DNA-free genome editing for ZmPLA1gene via targeting immature embryos in tropical maize

Autor: Rangari, Sagar Krushnaji, Kaur Sudha, Manjot, Kaur, Harjot, Uppal, Nidhi, Singh, Gagandeep, Vikal, Yogesh, Sharma, Priti
Zdroj: GM Crops & Food; December 2023, Vol. 14 Issue: 1 p1-7, 7p
Abstrakt: ABSTRACTDoubled haploid (DH) production accelerates the development of homozygous lines in a single generation. In maize, haploids are widely produced by the use of haploid inducer Stock 6, earlier reported in 1959. Three independent studies reported haploid induction in maize which is triggered due to a 4 bp frame-shift mutation in matrilineal(ZmPLA1) gene. The present study was focused on the generation of mutants for ZmPLA1gene in maize inbred line LM13 through site-directed mutagenesis via CRISPR/Cas9-mediated ribonucleoprotein (RNP) complex method to increase the haploid induction rate. Three single guide RNAs (sgRNAs) for the ZmPLA1gene locus were used for transforming the 14 days old immature embryos via bombardment. 373 regenerated plants were subjected to mutation detection followed by Sanger’s sequencing. Out of three putative mutants identified, one mutant depicted one base pair substitution and one base pair deletion at the target site.
Databáze: Supplemental Index