Overexpression of the Zygophyllum xanthoxylum Aquaporin, ZxPIP1;3, Promotes Plant Growth and Stress Tolerance.

Drought and salinity can result in cell dehydration and water unbalance in plants, which seriously diminish plant growth and development. Cellular water homeostasis maintained by aquaporin is one of the important strategies for plants to cope with these two stresses. In this study, a stress-induced aquaporin, ZxPIP1;3, belonging to the PIP1 subgroup, was identified from the succulent xerophyte Zygophyllum xanthoxylum. The subcellular localization showed that ZxPIP1;3-GFP was located in the plasma membrane. The overexpression of ZxPIP1;3 in Arabidopsis prompted plant growth under favorable condition. In addition, it also conferred salt and drought tolerance with better water status as well as less ion toxicity and membrane injury, which led to more efficient photosynthesis and improved growth vigor via inducing stress-related responsive genes. This study reveals the molecular mechanisms of xerophytes' stress tolerance and provides a valuable candidate that could be used in genetic engineering to improve crop growth and stress tolerance.

Edaphic factors determining the colonization of semiarid mine tailings by a ruderal shrub and two tree plant species: Implications for phytomanagement.

Phytomanagement has been considered a feasible technique to decrease the environmental risks associated to mine tailings and its implementation relies on a suitable plant species selection. The goal of this study was to identify the edaphic factors, including microbiology, affecting the establishment of plant species with contrasting growth patterns during the phytomanagement of mine tailings. For this purpose, a comprehensive rhizosphere characterization was performed in an early ruderal colonizer, Zygophyllum fabago and two late successional tree species, Pinus halepensis and Tetraclinis articulata, growing at a mine tailings pile in southeast Spain. The neutral pH of the tailings determined low 0.01 M CaCl2 metal extractable concentrations (e.g. <10 µg kg-1 Pb and Cd). Thus, other soil properties different from metal concentrations resulted more determining to explain plant establishment. Results revealed that Z. fabago selectively colonized tailings patches characterized by high salinity (3.5 dS m-1) and high silt percentages (42%), showing a specific halotolerant rhizospheric microbial composition, such as the bacterial Sphingomonadales and Cytophagales orders and the fungal Pleosporales and Hyprocreales orders. The two tree species grew at moderate salinity areas of the tailings pile (1.7 dS m-1) with high sand percentages (85%), where Actinomycetales was the most abundant bacterial order (>10% abundance). The contrasting mycorrhizal behaviour of both tree species (ectomycorrhizal for P. halepensis and endomycorrhizal for T. articulata) could explain the differences found between their fungal rhizospheric composition. In terms of phytomanagement, the selective plant species colonization following specific soil patches at mine tailings would increase their biodiversity and resilience against environmental stressors.

Effect of phosphorus efficiency on elemental stoichiometry of two shrubs.

Phosphorus (P) is an important nutrient that can restrict plant growth. However, the influence of P deficiency on elemental homeostasis and application of the growth rate hypothesis in higher plants remain to be assessed. Two shrubs, Zygophyllum xanthoxylum and Nitraria tangutorum, were used as experiment material and subjected to five P addition treatments: 0, 17.5, 35.0, 52.5 and 70.0 mg P·kg-1 soil. The biomass and relative growth rate of Z. xanthoxylum did not change with altered P supply. There was no significant difference in P concentration among the treatments for Z. xanthoxylum, but N. tangutorum showed an upward trend. The P stoichiometric homeostasis of Z. xanthoxylum was higher than that of N. tangutorum. For Z. xanthoxylum, available P in the rhizosphere improved significantly under extreme P deficiency conditions, and P concentrations in all treatments were lower than in N. tangutorum, showing that Z. xanthoxylum had stronger P absorption and P utilization capacity. No relationships between growth rate and C:N:P ratios were found in Z. xanthoxylum. The strong P efficiency, and high and stable dry matter accumulation, are likely contributors in maintaining stoichiometric homeostasis. In addition, the relatively high biomass accumulation and high P utilization efficiency for Z. xanthoxylum does not support the growth rate hypothesis for this species.

Complete Genome Sequence of Paenibacillus sp. JZ16, a Plant Growth Promoting Root Endophytic Bacterium of the Desert Halophyte Zygophyllum Simplex.

Paenibacillus sp. JZ16 is a gram-positive, rod-shaped, motile root endophytic bacterium of the pioneer desert halophytic plant Zygophyllum simplex. JZ16 was previously shown to promote salinity stress tolerance in Arabidopsis thaliana and possesses a highly motile phenotype on nutrient agar. JZ16 genome sequencing using PacBio generated 82,236 reads with a mean insert read length of 11,432 bp and an estimated genome coverage of 127X, resulting in a chromosome of 7,421,843 bp with a GC content of 49.25% encoding 6710 proteins, 8 rRNA operons, 117 ncRNAs and 73 tRNAs. Whole-genome sequencing analysis revealed a potentially new species for JZ16. Functional analysis revealed the presence of a number of enzymes involved in the breakdown of plant-based polymers. JZ16 could be of potential use in agricultural applications for promoting biotic and abiotic stress tolerance and for biotechnological processes (e.g., as biocatalysts for biofuel production). The culture-dependent collection of bacterial endophytes from desert plants combined with genome sequence mining provides new opportunities for industrial applications.

