Transient expression of artificial microRNAs targeting Grapevine fanleaf virus and evidence for RNA silencing in grapevine somatic embryos.
Identifieur interne : 000610 ( Main/Exploration ); précédent : 000609; suivant : 000611Transient expression of artificial microRNAs targeting Grapevine fanleaf virus and evidence for RNA silencing in grapevine somatic embryos.
Auteurs : Noémie S. Jelly [France] ; Paul Schellenbaum ; Bernard Walter ; Pascale MaillotSource :
- Transgenic research [ 1573-9368 ] ; 2012.
Descripteurs français
- KwdFr :
- ARN des plantes (génétique), ARN viral (génétique), Agrobacterium tumefaciens (génétique), Arabidopsis (génétique), Glucuronidase (génétique), Glucuronidase (métabolisme), Interférence par ARN (MeSH), Nepovirus (génétique), Protéines à fluorescence verte (métabolisme), Techniques de transfert de gènes (MeSH), Vecteurs génétiques (MeSH), Vitis (croissance et développement), Vitis (embryologie), Vitis (génétique), microARN (physiologie).
- MESH :
- croissance et développement : Vitis.
- embryologie : Vitis.
- génétique : ARN des plantes, ARN viral, Agrobacterium tumefaciens, Arabidopsis, Glucuronidase, Nepovirus, Vitis.
- métabolisme : Glucuronidase, Protéines à fluorescence verte.
- physiologie : microARN.
- Interférence par ARN, Techniques de transfert de gènes, Vecteurs génétiques.
English descriptors
- KwdEn :
- Agrobacterium tumefaciens (genetics), Arabidopsis (genetics), Gene Transfer Techniques (MeSH), Genetic Vectors (MeSH), Glucuronidase (genetics), Glucuronidase (metabolism), Green Fluorescent Proteins (metabolism), MicroRNAs (physiology), Nepovirus (genetics), RNA Interference (MeSH), RNA, Plant (genetics), RNA, Viral (genetics), Vitis (embryology), Vitis (genetics), Vitis (growth & development).
- MESH :
- chemical , genetics : Glucuronidase, RNA, Plant, RNA, Viral.
- embryology : Vitis.
- genetics : Agrobacterium tumefaciens, Arabidopsis, Nepovirus, Vitis.
- growth & development : Vitis.
- chemical , metabolism : Glucuronidase, Green Fluorescent Proteins.
- chemical , physiology : MicroRNAs.
- Gene Transfer Techniques, Genetic Vectors, RNA Interference.
Abstract
Grapevines are affected worldwide by viruses that compromise fruit yield and quality. Grapevine fanleaf virus (GFLV) causes fanleaf degeneration disease, a major threat to grapevine production. Transgenic approaches exploiting the RNA silencing machinery have proven suitable for engineering viral resistance in several crop species. However, the artificial microRNA (amiRNA)-based strategy has not yet been reported in grapevine. We developed two amiRNA precursors (pre-amiRNAs) targeting the coat protein (CP) gene of GFLV and characterised their functionality in grapevine somatic embryos. To create these pre-amiRNAs, natural pre-miR319a of Arabidopsis thaliana was modified by overlapping PCR in order to replace miR319a with two amiRNAs targeting different regions of the CP gene: amiR(CP)-1 or amiR(CP)-2. Transient expression of these two pre-amiRNA constructs was tested in grapevine somatic embryos after co-cultivation with Agrobacterium tumefaciens. Expression of amiR(CP)-1 and amiR(CP)-2 was detected in plant tissues by an endpoint stem-loop RT-PCR as early as 1 day after a 48-h co-cultivation, indicating active processing of pre-amiRNAs by the plant machinery. In parallel, GUS-sensor constructs (G(CP)-1 and G(CP)-2) were obtained by fusing the target sequence of amiR(CP)-1 or amiR(CP)-2 to the 3' terminus of the GUS gene. Co-transformation assays with GUS-sensors and the pre-amiRNA constructs provided evidence for in vivo recognition and cleavage of the 21-nt target sequence of GUS-sensors by the corresponding amiRNA. This is the first report of amiRNA ectopic expression in grapevine. The constructs we developed could be useful for engineering GFLV-resistant grapes in the future.
