Agrobacterium-mediated transformation of thin cell layer explants of Scutellaria ocmulgee small: a rare plant with anti-tumor properties
Scutellaria ocmulgee (Ocmulgee skullcap) is a rare plant species with medicinal and ornamental value and requires immediate conservation. We report here the first protocol for plant regeneration and Agrobacterium -mediated transformation of S. ocmulgee using leaf and shoot-derived transverse thin ce...
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| Veröffentlicht in: | Plant cell, tissue and organ culture tissue and organ culture, 2016-10, Vol.127 (1), p.57-69 |
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| creator | Vaidya, Brajesh N. Jackson, Carissa L. Perry, Zachary D. Dhekney, Sadanand A. Joshee, Nirmal |
| description | Scutellaria ocmulgee
(Ocmulgee skullcap) is a rare plant species with medicinal and ornamental value and requires immediate conservation. We report here the first protocol for plant regeneration and
Agrobacterium
-mediated transformation of
S. ocmulgee
using leaf and shoot-derived transverse thin cell layer (tTCL) explants. The effect of MS and B5 salts in combination with varying levels of growth regulators and two carbon sources on shoot proliferation and plant regeneration were studied. Among the various media treatment combinations, the best shoot induction response was observed from leaf-derived tTCL explants on B5 medium supplemented with 6- benzyl aminopurine (BAP), 1-naphthalene acetic acid (NAA), and sucrose whereas shoot-derived tTCL explants produced the maximum number of shoots also on B5 medium supplemented with either BAP or thidiazuron, NAA and maltose. Shoots obtained from both types of explants were rooted on MS medium containing 5.0 µM indole butyric acid.
Agrobacterium
-mediated genetic transformation protocol was optimized using leaf tTCL explants. A binary plasmid pq35SGR harboring the β glucuronidase and green fluorescent protein/neomycin phosphotransferase II fusion reporter gene was used to optimize transformation parameters. Explants were co-cultivated with
Agrobacterium tumefaciens
EHA105 and then transferred to shoot induction and regeneration medium to generate transgenic plants. Stable gene expression was observed in transgenic cultures and plants. The presence and integration of transgenes was confirmed by polymerase chain reaction, reverse transcriptase-polymerase chain reaction and Southern blot hybridization. |
| doi_str_mv | 10.1007/s11240-016-1029-y |
| format | Article |
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(Ocmulgee skullcap) is a rare plant species with medicinal and ornamental value and requires immediate conservation. We report here the first protocol for plant regeneration and
Agrobacterium
-mediated transformation of
S. ocmulgee
using leaf and shoot-derived transverse thin cell layer (tTCL) explants. The effect of MS and B5 salts in combination with varying levels of growth regulators and two carbon sources on shoot proliferation and plant regeneration were studied. Among the various media treatment combinations, the best shoot induction response was observed from leaf-derived tTCL explants on B5 medium supplemented with 6- benzyl aminopurine (BAP), 1-naphthalene acetic acid (NAA), and sucrose whereas shoot-derived tTCL explants produced the maximum number of shoots also on B5 medium supplemented with either BAP or thidiazuron, NAA and maltose. Shoots obtained from both types of explants were rooted on MS medium containing 5.0 µM indole butyric acid.
