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Targeted gene mutation in tetraploid potato through transient TALEN expression in protoplasts

Paper ID Volume ID Publish Year Pages File Format Full-Text
22807 43392 2015 8 PDF Available
Title
Targeted gene mutation in tetraploid potato through transient TALEN expression in protoplasts
Abstract

•A successful pipeline for site-directed mutagenesis in tetraploid potato was developed.•A transient TALEN expression system based on potato protoplasts was employed.•Protoplast-derived calli and shoots, carrying mutations at the expected target site, were obtained.

Potato is the third largest food crop in the world, however, the high degree of heterozygosity, the tetrasomic inheritance and severe inbreeding depression are major difficulties for conventional potato breeding. The rapid development of modern breeding methods offers new possibilities to enhance breeding efficiency and precise improvement of desirable traits. New site-directed mutagenesis techniques that can directly edit the target genes without any integration of recombinant DNA are especially favorable. Here we present a successful pipeline for site-directed mutagenesis in tetraploid potato through transient TALEN expression in protoplasts. The transfection efficiency of protoplasts was 38–39% and the site-directed mutation frequency was 7–8% with a few base deletions as the predominant type of mutation. Among the protoplast-derived calli, 11–13% showed mutations and a similar frequency (10%) was observed in the regenerated shoots. Our results indicate that the site-directed mutagenesis technology could be used as a new breeding method in potato as well as for functional analysis of important genes to promote sustainable potato production.

Keywords
SDM, site-directed mutagenesis; DSB, double strand break; HR, homologous recombination; NHEJ, non-homologous end joining; TALEN, transcription activator-like effector nuclease; ALS, acetolactate synthasePotato; Site-directed mutagenesis; TALEN; Gene editi
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Targeted gene mutation in tetraploid potato through transient TALEN expression in protoplasts
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 204, 20 June 2015, Pages 17–24
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us