Efficient Agrobacterium-Mediated Transformation of Hybrid Poplar Populus davidiana Dode x Populus bollena Lauche.
Identifieur interne : 002714 ( Main/Exploration ); précédent : 002713; suivant : 002715Efficient Agrobacterium-Mediated Transformation of Hybrid Poplar Populus davidiana Dode x Populus bollena Lauche.
Auteurs : Xue Han [République populaire de Chine] ; Shurong Ma ; Xianghui Kong ; Tetsuo Takano ; Shenkui LiuSource :
- International journal of molecular sciences [ 1422-0067 ] ; 2013.
Abstract
Poplar is a model organism for high in vitro regeneration in woody plants. We have chosen a hybrid poplar Populus davidiana Dode x Populus bollena Lauche. By optimizing the Murashige and Skoog medium with (0.3 mg/L) 6-benzylaminopurine and (0.08 mg/L) naphthaleneacetic acid, we have achieved the highest frequency (90%) for shoot regeneration from poplar leaves. It was also important to improve the transformation efficiency of poplar for genetic breeding and other applications. In this study, we found a significant improvement of the transformation frequency by controlling the leaf age. Transformation efficiency was enhanced by optimizing the Agrobacterium concentration (OD(600) = 0.8-1.0) and an infection time (20-30 min). According to transmission electron microscopy observations, there were more Agrobacterium invasions in the 30-day-old leaf explants than in 60-day-old and 90-day-old explants. Using the green fluorescent protein (GFP) marker, the expression of MD-GFP fusion proteins in the leaf, shoot, and root of hybrid poplar P. davidiana Dode x Populus. bollena Lauche was visualized for confirmation of transgene integration. Southern and Northern blot analysis also showed the integration of T-DNA into the genome and gene expression of transgenic plants. Our results suggest that younger leaves had higher transformation efficiency (~30%) than older leaves (10%).
DOI: 10.3390/ijms14022515
PubMed: 23354481
PubMed Central: PMC3587999
Affiliations:
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We have chosen a hybrid poplar Populus davidiana Dode x Populus bollena Lauche. By optimizing the Murashige and Skoog medium with (0.3 mg/L) 6-benzylaminopurine and (0.08 mg/L) naphthaleneacetic acid, we have achieved the highest frequency (90%) for shoot regeneration from poplar leaves. It was also important to improve the transformation efficiency of poplar for genetic breeding and other applications. In this study, we found a significant improvement of the transformation frequency by controlling the leaf age. Transformation efficiency was enhanced by optimizing the Agrobacterium concentration (OD(600) = 0.8-1.0) and an infection time (20-30 min). According to transmission electron microscopy observations, there were more Agrobacterium invasions in the 30-day-old leaf explants than in 60-day-old and 90-day-old explants. Using the green fluorescent protein (GFP) marker, the expression of MD-GFP fusion proteins in the leaf, shoot, and root of hybrid poplar P. davidiana Dode x Populus. bollena Lauche was visualized for confirmation of transgene integration. Southern and Northern blot analysis also showed the integration of T-DNA into the genome and gene expression of transgenic plants. Our results suggest that younger leaves had higher transformation efficiency (~30%) than older leaves (10%).</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">23354481</PMID><DateCompleted><Year>2013</Year><Month>01</Month><Day>29</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Print">1422-0067</ISSN><JournalIssue CitedMedium="Print"><Volume>14</Volume><Issue>2</Issue><PubDate><Year>2013</Year><Month>Jan</Month><Day>25</Day></PubDate></JournalIssue><Title>International journal of molecular sciences</Title><ISOAbbreviation>Int J Mol Sci</ISOAbbreviation></Journal><ArticleTitle>Efficient Agrobacterium-Mediated Transformation of Hybrid Poplar Populus davidiana Dode x Populus bollena Lauche.</ArticleTitle><Pagination><MedlinePgn>2515-28</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.3390/ijms14022515</ELocationID><Abstract><AbstractText>Poplar is a model organism for high in vitro regeneration in woody plants. We have chosen a hybrid poplar Populus davidiana Dode x Populus bollena Lauche. By optimizing the Murashige and Skoog medium with (0.3 mg/L) 6-benzylaminopurine and (0.08 mg/L) naphthaleneacetic acid, we have achieved the highest frequency (90%) for shoot regeneration from poplar leaves. It was also important to improve the transformation efficiency of poplar for genetic breeding and other applications. In this study, we found a significant improvement of the transformation frequency by controlling the leaf age. Transformation efficiency was enhanced by optimizing the Agrobacterium concentration (OD(600) = 0.8-1.0) and an infection time (20-30 min). According to transmission electron microscopy observations, there were more Agrobacterium invasions in the 30-day-old leaf explants than in 60-day-old and 90-day-old explants. Using the green fluorescent protein (GFP) marker, the expression of MD-GFP fusion proteins in the leaf, shoot, and root of hybrid poplar P. davidiana Dode x Populus. bollena Lauche was visualized for confirmation of transgene integration. Southern and Northern blot analysis also showed the integration of T-DNA into the genome and gene expression of transgenic plants. Our results suggest that younger leaves had higher transformation efficiency (~30%) than older leaves (10%).</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Han</LastName><ForeName>Xue</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin Hexing Road, Harbin 150040, China. shenkuiliu@nefu.edu.cn.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ma</LastName><ForeName>Shurong</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Kong</LastName><ForeName>Xianghui</ForeName><Initials>X</Initials></Author><Author ValidYN="Y"><LastName>Takano</LastName><ForeName>Tetsuo</ForeName><Initials>T</Initials></Author><Author 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