The Rg1 allele as a valuable tool for genetic transformation of the tomato 'Micro-Tom' model system


Autoria(s): PINO, Lilian E.; LOMBARDI-CRESTANA, Simone; AZEVEDO, Mariana S.; SCOTTON, Danielle C.; BORGO, Lucelia; QUECINI, Vera; FIGUEIRA, Antonio; PERES, Lazaro E. P.
Contribuinte(s)

UNIVERSIDADE DE SÃO PAULO

Data(s)

17/04/2012

17/04/2012

2010

Resumo

Background: 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. Results: Here we report the development of a MT near-isogenic genotype harboring the allele Rg1 (MT-Rg1), which greatly improves tomato in vitro regeneration. Regeneration was further improved in MT by including a two-day incubation of cotyledonary explants onto medium containing 0.4 mu M 1-naphthaleneacetic acid (NAA) before cytokinin treatment. Both strategies allowed the use of 5 mu M 6-benzylaminopurine (BAP), a cytokinin 100 times less expensive than zeatin. The use of MT-Rg1 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 Agrobacterium infection to transgenic plant acclimatization. Conclusions: 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.

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[02/00329-8]

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[04/15268-0]

Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq [475494/03-2]

Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq [308075/03-0]

Identificador

PLANT METHODS, v.6, 2010

1746-4811

http://producao.usp.br/handle/BDPI/14541

10.1186/1746-4811-6-23

http://dx.doi.org/10.1186/1746-4811-6-23

Idioma(s)

eng

Publicador

BIOMED CENTRAL LTD

Relação

Plant Methods

Direitos

openAccess

Copyright BIOMED CENTRAL LTD

Palavras-Chave #AGROBACTERIUM-MEDIATED TRANSFORMATION #SHOOT REGENERATION #FUNCTIONAL GENOMICS #IN-VITRO #PLANTS #TUMEFACIENS #PROTOCOL #GENOTYPE #AUXIN #BIOSYNTHESIS #Plant Sciences
Tipo

article

original article

publishedVersion