Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx. (muscadine grape).
Identifieur interne : 000684 ( Main/Corpus ); précédent : 000683; suivant : 000685Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx. (muscadine grape).
Auteurs : S A Dhekney ; Z T Li ; M. Dutt ; D J GraySource :
- Plant cell reports [ 0721-7714 ] ; 2008.
English descriptors
- KwdEn :
- Blotting, Southern (MeSH), Cells, Cultured (MeSH), Kanamycin (pharmacology), Kanamycin Kinase (genetics), Kanamycin Kinase (metabolism), Plants, Genetically Modified (drug effects), Plants, Genetically Modified (genetics), Plants, Genetically Modified (physiology), Polymerase Chain Reaction (MeSH), Regeneration (drug effects), Regeneration (genetics), Regeneration (physiology), Rhizobium (genetics), Transformation, Genetic (MeSH), Vitis (cytology), Vitis (embryology), Vitis (genetics).
- MESH :
- chemical , genetics : Kanamycin Kinase.
- chemical , metabolism : Kanamycin Kinase.
- chemical , pharmacology : Kanamycin.
- cytology : Vitis.
- drug effects : Plants, Genetically Modified, Regeneration.
- embryology : Vitis.
- genetics : Plants, Genetically Modified, Regeneration, Rhizobium, Vitis.
- physiology : Plants, Genetically Modified, Regeneration.
- Blotting, Southern, Cells, Cultured, Polymerase Chain Reaction, Transformation, Genetic.
Abstract
A method to produce transgenic plants of Vitis rotundifolia was developed. Embryogenic cultures were initiated from leaves of in vitro grown shoot cultures and used as target tissues for Agrobacterium-mediated genetic transformation. A green fluorescent protein/neomycin phosphotransferase II (gfp/nptII) fusion gene that allowed for simultaneous selection of transgenic cells based on GFP fluorescence and kanamycin resistance was used to optimize parameters influencing genetic transformation. It was determined that both proembryonal masses (PEM) and mid-cotyledonary stage somatic embryos (SE) were suitable target tissues for co-cultivation with Agrobacterium as evidenced by transient GFP expression. Kanamycin at 100 mg l(-1) in the culture medium was effective in suppression of non-transformed tissue and permitting the growth and development of transgenic cells, compared to 50 or 75 mg l(-1), which permitted the proliferation of more non-transformed cells. Transgenic plants of "Alachua" and "Carlos" were recovered after secondary somatic embryogenesis from primary SE explants co-cultivated with Agrobacterium. The presence and stable integration of transgenes in transgenic plants was confirmed by PCR and Southern blot hybridization. Transgenic plants exhibited uniform GFP expression in cells of all plant tissues and organs including leaves, stems, roots, inflorescences and the embryo and endosperm of developing berries.
