Inhibition of NPR1 Leads to Shoot Growth Improvement under Low-Calcium Conditions in Arabidopsis.

Under low-Ca conditions, plants accumulate salicylic acid (SA) and induce SA-responsive genes. However, the relationship between SA and low-Ca tolerance remains unclear. Here, we demonstrated that the inhibition or suppression of nonexpressor of pathogenesis-related 1 (NPR1) activity, a major regulator of the SA signaling pathway in the defense response, improves shoot growth under low-Ca conditions. Furthermore, mutations in phytoalexin-deficient 4 (PAD4) or enhanced disease susceptibility 1 (EDS1), which are upstream regulators of NPR1, improved shoot growth under low-Ca conditions, suggesting that NPR1 suppressed growth under low-Ca conditions. In contrast, growth of SA induction-deficient 2-2 (sid2-2), which is an SA-deficient mutant, was sensitive to low Ca levels, suggesting that SA accumulation by SID2 was not related to growth inhibition under low-Ca conditions. Additionally, npr1-1 showed low-Ca tolerance, and the application of tenoxicam-an inhibitor of the NPR1-mediated activation of gene expression-also improved shoot growth under low Ca conditions. The low-Ca tolerance of double mutants pad4-1, npr1-1 and eds1-22 npr1-1 was similar to that of the single mutants, suggesting that PAD4 and EDS1 are involved in the same genetic pathway in suppressing growth under low-Ca conditions as NPR1. Cell death and low-Ca tolerance did not correlate among the mutants, suggesting that growth improvement in the mutants was not due to cell death inhibition. In conclusion, we revealed that NPR1 suppresses plant growth under low-Ca conditions and that the other SA-related genes influence plant growth and cell death.

Role of GSL8 in low calcium tolerance in Arabidopsis thaliana.

Calcium (Ca) deficiency affects the yields and quality of agricultural products. Susceptibility to Ca deficiency varies among crops and cultivars; however, its genetic basis remains largely unknown. Genes required for low Ca tolerance in Arabidopsis thaliana have been identified. In this study, we identified a novel gene required for low Ca tolerance in A. thaliana. We isolated a mutant sensitive to low Ca concentrations and identified Glucan synthase-like (GSL) 8 as a gene responsible for low Ca tolerance. GSL8 is a paralog of the previously identified low Ca tolerance gene GSL10, which encodes ß-1,3 glucan(callose) synthase. Under low Ca conditions, the shoot growth of gsl8 mutants were inhibited compared to wild-type plants. A grafting experiment indicated that the shoot, but not root, genotype was important for the shoot growth phenotype. The ectopic accumulation of callose under low Ca conditions was reduced in gsl8 mutants. We further investigated the interaction between GSL8 and GSL10 by testing the gsl8 gsl10 double mutant for sensitivity to low Ca concentrations. The double mutant exhibited a more severe phenotype than the single mutant under 0.3 mM Ca, indicating additive effects of GSL8 and GSL10 with respect to low Ca tolerance. These results establish that GSL genes are required for low Ca tolerance in A. thaliana.

Arabidopsis Glucan Synthase-Like1 (GSL1) Is Required for Tolerance to Low-Calcium Conditions and Exhibits a Function Comparable to GSL10.

Crops that exhibit symptoms of calcium (Ca) deficiency constitute a major agricultural problem. Molecular breeding of resistant cultivars is a promising method for overcoming this problem. However, the involved genes must first be identified. Here, we show that the glucan synthase-like (GSL) 1 gene is essential for low-Ca tolerance in Arabidopsis thaliana. GSL1 is homologous to GSL10, which we previously showed was essential for low-Ca tolerance. Under low-Ca conditions, gsl1 mutants exhibit reduced growth and the onset of necrosis in new leaves. These symptoms are typical of Ca-deficient crops. A grafting experiment suggested that the shoot genotype, but not the root genotype, was important for the suppression of shoot necrosis. The ectopic accumulation of callose under low-Ca conditions was significantly reduced in gsl1 mutants compared with wild-type plants. Because the corresponding single-mutant phenotypes are similar, we investigated the interaction between GSL1 and GSL10 by testing the gsl1 gsl10 double mutant for sensitivity to low-Ca conditions. The double mutant exhibited a more severe phenotype than did the single mutants, indicating that the effects of GSL1 and GSL10 on low-Ca tolerance are additive. Because GSL genes are highly conserved within the plant kingdom, the GSL loci may be useful for breeding low-Ca tolerant crops.

Multisensory Proximity and Transition Cues for Improving Target Awareness in Narrow Field of View Augmented Reality Displays.

