The Solanum tuberosum GBSSI gene: a target for assessing gene and base editing in tetraploid potato

Abstract

The StGBSSI gene was successfully and precisely edited in the tetraploid potato using gene and base editing strategies, leading to plants with impaired amylose biosynthesis.

CRISPR-induced indels and base editing using theStaphylococcus aureusCas9 in potato

“…Taken together and compared to our previous work on genome editing in potato [17], our results show that SaCas9 constitutes an alternative to the classical SpCas9. As previous data showed that SaCas9…”

“…Binary plasmids described above were transferred into Agrobacterium C58pMP90 strain by heat shock. Agrobacterium-mediated stable plant transformation and plant regeneration were performed on explants of the Desiree cultivar, as previously described [17]. Plant tissues were cultured on 50 mg/L kanamycin, and regenerated plants were then transferred to a culture medium containing 50 mg/L kanamycin or tested for the presence of the T-DNA by PCR (S2 Table 1).…”

“…Genomic DNA from control and regenerated plants was extracted using the NucleoSpin Plant II kit (Macherey-Nagel, Germany) according to the manufacturer's instructions. HRM analysis was performed using the High Resolution Melting Master (Roche Applied Science, Germany) on the LightCycler® 480 II system (Roche Applied Science, Germany) (S2 Table 1), as previously described [17]. Plants harboring a HRM mutated profile were then Sanger sequenced (Genoscreen, France) (S2 Table 1).…”

“…The cultivated and tetraploid potato (Solanum tuberosum) received much attention for genome editing in the last few years by several groups. These achievements allowed to produce plants with new agronomic traits with gene knockout and/or base editing approaches, such as the production of tubers with low levels of amylose [15][16][17] or tubers with improved resistance to harvest and post-harvest procedures [18]. However, all the studies on potato used the classical or engineered variants [19] of SpCas9 so far, pointing out to the necessity to broaden the CRISPR toolbox for this species that constitutes one of the most important crops for food production worldwide.…”

Prime editing is achievable in the tetraploid potato, but needs improvement

“…Here, we show for the first time that PPE2 strategy developed for dicot species is able to edit an endogenous gene in the cultivated tetraploid potato. Compared to our previous work on cytosine base editing in potato and Arabidopsis (Bastet et al, 2019;Veillet et al, 2019b;Veillet et al, 2019a;Veillet et al, 2020), the efficiency of our PPE strategy appears to be limited, as already observed for cereals. Once improved, the prime editing technology, by largely expanding the range of targeted base substitutions, could make precision breeding amenable in polyploid and vegetatively propagated crops such as potato.…”