Development and characterization of an oat TILLING-population and identification of mutations in lignin and beta-glucan biosynthesis genes
Artikel i vetenskaplig tidskrift, 2010

Background: Oat, Avena sativa is the sixth most important cereal in the world. Presently oat is mostly used as feed for animals. However, oat also has special properties that make it beneficial for human consumption and has seen a growing importance as a food crop in recent decades. Increased demand for novel oat products has also put pressure on oat breeders to produce new oat varieties with specific properties such as increased or improved beta-glucan-, antioxidant-and omega-3 fatty acid levels, as well as modified starch and protein content. To facilitate this development we have produced a TILLING (Targeting Induced Local Lesions IN Genomes) population of the spring oat cultivar SW Belinda. Results: Here a population of 2600 mutagenised M2 lines, producing 2550 M3 seed lots were obtained. The M2 population was initially evaluated by visual inspection and a number of different phenotypes were seen ranging from dwarfs to giants, early flowering to late flowering, leaf morphology and chlorosis. Phloroglucinol/HCl staining of M3 seeds, obtained from 1824 different M2 lines, revealed a number of potential lignin mutants. These were later confirmed by quantitative analysis. Genomic DNA was prepared from the M2 population and the mutation frequency was determined. The estimated mutation frequency was one mutation per 20 kb by RAPD-PCR fingerprinting, one mutation per 38 kb by MALDI-TOF analysis and one mutation per 22.4 kb by DNA sequencing. Thus, the overall mutation frequency in the population is estimated to be one mutation per 20-40 kb, depending on if the method used addressed the whole genome or specific genes. During the investigation, 6 different mutations in the phenylalanine ammonia-lyase (AsPAL1) gene and 10 different mutations in the cellulose synthase-like (AsCslF6) beta-glucan biosynthesis gene were identified. Conclusion: The oat TILLING population produced in this work carries, on average, hundreds of mutations in every individual gene in the genome. It will therefore be an important resource in the development of oat with specific characters. The population (M5) will be available for academic research via Nordgen http://www.nordgen.org as soon as enough seeds are obtained.

barley

avena-sativa

induced point mutations

discovery

reverse genetics

genomics

chemically-induced mutations

arabidopsis

plant functional

fiber digestibility

lotus-japonicus

Författare

Aakash Chawade

Göteborgs universitet

Per Sikora

Göteborgs universitet

Marcus Bräutigam

Göteborgs universitet

Mikael Larsson

SuMo Biomaterials

Chalmers, Kemi- och bioteknik, Farmaceutisk teknologi

Vivekanand Vivekanand

Göteborgs universitet

M. A. Nakash

CropTailorAB

Tingsu Chen

Göteborgs universitet

Olof Olsson

Göteborgs universitet

BMC Plant Biology

1471-2229 (ISSN)

Vol. 10 86- 86

Ämneskategorier

Biokemi och molekylärbiologi

Kemi

Genetik

DOI

10.1186/1471-2229-10-86

Mer information

Skapat

2017-10-07