Effective selection system for generating marker-free transgenic plants independent of sexual crossing

Sugita, Koichi; Matsunaga, Etsuko; Ebinuma, Hiroyasu

doi:10.1007/s002990050688

Cited by 90 publications

(63 citation statements)

References 23 publications

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“…A rol-type MAT binary vector plasmid, pMAT101, used in this study was constructed by Sugita et al (1999), and kindly provided by Nippon Paper Industries, Japan (Figure 1). The pMAT101 plasmid contains a 'hit-and-run' cassette, in which the b-glucuronidase (gus) gene under the control of CaMV 35S promoter, the rol genes (rol A, B, C and D genes) of Ri T-DNA and the recombinase gene (R) with the 35S promoter are located between directly oriented recombination site (RS) sequences (Ebinuma et al 1997).…”

Section: Bacterial Strain and Vector Plasmidmentioning

confidence: 99%

Generation of phenotypically normal marker-free transgenic plants of Kalanchoe blossfeldiana through hairy root induction

Thirukkumarana

Ntui

Khan

et al. 2010

Plant Biotechnology

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Section: Bacterial Strain and Vector Plasmidmentioning

confidence: 99%

Generation of phenotypically normal marker-free transgenic plants of Kalanchoe blossfeldiana through hairy root induction

Thirukkumarana

Ntui

Khan

et al. 2010

Plant Biotechnology

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“…SR1 plants was performed as previously described (Sugita et al 1999). After co-cultivation of leaf segments with A. tumefaciens LBA4404 (Hoekema et al 1983) containing the appropriate plasmid on solid hormone-free MS medium containing 50 mg/1 acetosyringone for 3 days, the explants were transferred to solid SIM medium (MS medium containing 1 mg/l benzyladenine, 0.1 mg/l naphthalene acetic acid and 100 mg/l kanamycin) containing 500 mg/1 ticarcillin to prevent further bacterial growth.…”

Section: Transgenic Plantsmentioning

confidence: 99%

The transposon Tip100 from the common morning glory is an autonomous element that can transpose in tobacco plants

et al. 2002

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The mutable flaked or a (flaked) (a(f)) line of the common morning glory (Ipomoea purpurea) displays white flowers with colored flakes, and the a(f) mutation is caused by the insertion of a transposable element named Tip100 into the CHS-D gene for anthocyanin biosynthesis. The 3.9-kb Tip100 element belongs to the Ac/Ds family and contains an ORF encoding a polypeptide of 808 amino acids. The frequency and timing of flower variegation vary in different a(f) lines, and a genetic element termed Modulator has been postulated to affect the variegation pattern. Since the pattern of flower variegation is determined by the frequency and timing of excision of Tip100 from the CHS-D gene, we wished to determine whether Tip100 is an autonomous element that is itself capable of transposition in a heterologous host. To do this, we introduced the element into the genome of tobacco plants by Agrobacterium-mediated transformation. The intact Tip100 element was able to excise from its original position in the chromosome and reinsert into new sites in the tobacco genome, whereas an internal deletion derivative was not. Based on these results, we conclude that Tip100 is an autonomous element. We also discuss the nature of the putative Modulator element affecting flower and leaf variegation in various mutable lines of the morning glory.

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“…Most of these marker genes are isolated from microorganisms and are heterologous to plants. Therefore, transgenic plants that are resistant to antibiotics or herbicides may possibly be hazardous to human health and/or the environment (Sugita et al, 1999;Daniell et al, 2001). Methods have recently been developed to eliminate resistance marker genes, such as co-transformation, site-specific recombination systems, and the use of transposons or homologous recombination.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of an endo-1,4-β-glucanase V gene (EGV) from Trichoderma reesei leads to the accumulation of cellulase activity in transgenic rice

Li¹,

F²,

Yf³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. The ectopic expression of cellulase in biomass can reduce the cost of biofuel conversion. This trait modification technique is highly beneficial for biofuel production. In this study, we isolated an endo-1,4-beta-glucanase gene (EGV) from Trichoderma reesei and inserted this gene downstream of a fragment encoding the signal peptide Apo-SP in a modified pCAMBIA1301 vector to obtain an Apo-SP and AsRed fusion protein. Transient expression of this fusion protein in onion epidermal cells showed that the Apo-SP signal was localized to the plastids. EGV transgenic rice plants that did not carry screening marker genes were obtained through overexpression of the pDTB double T-DNA vector. Western blotting showed that EGV was expressed in the dry straw of T 0 generation transgenic rice plants and in fresh leaves of the T 1 generation. More importantly, our results also showed that the peptide product of EGV in the transgenic plants folded correctly and was capable of digesting the cellulase substrate CMC. Additionally, cellulase activity remained stable in

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Effective selection system for generating marker-free transgenic plants independent of sexual crossing

Cited by 90 publications

References 23 publications

Generation of phenotypically normal marker-free transgenic plants of Kalanchoe blossfeldiana through hairy root induction

Generation of phenotypically normal marker-free transgenic plants of Kalanchoe blossfeldiana through hairy root induction

The transposon Tip100 from the common morning glory is an autonomous element that can transpose in tobacco plants

Overexpression of an endo-1,4-β-glucanase V gene (EGV) from Trichoderma reesei leads to the accumulation of cellulase activity in transgenic rice

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