Agrobacterium-mediated transformation of switchgrass and inheritance of the transgenes

Yajun Xi; Chunxiang Fu; Yaxin Ge; Rangaraj Nandakumar; Hiroshi Hisano; Joseph Bouton; Zeng Yu Wang

doi:10.1007/s12155-009-9049-7

Agrobacterium-mediated transformation of switchgrass and inheritance of the transgenes

Yajun Xi, Chunxiang Fu, Yaxin Ge, Rangaraj Nandakumar, Hiroshi Hisano, Joseph Bouton, Zeng Yu Wang

Research output: Contribution to journal › Article › peer-review

65 Citations (Scopus)

Abstract

Switchgrass (Panicum virgatum L.) has been developed into an important biofuel crop. Embryogenic calli induced from caryopses or inflorescences of the lowland switchgrass cultivar Alamo were used for Agrobacterium-mediated transformation. A chimeric hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin as the selection agent. Embryogenic calli were infected with Agrobacterium tumefaciens strain EHA105. Calli resistant to hygromycin were obtained after 5 to 8 weeks of selection. Soil-grown transgenic switchgrass plants were obtained 4 to 5 months after Agrobacterium infection. The transgenic nature of the regenerated plants was demonstrated by PCR, Southern blot hybridization analysis, and GUS staining. T1 progeny were obtained after reciprocal crosses between transgenic and untransformed control plants. Molecular analyses of the T1 progeny revealed various patterns of segregation. Transgene silencing was observed in the progeny with multiple inserts. Interestingly, reversal of the expression of the silenced transgene was found in segregating progeny with a single insert.

Original language	English
Pages (from-to)	275-283
Number of pages	9
Journal	Bioenergy Research
Volume	2
Issue number	4
DOIs	https://doi.org/10.1007/s12155-009-9049-7
Publication status	Published - Nov 2009
Externally published	Yes

Keywords

Agrobacterium
Biofuel crop
Panicum virgatum
Switchgrass
Transgene inheritance
Transgenic plant

ASJC Scopus subject areas

Renewable Energy, Sustainability and the Environment
Agronomy and Crop Science
Energy (miscellaneous)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1007/s12155-009-9049-7

Cite this

@article{067c5d9d93ef422280d0d6b3b9447700,

title = "Agrobacterium-mediated transformation of switchgrass and inheritance of the transgenes",

abstract = "Switchgrass (Panicum virgatum L.) has been developed into an important biofuel crop. Embryogenic calli induced from caryopses or inflorescences of the lowland switchgrass cultivar Alamo were used for Agrobacterium-mediated transformation. A chimeric hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin as the selection agent. Embryogenic calli were infected with Agrobacterium tumefaciens strain EHA105. Calli resistant to hygromycin were obtained after 5 to 8 weeks of selection. Soil-grown transgenic switchgrass plants were obtained 4 to 5 months after Agrobacterium infection. The transgenic nature of the regenerated plants was demonstrated by PCR, Southern blot hybridization analysis, and GUS staining. T1 progeny were obtained after reciprocal crosses between transgenic and untransformed control plants. Molecular analyses of the T1 progeny revealed various patterns of segregation. Transgene silencing was observed in the progeny with multiple inserts. Interestingly, reversal of the expression of the silenced transgene was found in segregating progeny with a single insert.",

keywords = "Agrobacterium, Biofuel crop, Panicum virgatum, Switchgrass, Transgene inheritance, Transgenic plant",

author = "Yajun Xi and Chunxiang Fu and Yaxin Ge and Rangaraj Nandakumar and Hiroshi Hisano and Joseph Bouton and Wang, {Zeng Yu}",

note = "Funding Information: Acknowledgments We thank Frank Hardin and Jackie Kelly for critical reading of the manuscript. The work was supported by the US Department of Agriculture and US Department of Energy Biomass Initiative (project no. 2009-10003-05140), the BioEnergy Science Center, and the Samuel Roberts Noble Foundation. The BioEnergy Science Center is supported by the Office of Biological and Environmental Research in the DOE Office of Science. This report was prepared as an account of work partly sponsored by the US Government. Neither the US Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product or process disclosed or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favoring by the US Government or any agency thereof. The views and opinions of the authors expressed herein do not necessarily reflect those of the US Government or any agency thereof. Copyright: Copyright 2012 Elsevier B.V., All rights reserved.",

year = "2009",

month = nov,

doi = "10.1007/s12155-009-9049-7",

language = "English",

volume = "2",

pages = "275--283",

journal = "Bioenergy Research",

issn = "1939-1234",

publisher = "Springer New York",

number = "4",

}

TY - JOUR

T1 - Agrobacterium-mediated transformation of switchgrass and inheritance of the transgenes

