Expression and Promoter Methylation of the Genes Encoding the Mitochondrial and Cytosolic Forms of Fumarase in Sunflower (Helianthus annuus L.) Leaves Depending on Light Regime and Salinity

The expression of two genes, Fum1 and Fum2, encoding the mitochondrial and cytosolic forms of fumarase (EC 4.2.1.2); the methylation of individual CpGs of their promoters; and fumarase activity were studied in sunflower (Helianthus annuus L.) leaves depending on irradiation and salinity. Fumarase activity was twice as high in darkness compared to irradiation by white light and red light, while far-red light applied after darkness or after red light reverted the activity to the values in darkness, which indicates the involvement of phytochrome. Using qRT-PCR, it was demonstrated that this corresponded to the pattern of expression of the Fum1 gene, while the expression of the Fum2 gene was higher upon irradiation by white and red light, and lower in darkness and under far-red light. Under the application of 150 mM NaCl for 1, 3, 6, 12, and 24 h, fumarase activity increased fivefold from the start of incubation to 6 h, and then decreased after 12 h. These changes were associated with the transcriptional regulation of the Fum1 and Fum2 genes. Changes in the methylation status of the analyzed CpGs in their gene promoters, detected via semi-quantitative methylation-specific PCR, were associated with differences in their expression. The higher methylation levels of the analyzed CpGs in the Fum1 gene promoter under different light conditions and in the Fum2 gene promoter under salinity corresponded to low levels of their transcripts in sunflower leaves. It is suggested that the mitochondrial and cytosolic forms of fumarase are regulated by light and salinity at the gene expression level, presumably through changes in the methylation status of individual CpGs in their promoters.