DOI: 10.1007/s11248-012-9611-5
PubMed: 22427113
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Jelly</name><affiliation wicri:level="3"><nlm:affiliation>Laboratoire Vigne, Biotechnologies et Environnement, Université de Haute-Alsace, 33 rue de Herrlisheim, 68008 Colmar, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire Vigne, Biotechnologies et Environnement, Université de Haute-Alsace, 33 rue de Herrlisheim, 68008 Colmar</wicri:regionArea><placeName><region type="region" nuts="2">Grand Est</region><region type="old region" nuts="2">Alsace (région administrative)</region><settlement type="city">Colmar</settlement></placeName></affiliation></author><author><name sortKey="Schellenbaum, Paul" sort="Schellenbaum, Paul" uniqKey="Schellenbaum P" first="Paul" last="Schellenbaum">Paul Schellenbaum</name></author><author><name sortKey="Walter, Bernard" sort="Walter, Bernard" uniqKey="Walter B" first="Bernard" last="Walter">Bernard Walter</name></author><author><name sortKey="Maillot, Pascale" sort="Maillot, Pascale" uniqKey="Maillot P" first="Pascale" last="Maillot">Pascale Maillot</name></author></analytic><series><title level="j">Transgenic research</title><idno type="eISSN">1573-9368</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (genetics)</term><term>Arabidopsis (genetics)</term><term>Gene Transfer Techniques (MeSH)</term><term>Genetic Vectors (MeSH)</term><term>Glucuronidase (genetics)</term><term>Glucuronidase (metabolism)</term><term>Green Fluorescent Proteins (metabolism)</term><term>MicroRNAs (physiology)</term><term>Nepovirus (genetics)</term><term>RNA Interference (MeSH)</term><term>RNA, Plant (genetics)</term><term>RNA, Viral (genetics)</term><term>Vitis (embryology)</term><term>Vitis (genetics)</term><term>Vitis (growth & development)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN des plantes (génétique)</term><term>ARN viral (génétique)</term><term>Agrobacterium tumefaciens (génétique)</term><term>Arabidopsis (génétique)</term><term>Glucuronidase (génétique)</term><term>Glucuronidase (métabolisme)</term><term>Interférence par ARN (MeSH)</term><term>Nepovirus (génétique)</term><term>Protéines à fluorescence verte (métabolisme)</term><term>Techniques de transfert de gènes (MeSH)</term><term>Vecteurs génétiques (MeSH)</term><term>Vitis (croissance et développement)</term><term>Vitis (embryologie)</term><term>Vitis (génétique)</term><term>microARN (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Glucuronidase</term><term>RNA, Plant</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Vitis</term></keywords><keywords scheme="MESH" qualifier="embryologie" xml:lang="fr"><term>Vitis</term></keywords><keywords scheme="MESH" qualifier="embryology" xml:lang="en"><term>Vitis</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Arabidopsis</term><term>Nepovirus</term><term>Vitis</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Vitis</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN des plantes</term><term>ARN viral</term><term>Agrobacterium tumefaciens</term><term>Arabidopsis</term><term>Glucuronidase</term><term>Nepovirus</term><term>Vitis</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glucuronidase</term><term>Green Fluorescent Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glucuronidase</term><term>Protéines à fluorescence verte</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>microARN</term></keywords><keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>MicroRNAs</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Transfer Techniques</term><term>Genetic Vectors</term><term>RNA Interference</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Interférence par ARN</term><term>Techniques de transfert de gènes</term><term>Vecteurs génétiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Grapevines are affected worldwide by viruses that compromise fruit yield and quality. Grapevine fanleaf virus (GFLV) causes fanleaf degeneration disease, a major threat to grapevine production. Transgenic approaches exploiting the RNA silencing machinery have proven suitable for engineering viral resistance in several crop species. However, the artificial microRNA (amiRNA)-based strategy has not yet been reported in grapevine. We developed two amiRNA precursors (pre-amiRNAs) targeting the coat protein (CP) gene of GFLV and characterised their functionality in grapevine somatic embryos. To create these pre-amiRNAs, natural pre-miR319a of Arabidopsis thaliana was modified by overlapping PCR in order to replace miR319a with two amiRNAs targeting different regions of the CP gene: amiR(CP)-1 or amiR(CP)-2. Transient expression of these two pre-amiRNA constructs was tested in grapevine somatic embryos after co-cultivation with Agrobacterium tumefaciens. Expression of amiR(CP)-1 and amiR(CP)-2 was detected in plant tissues by an endpoint stem-loop RT-PCR as early as 1 day after a 48-h co-cultivation, indicating active processing of pre-amiRNAs by the plant machinery. In parallel, GUS-sensor constructs (G(CP)-1 and G(CP)-2) were obtained by fusing the target sequence of amiR(CP)-1 or amiR(CP)-2 to the 3' terminus of the GUS gene. Co-transformation assays with GUS-sensors and the pre-amiRNA constructs provided evidence for in vivo recognition and cleavage of the 21-nt target sequence of GUS-sensors by the corresponding amiRNA. This is the first report of amiRNA ectopic expression in grapevine. The constructs we developed could be useful for engineering GFLV-resistant grapes in the future.