GUS expression in sweet oranges (Citrus sinensis L. Osbeck) driven by three different phloem-specific promoters.
Identifieur interne : 000596 ( PubMed/Checkpoint ); précédent : 000595; suivant : 000597GUS expression in sweet oranges (Citrus sinensis L. Osbeck) driven by three different phloem-specific promoters.
Auteurs : Luzia Yuriko Miyata [Brésil] ; Ricardo Harakava ; Liliane Cristina Lib Rio Stipp ; Beatriz Madalena Januzzi Mendes ; Beatriz Appezzato-Da-Gl Ria ; Francisco De Assis Alves Mourão FilhoSource :
- Plant cell reports [ 1432-203X ] ; 2012.
English descriptors
- KwdEn :
- Arabidopsis (genetics), Arabidopsis Proteins (genetics), Citrus sinensis (cytology), Citrus sinensis (genetics), Citrus sinensis (metabolism), Cloning, Molecular, Gene Expression Regulation, Plant, Glucuronidase (genetics), Glucuronidase (metabolism), Membrane Transport Proteins (genetics), Organ Specificity, Phloem (genetics), Phloem (metabolism), Plant Diseases (microbiology), Plant Leaves (genetics), Plant Leaves (metabolism), Plant Lectins (genetics), Plant Proteins (genetics), Plant Stems (genetics), Plant Stems (metabolism), Plants, Genetically Modified, Promoter Regions, Genetic (genetics), Regeneration, Rhizobiaceae (physiology), Transformation, Genetic.
- MESH :
- chemical , genetics : Arabidopsis Proteins, Glucuronidase, Membrane Transport Proteins, Plant Lectins, Plant Proteins.
- cytology : Citrus sinensis.
- genetics : Arabidopsis, Citrus sinensis, Phloem, Plant Leaves, Plant Stems, Promoter Regions, Genetic.
- metabolism : Citrus sinensis, Glucuronidase, Phloem, Plant Leaves, Plant Stems.
- microbiology : Plant Diseases.
- physiology : Rhizobiaceae.
- Cloning, Molecular, Gene Expression Regulation, Plant, Organ Specificity, Plants, Genetically Modified, Regeneration, Transformation, Genetic.
Abstract
Huanglongbing (HLB) is associated with Candidatus Liberibacter spp., endogenous, sieve tube-restricted bacteria that are transmitted by citrus psyllid insect vectors. Transgenic expression in the phloem of specific genes that might affect Ca. Liberibacter spp. growth and development may be an adequate strategy to improve citrus resistance to HLB. To study specific phloem gene expression in citrus, we developed three different binary vector constructs with expression cassettes bearing the β-glucuronidase (GUS) reporter gene (uidA) under the control of one of the three different promoters: Citrus phloem protein 2 (CsPP2), Arabidopsis thaliana phloem protein 2 (AtPP2), and Arabidopsis thaliana sucrose transporter 2 (AtSUC2). Transgenic lines of 'Hamlin', 'Pera', and 'Valencia' sweet oranges [Citrus sinensis (L.) Osbeck] were produced via Agrobacterium tumefaciens transformation. The epicotyl segments collected from in vitro germinated seedlings were used as explants. The gene nptII, which confers resistance to the antibiotic kanamycin, was used for selection. The transformation efficiency was expressed as the number of GUS-positive shoots over the total number of explants and varied from 1.54 to 6.08 % among the three cultivars and three constructs studied. Several lines of the three sweet orange cultivars analyzed using PCR and Southern blot analysis were genetically transformed with the three constructs evaluated. The histological GUS activity in the leaves indicates that the uidA gene was preferentially expressed in the phloem, which suggests that the use of the three promoters might be adequate for producing HLB-resistant transgenic sweet oranges. The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters. Key message The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters.
