Efficient shoot regeneration from hairy roots of Antirrhinum majus L. transformed by the rol type MAT vector system

Cui, Ming; Takayanagi, Kenji; Kamada, Hiroshi; Nishimura, Shigeo; Handa, Takashi

doi:10.1007/s002990000277

Cited by 37 publications

(22 citation statements)

References 22 publications

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“…After a few weeks (to months), spontaneous regeneration may occur from the green segments of the hairy roots. Transformed wild-type shoots can be distinguished easily from the rol gene-containing shoots because the latter have wrinkled leaves, no apical dominance, and shortened internodes (Ebinuma et al 1997b;Cui et al 2000Cui et al , 2001. Unfortunately, chimeric transgenic shoots that are difficult to discriminate are also observed.…”

Section: The Multi-auto Transformation (Mat) Vector Systemmentioning

confidence: 96%

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Agrobacterium rhizogenes-Mediated Transformation of Plants

Velde¹,

Karimi²,

Herder³

et al. 2003

Genetic Transformation of Plants

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Section: The Multi-auto Transformation (Mat) Vector Systemmentioning

confidence: 96%

“…The system has been successfully used for tobacco and Antirrhinum majus (Cui et al 2000(Cui et al ,2001. In tobacco, roots have to be transferred to shootinducing medium to grow shoots.…”

Section: The Multi-auto Transformation (Mat) Vector Systemmentioning

confidence: 99%

Agrobacterium rhizogenes-Mediated Transformation of Plants

Velde¹,

Karimi²,

Herder³

et al. 2003

Genetic Transformation of Plants

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“…We show that the hairy root phenotype, which is regarded as being undesirable in standard transformation experiments (Ebinuma et al 1997;Cui et al 2001), is beneficial if the goal of the transformation is to rapidly produce and analyze significant quantities of the recombinant protein.…”

Section: Introductionmentioning

confidence: 99%

“…Cotransformation using a mixture of A. rhizogenes and A. tumefaciens cultures harboring the binary expression vector (Torregroza and Bouquet 1997) did not overcome these limitations, while the direct integration of the expression cassette into A. rhizogenes T-DNA via homologous recombination (Stougaard et al 1987) proved cumbersome and inefficient. Finally, a removal MAT vector system combined with a positive rooting marker consisting of four genes-rolA, rolB, rolC, and rolD (Ebinuma et al 1997)-was exploited for the transformation and regeneration of marker-free transgenic snapdragon plants, with only 3% of regenerated shoots reported suitable for the study (Cui et al 2001).…”

Section: Introductionmentioning

confidence: 99%

A quick and efficient system for antibiotic-free expression of heterologous genes in tobacco roots

et al. 2004

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Requirement for antibiotic-resistance selection markers and difficulty in identifying transgenes with the highest expression levels remain the major obstacles for rapid production of recombinant proteins in plants. An alternative approach to producing transgenic plants free of antibiotic-resistance markers is the phenotypic-based selection with root-proliferation genes (rol genes) of Agrobacterium rhizogenes. By using Agrobacterium tumefaciens harboring the pRYG transformation vector with a cluster of rol genes linked to a heterologous gene of interest, we have developed a rapid transformation tool using hairy root formation as a selection marker. The expression of beta-glucuronidase in newly induced transgenic tobacco roots could be detected as early as 12 days after inoculation. Higher levels of transgene expression in the roots correlated positively with the rates of root elongation on hormone-free medium and thus could be used for positive selection. When tobacco plants were transformed with pRYG harboring the expression cassette for secreted alkaline phosphatase (SEAP), the release of SEAP from roots of the fully regenerated transgenic plants could be quantified at rates as high as 28 microg/g root dry weight per day.

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“…In rol-type MAT vector system, on the other hand, the possibility of nontransgenic escapes might be minimized since transgenic tissues could be obtained as hairy roots as a transgenic event. However, successful transformation using rol-type MAT vector has been reported only in Antirrhinum majus (Cui et al 2000(Cui et al , 2001) and needs to be evaluated in other plant species.…”

mentioning

confidence: 99%