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22427113</PMID><DateCompleted><Year>2013</Year><Month>05</Month><Day>14</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1573-9368</ISSN><JournalIssue CitedMedium="Internet"><Volume>21</Volume><Issue>6</Issue><PubDate><Year>2012</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Transgenic research</Title><ISOAbbreviation>Transgenic Res</ISOAbbreviation></Journal><ArticleTitle>Transient expression of artificial microRNAs targeting Grapevine fanleaf virus and evidence for RNA silencing in grapevine somatic embryos.</ArticleTitle><Pagination><MedlinePgn>1319-27</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s11248-012-9611-5</ELocationID><Abstract><AbstractText>Grapevines are affected worldwide by viruses that compromise fruit yield and quality. Grapevine fanleaf virus (GFLV) causes fanleaf degeneration disease, a major threat to grapevine production. Transgenic approaches exploiting the RNA silencing machinery have proven suitable for engineering viral resistance in several crop species. However, the artificial microRNA (amiRNA)-based strategy has not yet been reported in grapevine. We developed two amiRNA precursors (pre-amiRNAs) targeting the coat protein (CP) gene of GFLV and characterised their functionality in grapevine somatic embryos. To create these pre-amiRNAs, natural pre-miR319a of Arabidopsis thaliana was modified by overlapping PCR in order to replace miR319a with two amiRNAs targeting different regions of the CP gene: amiR(CP)-1 or amiR(CP)-2. Transient expression of these two pre-amiRNA constructs was tested in grapevine somatic embryos after co-cultivation with Agrobacterium tumefaciens. Expression of amiR(CP)-1 and amiR(CP)-2 was detected in plant tissues by an endpoint stem-loop RT-PCR as early as 1 day after a 48-h co-cultivation, indicating active processing of pre-amiRNAs by the plant machinery. In parallel, GUS-sensor constructs (G(CP)-1 and G(CP)-2) were obtained by fusing the target sequence of amiR(CP)-1 or amiR(CP)-2 to the 3' terminus of the GUS gene. Co-transformation assays with GUS-sensors and the pre-amiRNA constructs provided evidence for in vivo recognition and cleavage of the 21-nt target sequence of GUS-sensors by the corresponding amiRNA. This is the first report of amiRNA ectopic expression in grapevine. The constructs we developed could be useful for engineering GFLV-resistant grapes in the future.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Jelly</LastName><ForeName>Noémie S</ForeName><Initials>NS</Initials><AffiliationInfo><Affiliation>Laboratoire Vigne, Biotechnologies et Environnement, Université de Haute-Alsace, 33 rue de Herrlisheim, 68008 Colmar, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Schellenbaum</LastName><ForeName>Paul</ForeName><Initials>P</Initials></Author><Author ValidYN="Y"><LastName>Walter</LastName><ForeName>Bernard</ForeName><Initials>B</Initials></Author><Author ValidYN="Y"><LastName>Maillot</LastName><ForeName>Pascale</ForeName><Initials>P</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2012</Year><Month>03</Month><Day>17</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Transgenic Res</MedlineTA><NlmUniqueID>9209120</NlmUniqueID><ISSNLinking>0962-8819</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D035683">MicroRNAs</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018749">RNA, Plant</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C485184">enhanced green fluorescent protein</NameOfSubstance></Chemical><Chemical><RegistryNumber>147336-22-9</RegistryNumber><NameOfSubstance UI="D049452">Green Fluorescent Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.31</RegistryNumber><NameOfSubstance 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last="Schellenbaum">Paul Schellenbaum</name><name sortKey="Walter, Bernard" sort="Walter, Bernard" uniqKey="Walter B" first="Bernard" last="Walter">Bernard Walter</name></noCountry><country name="France"><region name="Grand Est"><name sortKey="Jelly, Noemie S" sort="Jelly, Noemie S" uniqKey="Jelly N" first="Noémie S" last="Jelly">Noémie S. 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{{Explor lien
|wiki= Bois
|area= GrapevineDiseaseV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:22427113
|texte= Transient expression of artificial microRNAs targeting Grapevine fanleaf virus and evidence for RNA silencing in grapevine somatic embryos.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:22427113" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a GrapevineDiseaseV1
| This area was generated with Dilib version V0.6.37. |  |