Improved Agrobacterium -mediated transformation of cowpea via sonication and vacuum infiltration
Identifieur interne : 002552 ( Main/Exploration ); précédent : 002551; suivant : 002553Improved Agrobacterium -mediated transformation of cowpea via sonication and vacuum infiltration
Auteurs : Souvika Bakshi [Inde] ; Ayan Sadhukhan [Inde] ; Sagarika Mishra [Inde] ; Lingaraj Sahoo [Inde]Source :
- Plant Cell Reports [ 0721-7714 ] ; 2011-12-01.
English descriptors
Abstract
Abstract: An improved method of Agrobacterium-mediated transformation of cowpea was developed employing both sonication and vacuum infiltration treatments. 4 day-old cotyledonary nodes were used as explants for co-cultivation with Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pSouv-cry1Ac. Among the different injury treatments, vacuum infiltration and their combination treatments tested, sonication for 20 s followed by vacuum infiltration for 5 min with A. tumefaciens resulted in highest transient GUS expression efficiency (93% explants expressing GUS at regenerating sites). After 3 days of co-cultivation, the explants were cultured in 150 mg/l kanamycin-containing selection medium and putative transformed plants were recovered. The presence, integration and expression of nptII and cry1Ac genes in T0 transgenic plants were confirmed by polymerase chain reaction (PCR), genomic Southern and qualitative reverse transcription (RT)-PCR analysis. Western blot hybridization and enzyme-linked immunosorbent assay (ELISA) detected and demonstrated the accumulation of Cry1Ac protein in transgenic plants. The cry1Ac gene transmitted in a Mendelian fashion. The stable transformation efficiency increased by 88.4% using both sonication-assisted Agrobacterium-mediated transformation (SAAT) and vacuum infiltration than simple Agrobacterium-mediated transformation in cowpea.
Url:
DOI: 10.1007/s00299-011-1133-8
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000A54
- to stream Istex, to step Curation: 000A54
- to stream Istex, to step Checkpoint: 000481
- to stream Main, to step Merge: 002577
- to stream Main, to step Curation: 002552
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Improved Agrobacterium -mediated transformation of cowpea via sonication and vacuum infiltration</title><author><name sortKey="Bakshi, Souvika" sort="Bakshi, Souvika" uniqKey="Bakshi S" first="Souvika" last="Bakshi">Souvika Bakshi</name></author><author><name sortKey="Sadhukhan, Ayan" sort="Sadhukhan, Ayan" uniqKey="Sadhukhan A" first="Ayan" last="Sadhukhan">Ayan Sadhukhan</name></author><author><name sortKey="Mishra, Sagarika" sort="Mishra, Sagarika" uniqKey="Mishra S" first="Sagarika" last="Mishra">Sagarika Mishra</name></author><author><name sortKey="Sahoo, Lingaraj" sort="Sahoo, Lingaraj" uniqKey="Sahoo L" first="Lingaraj" last="Sahoo">Lingaraj Sahoo</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:DAC1E8C57D248B29FEAAE9AE60D836F759C9433D</idno><date when="2011" year="2011">2011</date><idno type="doi">10.1007/s00299-011-1133-8</idno><idno type="url">https://api.istex.fr/ark:/67375/VQC-T185RNLS-V/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000A54</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000A54</idno><idno type="wicri:Area/Istex/Curation">000A54</idno><idno type="wicri:Area/Istex/Checkpoint">000481</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000481</idno><idno type="wicri:doubleKey">0721-7714:2011:Bakshi S:improved:agrobacterium:mediated</idno><idno type="wicri:Area/Main/Merge">002577</idno><idno type="wicri:Area/Main/Curation">002552</idno><idno type="wicri:Area/Main/Exploration">002552</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Improved Agrobacterium -mediated transformation of cowpea via sonication and vacuum infiltration</title><author><name sortKey="Bakshi, Souvika" sort="Bakshi, Souvika" uniqKey="Bakshi S" first="Souvika" last="Bakshi">Souvika Bakshi</name><affiliation wicri:level="1"><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biotechnology, Indian Institute of Technology Guwahati, 781039, Guwahati, Assam</wicri:regionArea><wicri:noRegion>Assam</wicri:noRegion></affiliation></author><author><name sortKey="Sadhukhan, Ayan" sort="Sadhukhan, Ayan" uniqKey="Sadhukhan A" first="Ayan" last="Sadhukhan">Ayan Sadhukhan</name><affiliation