Highly efficient visual selection of transgenic rice plants using green fluorescent protein or anthocyanin synthetic genes

Hiroaki Saika, Wataru Sakamoto, Masahiko Maekawa, Seiichi Toki

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Visual selection enables transformation in plants that are sensitive to the stress of antibiotic selection. Furthermore, we proposed a method of isolating transformed cells in which the gene of interest is expressed at high level using visual selection. However, visual selection was inefficient and laborious because of the technical difficulties involved in clonal propagation of transformed calli selected using GFP (9.4-20.5% in the model japonica rice variety Nipponbare in our previous report). Very recently, we have discovered that an indica model cultivar, Kasalath, is very highly competent for Agrobacterium-mediated transformation compared to Nipponbare, suggesting that visual selection would be achieved more efficiently in Kasalath than in Nipponbare. We confirmed that the number of transformed cells emitting green fluorescence in Agrobacterium co-cultivated callus of Kasalath under no antibiotic pressure was >50-fold higher than the number obtained in Nipponbare. We succeeded in clonal propagation of GFP emitting cells at a frequency of 62.2-85% in Kasalath. Moreover, we showed that anthocyanin, that is an intrinsic pigmentation and thus might be more acceptable to consumers, can be used as an intrinsic marker in visual selection of rice. Thus, we propose that combination of the use of cultivar which has high transformation competency and visual selection system could enhance efficient production of transgenic rice expressing foreign gene product at higher level.

Original languageEnglish
Pages (from-to)107-110
Number of pages4
JournalPlant Biotechnology
Volume28
Issue number1
DOIs
Publication statusPublished - 2011

Fingerprint

Synthetic Genes
synthetic genes
Anthocyanins
Genetically Modified Plants
Green Fluorescent Proteins
green fluorescent protein
anthocyanins
Agrobacterium
Bony Callus
genetically modified organisms
rice
callus
antibiotics
Anti-Bacterial Agents
Pigmentation
cultivars
cells
pigmentation
Genes
genes

Keywords

  • Agrobacterium
  • Indica rice
  • Transformation frequency
  • Visible selection

ASJC Scopus subject areas

  • Plant Science
  • Biotechnology
  • Agronomy and Crop Science

Cite this

Highly efficient visual selection of transgenic rice plants using green fluorescent protein or anthocyanin synthetic genes. / Saika, Hiroaki; Sakamoto, Wataru; Maekawa, Masahiko; Toki, Seiichi.

In: Plant Biotechnology, Vol. 28, No. 1, 2011, p. 107-110.

Research output: Contribution to journalArticle

@article{25b05ab0f8e64d21a1308da411c9c15a,
title = "Highly efficient visual selection of transgenic rice plants using green fluorescent protein or anthocyanin synthetic genes",
abstract = "Visual selection enables transformation in plants that are sensitive to the stress of antibiotic selection. Furthermore, we proposed a method of isolating transformed cells in which the gene of interest is expressed at high level using visual selection. However, visual selection was inefficient and laborious because of the technical difficulties involved in clonal propagation of transformed calli selected using GFP (9.4-20.5{\%} in the model japonica rice variety Nipponbare in our previous report). Very recently, we have discovered that an indica model cultivar, Kasalath, is very highly competent for Agrobacterium-mediated transformation compared to Nipponbare, suggesting that visual selection would be achieved more efficiently in Kasalath than in Nipponbare. We confirmed that the number of transformed cells emitting green fluorescence in Agrobacterium co-cultivated callus of Kasalath under no antibiotic pressure was >50-fold higher than the number obtained in Nipponbare. We succeeded in clonal propagation of GFP emitting cells at a frequency of 62.2-85{\%} in Kasalath. Moreover, we showed that anthocyanin, that is an intrinsic pigmentation and thus might be more acceptable to consumers, can be used as an intrinsic marker in visual selection of rice. Thus, we propose that combination of the use of cultivar which has high transformation competency and visual selection system could enhance efficient production of transgenic rice expressing foreign gene product at higher level.",
keywords = "Agrobacterium, Indica rice, Transformation frequency, Visible selection",
author = "Hiroaki Saika and Wataru Sakamoto and Masahiko Maekawa and Seiichi Toki",
year = "2011",
doi = "10.5511/plantbiotechnology.10.1104a",
language = "English",
volume = "28",
pages = "107--110",
journal = "Plant Biotechnology",
issn = "1342-4580",
publisher = "Japanese Society for Plant Cell and Molecular Biology",
number = "1",

