Cesium Toxicity Alters MicroRNA Processing and AGO1 Expressions in Arabidopsis thaliana

MicroRNAs (miRNAs) are short RNA fragments that play important roles in controlled gene silencing, thus regulating many biological processes in plants. Recent studies have indicated that plants modulate miRNAs to sustain their survival in response to a variety of environmental stimuli, such as biotic stresses, cold, drought, nutritional starvation, and toxic heavy metals. Cesium and radio-cesium contaminations have arisen as serious problems that both impede plant growth and enter the food chain through contaminated plants. Many studies have been performed to define plant responses against cesium intoxication. However, the complete profile of miRNAs in plants during cesium intoxication has not been established. Here we show the differential expression of the miRNAs that are mostly down-regulated during cesium intoxication. Furthermore, we found that cesium toxicity disrupts both the processing of pri-miRNAs and AGONOUTE 1 (AGO1)-mediated gene silencing. AGO 1 seems to be especially destabilized by cesium toxicity, possibly through a proteolytic regulatory pathway. Our study presents a comprehensive profile of cesium-responsive miRNAs, which is distinct from that of potassium, and suggests two possible mechanisms underlying the cesium toxicity on miRNA metabolism.     

SUMO E3 Ligase AtMMS21 Regulates Drought Tolerance in Arabidopsis thaliana

Post-translational modifications of proteins by small ubiquitin-like modifiers (SUMOs) play crucial roles in plant growth and development, and in stress responses. The MMS21 is a newly-identified Arabidopsis thaliana L. SUMO E3 ligase gene aside from the SIZ1, and its function requires further elucidation. Here, we show that MMS21 deficient plants display improved drought tolerance, and constitutive expression of MMS21 reduces drought tolerance. The expression of MMS21 was reduced by abscisic acid (ABA), polyethylene glycol (PEG) or drought stress. Under drought conditions, mms21 mutants showed the highest survival rate and the slowest water loss, and accumulated a higher level of free proline compared to wild-type (WT) and MMS21 over-expression plants. Stomatal aperture, seed germination and cotyledon greening analysis indicated that mms21 was hypersensitive to ABA. Molecular genetic analysis revealed that MMS21 deficiency led to elevated expression of a series of ABA-mediated stress-responsive genes, including COR15A, RD22, and P5CS1 The ABA and drought-induced stress-responsive genes, including RAB18, RD29A and RD29B, were inhibited by constitutive expression of MMS21. Moreover, ABA-induced accumulation of SUMO-protein conjugates was blocked in the mms21 mutant. We thus conclude that MMS21 plays a role in the drought stress response, likely through regulation of gene expression in an ABA-dependent pathway.     

拟南芥耐盐、耐旱和抗寒的分子机理及其利用

非生物胁迫,尤其是盐分、干旱和寒冷是导致全球作物减产的主要原因.植物对环境胁迫的适应,依赖于与胁迫感知、信号转导、基因表达相关的分子级联网络的激活.因此,保护和维持细胞成分的结构和功能的基因工程可以增强植物对胁迫的耐受性.综述拟南芥对盐分、干旱和寒冷3种主要非生物胁迫因子耐受性的分子机理,以及相关机理和耐逆基因在改良作物耐逆性方面的应用.     

Hydrogen peroxide-induced gene expression in Arabidopsis thaliana

Hydrogen peroxide (H(2)O(2)) is generated in plants after exposure to a variety of biotic and abiotic stresses, and has been shown to induce a number of cellular responses. Previously, we showed that H(2)O(2) generated during plant-elicitor interactions acts as a signaling molecule to induce the expression of defense genes and initiate programmed cell death in Arabidopsis thaliana suspension cultures. Here, we report for the first time the identification by RNA differential display of four genes whose expression is induced by H(2)O(2). These include genes that have sequence homology to previously identified Arabidopsis genes encoding a late embryogenesis-abundant protein, a DNA-damage repair protein, and a serine/threonine kinase. Their putative roles in H(2)O(2)-induced defense responses are discussed.     

