Sugar Transporter Protein 1 (STP1) contributes to regulation of the genes involved in shoot branching via carbon partitioning in Arabidopsis.

We previously demonstrated that alterations in sugar partitioning affect the expression of genes involved in hormone biosynthesis and responses, including BRANCHED1 (BRC1), resulting in enhanced shoot branching in transgenic Arabidopsis plants overexpressing cyanobacterial fructose-1,6-bisphosphatase-II in the cytosol (AcF). The exogenous treatment of wild-type Arabidopsis plants with sugars showed the same transcript characteristics, indicating that sugars act as a signal for branching. We also found that the reductions induced in BRC1 expression levels in wild-type plants by the sugar treatments were suppressed in the knockout mutant of sugar transporter 1 (stp1-1). Intracellular sugar contents were similar in stp1-1 and wild-type plants following the sugar treatments, suggesting that STP1 acts as a factor for the regulation of shoot branching depending on extracellular sugar contents. Abbreviations: BRC1: BRABCHED1; FBP/SBPase: fructose-1,6-/sedoheptulose-1,7-bisphosphatase; Glc: glucose; HXK: hexokinase; SnRK1.1/AKIN10: SNF1-RELATED PROTEIN KINASE 1.1; Suc: sucrose; SnRK1: sucrose non-fermenting 1-related protein kinase; STP: sugar transporter protein.

Enhancements in sucrose biosynthesis capacity affect shoot branching in Arabidopsis.

We previously demonstrated that transgenic tobacco plants expressing cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase in the cytosol increased the number of lateral shoots and leaves at elevated CO2 levels. These findings suggest that alterations in carbon partitioning affect the development of shoot branching. In order to elucidate the underlying mechanisms at the molecular level, we generated transgenic Arabidopsis plants overexpressing cyanobacterial fructose-1,6-bisphosphatase-II in the cytosol (AcF). At elevated CO2 levels, the number of lateral shoots was significantly increased in AcF plants. Sucrose and hexose levels were also higher in AcF plants than in wild-type plants. The expression levels of MAX1, MAX4, YUCCA8, YUCCA9, and BRC1, which are involved in auxin or strigolactone biosynthesis and responses, were lower in AcF plants than in wild-type plants. These results suggest that alterations in sugar partitioning affect hormone metabolism and responses, resulting in enhanced shoot branching.

Enhanced photosynthetic capacity increases nitrogen metabolism through the coordinated regulation of carbon and nitrogen assimilation in Arabidopsis thaliana.

Plant growth and productivity depend on interactions between the metabolism of carbon and nitrogen. The sensing ability of internal carbon and nitrogen metabolites (the C/N balance) enables plants to regulate metabolism and development. In order to investigate the effects of an enhanced photosynthetic capacity on the metabolism of carbon and nitrogen in photosynthetically active tissus (source leaves), we herein generated transgenic Arabidopsis thaliana plants (ApFS) that expressed cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase in their chloroplasts. The phenotype of ApFS plants was indistinguishable from that of wild-type plants at the immature stage. However, as plants matured, the growth of ApFS plants was superior to that of wild-type plants. Starch levels were higher in ApFS plants than in wild-type plants at 2 and 5 weeks. Sucrose levels were also higher in ApFS plants than in wild-type plants, but only at 5 weeks. On the other hand, the contents of various free amino acids were lower in ApFS plants than in wild-type plants at 2 weeks, but were similar at 5 weeks. The total C/N ratio was the same in ApFS plants and wild-type plants, whereas nitrite levels increased in parallel with elevations in nitrate reductase activity at 5 weeks in ApFS plants. These results suggest that increases in the contents of photosynthetic intermediates at the early growth stage caused a temporary imbalance in the free-C/free-N ratio and, thus, the feedback inhibition of the expression of genes involved in the Calvin cycle and induction of the expression of those involved in nitrogen metabolism due to supply deficient free amino acids for maintenance of the C/N balance in source leaves of ApFS plants.

A gain-of-function mutation of plastidic invertase alters nuclear gene expression with sucrose treatment partially via GENOMES UNCOUPLED1-mediated signaling.

Plastid gene expression (PGE) is one of the signals that regulate the expression of photosynthesis-associated nuclear genes (PhANGs) via GENOMES UNCOUPLED1 (GUN1)-dependent retrograde signaling. We recently isolated Arabidopsis sugar-inducible cotyledon yellow-192 (sicy-192), a gain-of-function mutant of plastidic invertase, and showed that following the treatment of this mutant with sucrose, the expression of PhANGs as well as PGE decreased, suggesting that the sicy-192 mutation activates a PGE-evoked and GUN1-mediated retrograde pathway. To clarify the relationship between the sicy-192 mutation, PGE, and GUN1-mediated pathway, plastid and nuclear gene expression in a double mutant of sicy-192 and gun1-101, a null mutant of GUN1 was studied. Plastid-encoded RNA polymerase (PEP)-dependent PGE was markedly suppressed in the sicy-192 mutant by the sucrose treatment, but the suppression as well as cotyledon yellow phenotype was not mitigated by GUN1 disruption. Microarray analysis revealed that the altered expression of nuclear genes such as PhANG in the sucrose-treated sicy-192 mutant was largely dependent on GUN1. The present findings demonstrated that the sicy-192 mutation alters nuclear gene expression with sucrose treatment via GUN1, which is possibly followed by inhibiting PEP-dependent PGE, providing a new insight into the role of plastid sugar metabolism in nuclear gene expression.