Agrobacterium
-mediated genetic transformation protocol was optimized using leaf tTCL explants. A binary plasmid pq35SGR harboring the β glucuronidase and green fluorescent protein/neomycin phosphotransferase II fusion reporter gene was used to optimize transformation parameters. Explants were co-cultivated with
Agrobacterium tumefaciens
EHA105 and then transferred to shoot induction and regeneration medium to generate transgenic plants. Stable gene expression was observed in transgenic cultures and plants. The presence and integration of transgenes was confirmed by polymerase chain reaction, reverse transcriptase-polymerase chain reaction and Southern blot hybridization.</description><identifier>ISSN: 0167-6857</identifier><identifier>EISSN: 1573-5044</identifier><identifier>DOI: 10.1007/s11240-016-1029-y</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Acetic acid ; Agrobacterium ; Anticancer properties ; Benzyladenine ; Biomedical and Life Sciences ; Butyric acid ; Carbon sources ; Culture media ; Explants ; Fluorescence ; Fusion protein ; Gene expression ; Genetic transformation ; Green fluorescent protein ; Growth regulators ; Hybridization ; Indole-3-butyric acid ; Indoles ; Integration ; Leaves ; Life Sciences ; Maltose ; Medicinal plants ; Naphthalene ; Neomycin ; Neomycin phosphotransferase ; Original Article ; Ornamental plants ; Phosphotransferase ; Phosphotransferase II ; Plant Genetics and Genomics ; Plant Pathology ; Plant Physiology ; Plant Sciences ; Plant species ; Polymerase chain reaction ; Rare species ; Regeneration ; Regulators ; Scutellaria ocmulgee ; Shoots ; Sucrose ; Sugar ; Thidiazuron ; Transgenic plants ; Wildlife conservation</subject><ispartof>Plant cell, tissue and organ culture, 2016-10, Vol.127 (1), p.57-69</ispartof><rights>Springer Science+Business Media Dordrecht 2016</rights><rights>Copyright Springer Science & Business Media 2016</rights><rights>Plant Cell, Tissue and Organ Culture (PCTOC) is a copyright of Springer, (2016). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c344t-e52e9d82ad511e22cd7b8f85418c04b7a9c0618a23c766baff9a3f5964e4bdaf3</citedby><cites>FETCH-LOGICAL-c344t-e52e9d82ad511e22cd7b8f85418c04b7a9c0618a23c766baff9a3f5964e4bdaf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11240-016-1029-y$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11240-016-1029-y$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids></links><search><creatorcontrib>Vaidya, Brajesh N.</creatorcontrib><creatorcontrib>Jackson, Carissa L.</creatorcontrib><creatorcontrib>Perry, Zachary D.</creatorcontrib><creatorcontrib>Dhekney, Sadanand A.</creatorcontrib><creatorcontrib>Joshee, Nirmal</creatorcontrib><title>Agrobacterium-mediated transformation of thin cell layer explants of Scutellaria ocmulgee small: a rare plant with anti-tumor properties</title><title>Plant cell, tissue and organ culture</title><addtitle>Plant Cell Tiss Organ Cult</addtitle><description>Scutellaria ocmulgee
(Ocmulgee skullcap) is a rare plant species with medicinal and ornamental value and requires immediate conservation. We report here the first protocol for plant regeneration and
Agrobacterium
-mediated transformation of
S. ocmulgee
using leaf and shoot-derived transverse thin cell layer (tTCL) explants. The effect of MS and B5 salts in combination with varying levels of growth regulators and two carbon sources on shoot proliferation and plant regeneration were studied. Among the various media treatment combinations, the best shoot induction response was observed from leaf-derived tTCL explants on B5 medium supplemented with 6- benzyl aminopurine (BAP), 1-naphthalene acetic acid (NAA), and sucrose whereas shoot-derived tTCL explants produced the maximum number of shoots also on B5 medium supplemented with either BAP or thidiazuron, NAA and maltose. Shoots obtained from both types of explants were rooted on MS medium containing 5.0 µM indole butyric acid.
Agrobacterium
-mediated genetic transformation protocol was optimized using leaf tTCL explants. A binary plasmid pq35SGR harboring the β glucuronidase and green fluorescent protein/neomycin phosphotransferase II fusion reporter gene was used to optimize transformation parameters. Explants were co-cultivated with
Agrobacterium tumefaciens
