Abstract
Key message
A first creation of high oleic acid peanut varieties by using transcription activator-like effecter nucleases (TALENs) mediated targeted mutagenesis of Fatty Acid Desaturase 2 (FAD2).
Abstract
Transcription activator like effector nucleases (TALENs), which allow the precise editing of DNA, have already been developed and applied for genome engineering in diverse organisms. However, they are scarcely used in higher plant study and crop improvement, especially in allopolyploid plants. In the present study, we aimed to create targeted mutagenesis by TALENs in peanut. Targeted mutations in the conserved coding sequence of Arachis hypogaea fatty acid desaturase 2 (AhFAD2) were created by TALENs. Genetic stability of AhFAD2 mutations was identified by DNA sequencing in up to 9.52 and 4.11% of the regeneration plants at two different targeted sites, respectively. Mutation frequencies among AhFAD2 mutant lines were significantly correlated to oleic acid accumulation. Genetically, stable individuals of positive mutant lines displayed a 0.5–2 fold increase in the oleic acid content compared with non-transgenic controls. This finding suggested that TALEN-mediated targeted mutagenesis could increase the oleic acid content in edible peanut oil. Furthermore, this was the first report on peanut genome editing event, and the obtained high oleic mutants could serve for peanut breeding project.
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Acknowledgements
This study was supported by the Science and Technology Planning Project of Guangdong Province (2015B020231006, 2016B020201003 and 2016LM3161), the National Natural Science Foundation of China (31501246), and the Modern Agro-industry Technology Research System (CARS-14).
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HL, QL, and YH conceived the original screening and research plans. XC supervised the experiments. SW, XL performed most of the experiments. HL designed the experiments and analyzed the data. QL conceived the project and wrote the article with contributions of all the authors. SW, HL supervised and complemented the writing.
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Wen, S., Liu, H., Li, X. et al. TALEN-mediated targeted mutagenesis of fatty acid desaturase 2 (FAD2) in peanut (Arachis hypogaea L.) promotes the accumulation of oleic acid. Plant Mol Biol 97, 177–185 (2018). https://doi.org/10.1007/s11103-018-0731-z
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DOI: https://doi.org/10.1007/s11103-018-0731-z