Site- and species-specific climatic responses of two co-occurring shrubs in the temperate Alxa Desert Plateau, northwest China.

In the context of global precipitation anomalies and climate warming, the evolution of fragile desert ecosystems, which account for one-third of the world's land area, will become more complex. Studies of regional climate change and ecosystem response are important components of global climate change research, especially in arid desert regions. Zygophyllum xanthoxylum and Ammopiptanthus mongolicus are two dominant but endangered shrub species in the Alxa Desert in the arid region of central Asia. Using dendrochronological methods, we studied the response of radial growth of those two species to climate factors, and the adaptability of the two shrub populations under a regional warming trend. We found that radial growth of both shrubs was mainly affected by precipitation during the growing season. In additionally, along with the decrease of precipitation and the increase of temperature from east to west of Alxa desert Plateau, the limiting effect of drought during the growing season on radial growth increased. The climate response characteristics and changes between dry and wet periods exhibited spatial and temporal heterogeneity due to micro-level geomorphological factors. Under a regional climate warming trend, individual growth and population development of the two endangered shrubs will be adversely affected. In areas where these species are naturally distributed, populations will gradually become concentrated in micro-geomorphic regions with better soil moisture conditions, such as low-lying areas in the gullies that develop in alluvial fans. This finding has important scientific significance for understanding the development of the region's dominant shrub populations and protection of these and other endangered plants in arid desert areas.

Different mechanisms of the metalliferous Zygophyllum fabago shoots and roots to cope with Pb toxicity.

Lead (Pb) remains classified as a priority pollutant. Zygophyllum fabago is considered an early colonizer of heavy metal-polluted soils under semiarid conditions, but physiological mechanisms underlying this colonizing capacity remain unclear. In order to characterize Z. fabago plants' performance on Pb-contaminated soils, we evaluated how Pb influenced root and shoot growth, carbon metabolism, and oxidative status. For that, 30-day-old seedlings from one population colonizing a mine tailing ("Mercader") at Murcia (southeast Spain) were exposed to 500-µM Pb(NO3)2 for 1 week. Results showed that this high dose of Pb induced no plant mortality nor senescence, though promoting plant nanism. Besides the efficiency of roots to accumulate Pb, shoots also demonstrate a high efficiency to translocate and accumulate this metal. Pb exposure decreased Zn uptake to the aerial part and reduced net photosynthetic rate (A), RuBisCO activity, chlorophyll, and soluble sugar contents in shoots. Moreover, in shoots, Pb exposure increased the levels of O2- and decreased antioxidant capacity, culminating with a loss of cell membrane integrity (electrolyte leakage) and increased protein oxidation. Compared to controls, exposed roots had less Mn and Zn levels, and despite the rise in H2O2 levels, they were able to modulate non-protein thiols presenting a robust defense capacity. This capacity may support the roots' ability to maintain cell membrane integrity (electrolyte leakage) with regard to control. Principal component analysis (PCA) contributed to elucidate how this species adjusts physiological mechanisms to cope with Pb toxicity, showing that roots and shoots evolved different antioxidant defenses, which demonstrates the importance of organ specificity in the response of Z. fabago to heavy metals.

Evaluation of the environmental plasticity in the xerohalophyte Zygophyllum fabago L. for the phytomanagement of mine tailings in semiarid areas.

Phytomanagement by phytostabilisation of metal(loid)-enriched mine tailings in semiarid areas has been proposed as a suitable technique to promote a self-sustainable vegetal cover for decreasing the spread of polluted particles by erosion. The goal of this work was to evaluate the contribution of a pioneer plant species (Zygophyllum fabago) in ameliorating the soil conditions at two mine tailings piles located in a semiarid area in Southeast Spain. The ecophysiological performance of this plant species compared to a control population was assessed by analysing the nutritional and ecophysiological status. The presence of Z. fabago in mine tailings enhanced the soil microbial activity and increased the content of soil organic carbon within the rhizosphere (approx. 50% increasing). Metal(loid) concentrations in the tailings may play a minor role in the establishment of Z. fabago plants due to the low metal(loid) availability in the tailings (low CaCl2-extractable concentrations) and low uptake in the plants (e.g. up to 300 mg kg(-1) Zn in leaves). The lower δ13C and δ18O in the plants sampled at both tailings compared to the control ones may indicate softer stomatal regulation in relation to the control site plants and therefore lower WUE [corrected]. The Z. fabago plants may skip some energy-demanding mechanisms such as stomatal control and/or proline synthesis to overcome the environmental stresses posed at the tailings. The Z. fabago plants revealed high plasticity of the species for adapting to the low fertility soil conditions of the tailings and to overcome constraints associated to the dry season.

