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Shumbe, L.

Publications and source records attributed to Shumbe, L..

3 recordsLinked to original sources

Mutation of the VInv 5' UTR regulatory region reduces acrylamide levels in processed potato to reach EU food-safety standards

Protein-coding regions of genes have so far been the preferred targets for trait improvement using CRISPR-Cas9 genome editing in plants. Alteration of the reading frame can result in termination of transcription or translation, hence, loss of function of the encoded protein. 5 UTR sequences also represent a practical target region to alter gene expression and protein abundance. However, editing of 5 UTRs has so far been scarcely used to engineer trait in crop plants. Here, we demonstrate that insertion of a single adenine nucleotide mediated by Cas9 in the 5 UTR region of two alleles from the Vacuolar invertase gene (VInv) of the tetraploid potato variety Lady Rosetta (LaRo) is sufficient to substantially lower the content of reducing sugars in the potato under cold-storage conditions. Moreover, the acrylamide content generated during processing of the edited potato lines into crisps was more than three folds lower than the current EU-regulated maximum level of 750 g/kg in crisps. This gene-editing approach represents a durable strategy to improve food safety of potatoes in varieties widely preferred by the consumers and the industry.

plant biology↗

Differential DNA methylation in the Vinv promoter region controls Cold Induced Sweetening in potato

Control of potato sprouting is important to ensure constant supply of high-quality potato to the industry. Efficient control of sprouting can be achieved by chemical treatment or cold temperature. Recent bans on anti-sprouting molecules are prompting the use of cold storage in the potato value chain. Unfortunately, storage of potato at low temperatures is associated with cold induced sweetening (CIS) due to the induction of the vacuolar invertase gene under low temperatures. Because CIS is associated with the production of the potentially carcinogenic acrylamide in processed potatoes, concise knowledge on the regulatory mechanisms controlling the CIS-phenotype in potatoes is expected to help pave the way for the production of CIS-resistant potato varieties. Here, we dissect the promoters of the Vacuolar invertase (Vinv) genes from CIS-susceptible and CIS-resistant varieties to investigate their implication in CIS-phenotype determination. Using bisulfite sequencing and CRISPR-dCas9-DRM2-mediated de novo DNA methylation, we show that the CIS-resistant phenotype of Verdi, is in part due to hypermethylation of its Vinv promoter, more specifically in the 1.0-1.7kb region. Those findings open new perspectives to engineer CIS-resistant potatoes by genome and epigenome modifications.

plant biology↗

Asparagine accumulation in chicory storage roots is controlled by translocation and feedback regulation of asparagine biosynthesis in leaves

O_LIThe presence of acrylamide (AA), a potentially carcinogenic and neurotoxic compound, in food has become a major concern for public health. AA in plant-derived food mainly arises from the reaction of the amino acid asparagine (Asn) and reducing sugars during processing of foodstuffs at high temperature. C_LIO_LIUsing a selection of genotypes from the chicory germplasm we performed Asn measurements in storage roots and leaves to identify genotypes contrasting for Asn accumulation. We combined molecular analysis and grafting experiments to show that leaf to root translocation controls asparagine biosynthesis and accumulation in chicory storage roots. C_LIO_LIWe could demonstrate that Asn accumulation in storage roots depends on Asn biosynthesis and transport from the leaf, and that a negative feedback loop by Asn on CiASN1 expression impacts Asn biosynthesis in leaves. C_LIO_LIOur results provide a new model for asparagine biosynthesis in root crop species and highlight the importance of characterizing and manipulating asparagine transport to reduce AA content in processed plant-based foodstuffs. C_LI

plant biology↗