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CRISPR/Cas9-based gene-editing platform development by targeting genes encoding magnesium chelatase and phytoene desaturase in sugarbeet (Beta vulgaris L.)
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DOI：10.3389/fgeed.2026.1875376.png)
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Abstract
En 中文
Sugarbeet (Beta vulgaris ssp. vulgaris; L.); is a vital temperate crop; supplying nearly 40% of the world’s sugar. However; its high susceptibility to bacterial; fungal; and viral diseases creates an urgent need for improved; disease-resistant cultivars. The CRISPR/Cas9 system has rapidly advanced plant genetic engineering by enabling precise and targeted genome modifications. Our goal is to establish a gene-editing platform in sugarbeet to support future development of disease-resistant lines by targeting the candidate genes. In this study; we applied CRISPR/Cas9 to generate targeted mutations in two genes involved in chlorophyll biosynthesis and carotenoid-mediated leaf pigmentation: magnesium chelatase (Mg-chelatase) and phytoene desaturase (PDS). Two CRISPR/Cas9 constructs; each carrying an sgRNA targeting either Mg-chelatase or PDS; were developed and mobilized into Agrobacterium tumefaciens. A total of 233 and 200 hypocotyl explants were transformed with constructs targeting Mg-chelatase and PDS; resulting in regeneration efficiencies of 8% and 14% on kanamycin selection medium; respectively. Light green; yellow; variegated yellow-green; and albino phenotypes were observed among the putative transformants; whereas non-edited transformed lines resembled untransformed control plants. Targeted mutations; including insertions; deletions; and substitutions of nucleotides; were identified at both genomic loci; with editing efficiencies of 60.0% for Mg-chelatase and 68.75% for PDS underscoring the effectiveness of this approach in sugarbeet; a recalcitrant crop. Deletions ranged from 5 to 28 bp in Mg-chelatase and 2 to 21 bp in PDS; while insertion events consisted of single-base additions in Mg-chelatase edited lines and larger insertions of 7–16 bp in PDS mutants. The results demonstrate the successful deployment of CRISPR/Cas9 for targeted genome engineering in sugarbeet and establish a reliable platform for future gene-editing efforts aimed at enhancing resistance to a wide range of pathogens and diseases affecting the crop.
Keywords:
gene-editing
Beta vulgaris
phytoene desaturase
magnesium chelatase
sugarbeet
Journal
F
IF:
4.4
Papers:
82
Citations:
893
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Cited Papers