization cDNA library was constructed. A total of 454 cDNA clones were identified to be viral infection‐induced up‐regulated genes. The influence of Rop1 on the infection of maize by SCMV was investigated. The results showed that transient silencing of the
<i>ZmRop1</i>
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<i>Pennisetum mosaic virus</i>
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<p> These authors contributed equally to this work.</p>
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<i>Present address</i>
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<dateIssued encoding="w3cdtf">2012-09</dateIssued>
<copyrightDate encoding="w3cdtf">2012</copyrightDate>
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<languageTerm type="code" authority="iso639-2b">eng</languageTerm>
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<abstract lang="en">The expression of host genes can be altered during the process of viral infection. To investigate the viral infection‐induced up‐regulated gene expression changes of maize at different time intervals post‐inoculation with Sugarcane mosaic virus (SCMV), a suppression subtractive hybridization cDNA library was constructed. A total of 454 cDNA clones were identified to be viral infection‐induced up‐regulated genes. The influence of Rop1 on the infection of maize by SCMV was investigated. The results showed that transient silencing of the ZmRop1 gene through virus‐induced gene silencing enhanced the accumulation and systemic infection of SCMV and another potyvirus (Pennisetum mosaic virus) in maize plants, whereas transient over‐expression of ZmRop1 in maize protoplasts reduced SCMV accumulation. Furthermore, it was demonstrated that the heterologous expression of ZmRop1 impaired Potato virus X infection in Nicotiana benthamiana plants. These data suggest that ZmRop1 may play a role in plant defence responses to viral infection.</abstract>
<relatedItem type="host">
<titleInfo>
<title>Molecular Plant Pathology</title>
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<genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre>
<note type="content"> Fig. S1 Schematic diagram of pBMVA/G/Rop1 and pGFP‐ZmRop1s construction. (A) Schematic diagram of the C‐BMVA/G/Rop1 vector constructs, in which pF3‐5/13′A/G carries a specific 145‐bp PCR fragment of the ZmRop1 5′‐terminal untranslated region (UTR) (Rop1145). Black rectangles represent the viral sequence and grey rectangles represent Rop1145. The curly lines at both termini of each construct represent bacterial plasmid sequences. The clover‐like symbol represents the tRNA‐like structure at the 3′‐terminus of the viral RNA sequence. PshAI and SpeI are restriction sites for plasmid linearization prior to in vitro transcription, and HindIII in pF3‐5/13′A/G is used for Rop1145 insertion. T3 indicates the T3 promoter sequence (open rectangle) for in vitro transcription of viral sequences. (B) Schematic representation of four types of GFP‐ZmRop1 fusion construct. G, green fluorescent protein (GFP); CA, constitutively active; DN, dominant negative; 35 S, 35S promoter derived from Cauliflower mosaic virus; T, nopaline synthase (NOS) terminator derived from Agrobacterium tumefaciens. Fig. S2 Quantitative reverse transcription‐polymerase chain reaction (QRT‐PCR) analysis of the effects of Sugarcane mosaic virus (SCMV) infection on the relative mRNA levels of ZmRop4 (A), ZmRop9 (B) and ZmRop8 (C) in maize leaves. Each value is shown as the average of three independent experiments ± standard deviation (SD). IL, inoculated leaves; SL, systemic leaves. Different letters on the columns indicate a significant difference by Student's t‐test (P < 0.05). Fig. S3 The subcellular localization of ZmRop1. (A) Fluorescence micrographs of maize protoplasts transformed with four types of GFP‐ZmRop1 fusion construct using laser scanning confocal microscopy. Scale bar, 10 µm. pGRop1, pGFP‐ZmRop1; pGRop1CA, pGFP‐ZmRop1CA; pGRop1DN, pGFP‐ZmRop1DN; pGRop1mS198, pGFP‐ZmRop1mS198. CA, constitutively active, DN, dominant negative; C, cytoplasm; V, vacuole. (B) The expression levels of fusion proteins were detected by