Large Root Cortical Cell Size Improves Drought Tolerance in Maize

The objective of this study was to test the hypothesis that large cortical cell size (CCS) would improve drought tolerance by reducing root metabolic costs. Maize (Zea mays) lines contrasting in root CCS measured as cross-sectional area were grown under well-watered and water-stressed conditions in greenhouse mesocosms and in the field in the United States and Malawi. CCS varied among genotypes, ranging from 101 to 533 µm². In mesocosms, large CCS reduced respiration per unit of root length by 59%. Under water stress in mesocosms, lines with large CCS had between 21% and 27% deeper rooting (depth above which 95% of total root length is located in the soil profile), 50% greater stomatal conductance, 59% greater leaf CO₂ assimilation, and between 34% and 44% greater shoot biomass than lines with small CCS. Under water stress in the field, lines with large CCS had between 32% and 41% deeper rooting (depth above which 95% of total root length is located in the soil profile), 32% lighter stem water isotopic ratio of ¹⁸O to ¹⁶O signature, signifying deeper water capture, between 22% and 30% greater leaf relative water content, between 51% and 100% greater shoot biomass at flowering, and between 99% and 145% greater yield than lines with small cells. Our results are consistent with the hypothesis that large CCS improves drought tolerance by reducing the metabolic cost of soil exploration, enabling deeper soil exploration, greater water acquisition, and improved growth and yield under water stress. These results, coupled with the substantial genetic variation for CCS in diverse maize germplasm, suggest that CCS merits attention as a potential breeding target to improve the drought tolerance of maize and possibly other cereal crops.Suboptimal water availability is a primary constraint for terrestrial plants and a primary limitation to crop production. In developing countries, the problem of yield loss due to drought is most severe (Edmeades, 2008, 2013), and the problem will be further exacerbated in the future due to climate change (Burke et al., 2009; Schlenker and Lobell, 2010; Lobell et al., 2011a; IPCC, 2014; St. Clair and Lynch, 2010). The development of drought-tolerant crops is therefore an important goal for global agriculture. Breeding for drought adaptation using yield as a selection criterion is generally not efficient, since yield is an integration of complex mechanisms at different levels of organization affected by many elements of the phenotype and the environment interacting in complex and often unknown ways. Trait-based selection or ideotype breeding is generally a more efficient selection strategy, permitting the identification of useful sources of variation among lines that have poor agronomic adaptation, elucidation of genotype-environment interactions, and informed trait stacking (Lynch, 2007; Araus et al., 2008; Richards et al., 2010; Wasson et al., 2012; York et al., 2013; Lynch, 2014).Under drought stress, plants allocate more resources to root growth relative to shoot growth, which can enhance water acquisition (Sharp and Davies, 1979; Palta and Gregory, 1997; Lynch and Ho, 2005). The metabolic costs of soil exploration by root systems are significant and can exceed 50% of daily photosynthesis (Lambers et al., 2002). With a large root system, each unit of leaf area has more nonphotosynthetic tissue to sustain, which may reduce productivity by diverting resources from shoot and reproductive growth (Smucker, 1993; Nielsen et al., 2001; Boyer and Westgate, 2004). Genotypes with less costly root tissue could develop the extensive, deep root systems required to fully utilize soil water resources in drying soil without as much yield penalty. Therefore, root phenes that reduce the metabolic costs of soil exploration, thereby improving water acquisition, are likely to be valuable for improving drought tolerance (Lynch and Ho, 2005; Zhu et al., 2010; Lynch, 2011; Richardson et al., 2011; Jaramillo et al., 2013; Lynch 2014).Maize (Zea mays) is the principal global cereal. Maize production is facing major challenges as a result of the increasing frequency and intensity of drought (Tuberosa and Salvi, 2006), and this problem will likely be exacerbated by climate change (Lobell et al., 2011b). The Steep, Cheap, and Deep ideotype has been proposed for improving water and nitrogen acquisition