Development of EMS-induced Mutagenized Groundnut Population and Discovery of Point Mutations in the ahFAD2 and Ara h 1 Genes by TILLING

Karaman K., Kızıl S., Başak M., Uzun B., Yol E.

Journal Of Oleo Science, cilt.70, sa.11, ss.1-10, 2021 (SCI-Expanded)

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 70 Sayı: 11
  • Basım Tarihi: 2021
  • Doi Numarası: 10.5650/jos.ess21075
  • Dergi Adı: Journal Of Oleo Science
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, CAB Abstracts, Chemical Abstracts Core, Compendex, EMBASE, Food Science & Technology Abstracts, MEDLINE, Pollution Abstracts, Veterinary Science Database, Directory of Open Access Journals
  • Sayfa Sayıları: ss.1-10
  • Akdeniz Üniversitesi Adresli: Evet

Özet

Reducing allergenicity and increasing oleic content are important goals in groundnut breeding

studies. Ara h 1 is a major allergen gene and Delta(12)-fatty-acid desaturase (FAD2) is responsible for

converting oleic into linoleic acid. These genes have homoeologues with one copy in each subgenome,

identified as Ara h 1.01, Ara h 1.02, ahFAD2A and ahFAD2B in tetraploid groundnut. To alter functional

properties of these genes we have generated an Ethyl Methane Sulfonate (EMS) induced mutant population

to be used in Targeting Induced Local Lesions in Genomes (TILLING) approach. Seeds were exposed to two

EMS concentrations and the germination rates were calculated as 90.1% (1353 plants) for 0.4% and 60.4%

(906 plants) for 1.2% EMS concentrations in the M1 generation. Among the 1541 M2 mutants, 768 were

analyzed by TILLING using four homoeologous genes. Two heterozygous mutations were identified in the

ahFAD2B and ahFAD2A gene regions from 1.2% and 0.4% EMS-treated populations, respectively. The

mutation in ahFAD2B resulted in an amino acid change, which was serine to threonine predicted to be

tolerated according to SIFT analysis. The other mutation causing amino acid change, glycine to aspartic

acid was predicted to affect protein function in ahFAD2A. No mutations were detected in Ara h 1.01 and Ara

h 1.02 for both EMS-treatments after sequencing. We estimated the overall mutation rate to be 1 mutation

every 2139 kb. The mutation frequencies were also 1/317 kb for ahFAD2A in 0.4% EMS and 1/466 kb for

ahFAD2B in 1.2% EMS treatments. The results demonstrated that TILLING is a powerful tool to interfere

with gene function in crops and the mutagenized population developed in this study can be used as an

efficient reverse genetics tool for groundnut improvement and functional genomics.