OASIS, a CREB/ATF-family member, modulates UPR signalling in astrocytes.

Endoplasmic reticulum (ER) stress transducers IRE1, PERK and ATF6 are well known to transduce signals from the ER to the cytoplasm and nucleus when unfolded proteins are accumulated in the ER. Here, we identified OASIS as a novel ER stress transducer. OASIS is a basic leucine zipper (bZIP) transcription factor of the CREB/ATF family with a transmembrane domain that allows it to associate with the ER. The molecule is cleaved at the membrane in response to ER stress, and its cleaved amino-terminal cytoplasmic domain, which contains the bZIP domain, translocates into the nucleus where it activates the transcription of target genes that are mediated by ER stress-responsive and cyclic AMP-responsive elements. Intriguingly, OASIS was induced at the transcriptional level during ER stress in astrocytes of the central nervous system, but not in other cell types examined. Furthermore, overexpression of OASIS resulted in induction of BiP and suppression of ER-stress-induced cell death, whereas knockdown partially reduced BiP levels and led to ER stress in susceptible astrocytes. Our results reveal pivotal roles for OASIS in modulating the unfolded protein response in astrocytes, and the possibility that cell type-specific UPR signalling also exists in other cells.

Point mutation of a plastidic invertase inhibits development of the photosynthetic apparatus and enhances nitrate assimilation in sugar-treated Arabidopsis seedlings.

Because the photosynthetic apparatus contains a massive amount of nitrogen in plants, the regulation of its development by sugar signals is important to the maintenance of the carbon-nitrogen balance. In this study we isolated an Arabidopsis mutant (sicy-192) whose cotyledon greening was inhibited by treatments with sugars such as sucrose, glucose, and fructose. In the mutant, the gene encoding plastidic alkaline/neutral invertase (INV-E) was point-mutated at codon 294, with Tyr substituted for Cys (C294Y). Interestingly, the greening of cotyledons in the knock-out INV-E lines was not inhibited by treatment with the sugars. In addition, the knock-out INV-E lines expressing an INV-E:C294Y or INV-E:C294A gene had the same phenotype as sicy-192 mutants, whereas the lines expressing a wild-type INV-E gene had the same phenotype as wild-type plants. A recombinant INV-E:C294Y protein had the same enzymatic activity as a recombinant INV-E protein, suggesting that the Cys-294 residue of INV-E is important for its functions in the chloroplasts. On treatment with sucrose, the expression of photosynthesis-related genes was weaker in seedlings of mutant plants than wild-type seedlings, whereas the activity of nitrate reductase was stronger in the mutant plants than wild-type plants. These findings suggest that Cys-294 of INV-E is associated with the development of the photosynthetic apparatus and the assimilation of nitrogen in Arabidopsis seedlings to control the ratio of sucrose content to hexose content.

Increase in the activity of fructose-1,6-bisphosphatase in cytosol affects sugar partitioning and increases the lateral shoots in tobacco plants at elevated CO2 levels.

We generated transgenic tobacco plants with high levels of fructose-1,6-bisphosphatase expressing cyanobacterialfructose-1,6-/sedoheptulose-1,7-bisphosphatase in the cytosol. At ambient CO(2) levels (360 ppm), growth, photosynthetic activity, and fresh weight were unchanged but the sucrose/hexose/starch ratio was slightly altered in the transgenic plants compared with wild-type plants. At elevated CO(2) levels (1200 ppm), lateral shoot, leaf number, and fresh weight were significantly increased in the transgenic plants. Photosynthetic activity was also increased. Hexose accumulated in the upper leaves in the wild-type plants, while sucrose and starch accumulated in the lower leaves and lateral shoots in the transgenic plants. These findings suggest that cytosolic fructose-1,6-bisphosphatase contributes to the efficient conversion of hexose into sucrose, and that the change in carbon partitioning affects photosynthetic capacity and morphogenesis at elevated CO(2) levels.

New insights into the regulation of greening and carbon-nitrogen balance by sugar metabolism through a plastidic invertase.

Since the photosynthetic apparatus of plants contains a massive amount of nitrogen, the regulation of its development by sugar signals is important to the maintenance of the carbon-nitrogen balance. Recently, we isolated a new Arabidopsis mutant, sicy (sugar-inducible cotyledon yellow)-192, whose cotyledons were prevented from greening by treatment with sucrose. On treatment with sucrose, the expression of photosynthesis- and nitrogen assimilation-related genes was respectively weaker and stronger in the mutant seedlings than the wild-type seedlings. In the mutants, the gene encoding plastidic alkaline/neutral (A/N) invertase (INV-E) was point-mutated at codon 294, with Tyr substituted for Cys (C294Y). These findings provide new insights into the regulation of greening and carbon-nitrogen balance by sugar metabolism through INV-E in plastids. In this addendum, we describe the phenotypes of sicy-192 on treatment with sucrose in more detail, and propose a possible relationship among sugar metabolism through INV-E, plastid-to-nucleus retrograde signaling, and ethylene, a plant hormone, in the regulation of plant development and metabolism.