An improved method for Agrobacterium rhizogenes -mediated transformation of tomato suitable for the study of arbuscular
- PDF / 2,439,584 Bytes
- 12 Pages / 595.276 x 790.866 pts Page_size
- 59 Downloads / 204 Views
nt Methods Open Access
METHODOLOGY
An improved method for Agrobacterium rhizogenes‑mediated transformation of tomato suitable for the study of arbuscular mycorrhizal symbiosis Tania Ho‑Plágaro1, Raúl Huertas2, María I. Tamayo‑Navarrete1, Juan A. Ocampo1 and José M. García‑Garrido1*
Abstract Background: Solanum lycopersicum, an economically important crop grown worldwide, has been used as a model for the study of arbuscular mycorrhizal (AM) symbiosis in non-legume plants for several years and several cDNA array hybridization studies have revealed specific transcriptomic profiles of mycorrhizal tomato roots. However, a method to easily screen candidate genes which could play an important role during tomato mycorrhization is required. Results: We have developed an optimized procedure for composite tomato plant obtaining achieved through Agrobacterium rhizogenes-mediated transformation. This protocol involves the unusual in vitro culture of composite plants between two filter papers placed on the culture media. In addition, we show that DsRed is an appropriate molecular marker for the precise selection of cotransformed tomato hairy roots. S. lycopersicum composite plant hairy roots appear to be colonized by the AM fungus Rhizophagus irregularis in a manner similar to that of normal roots, and a modified construct useful for localizing the expression of promoters putatively associated with mycorrhization was developed and tested. Conclusions: In this study, we present an easy, fast and low-cost procedure to study AM symbiosis in tomato roots. Keywords: Transformation, Composite plants, Hairy roots, Tomato, Arbuscular mycorrhiza Background The Solanum lycopersicum (tomato) is an economically important crop grown worldwide. Tomato production accounts for 16% of global vegetable production and constitutes the largest vegetable category. The total tomato crop is forecast to reach 38.6 million tons in 2017 and is expected to continue growing (https://www.wptc.to/). Tomato crops are susceptible to many insects, bacteria and nematodes which cause significant reductions in fruit yields under current production practices [1]. Rhizosphere symbiotic microorganisms are increasingly *Correspondence: [email protected] 1 Department of Soil Microbiology and Symbiotic Systems, Estación Experimental del Zaidín (EEZ), CSIC, Calle Profesor Albareda n◦1, 18008 Granada, Spain Full list of author information is available at the end of the article
proposed as a possible alternative to overcoming these problems [2], with particular emphasis on arbuscular mycorrhizal (AM) fungi which form mutualistic symbiotic association with most higher plants. Although tomato growth is not very responsive to AM fungi [3], mycorrhization significantly reduces tomato infection and disease severity caused by several root pathogens [4–7] and is regarded as a potential beneficial crop management tool to reduce pathogen populations in the soil which positively impacts crop health, quality and production [8]. In addition to its cultivation and e
Data Loading...
