The transition from vegetative growth to reproduction is a major developmental event in plants. To maximise reproductive success, its timing is determined by complex interactions between environmental cues like the photoperiod, temperature and nutrient availability and internal genetic programs. While the photoperiod- and temperature- and gibberellic acid-signalling pathways have been subjected to extensive analysis, little is known about how nutrients regulate floral induction. This is partly because nutrient supply also has large effects on vegetative growth, making it difficult to distinguish primary and secondary influences on flowering. A growth system using glutamine supplementation was established to allow nitrate to be varied without a large effect on amino acid and protein levels, or the rate of growth. Under nitrate-limiting conditions, flowering was more rapid in neutral (12/12) or short (8/16) day conditions in C24, Col-0 and Laer. Low nitrate still accelerated flowering in late-flowering mutants impaired in the photoperiod, temperature, gibberellic acid and autonomous flowering pathways, in the fca co-2 ga1-3 triple mutant and in the ft-7 soc1-1 double mutant, showing that nitrate acts downstream of other known floral induction pathways. Several other abiotic stresses did not trigger flowering in fca co-2 ga1-3, suggesting that nitrate is not acting via general stress pathways. Low nitrate did not further accelerate flowering in long days (16/8) or in 35S::CO lines, and did override the late-flowering phenotype of 35S::FLC lines. We conclude that low nitrate induces flowering via a novel signalling pathway that acts downstream of, but interacts with, the known floral induction pathways.Electronic supplementary materialThe online version of this article (doi:10.1007/s00425-010-1316-5) contains supplementary material, which is available to authorized users.

Freshly cut discs from growing potato tubers were incubated for 3 h with 10 mM orotate or 10 mM uridine. Control discs incubated without precursors showed a 30-40% decrease of uridine nucleotides, but not of adenine nucleotides. Orotate- and uridine-feeding led to a 1.5- to 2-fold increase in the levels of uridine nucleotides compared with control discs, and a 15-30% increase compared with the original values in intact tubers, but did not alter the levels of adenine nucleotides. Between 70-80% of the uridine nucleotides were present as UDPglucose, 15-25% as UTP, and 2-3% as UDP. The increase of uridine nucleotides involved a similar relative increase of UDPglucose, UTP and UDP. It was accompanied by a slight stimulation of the rate of [(14)C]sucrose uptake, a 2-fold stimulation of the rate at which the [(14)C]sucrose was subsequently metabolised, a small increase in the levels of hexose phosphates, glycerate-3-phospate and ADPglucose, and a 30% shift in the allocation of the metabolised label in favour of starch synthesis, resulting in a 2.4-fold stimulation of the rate of starch synthesis. Orotate led to a similar increase of uridine nucleotide levels in the presence of [(14)C]glucose, but did not significantly alter the rate of glucose uptake and metabolism to starch, nor did it increase the rate of sucrose resynthesis. The levels of uridine nucleotides were high in tubers on 6 to 10-week-old potato plants, and declined in tubers on 12 to 15-week-old plants. Comparison with the effect of the uridine nucleotide level in discs shows that the high levels of uridine nucleotides in tubers on young plants will play an important role in determining the rate at which sucrose can be converted to starch, and that the level of uridine nucleotides is probably co-limiting for sucrose-starch conversions in tubers on older plants.

To investigate the importance of the overall size of the total adenine nucleotide pool for the regulation of primary metabolism in growing potato tubers, freshly cut discs were provided with zero or 2 mM adenine in the presence of 1 or 100 mM [U-14C]glucose or 100 mM [U-14C]sucrose in the presence and absence of 20 mM orthophosphate (Pi). Adenine led to a 150-250% increase of the total adenine nucleotide pool, which included an increase of ADP, a larger increase of ATP and an increase of the ATP:ADP ratio. There was a 50-100% increase of ADP-glucose (ADPGlc), and starch synthesis was stimulated. Respiratory oxygen uptake was stimulated, and the levels of glycerate-3-phosphate, phosphoenolpyruvate and alpha-ketoglutarate decreased. The response to adenine was not modified by Pi. It is proposed that increased ATP stimulates ADPGlc pyrophosphorylase, leading to a higher rate of starch synthesis. The impact on starch synthesis is constrained, however, because increased ADP can lead to a stimulation of respiration and decline of glycerate-3-phosphate, which will inhibit ADPGlc pyrophosphorylase. The quantitative impact depends on the conditions. In the presence of 1 mM glucose, the levels of phosphorylated intermediates and the rate of starch synthesis were low. Adenine led to a relatively large stimulation of respiration, but only a small stimulation of starch synthesis. In the presence of 100 mM glucose, discs contained high levels of phosphorylated intermediates, low ATP:ADP ratios (< 3) and low rates of starch synthesis (< 20% of the metabolised glucose). Adenine led to marked increase of ATP and 2- to 4-fold stimulation of starch synthesis. Discs incubated with 100 mM sucrose already had high ATP:ADP ratios (> 8) and high rates of starch synthesis (> 50% of the metabolised sucrose). Adenine led to a further increase, but the stimulation was less marked than in high glucose. These results have implications for the function of nucleotide cofactors in segregating sucrose mobilisation and respiration, and the need for energy conservation during sugar-starch conversions.

The marine sponge-derived fungus Auxarthron reticulatum produces the cannabinoid receptor antagonist amauromine (1). Recultivation of the fungus to obtain further amounts for more detailed pharmacological evaluation of 1 additionally yielded the novel triterpene glycoside auxarthonoside (2), bearing, in nature, a rather rare sugar moiety, i.e., N-acetyl-6-methoxy-glucosamine. Amauromine (1), which inhibited cannabinoid CB1 receptors (Ki 0.178 µM) also showed antagonistic activity at the cannabinoid-like orphan receptor GPR18 (IC50 3.74 µM). The diketopiperazine 1 may thus serve as a lead structure for the development of more potent and selective GPR18 antagonists, which are required to study the orphan receptor's potential as a new drug target. Despite the execution of many biological assays, to date, no bioactivity could be found for auxarthonoside (2).