title: Improving cold storage and processing traits in potato through targeted gene knockout.
abstract: Cold storage of potato tubers is commonly used to reduce sprouting and extend postharvest shelf life. However, cold temperature stimulates the accumulation of reducing sugars in potato tubers. Upon high-temperature processing, these reducing sugars react with free amino acids, resulting in brown, bitter-tasting products and elevated levels of acrylamide--a potential carcinogen. To minimize the accumulation of reducing sugars, RNA interference (RNAi) technology was used to silence the vacuolar invertase gene (VInv), which encodes a protein that breaks down sucrose to glucose and fructose. Because RNAi often results in incomplete gene silencing and requires the plant to be transgenic, here we used transcription activator-like effector nucleases (TALENs) to knockout VInv within the commercial potato variety, Ranger Russet. We isolated 18 plants containing mutations in at least one VInv allele, and five of these plants had mutations in all VInv alleles. Tubers from full VInv-knockout plants had undetectable levels of reducing sugars, and processed chips contained reduced levels of acrylamide and were lightly coloured. Furthermore, seven of the 18 modified plant lines appeared to contain no TALEN DNA insertions in the potato genome. These results provide a framework for using TALENs to quickly improve traits in commercially relevant autotetraploid potato lines.
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Clasen
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Stoddard
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Song
authors.initials: S
authors.lastname: Luo
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Zachary L
authors.initials: ZL
authors.lastname: Demorest
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Li
authors.affiliation: Cellectis SA, Paris, France.
authors.forename: Frederic
authors.initials: F
authors.lastname: Cedrone
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Redeat
authors.initials: R
authors.lastname: Tibebu
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Shawn
authors.initials: S
authors.lastname: Davison
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Erin E
authors.initials: EE
authors.lastname: Ray
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Aurelie
authors.initials: A
authors.lastname: Daulhac
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Coffman
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Ann
authors.initials: A
authors.lastname: Yabandith
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Adam
authors.initials: A
authors.lastname: Retterath
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: William
authors.initials: W
authors.lastname: Haun
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Baltes
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Luc
authors.initials: L
authors.lastname: Mathis
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.affiliation: Cellectis plant sciences Inc., New Brighton, MN, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
mesh_terms: D020106:Acrylamide / Q000032:analysis; D001483:Base Sequence; D002241:Carbohydrates / Q000032:analysis; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D017343:Genes, Plant; D009154:Mutation / Q000235:genetics; D030821:Plants, Genetically Modified; D011198:Solanum tuberosum / Q000235:genetics*; D000069896:Transcription Activator-Like Effector Nucleases / Q000378:metabolism; D014617:Vacuoles / Q000201:enzymology; D043324:beta-Fructofuranosidase / Q000235:genetics
chemical_list: D002241:Carbohydrates; D020106:Acrylamide; D000069896:Transcription Activator-Like Effector Nucleases; D043324:beta-Fructofuranosidase
keywords: acrylamide reduction; cold-induced sweetening; gene editing; potato; transcription activator-like effector nucleases; vacuolar invertase
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