DOI: 10.1007/s00299-008-0512-2
PubMed: 18256837
Links to Exploration step
pubmed:18256837Le document en format XML
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<author><name sortKey="Dhekney, S A" sort="Dhekney, S A" uniqKey="Dhekney S" first="S A" last="Dhekney">S A Dhekney</name>
<affiliation><nlm:affiliation>Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL 32703-8504, USA.</nlm:affiliation>
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<author><name sortKey="Li, Z T" sort="Li, Z T" uniqKey="Li Z" first="Z T" last="Li">Z T Li</name>
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<author><name sortKey="Dutt, M" sort="Dutt, M" uniqKey="Dutt M" first="M" last="Dutt">M. Dutt</name>
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<author><name sortKey="Gray, D J" sort="Gray, D J" uniqKey="Gray D" first="D J" last="Gray">D J Gray</name>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx. (muscadine grape).</title>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Blotting, Southern (MeSH)</term>
<term>Cells, Cultured (MeSH)</term>
<term>Kanamycin (pharmacology)</term>
<term>Kanamycin Kinase (genetics)</term>
<term>Kanamycin Kinase (metabolism)</term>
<term>Plants, Genetically Modified (drug effects)</term>
<term>Plants, Genetically Modified (genetics)</term>
<term>Plants, Genetically Modified (physiology)</term>
<term>Polymerase Chain Reaction (MeSH)</term>
<term>Regeneration (drug effects)</term>
<term>Regeneration (genetics)</term>
<term>Regeneration (physiology)</term>
<term>Rhizobium (genetics)</term>
<term>Transformation, Genetic (MeSH)</term>
<term>Vitis (cytology)</term>
<term>Vitis (embryology)</term>
<term>Vitis (genetics)</term>
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<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Kanamycin Kinase</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Kanamycin Kinase</term>
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<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Kanamycin</term>
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<keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Plants, Genetically Modified</term>
<term>Regeneration</term>
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<keywords scheme="MESH" qualifier="embryology" xml:lang="en"><term>Vitis</term>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Plants, Genetically Modified</term>
<term>Regeneration</term>
<term>Rhizobium</term>
<term>Vitis</term>
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<keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Plants, Genetically Modified</term>
<term>Regeneration</term>
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<keywords scheme="MESH" xml:lang="en"><term>Blotting, Southern</term>
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<front><div type="abstract" xml:lang="en">A method to produce transgenic plants of Vitis rotundifolia was developed. Embryogenic cultures were initiated from leaves of in vitro grown shoot cultures and used as target tissues for Agrobacterium-mediated genetic transformation. A green fluorescent protein/neomycin phosphotransferase II (gfp/nptII) fusion gene that allowed for simultaneous selection of transgenic cells based on GFP fluorescence and kanamycin resistance was used to optimize parameters influencing genetic transformation. It was determined that both proembryonal masses (PEM) and mid-cotyledonary stage somatic embryos (SE) were suitable target tissues for co-cultivation with Agrobacterium as evidenced by transient GFP expression. Kanamycin at 100 mg l(-1) in the culture medium was effective in suppression of non-transformed tissue and permitting the growth and development of transgenic cells, compared to 50 or 75 mg l(-1), which permitted the proliferation of more non-transformed cells. Transgenic plants of "Alachua" and "Carlos" were recovered after secondary somatic embryogenesis from primary SE explants co-cultivated with Agrobacterium. The presence and stable integration of transgenes in transgenic plants was confirmed by PCR and Southern blot hybridization. Transgenic plants exhibited uniform GFP expression in cells of all plant tissues and organs including leaves, stems, roots, inflorescences and the embryo and endosperm of developing berries.</div>
</front>
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<Title>Plant cell reports</Title>
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<ArticleTitle>Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx. (muscadine grape).</ArticleTitle>
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<Abstract><AbstractText>A method to produce transgenic plants of Vitis rotundifolia was developed. Embryogenic cultures were initiated from leaves of in vitro grown shoot cultures and used as target tissues for Agrobacterium-mediated genetic transformation. A green fluorescent protein/neomycin phosphotransferase II (gfp/nptII) fusion gene that allowed for simultaneous selection of transgenic cells based on GFP fluorescence and kanamycin resistance was used to optimize parameters influencing genetic transformation. It was determined that both proembryonal masses (PEM) and mid-cotyledonary stage somatic embryos (SE) were suitable target tissues for co-cultivation with Agrobacterium as evidenced by transient GFP expression. Kanamycin at 100 mg l(-1) in the culture medium was effective in suppression of non-transformed tissue and permitting the growth and development of transgenic cells, compared to 50 or 75 mg l(-1), which permitted the proliferation of more non-transformed cells. Transgenic plants of "Alachua" and "Carlos" were recovered after secondary somatic embryogenesis from primary SE explants co-cultivated with Agrobacterium. The presence and stable integration of transgenes in transgenic plants was confirmed by PCR and Southern blot hybridization. Transgenic plants exhibited uniform GFP expression in cells of all plant tissues and organs including leaves, stems, roots, inflorescences and the embryo and endosperm of developing berries.</AbstractText>
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