The <it>Rg1 </it>allele as a valuable tool for genetic transformation of the tomato 'Micro-Tom' model system
2010
Quecini Vera | Borgo Lucélia | Scotton Danielle C | Azevedo Mariana S | Lombardi-Crestana Simone | Pino Lilian E | Figueira Antonio | Peres Lázaro EP
<p>Abstract</p> <p>Background</p> <p>The cultivar Micro-Tom (MT) is regarded as a model system for tomato genetics due to its short life cycle and miniature size. However, efforts to improve tomato genetic transformation have led to protocols dependent on the costly hormone zeatin, combined with an excessive number of steps.</p> <p>Results</p> <p>Here we report the development of a MT near-isogenic genotype harboring the allele <it>Rg1 </it>(MT-<it>Rg1</it>), which greatly improves tomato <it>in vitro </it>regeneration. Regeneration was further improved in MT by including a two-day incubation of cotyledonary explants onto medium containing 0.4 μM 1-naphthaleneacetic acid (NAA) before cytokinin treatment. Both strategies allowed the use of 5 μM 6-benzylaminopurine (BAP), a cytokinin 100 times less expensive than zeatin. The use of MT-<it>Rg1 </it>and NAA pre-incubation, followed by BAP regeneration, resulted in high transformation frequencies (near 40%), in a shorter protocol with fewer steps, spanning approximately 40 days from <it>Agrobacterium </it>infection to transgenic plant acclimatization.</p> <p>Conclusions</p> <p>The genetic resource and the protocol presented here represent invaluable tools for routine gene expression manipulation and high throughput functional genomics by insertional mutagenesis in tomato.</p>
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