First TILLING platform in Cucurbita pepo: a new mutant resource for gene function and crop improvement.
Although the availability of genetic and genomic resources for Cucurbita pepo has increased significantly, functional genomic resources are still limited for this crop. In this direction, we have developed a high throughput reverse genetic tool: the first TILLING (Targeting Induced Local Lesions IN Genomes) resource for this species. Additionally, we have used this resource to demonstrate that the previous EMS mutant population we developed has the highest mutation density compared with other cucurbits mutant populations. The overall mutation density in this first C. pepo TILLING platform was estimated to be 1/133 Kb by screening five additional genes. In total, 58 mutations confirmed by sequencing were identified in the five targeted genes, thirteen of which were predicted to have an impact on the function of the protein. The genotype/phenotype correlation was studied in a peroxidase gene, revealing that the phenotype of seedling homozygous for one of the isolated mutant alleles was albino. These results indicate that the TILLING approach in this species was successful at providing new mutations and can address the major challenge of linking sequence information to biological function and also the identification of novel variation for crop breeding.
IFAPA Centro La Mojonera, Camino de San Nicolás, 1, 04745 La Mojonera, Almería, Spain.
Nelly
N
Vicente-Dólera
INRA-URGV, UMR1165, Unité de Recherche en Génomique Végétale, Saclay Plant Sciences, Evry, France.
Christelle
C
Troadec
IFAPA Centro La Mojonera, Camino de San Nicolás, 1, 04745 La Mojonera, Almería, Spain.
Manuel
M
Moya
IFAPA Centro La Mojonera, Camino de San Nicolás, 1, 04745 La Mojonera, Almería, Spain.
Mercedes
M
del Río-Celestino
IFAPA Centro La Mojonera, Camino de San Nicolás, 1, 04745 La Mojonera, Almería, Spain.
Teresa
T
Pomares-Viciana
INRA-URGV, UMR1165, Unité de Recherche en Génomique Végétale, Saclay Plant Sciences, Evry, France.
Abdelhafid
A
Bendahmane
Institute for the Conservation and Breeding of Agricultural Biodiversity (COMAV-UPV), Universitat Politécnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain.
Belén
B
Picó
IFAPA Centro Alameda del Obispo, Avd. Menéndez Pidal s/n, 14004, Córdoba, Spain.
Belén
B
Román
IFAPA Centro La Mojonera, Camino de San Nicolás, 1, 04745 La Mojonera, Almería, Spain.
Pedro
P
Gómez
D018556:Crops, Agricultural; D028464:Cucurbita / Q000187:drug effects / Q000235:genetics*; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D017343:Genes, Plant; D056726:Genetic Association Studies; D005821:Genetic Techniques; D018745:Genome, Plant; D016296:Mutagenesis; D009154:Mutation; D059645:Mutation Rate; D010544:Peroxidases / Q000235:genetics; D015203:Reproducibility of Results
D005020:Ethyl Methanesulfonate; D010544:Peroxidases; C120494:anionic peroxidase