Augmented reality applications allow users to enrich their real surroundings with additional digital content. However, due to the limited field of view of augmented reality devices, it can sometimes be difficult to become aware of newly emerging information inside or outside the field of view. Typical visual conflicts like clutter and occlusion of augmentations occur and can be further aggravated especially in the context of dense information spaces. In this article, we evaluate how multisensory cue combinations can improve the awareness for moving out-of-view objects in narrow field of view augmented reality displays. We distinguish between proximity and transition cues in either visual, auditory or tactile manner. Proximity cues are intended to enhance spatial awareness of approaching out-of-view objects while transition cues inform the user that the object just entered the field of view. In study 1, user preference was determined for 6 different cue combinations via forced-choice decisions. In study 2, the 3 most preferred modes were then evaluated with respect to performance and awareness measures in a divided attention reaction task. Both studies were conducted under varying noise levels. We show that on average the Visual-Tactile combination leads to 63% and Audio-Tactile to 65% faster reactions to incoming out-of-view augmentations than their Visual-Audio counterpart, indicating a high usefulness of tactile transition cues. We further show a detrimental effect of visual and audio noise on performance when feedback included visual proximity cues. Based on these results, we make recommendations to determine which cue combination is appropriate for which application.

An SMU Splicing Factor Complex Within Nuclear Speckles Contributes to Magnesium Homeostasis in Arabidopsis.

Mg2+ is among the most abundant divalent cations in living cells. In plants, investigations on magnesium (Mg) homeostasis are restricted to the functional characterization of Mg2+ transporters. Here, we demonstrate that the splicing factors SUPPRESSORS OF MEC-8 AND UNC-52 1 (SMU1) and SMU2 mediate Mg homeostasis in Arabidopsis (Arabidopsis thaliana). A low-Mg sensitive Arabidopsis mutant was isolated, and the causal gene was identified as SMU1 Disruption of SMU2, a protein that can form a complex with SMU1, resulted in a similar low-Mg sensitive phenotype. In both mutants, an Mg2+ transporter gene, Mitochondrial RNA Splicing 2 (MRS2-7), showed altered splicing patterns. Genetic evidence indicated that MRS2-7 functions in the same pathway as SMU1 and SMU2 for low-Mg adaptation. In contrast with previous results showing that the SMU1-SMU2 complex is the active form in RNA splicing, MRS2-7 splicing was promoted in the smu2 mutant overexpressing SMU1, indicating that complex formation is not a prerequisite for the splicing. We found here that formation of the SMU1-SMU2 complex is an essential step for their compartmentation in the nuclear speckles, a type of nuclear body enriched with proteins that participate in various aspects of RNA metabolism. Taken together, our study reveals the involvement of the SMU splicing factors in plant Mg homeostasis and provides evidence that complex formation is required for their intranuclear compartmentation.

Callose Synthesis Suppresses Cell Death Induced by Low-Calcium Conditions in Leaves.

Despite the importance of preventing calcium (Ca) deficiency disorders in agriculture, knowledge of the molecular mechanisms underlying plant adaptations to low-Ca conditions is limited. In this study, we provide evidence for a crucial involvement of callose synthesis in the survival of Arabidopsis (Arabidopsis thaliana) under low-Ca conditions. A mutant sensitive to low-Ca conditions, low calcium sensitive3 (lcs3), exhibited high levels of cell death in emerging leaves and had defects in its expanding true leaves under low-Ca conditions. Further analyses showed that the causal mutation was located in a putative ß-1,3-glucan (callose) synthase gene, GLUCAN SYNTHASE-LIKE10 (GSL10). Yeast complementation assay results showed that GSL10 encodes a functional callose synthase. Ectopic callose significantly accumulated in wild-type plants under low-Ca conditions, but at a low level in lcs3 The low-Ca sensitivity of lcs3 was phenocopied by the application of callose synthase inhibitors in wild-type plants, which resulted in leaf expansion failure, cell death, and reduced ectopic callose levels under low-Ca conditions. Transcriptome analyses showed that the expression of genes related to cell wall and defense responses was altered in both wild-type plants under low-Ca conditions and in lcs3 under normal-Ca conditions, suggesting that GSL10 is required for the alleviation of both cell wall damage and defense responses caused by low Ca levels. These results suggest that callose synthesis is essential for the prevention of cell death under low-Ca conditions and plays a key role in plants' survival strategies under low-Ca conditions.

The Arabidopsis Mg Transporter, MRS2-4, is Essential for Mg Homeostasis Under Both Low and High Mg Conditions.

Magnesium (Mg) is an essential macronutrient, functioning as both a cofactor of many enzymes and as a component of Chl. Mg is abundant in plants; however, further investigation of the Mg transporters involved in Mg uptake and distribution is needed. Here, we isolated an Arabidopsis thaliana mutant sensitive to high calcium (Ca) conditions without Mg supplementation. The causal gene of the mutant encodes MRS2-4, an Mg transporter.MRS2-4 single mutants exhibited growth defects under low Mg conditions, whereas an MRS2-4 and MRS2-7 double mutant exhibited growth defects even under normal Mg concentrations. Under normal Mg conditions, the Mg concentration of the MRS2-4 mutant was lower than that of the wild type. The transcriptome profiles of mrs2-4-1 mutants under normal conditions were similar to those of wild-type plants grown under low Mg conditions. In addition, both mrs2-4 and mrs2-7 mutants were sensitive to high levels of Mg. These results indicate that both MRS2-4 and MRS2-7 are essential for Mg homeostasis, even under normal and high Mg conditions. MRS2-4-green fluorescent protein (GFP) was mainly detected in the endoplasmic reticulum. These results indicate that these two MRS2 transporter genes are essential for the ability to adapt to a wide range of environmental Mg concentrations.