EHA105 and then transferred to shoot induction and regeneration medium to generate transgenic plants. Stable gene expression was observed in transgenic cultures and plants. The presence and integration of transgenes was confirmed by polymerase chain reaction, reverse transcriptase-polymerase chain reaction and Southern blot hybridization.</description><subject>Acetic acid</subject><subject>Agrobacterium</subject><subject>Anticancer properties</subject><subject>Benzyladenine</subject><subject>Biomedical and Life Sciences</subject><subject>Butyric acid</subject><subject>Carbon sources</subject><subject>Culture media</subject><subject>Explants</subject><subject>Fluorescence</subject><subject>Fusion protein</subject><subject>Gene expression</subject><subject>Genetic transformation</subject><subject>Green fluorescent protein</subject><subject>Growth regulators</subject><subject>Hybridization</subject><subject>Indole-3-butyric acid</subject><subject>Indoles</subject><subject>Integration</subject><subject>Leaves</subject><subject>Life Sciences</subject><subject>Maltose</subject><subject>Medicinal plants</subject><subject>Naphthalene</subject><subject>Neomycin</subject><subject>Neomycin phosphotransferase</subject><subject>Original Article</subject><subject>Ornamental plants</subject><subject>Phosphotransferase</subject><subject>Phosphotransferase II</subject><subject>Plant Genetics and Genomics</subject><subject>Plant Pathology</subject><subject>Plant Physiology</subject><subject>Plant Sciences</subject><subject>Plant species</subject><subject>Polymerase chain reaction</subject><subject>Rare species</subject><subject>Regeneration</subject><subject>Regulators</subject><subject>Scutellaria ocmulgee</subject><subject>Shoots</subject><subject>Sucrose</subject><subject>Sugar</subject><subject>Thidiazuron</subject><subject>Transgenic plants</subject><subject>Wildlife conservation</subject><issn>0167-6857</issn><issn>1573-5044</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNp9kUFv1DAQhS1EpS6FH9CbJc4G27Edh1tVQUGqxAE4WxNnvHWVxIvtCPYf9GfjZTlwgdOM9L73RppHyLXgbwTn_dsihFSccWGY4HJgx2dkJ3TfMc2Vek52TeiZsbq_JC9KeeScm06JHXm62ec0gq-Y47awBacIFSdaM6wlpLxAjWmlKdD6EFfqcZ7pDEfMFH8eZlhrOWlf_FabAjkCTX7Z5j0iLQvM8zsKNENG-humP2J9oG2JrG5LyvSQ0wFzjVhekosAc8FXf-YV-fbh_dfbj-z-892n25t75julKkMtcZishEkLgVL6qR9tsFoJ67kaexg8N8KC7HxvzAghDNAFPRiFapwgdFfk9Tm3nf6-YanuMW15bSedlHrobG-t-R8lrOXWDJp3jRJnyudUSsbgDjkukI9OcHeqxZ1rce377lSLOzaPPHtKY9c95r-S_2n6BbsCkyY</recordid><startdate>20161001</startdate><enddate>20161001</enddate><creator>Vaidya, Brajesh N.</creator><creator>Jackson, Carissa L.</creator><creator>Perry, Zachary D.</creator><creator>Dhekney, Sadanand A.</creator><creator>Joshee, Nirmal</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X2</scope><scope>8FE</scope><scope>8FH</scope><scope>8FK</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>LK8</scope><scope>M0K</scope><scope>M7P</scope><scope>PHGZM</scope><scope>PHGZT</scope><scope>PKEHL</scope><scope>PQEST</scope><scope>PQGLB</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope></search><sort><creationdate>20161001</creationdate><title>Agrobacterium-mediated transformation of thin cell layer explants of Scutellaria ocmulgee small: a rare plant with anti-tumor properties</title><author>Vaidya, Brajesh N. ; Jackson, Carissa L. ; Perry, Zachary D. ; Dhekney, Sadanand A. ; Joshee, Nirmal</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c344t-e52e9d82ad511e22cd7b8f85418c04b7a9c0618a23c766baff9a3f5964e4bdaf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Acetic acid</topic><topic>Agrobacterium</topic><topic>Anticancer properties</topic><topic>Benzyladenine</topic><topic>Biomedical and Life Sciences</topic><topic>Butyric acid</topic><topic>Carbon sources</topic><topic>Culture media</topic><topic>Explants</topic><topic>Fluorescence</topic><topic>Fusion protein</topic><topic>Gene expression</topic><topic>Genetic transformation</topic><topic>Green fluorescent protein</topic><topic>Growth regulators</topic><topic>Hybridization</topic><topic>Indole-3-butyric acid</topic><topic>Indoles</topic><topic>Integration</topic><topic>Leaves</topic><topic>Life Sciences</topic><topic>Maltose</topic><topic>Medicinal plants</topic><topic>Naphthalene</topic><topic>Neomycin</topic><topic>Neomycin phosphotransferase</topic><topic>Original Article</topic><topic>Ornamental plants</topic><topic>Phosphotransferase</topic><topic>Phosphotransferase II</topic><topic>Plant Genetics and Genomics</topic><topic>Plant Pathology</topic><topic>Plant Physiology</topic><topic>Plant Sciences</topic><topic>Plant species</topic><topic>Polymerase chain reaction</topic><topic>Rare species</topic><topic>Regeneration</topic><topic>Regulators</topic><topic>Scutellaria ocmulgee</topic><topic>Shoots</topic><topic>Sucrose</topic><topic>Sugar</topic><topic>Thidiazuron</topic><topic>Transgenic plants</topic><topic>Wildlife conservation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Vaidya, Brajesh N.