}

TY - JOUR

T1 - Highly efficient visual selection of transgenic rice plants using green fluorescent protein or anthocyanin synthetic genes

AU - Saika, Hiroaki

AU - Sakamoto, Wataru

AU - Maekawa, Masahiko

AU - Toki, Seiichi

PY - 2011

Y1 - 2011

N2 - Visual selection enables transformation in plants that are sensitive to the stress of antibiotic selection. Furthermore, we proposed a method of isolating transformed cells in which the gene of interest is expressed at high level using visual selection. However, visual selection was inefficient and laborious because of the technical difficulties involved in clonal propagation of transformed calli selected using GFP (9.4-20.5% in the model japonica rice variety Nipponbare in our previous report). Very recently, we have discovered that an indica model cultivar, Kasalath, is very highly competent for Agrobacterium-mediated transformation compared to Nipponbare, suggesting that visual selection would be achieved more efficiently in Kasalath than in Nipponbare. We confirmed that the number of transformed cells emitting green fluorescence in Agrobacterium co-cultivated callus of Kasalath under no antibiotic pressure was >50-fold higher than the number obtained in Nipponbare. We succeeded in clonal propagation of GFP emitting cells at a frequency of 62.2-85% in Kasalath. Moreover, we showed that anthocyanin, that is an intrinsic pigmentation and thus might be more acceptable to consumers, can be used as an intrinsic marker in visual selection of rice. Thus, we propose that combination of the use of cultivar which has high transformation competency and visual selection system could enhance efficient production of transgenic rice expressing foreign gene product at higher level.

AB - Visual selection enables transformation in plants that are sensitive to the stress of antibiotic selection. Furthermore, we proposed a method of isolating transformed cells in which the gene of interest is expressed at high level using visual selection. However, visual selection was inefficient and laborious because of the technical difficulties involved in clonal propagation of transformed calli selected using GFP (9.4-20.5% in the model japonica rice variety Nipponbare in our previous report). Very recently, we have discovered that an indica model cultivar, Kasalath, is very highly competent for Agrobacterium-mediated transformation compared to Nipponbare, suggesting that visual selection would be achieved more efficiently in Kasalath than in Nipponbare. We confirmed that the number of transformed cells emitting green fluorescence in Agrobacterium co-cultivated callus of Kasalath under no antibiotic pressure was >50-fold higher than the number obtained in Nipponbare. We succeeded in clonal propagation of GFP emitting cells at a frequency of 62.2-85% in Kasalath. Moreover, we showed that anthocyanin, that is an intrinsic pigmentation and thus might be more acceptable to consumers, can be used as an intrinsic marker in visual selection of rice. Thus, we propose that combination of the use of cultivar which has high transformation competency and visual selection system could enhance efficient production of transgenic rice expressing foreign gene product at higher level.

KW - Agrobacterium

KW - Indica rice

KW - Transformation frequency

KW - Visible selection

UR - http://www.scopus.com/inward/record.url?scp=79955149226&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79955149226&partnerID=8YFLogxK

U2 - 10.5511/plantbiotechnology.10.1104a

DO - 10.5511/plantbiotechnology.10.1104a

M3 - Article

VL - 28

SP - 107

EP - 110

JO - Plant Biotechnology

JF - Plant Biotechnology

SN - 1342-4580

IS - 1

ER -