DOI: 10.1007/s00299-012-1312-2
PubMed: 22801867
Affiliations:
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Osbeck) driven by three different phloem-specific promoters.</title><author><name sortKey="Miyata, Luzia Yuriko" sort="Miyata, Luzia Yuriko" uniqKey="Miyata L" first="Luzia Yuriko" last="Miyata">Luzia Yuriko Miyata</name><affiliation wicri:level="4"><nlm:affiliation>Departamento de Produção Vegetal, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, 13418-900, Brazil.</nlm:affiliation><country xml:lang="fr">Brésil</country><wicri:regionArea>Departamento de Produção Vegetal, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, 13418-900</wicri:regionArea><orgName type="university">Université de São Paulo</orgName><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Harakava, Ricardo" sort="Harakava, Ricardo" uniqKey="Harakava R" first="Ricardo" last="Harakava">Ricardo Harakava</name></author><author><name sortKey="Stipp, Liliane Cristina Lib Rio" sort="Stipp, Liliane Cristina Lib Rio" uniqKey="Stipp L" first="Liliane Cristina Lib Rio" last="Stipp">Liliane Cristina Lib Rio Stipp</name></author><author><name sortKey="Mendes, Beatriz Madalena Januzzi" sort="Mendes, Beatriz Madalena Januzzi" uniqKey="Mendes B" first="Beatriz Madalena Januzzi" last="Mendes">Beatriz Madalena Januzzi Mendes</name></author><author><name sortKey="Appezzato Da Gl Ria, Beatriz" sort="Appezzato Da Gl Ria, Beatriz" uniqKey="Appezzato Da Gl Ria B" first="Beatriz" last="Appezzato-Da-Gl Ria">Beatriz Appezzato-Da-Gl Ria</name></author><author><name sortKey="De Assis Alves Mourao Filho, Francisco" sort="De Assis Alves Mourao Filho, Francisco" uniqKey="De Assis Alves Mourao Filho F" first="Francisco" last="De Assis Alves Mourão Filho">Francisco De Assis Alves Mourão Filho</name></author></analytic><series><title level="j">Plant cell reports</title><idno type="eISSN">1432-203X</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Arabidopsis (genetics)</term><term>Arabidopsis Proteins (genetics)</term><term>Citrus sinensis (cytology)</term><term>Citrus sinensis (genetics)</term><term>Citrus sinensis (metabolism)</term><term>Cloning, Molecular</term><term>Gene Expression Regulation, Plant</term><term>Glucuronidase (genetics)</term><term>Glucuronidase (metabolism)</term><term>Membrane Transport Proteins (genetics)</term><term>Organ Specificity</term><term>Phloem (genetics)</term><term>Phloem (metabolism)</term><term>Plant Diseases (microbiology)</term><term>Plant Leaves (genetics)</term><term>Plant Leaves (metabolism)</term><term>Plant Lectins (genetics)</term><term>Plant Proteins (genetics)</term><term>Plant Stems (genetics)</term><term>Plant Stems (metabolism)</term><term>Plants, Genetically Modified</term><term>Promoter Regions, Genetic (genetics)</term><term>Regeneration</term><term>Rhizobiaceae (physiology)</term><term>Transformation, Genetic</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Arabidopsis Proteins</term><term>Glucuronidase</term><term>Membrane Transport Proteins</term><term>Plant Lectins</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Citrus sinensis</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Arabidopsis</term><term>Citrus sinensis</term><term>Phloem</term><term>Plant Leaves</term><term>Plant Stems</term><term>Promoter Regions, Genetic</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Citrus sinensis</term><term>Glucuronidase</term><term>Phloem</term><term>Plant Leaves</term><term>Plant Stems</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Plant Diseases</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Rhizobiaceae</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cloning, Molecular</term><term>Gene Expression Regulation, Plant</term><term>Organ Specificity</term><term>Plants, Genetically Modified</term><term>Regeneration</term><term>Transformation, Genetic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Huanglongbing (HLB) is associated with Candidatus Liberibacter spp., endogenous, sieve tube-restricted bacteria that are transmitted by citrus psyllid insect vectors. Transgenic expression in the phloem of specific genes that might affect Ca. Liberibacter spp. growth and development may be an adequate strategy to improve citrus resistance to HLB. To study specific