Cold treatment prior to bolting transforms sugar beet taproots from sucrose-storing sink organs to sucrose-mobilizing source organs.

Background: DNA methylation is thought to influence the expression of genes, especially in response to changing environmental conditions and developmental changes. Sugar beet (Beta vulgaris ssp. vulgaris), and other biennial or perennial plants are inevitably exposed to fluctuating temperatures throughout their lifecycle and might even require such stimulus to acquire floral competence. Therefore, plants such as beets, need to fine-tune their epigenetic makeup to ensure phenotypic plasticity towards changing environmental conditions while at the same time steering essential developmental processes. Different crop species may show opposing reactions towards the same abiotic stress, or, vice versa, identical species may respond differently depending on the specific kind of stress. Results: In this study, we investigated common effects of cold treatment on genome-wide DNA methylation and gene expression of two Beta vulgaris accessions via multi-omics data analysis. Cold exposure resulted in a pronounced reduction of DNA methylation levels, which particularly affected methylation in CHH context (and to a lesser extent CHG) and was accompanied by transcriptional downregulation of the chromomethyltransferase CMT2 and strong upregulation of several genes mediating active DNA demethylation. Conclusion: Integration of methylomic and transcriptomic data revealed that, rather than methylation having directly influenced expression, epigenetic modifications correlated with changes in expression of known players involved in DNA (de)methylation. In particular, cold triggered upregulation of genes putatively contributing to DNA demethylation via the ROS1 pathway. Our observations suggest that these transcriptional responses precede the cold-induced global DNA-hypomethylation in non-CpG, preparing beets for additional transcriptional alterations necessary for adapting to upcoming environmental changes.

Despite a high sucrose accumulation in its taproot vacuoles, sugar beet (Beta vulgaris subsp. vulgaris) is sensitive to freezing. Earlier, a taproot‐specific accumulation of raffinose was shown to have beneficial effects on the freezing tolerance of the plant. However, synthesis of raffinose and other oligosaccharides of the raffinose family depends on the availability of myo‐inositol. Since inositol and inositol‐metabolising enzymes reside in different organelles, functional inositol metabolism and raffinose synthesis depend on inositol transporters. We identified five homologues of putative inositol transporters in the sugar beet genome, two of which, BvINT1;1 and BvINT1;2, are localised at the tonoplast. Among these, only the transcript of BvINT1;1 is highly upregulated in sugar beet taproots under cold. BvINT1;1 exhibits a high transport specificity for inositol and sugar beet mutants lacking functional BvINT1;1 contain increased inositol levels, likely accumulating in the vacuole, and decreased raffinose contents under cold treatment. Due to the quenching capacity of raffinose for Reactive Oxygen Species (ROS), which accumulate under cold stress, bvint1;1 sugar beet plants show increased expression of both, ROS marker genes and detoxifying enzymes. Based on these findings, we conclude that the vacuolar inositol transporter BvINT1;1 is contributing to ROS‐homoeostasis in the cold metabolism of sugar beet.

Sugar beet ( Beta vulgaris subsp. vulgaris ) is the exclusive source of sugar in the form of sucrose in temperate climate zones. There, sugar beet is grown as an annual crop from spring to autumn because of the damaging effect of freezing temperatures to taproot tissue. Natural and breeded varieties display variance in the degree of tolerance to freezing temperatures and genotypes with elevated tolerance to freezing have been isolated. Here we compare initial responses to frost between genotypes with either low and high winter survival rates. The selected genotypes differed in the severity of frost injury. We combined transcriptomic and metabolite analyses of leaf- and taproot tissues from such genotypes to elucidate mechanisms of the early freezing response and to dissect genotype- and tissue-dependent responses. Freezing temperatures induced drastic downregulation of photosynthesis-related genes in leaves but upregulation of genes related to minor carbohydrate metabolism, particularly of genes involved in raffinose metabolism in both, leaf and taproot tissue. In agreement with this, it has been revealed that raffinose and the corresponding intermediates, inositol and galactinol, increased markedly in these tissues. We found that genotypes with improved tolerance to freezing, showed higher accumulation of raffinose in a defined interior region within the upper part of the taproot, the pith, representing the tissue most susceptible to freeze damages. This accumulation was accompanied by specific upregulation of raffinose synthesizing enzymes in taproots, suggesting a protective role for raffinose and its precursors for freezing damage in sugar beet.