The causal mutation leading to sweetness in modern white lupin cultivars.

Lupins are high-protein crops that are rapidly gaining interest as hardy alternatives to soybean; however, they accumulate antinutritional alkaloids of the quinolizidine type (QAs). Lupin domestication was enabled by the discovery of genetic loci conferring low QA levels (sweetness), but the precise identity of the underlying genes remains uncertain. We show that pauper, the most common sweet locus in white lupin, encodes an acetyltransferase (AT) unexpectedly involved in the early QA pathway. In pauper plants, a single-nucleotide polymorphism (SNP) strongly impairs AT activity, causing pathway blockage. We corroborate our hypothesis by replicating the pauper chemotype in narrow-leafed lupin via mutagenesis. Our work adds a new dimension to QA biosynthesis and establishes the identity of a lupin sweet gene for the first time, thus facilitating lupin breeding and enabling domestication of other QA-containing legumes.

FLC expression is down-regulated by cold treatment in Diplotaxis tenuifolia (wild rocket), but flowering time is unaffected.

Wild rocket (Diplotaxis tenuifolia) has become a very popular salad leaf due to its peppery taste. It is part of the Brassicaceae family and thus has a high level of homology at the DNA level to other Brassica species including Arabidopsis thaliana. The vernalization and photoperiodic requirements of wild rocket have not been reported to date. Photoperiodic experiments described here demonstrate that rocket is a facultative long day plant. To investigate the vernalization requirement, both seed and young plants were given vernalization treatments at 4°C for different lengths of time. A rocket homologue of FLOWERING LOCUS C (DtFLC) was isolated and shown to functionally complement the Arabidopsis FRI+flc3 null mutant. Whilst the expression of DtFLC was significantly reduced after just one week of cold treatment, cold treatments of two to eight weeks had no significant effect on bolting time of wild rocket indicating that rocket does not have a vernalization requirement. These findings illustrate that important fundamental differences can exist between model and crop plant species, such as in this case where down-regulation of DtFLC expression does not enable earlier flowering in wild rocket as it does in Arabidopsis and many other Brassica species.

Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots.

BACKGROUND: Tremendous progress has been made in understanding the functions of the GLUTAMATE RECEPTOR-LIKE (GLR) family in Arabidopsis. Still, the functions of OsGLRs in rice, especially the ion channel activities, are largely unknown. RESULTS: Using the aequorin-based luminescence imaging system, we screened the specificity of amino acids involved in the induction of Ca(2+) flux in rice roots. Of all the amino acids tested, glutamate (Glu) was the only one to trigger Ca(2+) flux significantly in rice roots. Detailed analysis showed a dose response of Ca(2+) increase to different concentrations of Glu. In addition, the Ca(2+) spike response to Glu was rapid, within 20 s after the application. A desensitization assay and pharmacological tests showed that the Glu-triggered Ca(2+) flux is mediated by OsGLRs. Whole genome analysis identified 13 OsGLR genes in rice, and these genes have various expression patterns in different tissues. Subcellular localization studies showed that all the OsGLRs examined are likely localized to the plasma membrane. Bacteria growth assays showed that at least OsGLR2.1 and OsGLR3.2 have the potential to mediate ion uptake in bacteria. Further analysis using Fura-2-based Ca(2+) imaging revealed a Glu-triggered Ca(2+) increase in OsGLR2.1-expressing human embryonic kidney (HEK) cells. CONCLUSIONS: Our work provides a molecular basis for investigating mechanisms of Glu-triggered Ca(2+) flux in rice.

Enlightenment on the aequorin-based platform for screening Arabidopsis stress sensory channels related to calcium signaling.

Free calcium ions (Ca(2+)) are an important signal molecule in response to a large array of external stimuli encountered by plants. Using the aequorin-based Ca(2+) recording system, tremendous progress has been made in understanding the Ca(2+) responses to biotic or abiotic stresses in dicotyledonous Arabidopsis. However, due to the lack of a similar detection system, little information has been obtained from the monocotyledonous rice (Oryza sativa). Recombinant aequorin has been introduced into rice, and the Ca(2+) responses to NaCl and H2O2 in rice roots were characterized. Although rice calcium signal sensor research has just started, the transgenic rice expressing aequorin provides a good platform to study rice adapted to different environmental conditions.

Aequorin-based luminescence imaging reveals differential calcium signalling responses to salt and reactive oxygen species in rice roots.

It is well established that both salt and reactive oxygen species (ROS) stresses are able to increase the concentration of cytosolic free Ca(2+) ([Ca(2+)]i), which is caused by the flux of calcium (Ca(2+)). However, the differences between these two processes are largely unknown. Here, we introduced recombinant aequorin into rice (Oryza sativa) and examined the change in [Ca(2+)]i in response to salt and ROS stresses. The transgenic rice harbouring aequorin showed strong luminescence in roots when treated with exogenous Ca(2+). Considering the histological differences in roots between rice and Arabidopsis, we reappraised the discharging solution, and suggested that the percentage of ethanol should be 25%. Different concentrations of NaCl induced immediate [Ca(2+)]i spikes with the same durations and phases. In contrast, H2O2 induced delayed [Ca(2+)]i spikes with different peaks according to the concentrations of H2O2. According to the Ca(2+) inhibitor research, we also showed that the sources of Ca(2+) induced by NaCl and H2O2 are different. Furthermore, we evaluated the contribution of [Ca(2+)]i responses in the NaCl- and H2O2-induced gene expressions respectively, and present a Ca(2+)- and H2O2-mediated molecular signalling model for the initial response to NaCl in rice.