Effects of the repression of GIGANTEA gene StGI.04 on the potato leaf transcriptome and the anthocyanin content of tuber skin.

BACKGROUND: GIGANTEA (GI) is a plant-specific, circadian clock-regulated, nuclear protein with pleiotropic functions found in many plant species. This protein is involved in flowering, circadian clock control, chloroplast biogenesis, carbohydrate metabolism, stress responses, and volatile compound synthesis. In potato (Solanum tuberosum L.), its only role appears to be tuber initiation; however, based on findings in other plant species, we hypothesised that the function of GI in potatoes is not restricted only to tuberisation. RESULTS: To test this hypothesis, the expression of a GI gene in the commercial potato cultivar 'Désirée' was repressed, and the effects of repression at morphological and transcriptome level were investigated. Previously, two copies of GI genes in potato were found. A construct to reduce the mRNA levels of one of these genes (StGI.04) was assembled, and the effects of antisense repression were studied in greenhouse-grown plants. The highest level of repression reached around 50%. However, this level did not influence tuber formation and yield but did cause a reduction in tuber colour. Using high-performance liquid chromatography (HPLC), significant reductions in cyanidin 3,5-di-O-glucoside and pelargonidin 3,5-di-O-glucoside contents of tuber peels were detected. Anthocyanins are synthesized through a branch of the phenylpropanoid pathway. The transcriptome analysis indicated down-regulation in the expression of PHENYLALANINE AMMONIA LYASE (PAL), the LEUCOANTHOCYANIDIN OXIDISING enzyme gene LDOX, and the MYB-RELATED PROTEIN Hv1 (MYB-Hv1), a transcription factor coding gene, which is presumably involved in the regulation of flavonoid biosynthesis, in the leaves of a selected StGI.04-repressed line. Furthermore, alterations in expression of genes affecting the circadian clock, flowering, starch synthesis, and stress responses were detected in the leaves of the selected StGI.04-repressed line. CONCLUSIONS: We tested the effects of antisense repression of StGI.04 expression in potatoes and found that as with GI in other plant species, it influences the expression of the key genes of the circadian clock, flowering, starch synthesis, and stress responses. Furthermore, we detected a novel function of a GI gene in influencing the anthocyanin synthesis and potato tuber skin colour.

In Silico Characterization and Expression Analysis of GIGANTEA Genes in Potato.

GIGANTEA (GI) genes are ubiquitous in the plant kingdom and are involved in diverse processes from flowering during stress responses to tuberization; the latter occurs in potato (Solanum tuberosum L.). GI genes have a diurnal cycle of expression; however, no details on the regulation of GI gene expression in potato have been reported thus far. The aim of our work was the analysis of the GI promoter sequence and studying GI expression in different organs and under abiotic stress conditions in potato. Two GI genes homologous to Arabidopsis GI located on chromosomes 4 and 12 (StGI.04 and StGI.12) were identified in the genome-sequenced potato S. phureja. The GI promoter regions of the commercial potato cultivar 'Désirée' were cloned and found to be almost identical to the S. phureja GI promoter sequence. More than ten TF families binding to the GI promoters were predicted. EVENING ELEMENT and ABSCISIC ACID RESPONSE ELEMENT LIKE elements related to circadian regulation and a binding site for POTATO HOMEOBOX 20 presumably involved in tuber initiation were detected in both GI promoters. However, the locations of these elements and several other cis-acting regulatory elements as well as the organ-specific expression and responses of the genes to abiotic stresses and abscisic acid were different. Thus, we presume that the function of StGI.04 and StGI.12 are at least partially different. This study lays foundation for further investigation of the roles of GI genes in potato.

Metabolite analysis of tubers and leaves of two potato cultivars and their grafts.

Grafting experiments have shown that photoperiod-dependent induction of tuberisation in potato (Solanum tuberosum L.) is controlled by multiple overlapping signals, including mobile proteins, mRNAs, miRNAs and phytohormones. The effect of vegetative organs and tubers at metabolite level and vice versa, however, has not been studied in detail in potato. To unravel the influence of vegetative organs on the primary polar metabolite content of potato tubers and the effect of tuberisation on the metabolite content of leaves grafting experiments were carried out. Two potato cultivars, Hópehely (HP) and White Lady (WL), were homo- and hetero-grafted, and the effects of grafting were investigated in comparison to non-grafted controls. Non-targeted metabolite analysis using gas chromatography-mass spectrometry showed that the major difference between HP and WL tubers is in sucrose concentration. The sucrose level was higher in HP than in WL tubers and was not changed by grafting, suggesting that the sucrose concentration of tubers is genetically determined. The galactinol level was 8-fold higher in the WL leaves than in the HP leaves and, unlike the sucrose concentration of tubers, was altered by grafting. A positive correlation between the growth rate of the leaves and the time of tuber initiation was detected. The time of tuber initiation was delayed in the WL rootstocks by HP scions and shortened in the HP rootstocks by WL scions, supporting the previous finding that tuberisation is triggered by source-derived mobile signals.