Silicon improves photosynthesis and strengthens enzyme activities in the C3 succulent xerophyte Zygophyllum xanthoxylum under drought stress.

One main strategic adaptive mechanism adopted by succulent xerophyte species, resistance to drought stress is absorbing and accumulating large amounts of sodium (Na+) from poor and dry soil which was stored in photosynthesizing branches as well as leaves as major osmoregulators, while still accumulating and storing a great deal of silicon (Si) in roots to resist to arid environments. To understand the possible adaptive strategies underlying how Si accumulation stimulates growth and ameliorates the adverse environmental impacts of drought stress on the C3 succulent xerophyte Zygophyllum xanthoxylum, plants grown for 3 weeks were suffered different K2SiO3 concentrations (1.5-7.5mM) (3-15mM KCl as control) treatments in sand culture experiments. Plants were also treated with different osmotic stresses caused by polyethylene glycol (PEG 6000) and drought stress (maintain water content about 30% of field water capacity) (30% of FWC) with or without additional 2.5mMK2SiO3 (5mMKCl as control) treatment in sand culture and pot experiments, respectively. We found that 2.5mMK2SiO3 (5mMKCl as control) resulted in optimal plant growth and alleviated adverse influences of drought stress on Z. xanthoxylum, by strengthening the activities of superoxide dismutase, peroxidase and catalase, reducing membrane lipid peroxidation and decreasing soluble sugar and free proline concentrations, concomitantly, increasing tissue water content, leaf area and chlorophyll a concentration. The result of ion analysis indicated that the Si absorption of Z. xanthoxylum was markedly induced by drought stress and that the 2.5mMK2SiO3 (5mMKCl as control) treatment significantly increased the aboveground and root Si concentration under different osmotic stresses and 30% of field water capacity compared with the drought and drought with 5mMKCl treatments. Although the K+ concentration in root in the drought with 2.5mMK2SiO3 treatment was no significant changes compared with the drought treatment, K+ concentration in aboveground and root in drought with 2.5mMK2SiO3 treatment were significantly decreased by 42% and 65.2% compared with drought with 5mMKCl treatment under 30% of FWC, indicating that Si replaced the function of K+, thus stimulating the growth and mitigating adverse effects of Z. xanthoxylum under water deficit. These findings showed that the positive roles of Si in the drought tolerance of Z. xanthoxylum might be due to the ability of plant to accumulate a great quantity of Si and utilize it as an osmoregulator to copy with water deficit, which was coupled with an obvious improvement in photosynthetic activity and anti-oxidative enzyme activities.

ZxNHX controls Na⁺ and K⁺ homeostasis at the whole-plant level in Zygophyllum xanthoxylum through feedback regulation of the expression of genes involved in their transport.

BACKGROUND AND AIMS: In order to cope with arid environments, the xerohalophyte Zygophyllum xanthoxylum efficiently compartmentalizes Na(+) into vacuoles, mediated by ZxNHX, and maintains stability of K(+) in its leaves. However, the function of ZxNHX in controlling Na(+) and K(+) homeostasis at the whole-plant level remains unclear. In this study, the role of ZxNHX in regulating the expression of genes involved in Na(+) and K(+) transport and spatial distribution was investigated. METHODS: The role of ZxNHX in maintaining Na(+) and K(+) homeostasis in Z. xanthoxylum was studied using post-transcriptional gene silencing via  Agrobacterium-mediated transformation. Transformed plants were grown with or without 50 mm NaCl, and expression levels and physiological parameters were measured. KEY RESULTS: It was found that 50 mm NaCl induced a 620 % increase in transcripts of ZxSOS1 but only an 80 % increase in transcripts of ZxHKT1;1 in roots of wild-type (WT) plants. Consequently, the ability of ZxSOS1 to transport Na(+) exceeded that of ZxHKT1;1, and Na(+) was loaded into the xylem by ZxSOS1 and delivered to the shoots. However, in a ZxNHX-silenced line (L7), the capacity to sequester Na(+) into vacuoles of leaves was weakened, which in turn regulated long-distance Na(+) transport from roots to shoots. In roots of L7, NaCl (50 mm) increased transcripts of ZxSOS1 by only 10 %, whereas transcripts of ZxHKT1;1 increased by 53 %. Thus, in L7, the transport ability of ZxHKT1;1 for Na(+) outweighed that of ZxSOS1. Na(+) was unloaded from the xylem stream, consequently reducing Na(+) accumulation and relative distribution in leaves, but increasing the relative distribution of Na(+) in roots and the net selective transport capacity for K(+) over Na(+) from roots to shoots compared with the WT. Silencing of ZxNHX also triggered a downregulation of  ZxAKT1 and ZxSKOR in roots, resulting in a significant decrease in K(+) accumulation in all the tissues in plants grown in 50 mm NaCl. These changes led to a significant reduction in osmotic adjustment, and thus an inhibition of growth in ZxNHX-silenced lines. CONCLUSIONS: The results suggest that ZxNHX is essential for controlling Na(+), K(+) uptake, long-distance transport and their homeostasis at whole-plant level via feedback regulation of the expression of genes involved in Na(+), K(+) transport. The net result is the maintenance of the characteristic salt accumulation observed in Z. xanthoxylum and the regulation of its normal growth. A model is proposed for the role of ZxNHX in regulating the Na(+) transport system in Z. xanthoxylum under saline conditions.