</creatorcontrib><creatorcontrib>Jackson, Carissa L.</creatorcontrib><creatorcontrib>Perry, Zachary D.</creatorcontrib><creatorcontrib>Dhekney, Sadanand A.</creatorcontrib><creatorcontrib>Joshee, Nirmal</creatorcontrib><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Agricultural Science Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection (ProQuest)</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Biological Science Database</collection><collection>ProQuest Central (New)</collection><collection>ProQuest One Academic (New)</collection><collection>ProQuest One Academic Middle East (New)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Applied & Life Sciences</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><jtitle>Plant cell, tissue and organ culture</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Vaidya, Brajesh N.</au><au>Jackson, Carissa L.</au><au>Perry, Zachary D.</au><au>Dhekney, Sadanand A.</au><au>Joshee, Nirmal</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Agrobacterium-mediated transformation of thin cell layer explants of Scutellaria ocmulgee small: a rare plant with anti-tumor properties</atitle><jtitle>Plant cell, tissue and organ culture</jtitle><stitle>Plant Cell Tiss Organ Cult</stitle><date>2016-10-01</date><risdate>2016</risdate><volume>127</volume><issue>1</issue><spage>57</spage><epage>69</epage><pages>57-69</pages><issn>0167-6857</issn><eissn>1573-5044</eissn><abstract>Scutellaria ocmulgee
(Ocmulgee skullcap) is a rare plant species with medicinal and ornamental value and requires immediate conservation. We report here the first protocol for plant regeneration and
Agrobacterium
-mediated transformation of
S. ocmulgee
using leaf and shoot-derived transverse thin cell layer (tTCL) explants. The effect of MS and B5 salts in combination with varying levels of growth regulators and two carbon sources on shoot proliferation and plant regeneration were studied. Among the various media treatment combinations, the best shoot induction response was observed from leaf-derived tTCL explants on B5 medium supplemented with 6- benzyl aminopurine (BAP), 1-naphthalene acetic acid (NAA), and sucrose whereas shoot-derived tTCL explants produced the maximum number of shoots also on B5 medium supplemented with either BAP or thidiazuron, NAA and maltose. Shoots obtained from both types of explants were rooted on MS medium containing 5.0 µM indole butyric acid.
Agrobacterium
-mediated genetic transformation protocol was optimized using leaf tTCL explants. A binary plasmid pq35SGR harboring the β glucuronidase and green fluorescent protein/neomycin phosphotransferase II fusion reporter gene was used to optimize transformation parameters. Explants were co-cultivated with
Agrobacterium tumefaciens
EHA105 and then transferred to shoot induction and regeneration medium to generate transgenic plants. Stable gene expression was observed in transgenic cultures and plants. The presence and integration of transgenes was confirmed by polymerase chain reaction, reverse transcriptase-polymerase chain reaction and Southern blot hybridization.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><doi>10.1007/s11240-016-1029-y</doi><tpages>13</tpages></addata></record> |
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| subjects | Acetic acid Agrobacterium Anticancer properties Benzyladenine Biomedical and Life Sciences Butyric acid Carbon sources Culture media Explants Fluorescence Fusion protein Gene expression Genetic transformation Green fluorescent protein Growth regulators Hybridization Indole-3-butyric acid Indoles Integration Leaves Life Sciences Maltose Medicinal plants Naphthalene Neomycin Neomycin phosphotransferase Original Article Ornamental plants Phosphotransferase Phosphotransferase II Plant Genetics and Genomics Plant Pathology Plant Physiology Plant Sciences Plant species Polymerase chain reaction Rare species Regeneration Regulators Scutellaria ocmulgee Shoots Sucrose Sugar Thidiazuron Transgenic plants Wildlife conservation |
| title | Agrobacterium-mediated transformation of thin cell layer explants of Scutellaria ocmulgee small: a rare plant with anti-tumor properties |
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