PJB-2019-767
Highly Efficient Targeted Mutagenesis of MaPDS in Dongguan Dajiao Banana via the CRISPR/Cas9 System
Chunhua Hu
Abstract
Banana and plantain (Musa spp.) are important staple and fruit crops for millions of people in the tropical and subtropical regions of the world, however, most cultivated bananas are triploid, with high levels of male and female infertility, which makes itself difficult to be genetically improved. The invention and wide application of CRISPR-Cas9 technology bring new opportunities for targeted genome editing and modification for banana improvement. Using the phytoene desaturase gene 8 (MaPDS) as the target gene, we genome edited Dajiao banana (Musa spp.ABB group). In this research, we designed three guide RNAs targeting to edit PDS gene, which were transformed into Dajiao banana embryogenic suspension cells mediated by Agrobacterium tumefaciens, and we obtained twenty-five independent hygromycin resistant regeneration lines, and among them, 22 lines showed albino phenotype, and the mutation rate was 88%. The sequencing results showed different types of mutation at the locus was found, such as base insertion, transformation and transversion. In this study, site-directed knockout of endogenous MaPDS was successfully carried out in Dajiao banana, and mutant lines with site-directed knockout were obtained, which laid a foundation for the further application of gene editing technology in genetic improvement of banana.
To Cite this article:
Download