by maize when these resources are limited (Lynch, 2013). This ideotype consists of root architectural, anatomical, and physiological traits that may increase rooting depth and thereby improve water acquisition from drying soils. Anatomical phenes could influence the metabolic cost of soil exploration by changing the proportion of respiring and nonrespiring root tissue and affecting the metabolic cost of tissue construction and maintenance, which is an important limitation to root growth and plant development under edaphic stress. Specific anatomical phenes that may contribute to rooting depth by reducing root metabolic costs include components of living cortical area (LCA; Jaramillo et al., 2013), including root cortical aerenchyma (RCA), cortical cell size (CCS), and cortical cell file number (Lynch, 2013).RCA consists of large air-filled lacunae that replace living cortical cells as a result of programmed cell death (Evans, 2004). Previous studies have demonstrated that RCA improves crop adaptation to edaphic stress by reducing the metabolic cost of soil exploration and exploitation (Fan et al., 2003; Zhu et al., 2010; Postma and Lynch, 2011a, Saengwilai et al., 2014a). RCA is associated with a disproportionate reduction of root respiration, thereby permitting greater root growth and acquisition of soil resources (Fan et al., 2003; Zhu et al., 2010). SimRoot modeling indicated that RCA can substantially increase the acquisition of nitrogen, phosphorus, and potassium in maize by reducing respiration and the nutrient content of root tissue (Postma and Lynch, 2011b). Under water stress in the field, maize genotypes with more RCA had deeper roots, better leaf water status, and 800% greater yield than genotypes with less RCA (Zhu et al., 2010). Under nitrogen stress in the field and in greenhouse mesocosms, maize genotypes with more RCA had greater rooting depth, greater nitrogen capture from deep soil strata, greater nitrogen content, greater leaf photosynthesis, greater biomass, and greater yield (Saengwilai et al., 2014a).LCA refers to the living portion of the cortex that remains after the formation of aerenchyma (Jaramillo et al., 2013). Recently, we reported that LCA is an important determinant of root metabolic cost and a better predictor of root respiration than RCA (Jaramillo et al., 2013). In that study, maize lines contrasting in LCA were grown under well-watered or water-stressed conditions in soil mesocosms, and LCA was associated with a reduction of specific root respiration. These results provided the impetus to investigate the relative contribution of each component of LCA to metabolic cost. Our focus here is on root CCS.Plant cell size varies substantially both among and within species (Sugimoto-Shirasu and Roberts, 2003). Cell size in a given species and tissue is under genetic control and results from the coordinated control of cell growth and cell division (Sablowski and Carnier Dornelas, 2014). The increased volume of individual cells is attributable to cytoplasmic growth and cell expansion (Marshall et al., 2012; Chevalier et al., 2014). Cytoplasmic growth is the net accumulation of macromolecules and cellular organelles, while cell expansion refers to increased cell volume caused by enlargement of the vacuole (Taiz, 1992; Sablowski and Carnier Dornelas, 2014). Lynch (2013) proposed that large CCS would decrease the metabolic costs of root growth and maintenance, both in terms of the carbon cost of root respiration and the nutrient content of living tissue, by increasing the ratio of vacuolar to cytoplasmic volume.The objective of this study was to test the hypothesis that large CCS would reduce specific root respiration (i.e. respiration per unit of root length), which under water stress would result in greater root growth, greater acquisition of subsoil water, better plant water status, and improved plant growth and yield. Diverse sets of genotypes (including landraces and recombinant inbred lines [RILs]) contrasting for CCS were evaluated under water stress and well-watered conditions in soil mesocosms in controlled environments, in the field in the United States using automated rainout shelters, and in the field in Malawi. Our results demonstrate that substantial variation for CCS exists in maize and that this variation has substantial effects on the metabolic cost of soil exploration and thereby water acquisition under drought.     