AU - Xi, Yajun

AU - Fu, Chunxiang

AU - Ge, Yaxin

AU - Nandakumar, Rangaraj

AU - Hisano, Hiroshi

AU - Bouton, Joseph

AU - Wang, Zeng Yu

N1 - Funding Information: Acknowledgments We thank Frank Hardin and Jackie Kelly for critical reading of the manuscript. The work was supported by the US Department of Agriculture and US Department of Energy Biomass Initiative (project no. 2009-10003-05140), the BioEnergy Science Center, and the Samuel Roberts Noble Foundation. The BioEnergy Science Center is supported by the Office of Biological and Environmental Research in the DOE Office of Science. This report was prepared as an account of work partly sponsored by the US Government. Neither the US Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product or process disclosed or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favoring by the US Government or any agency thereof. The views and opinions of the authors expressed herein do not necessarily reflect those of the US Government or any agency thereof. Copyright: Copyright 2012 Elsevier B.V., All rights reserved.

PY - 2009/11

Y1 - 2009/11

N2 - Switchgrass (Panicum virgatum L.) has been developed into an important biofuel crop. Embryogenic calli induced from caryopses or inflorescences of the lowland switchgrass cultivar Alamo were used for Agrobacterium-mediated transformation. A chimeric hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin as the selection agent. Embryogenic calli were infected with Agrobacterium tumefaciens strain EHA105. Calli resistant to hygromycin were obtained after 5 to 8 weeks of selection. Soil-grown transgenic switchgrass plants were obtained 4 to 5 months after Agrobacterium infection. The transgenic nature of the regenerated plants was demonstrated by PCR, Southern blot hybridization analysis, and GUS staining. T1 progeny were obtained after reciprocal crosses between transgenic and untransformed control plants. Molecular analyses of the T1 progeny revealed various patterns of segregation. Transgene silencing was observed in the progeny with multiple inserts. Interestingly, reversal of the expression of the silenced transgene was found in segregating progeny with a single insert.

AB - Switchgrass (Panicum virgatum L.) has been developed into an important biofuel crop. Embryogenic calli induced from caryopses or inflorescences of the lowland switchgrass cultivar Alamo were used for Agrobacterium-mediated transformation. A chimeric hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin as the selection agent. Embryogenic calli were infected with Agrobacterium tumefaciens strain EHA105. Calli resistant to hygromycin were obtained after 5 to 8 weeks of selection. Soil-grown transgenic switchgrass plants were obtained 4 to 5 months after Agrobacterium infection. The transgenic nature of the regenerated plants was demonstrated by PCR, Southern blot hybridization analysis, and GUS staining. T1 progeny were obtained after reciprocal crosses between transgenic and untransformed control plants. Molecular analyses of the T1 progeny revealed various patterns of segregation. Transgene silencing was observed in the progeny with multiple inserts. Interestingly, reversal of the expression of the silenced transgene was found in segregating progeny with a single insert.

KW - Agrobacterium

KW - Biofuel crop

KW - Panicum virgatum

KW - Switchgrass

KW - Transgene inheritance

KW - Transgenic plant

UR - http://www.scopus.com/inward/record.url?scp=70449567552&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70449567552&partnerID=8YFLogxK

U2 - 10.1007/s12155-009-9049-7

DO - 10.1007/s12155-009-9049-7

M3 - Article

AN - SCOPUS:70449567552

SN - 1939-1234

VL - 2

SP - 275

EP - 283

JO - Bioenergy Research

JF - Bioenergy Research

IS - 4

ER -

Agrobacterium-mediated transformation of switchgrass and inheritance of the transgenes

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this