Root-based inorganic carbon uptake increases the growth of Arabidopsis thaliana and changes transporter expression and nitrogen and sulfur metabolism.

Root-based uptake of inorganic carbon has been suggested as an additional carbon source. Our study aimed to characterize and understand the root-based uptake and fixation mechanisms and their impact on plant growth. 13C-labeled bicarbonate fed to Arabidopsis roots was assimilated into aspartic acid but mainly into sucrose, indicating that the added inorganic carbon was transported to the leaves. A hydroponic treatment was also established for A. thaliana using 2 mM NaHCO3 at pH 5.6, which enhanced the photosynthetic and growth parameters. According to transcriptome sequencing data, the observed enhancement in growth may be orchestrated by trehalose-6-phosphate signaling and supported by augmented nitrogen and sulfur assimilation. The analysis also revealed regulatory and transporter activities, including several nitrate (NRT2.1), and sulfate transporter (SULTR1;1 and SULTR1;2) candidates that could participate in bicarbonate uptake. Different transporters and carbon fixation mutants were assessed. Arabidopsis homologs of SLOW-TYPE ANION CHANNEL 1 (slah3) CARBONIC ANHYDRASE (ßca4), and SULFATE TRANSPORTER (sultr1;2) mutants were shown to be inferior to the bicarbonate-treated wild types in several growth and root ultrastructural parameters. Besides, aquaporin genes PIP1;3 and PIP2;6 could play a negative role in the carbon uptake by venting carbon dioxide out of the plant. The findings support the hypothesis that the inorganic carbon is taken up by the root anion channels, mostly transported up to the shoots by the xylem, and fixed there by RuBisCo after the conversion to CO2 by carbonic anhydrases. The process boosts photosynthesis and growth by providing an extra carbon supply.

Transcriptome, hormonal, and secondary metabolite changes in leaves of DEFENSE NO DEATH 1 (DND1) silenced potato plants.

DEFENSE NO DEATH 1 (DND1) is a cyclic nucleotide-gated ion channel protein. Earlier, it was shown that the silencing of DND1 in the potato (Solanum tuberosum L.) leads to resistance to late blight, powdery mildew, and gray mold diseases. At the same time, however, it can reduce plant growth and cause leaf necrosis. To obtain knowledge of the molecular events behind the pleiotropic effect of DND1 downregulation in the potato, metabolite and transcriptome analyses were performed on three DND1 silenced lines of the cultivar 'Désirée.' A massive increase in the salicylic acid content of leaves was detected. Concentrations of jasmonic acid and chlorogenic acid and their derivatives were also elevated. Expression of 1866 genes was altered in the same way in all three DND1 silenced lines, including those related to the synthesis of secondary metabolites. The activation of several alleles of leaf rust, late blight, and other disease resistance genes, as well as the induction of pathogenesis-related genes, was detected. WRKY and NAC transcription factor families were upregulated, whereas bHLHs were downregulated, indicating their central role in transcriptome changes. These results suggest that the maintenance of the constitutive defense state leads to the reduced growth of DND1 silenced potato plants.

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.

Global transcriptome and targeted metabolite analyses of roots reveal different defence mechanisms against Ralstonia solanacearum infection in two resistant potato cultivars.

Ralstonia solanacearum (Rs), the causal agent of bacterial wilt disease in an unusually wide range of host plants, including potato (Solanum tuberosum), is one of the most destructive phytopathogens that seriously reduces crop yields worldwide. Identification of defence mechanisms underlying bacterial wilt resistance is a prerequisite for biotechnological approaches to resistance breeding. Resistance to Rs has been reported only in a few potato landraces and cultivars. Our in vitro inoculation bioassays confirmed that the cultivars 'Calalo Gaspar' (CG) and 'Cruza 148' (CR) are resistant to Rs infection. Comparative transcriptome analyses of CG and CR roots, as well as of the roots of an Rs-susceptible cultivar, 'Désirée' (DES), were carried out two days after Rs infection, in parallel with their respective noninfected controls. In CR and DES, the upregulation of chitin interactions and cell wall-related genes was detected. The phenylpropanoid biosynthesis and glutathione metabolism pathways were induced only in CR, as confirmed by high levels of lignification over the whole stele in CR roots six days after Rs infection. At the same time, Rs infection greatly increased the concentrations of chlorogenic acid and quercetin derivatives in CG roots as it was detected using ultra-performance liquid chromatography - tandem mass spectrometry. Characteristic increases in the expression of MAP kinase signalling pathway genes and in the concentrations of jasmonic, salicylic, abscisic and indoleacetic acid were measured in DES roots. These results indicate different Rs defence mechanisms in the two resistant potato cultivars and a different response to Rs infection in the susceptible cultivar.