Western blotting using polyclonal antiserum against green fluorescent protein (GFP), as indicated. The Coomassie brilliant blue‐stained 8% sodium dodecylsulphate‐polyacrylamide gel (bottom) for different samples was used as a loading control. Table S1 List of up‐regulated gene transcripts post‐Sugarcane mosaic virus (SCMV) infection with BLAST hits against the National Center for Biotechnology Information (NCBI) GenBank nonredundant (nr) database. Fig. S1 Schematic diagram of pBMVA/G/Rop1 and pGFP‐ZmRop1s construction. (A) Schematic diagram of the C‐BMVA/G/Rop1 vector constructs, in which pF3‐5/13′A/G carries a specific 145‐bp PCR fragment of the ZmRop1 5′‐terminal untranslated region (UTR) (Rop1145). Black rectangles represent the viral sequence and grey rectangles represent Rop1145. The curly lines at both termini of each construct represent bacterial plasmid sequences. The clover‐like symbol represents the tRNA‐like structure at the 3′‐terminus of the viral RNA sequence. PshAI and SpeI are restriction sites for plasmid linearization prior to in vitro transcription, and HindIII in pF3‐5/13′A/G is used for Rop1145 insertion. T3 indicates the T3 promoter sequence (open rectangle) for in vitro transcription of viral sequences. (B) Schematic representation of four types of GFP‐ZmRop1 fusion construct. G, green fluorescent protein (GFP); CA, constitutively active; DN, dominant negative; 35 S, 35S promoter derived from Cauliflower mosaic virus; T, nopaline synthase (NOS) terminator derived from Agrobacterium tumefaciens. Fig. S2 Quantitative reverse transcription‐polymerase chain reaction (QRT‐PCR) analysis of the effects of Sugarcane mosaic virus (SCMV) infection on the relative mRNA levels of ZmRop4 (A), ZmRop9 (B) and ZmRop8 (C) in maize leaves. Each value is shown as the average of three independent experiments ± standard deviation (SD). IL, inoculated leaves; SL, systemic leaves. Different letters on the columns indicate a significant difference by Student's t‐test (P < 0.05). Fig. S3 The subcellular localization of ZmRop1. (A) Fluorescence micrographs of maize protoplasts transformed with four types of GFP‐ZmRop1 fusion construct using laser scanning confocal microscopy. Scale bar, 10 µm. pGRop1, pGFP‐ZmRop1; pGRop1CA, pGFP‐ZmRop1CA; pGRop1DN, pGFP‐ZmRop1DN; pGRop1mS198, pGFP‐ZmRop1mS198. CA, constitutively active, DN, dominant negative; C, cytoplasm; V, vacuole. (B) The expression levels of fusion proteins were detected by Western blotting using polyclonal antiserum against green fluorescent protein (GFP), as indicated. The Coomassie brilliant blue‐stained 8% sodium dodecylsulphate‐polyacrylamide gel (bottom) for different samples was used as a loading control. Table S1 List of up‐regulated gene transcripts post‐Sugarcane mosaic virus (SCMV) infection with BLAST hits against the National Center for Biotechnology Information (NCBI) GenBank nonredundant (nr) database. Fig. S1 Schematic diagram of pBMVA/G/Rop1 and pGFP‐ZmRop1s construction. (A) Schematic diagram of the C‐BMVA/G/Rop1 vector constructs, in which pF3‐5/13′A/G carries a specific 145‐bp PCR fragment of the ZmRop1 5′‐terminal untranslated region (UTR) (Rop1145). Black rectangles represent the viral sequence and grey rectangles represent Rop1145. The curly lines at both termini of each construct represent bacterial plasmid sequences. The clover‐like symbol represents the tRNA‐like structure at the 3′‐terminus of the viral RNA sequence. PshAI and SpeI are restriction sites for plasmid linearization prior to in vitro transcription, and HindIII in pF3‐5/13′A/G is used for Rop1145 insertion. T3 indicates the T3 promoter sequence (open rectangle) for in vitro transcription of viral sequences. (B) Schematic representation of four types of GFP‐ZmRop1 fusion construct. G, green fluorescent protein (GFP); CA, constitutively active; DN, dominant negative; 35 S, 35S promoter derived from Cauliflower mosaic virus; T, nopaline synthase (NOS) terminator derived from Agrobacterium tumefaciens. Fig. S2 Quantitative reverse transcription‐polymerase chain reaction (QRT‐PCR) analysis of the effects of Sugarcane mosaic virus (SCMV) infection on the relative mRNA