phloem gene expression in citrus, we developed three different binary vector constructs with expression cassettes bearing the β-glucuronidase (GUS) reporter gene (uidA) under the control of one of the three different promoters: Citrus phloem protein 2 (CsPP2), Arabidopsis thaliana phloem protein 2 (AtPP2), and Arabidopsis thaliana sucrose transporter 2 (AtSUC2). Transgenic lines of 'Hamlin', 'Pera', and 'Valencia' sweet oranges [Citrus sinensis (L.) Osbeck] were produced via Agrobacterium tumefaciens transformation. The epicotyl segments collected from in vitro germinated seedlings were used as explants. The gene nptII, which confers resistance to the antibiotic kanamycin, was used for selection. The transformation efficiency was expressed as the number of GUS-positive shoots over the total number of explants and varied from 1.54 to 6.08 % among the three cultivars and three constructs studied. Several lines of the three sweet orange cultivars analyzed using PCR and Southern blot analysis were genetically transformed with the three constructs evaluated. The histological GUS activity in the leaves indicates that the uidA gene was preferentially expressed in the phloem, which suggests that the use of the three promoters might be adequate for producing HLB-resistant transgenic sweet oranges. The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters. Key message The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22801867</PMID><DateCreated><Year>2012</Year><Month>10</Month><Day>16</Day></DateCreated><DateCompleted><Year>2013</Year><Month>03</Month><Day>21</Day></DateCompleted><DateRevised><Year>2012</Year><Month>10</Month><Day>16</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1432-203X</ISSN><JournalIssue CitedMedium="Internet"><Volume>31</Volume><Issue>11</Issue><PubDate><Year>2012</Year><Month>Nov</Month></PubDate></JournalIssue><Title>Plant cell reports</Title><ISOAbbreviation>Plant Cell Rep.</ISOAbbreviation></Journal><ArticleTitle>GUS expression in sweet oranges (Citrus sinensis L. Osbeck) driven by three different phloem-specific promoters.</ArticleTitle><Pagination><MedlinePgn>2005-13</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s00299-012-1312-2</ELocationID><Abstract><AbstractText>Huanglongbing (HLB) is associated with Candidatus Liberibacter spp., endogenous, sieve tube-restricted bacteria that are transmitted by citrus psyllid insect vectors. Transgenic expression in the phloem of specific genes that might affect Ca. Liberibacter spp. growth and development may be an adequate strategy to improve citrus resistance to HLB. To study specific phloem gene expression in citrus, we developed three different binary vector constructs with expression cassettes bearing the β-glucuronidase (GUS) reporter gene (uidA) under the control of one of the three different promoters: Citrus phloem protein 2 (CsPP2), Arabidopsis thaliana phloem protein 2 (AtPP2), and Arabidopsis thaliana sucrose transporter 2 (AtSUC2). Transgenic lines of 'Hamlin', 'Pera', and 'Valencia' sweet oranges [Citrus sinensis (L.) Osbeck] were produced via Agrobacterium tumefaciens transformation. The epicotyl segments collected from in vitro germinated seedlings were used as explants. The gene nptII, which confers resistance to the antibiotic kanamycin, was used for selection. The transformation efficiency was expressed as the number of GUS-positive shoots over the total number of explants and varied from 1.54 to 6.08 % among the three cultivars and three constructs studied. Several lines of the three sweet orange cultivars analyzed using PCR and Southern blot analysis were genetically transformed with the three constructs evaluated. The histological GUS activity in the leaves indicates that the uidA gene was preferentially expressed in the phloem, which suggests that the use of the three promoters might be adequate for producing HLB-resistant transgenic sweet oranges. The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters. Key message The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Miyata</LastName><ForeName>Luzia Yuriko</ForeName><Initials>LY</Initials><AffiliationInfo><Affiliation>Departamento de Produção Vegetal, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, 13418-900, Brazil.