wicri:level="1"><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biotechnology, Indian Institute of Technology Guwahati, 781039, Guwahati, Assam</wicri:regionArea><wicri:noRegion>Assam</wicri:noRegion></affiliation></author><author><name sortKey="Mishra, Sagarika" sort="Mishra, Sagarika" uniqKey="Mishra S" first="Sagarika" last="Mishra">Sagarika Mishra</name><affiliation wicri:level="1"><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biotechnology, Indian Institute of Technology Guwahati, 781039, Guwahati, Assam</wicri:regionArea><wicri:noRegion>Assam</wicri:noRegion></affiliation></author><author><name sortKey="Sahoo, Lingaraj" sort="Sahoo, Lingaraj" uniqKey="Sahoo L" first="Lingaraj" last="Sahoo">Lingaraj Sahoo</name><affiliation wicri:level="1"><country xml:lang="fr">Inde</country><wicri:regionArea>Department of Biotechnology, Indian Institute of Technology Guwahati, 781039, Guwahati, Assam</wicri:regionArea><wicri:noRegion>Assam</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Inde</country></affiliation></author></analytic><monogr></monogr><series><title level="j">Plant Cell Reports</title><title level="j" type="abbrev">Plant Cell Rep</title><idno type="ISSN">0721-7714</idno><idno type="eISSN">1432-203X</idno><imprint><publisher>Springer-Verlag</publisher><pubPlace>Berlin/Heidelberg</pubPlace><date type="published" when="2011-12-01">2011-12-01</date><biblScope unit="volume">30</biblScope><biblScope unit="issue">12</biblScope><biblScope unit="page" from="2281">2281</biblScope><biblScope unit="page" to="2292">2292</biblScope></imprint><idno type="ISSN">0721-7714</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0721-7714</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Cowpea</term><term>Sonication</term><term>Vacuum infiltration</term><term>Wounding</term><term>cry1Ac</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: An improved method of Agrobacterium-mediated transformation of cowpea was developed employing both sonication and vacuum infiltration treatments. 4 day-old cotyledonary nodes were used as explants for co-cultivation with Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pSouv-cry1Ac. Among the different injury treatments, vacuum infiltration and their combination treatments tested, sonication for 20 s followed by vacuum infiltration for 5 min with A. tumefaciens resulted in highest transient GUS expression efficiency (93% explants expressing GUS at regenerating sites). After 3 days of co-cultivation, the explants were cultured in 150 mg/l kanamycin-containing selection medium and putative transformed plants were recovered. The presence, integration and expression of nptII and cry1Ac genes in T0 transgenic plants were confirmed by polymerase chain reaction (PCR), genomic Southern and qualitative reverse transcription (RT)-PCR analysis. Western blot hybridization and enzyme-linked immunosorbent assay (ELISA) detected and demonstrated the accumulation of Cry1Ac protein in transgenic plants. The cry1Ac gene transmitted in a Mendelian fashion. The stable transformation efficiency increased by 88.4% using both sonication-assisted Agrobacterium-mediated transformation (SAAT) and vacuum infiltration than simple Agrobacterium-mediated transformation in cowpea.</div></front></TEI><affiliations><list><country><li>Inde</li></country></list><tree><country name="Inde"><noRegion><name sortKey="Bakshi, Souvika" sort="Bakshi, Souvika" uniqKey="Bakshi S" first="Souvika" last="Bakshi">Souvika Bakshi</name></noRegion><name sortKey="Mishra, Sagarika" sort="Mishra, Sagarika" uniqKey="Mishra S" first="Sagarika" last="Mishra">Sagarika Mishra</name><name sortKey="Sadhukhan, Ayan" sort="Sadhukhan, Ayan" uniqKey="Sadhukhan A" first="Ayan" last="Sadhukhan">Ayan Sadhukhan</name><name sortKey="Sahoo, Lingaraj" sort="Sahoo, Lingaraj" uniqKey="Sahoo L" first="Lingaraj" last="Sahoo">Lingaraj Sahoo</name><name sortKey="Sahoo, Lingaraj" sort="Sahoo, Lingaraj" uniqKey="Sahoo L" first="Lingaraj" last="Sahoo">Lingaraj Sahoo</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002552 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 002552 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:DAC1E8C57D248B29FEAAE9AE60D836F759C9433D
|texte= Improved Agrobacterium -mediated transformation of cowpea via sonication and vacuum infiltration
}}
| This area was generated with Dilib version V0.6.33. |  |