levels of ZmRop4 (A), ZmRop9 (B) and ZmRop8 (C) in maize leaves. Each value is shown as the average of three independent experiments ± standard deviation (SD). IL, inoculated leaves; SL, systemic leaves. Different letters on the columns indicate a significant difference by Student's t‐test (P < 0.05). Fig. S3 The subcellular localization of ZmRop1. (A) Fluorescence micrographs of maize protoplasts transformed with four types of GFP‐ZmRop1 fusion construct using laser scanning confocal microscopy. Scale bar, 10 µm. pGRop1, pGFP‐ZmRop1; pGRop1CA, pGFP‐ZmRop1CA; pGRop1DN, pGFP‐ZmRop1DN; pGRop1mS198, pGFP‐ZmRop1mS198. CA, constitutively active, DN, dominant negative; C, cytoplasm; V, vacuole. (B) The expression levels of fusion proteins were detected by Western blotting using polyclonal antiserum against green fluorescent protein (GFP), as indicated. The Coomassie brilliant blue‐stained 8% sodium dodecylsulphate‐polyacrylamide gel (bottom) for different samples was used as a loading control. Table S1 List of up‐regulated gene transcripts post‐Sugarcane mosaic virus (SCMV) infection with BLAST hits against the National Center for Biotechnology Information (NCBI) GenBank nonredundant (nr) database. Fig. S1 Schematic diagram of pBMVA/G/Rop1 and pGFP‐ZmRop1s construction. (A) Schematic diagram of the C‐BMVA/G/Rop1 vector constructs, in which pF3‐5/13′A/G carries a specific 145‐bp PCR fragment of the ZmRop1 5′‐terminal untranslated region (UTR) (Rop1145). Black rectangles represent the viral sequence and grey rectangles represent Rop1145. The curly lines at both termini of each construct represent bacterial plasmid sequences. The clover‐like symbol represents the tRNA‐like structure at the 3′‐terminus of the viral RNA sequence. PshAI and SpeI are restriction sites for plasmid linearization prior to in vitro transcription, and HindIII in pF3‐5/13′A/G is used for Rop1145 insertion. T3 indicates the T3 promoter sequence (open rectangle) for in vitro transcription of viral sequences. (B) Schematic representation of four types of GFP‐ZmRop1 fusion construct. G, green fluorescent protein (GFP); CA, constitutively active; DN, dominant negative; 35 S, 35S promoter derived from Cauliflower mosaic virus; T, nopaline synthase (NOS) terminator derived from Agrobacterium tumefaciens. Fig. S2 Quantitative reverse transcription‐polymerase chain reaction (QRT‐PCR) analysis of the effects of Sugarcane mosaic virus (SCMV) infection on the relative mRNA levels of ZmRop4 (A), ZmRop9 (B) and ZmRop8 (C) in maize leaves. Each value is shown as the average of three independent experiments ± standard deviation (SD). IL, inoculated leaves; SL, systemic leaves. Different letters on the columns indicate a significant difference by Student's t‐test (P < 0.05). Fig. S3 The subcellular localization of ZmRop1. (A) Fluorescence micrographs of maize protoplasts transformed with four types of GFP‐ZmRop1 fusion construct using laser scanning confocal microscopy. Scale bar, 10 µm. pGRop1, pGFP‐ZmRop1; pGRop1CA, pGFP‐ZmRop1CA; pGRop1DN, pGFP‐ZmRop1DN; pGRop1mS198, pGFP‐ZmRop1mS198. CA, constitutively active, DN, dominant negative; C, cytoplasm; V, vacuole. (B) The expression levels of fusion proteins were detected by Western blotting using polyclonal antiserum against green fluorescent protein (GFP), as indicated. The Coomassie brilliant blue‐stained 8% sodium dodecylsulphate‐polyacrylamide gel (bottom) for different samples was used as a loading control. Table S1 List of up‐regulated gene transcripts post‐Sugarcane mosaic virus (SCMV) infection with BLAST hits against the National Center for Biotechnology Information (NCBI) GenBank nonredundant (nr) database.Supporting Info Item: Supporting info item - Supporting info item - Supporting info item - Supporting info item - </note>
<subject>
<genre>article-category</genre>
<topic>Original articles</topic>
<topic>Original article</topic>
</subject>
<identifier type="ISSN">1464-6722</identifier>
<identifier type="eISSN">1364-3703</identifier>
<identifier type="DOI">10.1111/(ISSN)1364-3703</identifier>
<identifier type="PublisherID">MPP</identifier>
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