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Harakava</LastName><ForeName>Ricardo</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Stipp</LastName><ForeName>Liliane Cristina Libório</ForeName><Initials>LC</Initials></Author><Author ValidYN="Y"><LastName>Mendes</LastName><ForeName>Beatriz Madalena Januzzi</ForeName><Initials>BM</Initials></Author><Author ValidYN="Y"><LastName>Appezzato-da-Glória</LastName><ForeName>Beatriz</ForeName><Initials>B</Initials></Author><Author ValidYN="Y"><LastName>de Assis Alves Mourão Filho</LastName><ForeName>Francisco</ForeName><Initials>F</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>Jul</Month><Day>17</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Plant Cell Rep</MedlineTA><NlmUniqueID>9880970</NlmUniqueID><ISSNLinking>0721-7714</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D029681">Arabidopsis Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D026901">Membrane Transport Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C552547">PP2-A1 protein, Arabidopsis</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D037121">Plant Lectins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C078729">sucrose transport protein, plant</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.2.1.31</RegistryNumber><NameOfSubstance UI="D005966">Glucuronidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D017360" MajorTopicYN="N">Arabidopsis</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D029681" MajorTopicYN="N">Arabidopsis Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D032084" MajorTopicYN="N">Citrus sinensis</DescriptorName><QualifierName UI="Q000166" MajorTopicYN="N">cytology</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="N">Gene Expression Regulation, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005966" MajorTopicYN="N">Glucuronidase</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D026901" MajorTopicYN="N">Membrane Transport Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009928" MajorTopicYN="N">Organ Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D052585" MajorTopicYN="N">Phloem</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D037121" MajorTopicYN="N">Plant Lectins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010940" MajorTopicYN="N">Plant Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018547" MajorTopicYN="N">Plant Stems</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011401" MajorTopicYN="N">Promoter Regions, Genetic</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012038" MajorTopicYN="N">Regeneration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012230" MajorTopicYN="N">Rhizobiaceae</DescriptorName><QualifierName UI="Q000502" 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IdType="doi">10.1007/s00299-012-1312-2</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Brésil</li></country><region><li>État de São Paulo</li></region><settlement><li>São Paulo</li></settlement><orgName><li>Université de São Paulo</li></orgName></list><tree><noCountry><name sortKey="Appezzato Da Gl Ria, Beatriz" sort="Appezzato Da Gl Ria, Beatriz" uniqKey="Appezzato Da Gl Ria B" first="Beatriz" last="Appezzato-Da-Gl Ria">Beatriz Appezzato-Da-Gl Ria</name><name sortKey="De Assis Alves Mourao Filho, Francisco" sort="De Assis Alves Mourao Filho, Francisco" uniqKey="De Assis Alves Mourao Filho F" first="Francisco" last="De Assis Alves Mourão Filho">Francisco De Assis Alves Mourão Filho</name><name sortKey="Harakava, Ricardo" sort="Harakava, Ricardo" uniqKey="Harakava R" first="Ricardo" last="Harakava">Ricardo Harakava</name><name sortKey="Mendes, Beatriz Madalena Januzzi" sort="Mendes, Beatriz Madalena Januzzi" uniqKey="Mendes B" first="Beatriz Madalena Januzzi" last="Mendes">Beatriz Madalena Januzzi Mendes</name><name sortKey="Stipp, Liliane Cristina Lib Rio" sort="Stipp, Liliane Cristina Lib Rio" uniqKey="Stipp L" first="Liliane Cristina Lib Rio" last="Stipp">Liliane Cristina Lib Rio Stipp</name></noCountry><country name="Brésil"><region name="État de São Paulo"><name sortKey="Miyata, Luzia Yuriko" sort="Miyata, Luzia Yuriko" uniqKey="Miyata L" first="Luzia Yuriko" last="Miyata">Luzia Yuriko Miyata</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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