Transformation of androgenic-derived Festulolium plants (Lolium perenne L. × Festuca pratensis Huds.) by Agrobacterium tumefaciens

Yang Dong Guo; Hisano Hisano; Yoshiya Shimamoto; Toshihiko Yamada

doi:10.1007/s11240-008-9478-6

Transformation of androgenic-derived Festulolium plants (Lolium perenne L. × Festuca pratensis Huds.) by Agrobacterium tumefaciens

Yang Dong Guo, Hisano Hisano, Yoshiya Shimamoto, Toshihiko Yamada

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

10 Citations (Scopus)

Abstract

Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. × Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium "Bx351" were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and β-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling.

Original language	English
Pages (from-to)	219-227
Number of pages	9
Journal	Plant Cell, Tissue and Organ Culture
Volume	96
Issue number	2
DOIs	https://doi.org/10.1007/s11240-008-9478-6
Publication status	Published - Feb 2009
Externally published	Yes

Keywords

Androgenic callus
Anther culture
Dihaploid
GUS activity
Transgenic forage grass

ASJC Scopus subject areas

Horticulture

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10.1007/s11240-008-9478-6

Cite this

@article{10f20980cad44bc58fea9564ab672020,

title = "Transformation of androgenic-derived Festulolium plants (Lolium perenne L. × Festuca pratensis Huds.) by Agrobacterium tumefaciens",

abstract = "Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. × Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium {"}Bx351{"} were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and β-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling.",

keywords = "Androgenic callus, Anther culture, Dihaploid, GUS activity, Transgenic forage grass",

author = "Guo, {Yang Dong} and Hisano Hisano and Yoshiya Shimamoto and Toshihiko Yamada",

note = "Funding Information: Acknowledgments We thank Prof. K. Nakamura, the Graduate School of Bioagricultural Sciences, Nagoya University, Japan for providing the plasmid pIG121-Hm and Ms. M. Kawamura for technical assistance. We also thank Prof. M. O. Humphreys, the Institute of Grassland and Environmental Research, UK for careful critical reading of the manuscript. This work was partially supported by Grants from the Japanese Society for the Promotion of Science (JSPS, P02440) and Chinese government (NSFC30871821, 2008BAD B3B03) to Y.-D. Guo and from the Northern Advancement Center for Science & Technology to T. Yamada.",

year = "2009",

month = feb,

doi = "10.1007/s11240-008-9478-6",

language = "English",

volume = "96",

pages = "219--227",

journal = "Plant Cell, Tissue and Organ Culture",

issn = "0167-6857",

publisher = "Springer Netherlands",

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}

TY - JOUR

T1 - Transformation of androgenic-derived Festulolium plants (Lolium perenne L. × Festuca pratensis Huds.) by Agrobacterium tumefaciens

AU - Guo, Yang Dong

AU - Hisano, Hisano

AU - Shimamoto, Yoshiya

AU - Yamada, Toshihiko

N1 - Funding Information: Acknowledgments We thank Prof. K. Nakamura, the Graduate School of Bioagricultural Sciences, Nagoya University, Japan for providing the plasmid pIG121-Hm and Ms. M. Kawamura for technical assistance. We also thank Prof. M. O. Humphreys, the Institute of Grassland and Environmental Research, UK for careful critical reading of the manuscript. This work was partially supported by Grants from the Japanese Society for the Promotion of Science (JSPS, P02440) and Chinese government (NSFC30871821, 2008BAD B3B03) to Y.-D. Guo and from the Northern Advancement Center for Science & Technology to T. Yamada.

PY - 2009/2

Y1 - 2009/2

N2 - Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. × Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium "Bx351" were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and β-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling.

AB - Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. × Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium "Bx351" were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and β-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling.

KW - Androgenic callus

KW - Anther culture

KW - Dihaploid

KW - GUS activity

KW - Transgenic forage grass

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JO - Plant Cell, Tissue and Organ Culture

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ER -

Transformation of androgenic-derived Festulolium plants (Lolium perenne L. × Festuca pratensis Huds.) by Agrobacterium tumefaciens

Abstract

Keywords

ASJC Scopus subject areas

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