A new mutant genetic resource for tomato crop improvement by TILLING technology

Silvia Minoia,Olimpia D'Onofrio,Giovanni Sozio,Filomena Carriero,Francesco Cellini,Giuseppina Mosca,Angelo Petrozza,Abdelhafid Bendahmane,Florence Piron

doi:10.1186/1756-0500-3-69

Abstract

BackgroundIn the last decade, the availability of gene sequences of many plant species, including tomato, has encouraged the development of strategies that do not rely on genetic transformation techniques (GMOs) for imparting desired traits in crops. One of these new emerging technology is TILLING (Targeting Induced Local Lesions In Genomes), a reverse genetics tool, which is proving to be very valuable in creating new traits in different crop species.ResultsTo apply TILLING to tomato, a new mutant collection was generated in the genetic background of the processing tomato cultivar Red Setter by treating seeds with two different ethylemethane sulfonate doses (0.7% and 1%). An associated phenotype database, LycoTILL, was developed and a TILLING platform was also established. The interactive and evolving database is available online to the community for phenotypic alteration inquiries. To validate the Red Setter TILLING platform, induced point mutations were searched in 7 tomato genes with the mismatch-specific ENDO1 nuclease. In total 9.5 kb of tomato genome were screened and 66 nucleotide substitutions were identified. The overall mutation density was estimated and it resulted to be 1/322 kb and 1/574 kb for the 1% EMS and 0.7% EMS treatment respectively.ConclusionsThe mutation density estimated in our collection and its comparison with other TILLING populations demonstrate that the Red Setter genetic resource is suitable for use in high-throughput mutation discovery. The Red Setter TILLING platform is open to the research community and is publicly available via web for requesting mutation screening services.

Highlights

In the last decade, the availability of gene sequences of many plant species, including tomato, has encouraged the development of strategies that do not rely on genetic transformation techniques (GMOs) for imparting desired traits in crops
The genome sequencing program of many plant species [8,9,10], including tomato [11] has led to the availability of a large number of gene sequences in public databases which subsequently has encouraged the development of reverse genetics approaches
In the present paper we report the construction of a high-quality tomato genetic mutant reference collection which could be used for both forward and reverse genetic studies

Summary

Introduction

The availability of gene sequences of many plant species, including tomato, has encouraged the development of strategies that do not rely on genetic transformation techniques (GMOs) for imparting desired traits in crops. One of these new emerging technology is TILLING (Targeting Induced Local Lesions In Genomes), a reverse genetics tool, which is proving to be very valuable in creating new traits in different crop species. The genome sequencing program of many plant species [8,9,10], including tomato [11] has led to the availability of a large number of gene sequences in public databases which subsequently has encouraged the development of reverse genetics approaches. The strength and potency of this reverse genetic strategy has been validated by its successful application in both plants (Arabidopsis thaliana [16,17,18], pea [19], wheat [20], rice [21,22], barley [23], maize [24], soybean [25], Lotus japonicus [26], sorghum [27], tomato [28,29]) and animals (zebrafish [30], drosophila [31])

Methods

Results

Discussion

Conclusion