Heritable virus‐induced germline editing in tomato DOI Creative Commons
Youngbin Oh, Ugrappa Nagalakshmi, Douglas Dahlbeck

et al.

The Plant Journal, Journal Year: 2025, Volume and Issue: 122(1)

Published: March 31, 2025

SUMMARY Here, we report the successful implementation of heritable virus‐induced genome editing (VIGE) in tomato ( Solanum lycopersicum ). We generated three transgenic lines expressing Streptococcus pyogenes Cas9 (SpCas9) under control Cauliflower mosaic virus 35S (35S), S. ribosomal protein S5A Sl RPS5A), or YAO promoters YAO). These were tested for somatic and using tobacco rattle (TRV)‐based system carrying guide RNAs (gRNAs) fused with mobile RNA sequences. TRV gRNA targeted to Phytoene desaturase SlPDS ) Downy mildew resistance 6 SlDMR6 genes sequences showed significant efficiency all SpCas9. However, progenies from promoter‐driven SpCas9 infected sequence resulted monoallelic mutations a frequency 3%. Optimization environmental conditions, such as reduced light intensity, significantly increased frequencies, 0% 86% at 3% 100% , including biallelic mutations. findings underscore use appropriate express Cas nucleases optimized conditions enhance VIGE. Furthermore, our method enables generation mutants without additional tissue culture transformation once SpCas9‐expressing line is established.

Language: Английский

Heritable virus‐induced germline editing in tomato DOI Creative Commons
Youngbin Oh, Ugrappa Nagalakshmi, Douglas Dahlbeck

et al.

The Plant Journal, Journal Year: 2025, Volume and Issue: 122(1)

Published: March 31, 2025

SUMMARY Here, we report the successful implementation of heritable virus‐induced genome editing (VIGE) in tomato ( Solanum lycopersicum ). We generated three transgenic lines expressing Streptococcus pyogenes Cas9 (SpCas9) under control Cauliflower mosaic virus 35S (35S), S. ribosomal protein S5A Sl RPS5A), or YAO promoters YAO). These were tested for somatic and using tobacco rattle (TRV)‐based system carrying guide RNAs (gRNAs) fused with mobile RNA sequences. TRV gRNA targeted to Phytoene desaturase SlPDS ) Downy mildew resistance 6 SlDMR6 genes sequences showed significant efficiency all SpCas9. However, progenies from promoter‐driven SpCas9 infected sequence resulted monoallelic mutations a frequency 3%. Optimization environmental conditions, such as reduced light intensity, significantly increased frequencies, 0% 86% at 3% 100% , including biallelic mutations. findings underscore use appropriate express Cas nucleases optimized conditions enhance VIGE. Furthermore, our method enables generation mutants without additional tissue culture transformation once SpCas9‐expressing line is established.

Language: Английский

Citations

0