Repositorium
CRISPR/Cas9-induced Targeted Mutagenesis and Gene Replacement to Generate Long-shelf Life Tomato Lines
382
Journal Article / 2017
Yu, Qing-hui; Wang, Baike; Li, Ning; Tang, Yaping; Yang, Shengbao; Yang, Tao; Xu, Juan; Guo, Chunmiao; Yan, Peng; Wang, Qiang; Asmutola, Patiguli
Quickly and precisely gain genetically enhanced breeding elites with value-adding performance traits is desired by the crop breeders all the time. The present of gene editing technologies, especially the CRISPR/Cas9 system with the capacities of efficiency, versatility and multiplexing provides a reasonable expectation towards breeding goals. For exploiting possible application to accelerate the speed of process at breeding by CRISPR/Cas9 technology, in this study, the Agrobacterium tumefaciens-mediated CRISPR/Cas9 system transformation method was used for obtaining tomato ALC gene mutagenesis and replacement, in absence and presence of the homologous repair template. The average mutation frequency (72.73%) and low replacement efficiency (7.69%) were achieved in T0 transgenic plants respectively. None of homozygous mutation was detected in T0 transgenic plants, but one plant carry the heterozygous genes (Cas9/*-ALC/alc) was stably transmitted to T1 generations for segregation and genotyping. Finally, the desired alc homozygous mutants without T-DNA insertion (*/*-alc/alc) in T1 generations were acquired and further confirmed by genotype and phenotype characterization, with highlight of excellent storage performance, thus the recessive homozygous breeding elites with the character of long-shelf life were generated. Our results support that CRISPR/Cas9-induced gene replacement via HDR provides a valuable method for breeding elite innovation in tomato.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
881 |
Yu, Qing-hui China |
Solanum lycopersicum |
CRISPR/Cas9 ALC |
Improved storage time and prolong the shelf life SDN1 |
Market-oriented Abiotic stress tolerance |
882 |
Yu, Qing-hui China |
Solanum lycopersicum |
CRISPR/Cas9 ALC |
Improved storage time and prolong the shelf life SDN2 |
Basic research Basic research |