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Open Access Highly Accessed Research article

SMART – Sunflower Mutant population And Reverse genetic Tool for crop improvement

Anish PK Kumar1, Adnane Boualem234, Anjanabha Bhattacharya1, Seema Parikh1, Nirali Desai1, Andres Zambelli5, Alberto Leon5, Manash Chatterjee16* and Abdelhafid Bendahmane234

Author Affiliations

1 Bench Bio Pvt Ltd., c/o Jai Research Foundation, Vapi, Gujarat, 396195, India

2 INRA, UMR1165 Unité de Recherche en Génomique Végétale URGV, Evry, F-91057, France

3 UEVE, UMR Unité de Recherche en Génomique Végétale URGV, Evry, F-91057, France

4 CNRS, ERL8196 UMR Unité de Recherche en Génomique Végétale URGV, Evry, F-91057, France

5 Biotechnology Research Centre, Nutrisun Business Unit- Advanta Semillas SAIC, Belcarce, Argentina

6 National University of Ireland Galway (NUIG), Galway, Ireland

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BMC Plant Biology 2013, 13:38  doi:10.1186/1471-2229-13-38

Published: 5 March 2013

Abstract

Background

Sunflower (Helianthus annuus L.) is an important oilseed crop grown widely in various areas of the world. Classical genetic studies have been extensively undertaken for the improvement of this particular oilseed crop. Pertaining to this endeavor, we developed a “chemically induced mutated genetic resource for detecting SNP by TILLING” in sunflower to create new traits.

Results

To optimize the EMS mutagenesis, we first conducted a “kill curve” analysis with a range of EMS dose from 0.5% to 3%. Based on the observed germination rate, a 50% survival rate i.e. LD50, treatment with 0.6% EMS for 8 hours was chosen to generate 5,000 M2 populations, out of which, 4,763 M3 plants with fertile seed set. Phenotypic characterization of the 5,000 M2 mutagenised lines were undertaken to assess the mutagenesis quality and to identify traits of interest. In the M2 population, about 1.1% of the plants showed phenotypic variations. The sunflower TILLING platform was setup using Endo-1-nuclease as mismatch detection system coupled with an eight fold DNA pooling strategy. As proof-of-concept, we screened the M2 population for induced mutations in two genes related to fatty acid biosynthesis, FatA an acyl-ACP thioesterase and SAD the stearoyl-ACP desaturase and identified a total of 26 mutations.

Conclusion

Based on the TILLING of FatA and SAD genes, we calculated the overall mutation rate to one mutation every 480 kb, similar to other report for this crop so far. As sunflower is a plant model for seed oil biosynthesis, we anticipate that the developed genetic resource will be a useful tool to identify novel traits for sunflower crop improvement.

Keywords:
Sunflower; TILLING; Functional genomics; Oil quality; Fatty acids; Crop improvement