Hydrogen sulfide improves drought resistance in Arabidopsis thaliana

Hydrogen sulfide (H₂S) plays a crucial role in human and animal physiology. Its ubiquity and versatile properties have recently caught the attention of plant physiologists and biochemists. Two cysteine desulfhydrases (CDes), l-cysteine desulfhydrase and d-cysteine desulfhydrase, were identified as being mainly responsible for the degradation of cysteine in order to generate H₂S. This study investigated the expression regulation of these genes and their relationship to drought tolerance in Arabidopsis. First, the expression pattern of CDes in Arabidopsis was investigated. The expression levels of CDes gradually increased in an age-dependent manner. The expression of CDes was significantly higher in stems and cauline leaves than in roots, rosette leaves and flowers. Second, the protective effect of H₂S against drought was evaluated. The expression pattern of CDes was similar to the drought associated genes induced by dehydration, and H₂S fumigation was found to stimulate further the expression of drought associated genes. Drought also significantly induced increased H₂S production, a process that was reversed by re-watering. In addition, seedlings after treatment with NaHS (a H₂S donor) showed a higher survival rate and displayed a significant reduction in the size of the stomatal aperture compared to the control. These findings provide evidence that H₂S, as a gasotransmitter, improves drought resistance in Arabidopsis.     

Tolerance of Arabidopsis thaliana to the Allelochemical Protocatechualdehyde

We investigated the effects of the secondary metabolite protocatechualdehyde (PCA, 3,4-dihydroxybenzaldehyde) on stress markers, including fluorescence parameters and the concentrations of pigments, free radicals, protein, and lipid peroxides, in adult plants of Arabidopsis thaliana. The content of proline, carotenoids, and chlorophylls a and b peaked 9?h after administration of 3?mM PCA (the highest concentration used in this study), although malonyldialdehyde and dry mass contents peaked 24?h following PCA treatment. Leaf staining revealed peak production of O₂ ^? between 30 and 90?min post-treatment and peak production of H₂O₂ between 1 and 9?h post-treatment. Several effects, including the observed furling of leaf margins (leaf rolling), the increases in proline content and dry to fresh weight ratio, and the oxidative burst, are reminiscent of the plant response to drought. Early dehydration in PCA-treated plants was confirmed by decreases in leaf water potential, relative water content, and stomatal opening in the first hours of treatment. Thus, PCA seems to be either inducing water deficiency stress (probably through action in the roots) or directly triggering antidrought defenses. In either case, plants showed tolerance to the concentrations employed in this study, with most of the parameters observed having recovered control values within a week of treatment.     

Ectopic Expression of Riboflavin-binding Protein Gene TsRfBP Paradoxically Enhances Both Plant Growth and Drought Tolerance in Transgenic Arabidopsis thaliana

Riboflavin is the precursor of the coenzymes flavin monophosphate (FMN) and flavin adenine dinucleotide (FAD), which serve as indispensable redox cofactors in all plants. Numerous data indicate that riboflavin is involved in pathogen resistance but less data are available on abiotic stress tolerance. In this experiment, the overexpression of the riboflavin-binding protein resulted in an enhancement of vegetative growth and net photosynthetic rate, and an acceleration of floral transition in transgenic Arabidopsis thaliana REAT11 (containing less than half the normal levels of free riboflavin, FMN, and FAD) compared to wild-type Col-0 under nonstressed conditions. The effect of drought stress on the antioxidant response of Col-0 and REAT11 was compared, where 20- and 40-day-old grown plants were subjected to 10 % PEG 6000 treatment for 2 days. Stress conditions caused a significant increase in H₂O₂ accumulation, lipid peroxidation, and membrane permeability in Col-0 over that in REAT11. Greater activity levels of superoxide dismutase, ascorbate peroxidase, and glutathione reductase were observed in the leaves of REAT11 compared to those of Col-0. Significant increases in total ascorbate and glutathione content and higher ratios of ASC/DHA: (ASC and DHA are reduced and oxidized ascorbate, respectively) and GSH/GSSG: (GSH and GSSG are reduced and oxidized glutathione, respectively) were observed in the leaves of REAT11 compared to those in Col-0 under drought conditions. In addition, enhancement of free proline and soluble sugar accumulation was observed in REAT11 compared to Col-0 under stress. Our results suggest that a slight deficiency in free riboflavin can paradoxically induce both a higher vegetative growth rate and an enhanced tolerance to drought in transgenic plants. The “stress escape” hypothesis is proposed here to explain this interesting phenomenon.     

Overexpression of Tomato SIZ2 in Arabidopsis Improves Plant Salinity Tolerance

Small ubiquitin-like modifier (SUMO) conjugation to target proteins is an important post-translational modification, which regulates plant tolerance to biotic and abiotic stresses. SIZ1, a well-characterized SUMO E3 ligase, facilitates the conjugation of SUMO to target proteins. Here, a SIZ/PAIS-type protein SlSIZ2 was identified in tomato (Solanum lycopersicum) that is a homolog of AtSIZ1 and SlSIZ1. SlSIZ2 was expressed in tomato vegetative and reproductive tissues, and induced by ABA and NaCl. Nucleus-localized SlSIZ2 partially rescued atsiz1-2 dwarfism and also alleviated the sensitivity of atsiz1-2 to ABA and NaCl, suggesting the functional replacement of SlSIZ2 to AtSIZ1. Moreover, SlSIZ2-overexpressing Arabidopsis has higher cotyledon expansion rate, lateral root density and survival rate under salinity stress. These results suggested the contribution of SlSIZ2 to the tolerance of salinity stress.