Přehled o publikaci

Hairy root cultures are a versatile system with many applications, including the production of recombinant proteins and valuable secondary metabolites, as well as the functional characterisation of genes. We have developed efficient transformation protocols for the spring cultivars of Brassica napus and Brassica rapa using hairy roots. We have generated various reporter lines, including new fluorescent reporters that monitor the response of the plant hormones auxin and cytokinin to abiotic stress stimuli. Furthermore, we have developed a pipeline for gene editing and hairy root regeneration in B. napus, with the aim of creating transgene-free, gene-edited plants. Since oilseed rape is an allotetraploid species (AACC, 2n=38), mutating homoeologs from both subgenomes is necessary to overcome functional genetic redundancy. Using optimised CRISPR/Cas9 vectors, we generated multiple mutant lines in four B. napus TRYPTOPHAN AMINOTRANSFERASE (BnaTAA1) auxin biosynthetic genes. In conclusion, we have initiated a functional analysis of BnaTAA1 and a study of the auxin biosynthesis pathway in oilseed rape using hairy root cultures. We offer transformation services for selected oilseed rape cultivars to generate CRISPR-edited plants and reporter lines. Funding sources: GA23-06140S by the Czech Science Foundation and TANGENC (TowArds Next GENeration Crops) CZ.02.01.01/00/22_008/0004581 of the ERDF Programme Johannes Amos Comenius by MEYS.