Profiling of rutin-mediated alleviation of cadmium-induced oxidative stress in Zygophyllum fabago.

Zygophyllum fabago grows in arid, saline soil, or disturbed sites, such as former industrial or mining areas. This species is able to grow in coarse mineral substrates contaminated with heavy metals. To investigate the effects of the flavonoid rutin (Rtn) on certain heavy metal stress responses such as antioxidant defense systems and water status, seedlings were subjected to 100 and 200 µM CdCl2 treatment without or with 0.25 and 1 mM Rtn for 7 and 14 d (days). Cd stress decreased growth (RGR), water content (RWC), leaf osmotic potential (Ψ(Π)), and chlorophyll fluorescence, all of which could be partly alleviated by addition of Rtn. Activities of superoxide dismutase, peroxidase (POX), ascorbate peroxidase, and glutathione reductase increased within the first 7 d after exposure to Cd. However, failure of antioxidant defense in the scavenging of reactive oxygen species (ROS) was evidenced by an abnormal rise in superoxide anion radical ( O2(•-)) and hydrogen peroxide contents and a decline in hydroxyl radical (OH(•)) scavenging activity, resulting in enhancement of lipid peroxidation (TBARS) as a marker of Cd-induced oxidative stress. However, exogenously applied Rtn considerably improved the stress tolerance of plants via a reduction in Cd accumulation, modulation of POX activity, increase of proline (Pro) content, decrease in TBARS and ROS content and consequent lowering of oxidative damage of membrane. Overall, 0.25 and 1 mM Rtn could protect Z. fabago from the harmful effects of 100 µM Cd-induced oxidative stress throughout the experiment.

Syrian bean-caper (Zygophyllum fabago L.) improves organic matter and other properties of mine wastes deposits.

The omni-presence of Zygophyllum fabago L. (Syrian bean-caper) natural colonies in post mining areas prompted us to investigate its contributions to reclamation of mine wastes deposits in southeast Spain. Select plant-related (edaphic) characteristics and bio- and water soluble-Cd, Cu, Pb and Zn in rhizosphere of Z. fabago were compared to deposits one year since application of pig slurry and marble waste. Total N in rhizosphere increased up to a factor of 20X (339 vs 17 mg N kg(-1)) in El Gorguel and 27X (85 vs 3.1 mg N kg(-1)) in El Lirio sites. Organic matter accumulation in rhizosphere from litter and roots of Z. fabago increased organic C from 6.6 to 19.5 g kg(-1) in El Gorguel and from 2.1 to 5.7 g kg(-1) in El Lirio in one year. Dissolution of inorganic C takes place due to organic acids from root exudates of Z. fabago. Reduction in bio-available Cd, Cu, Pb, and Zn in rhizosphere of Z. fabago at El Lirio is attributed to increase in pH from 5.3 to 7.7 through marble waste addition, although increased cation exchange capacity may also have played a role. Addition of marble waste to encourage colonization by Z. fabago in acidic mine wastes deposits was recommended.

Sodium chloride improves photosynthesis and water status in the succulent xerophyte Zygophyllum xanthoxylum.

Zygophyllum xanthoxylum, a C(3) woody species, is a succulent xerophyte that is well adapted to arid environments. Our previous investigations showed that Na(+) has a positive effect on the growth of Z. xanthoxylum under drought conditions, which was closely related to high Na(+) accumulation in leaves. To reveal the physiological mechanisms underlying how Na(+) accumulation improves the drought resistance of Z. xanthoxylum, 3-week-old seedlings were treated with a series of additional external NaCl concentrations (5-150 mM) in sand culture experiments. Seedlings were also subjected to water deficit (30% of field water capacity) in the presence or absence of additional NaCl (50 mM) in pot experiments. The results indicated that 50 mM NaCl could mitigate deleterious impacts of water deficit on the growth of Z. xanthoxylum, by improving the relative water content, inducing a significant drop in leaf water potential and, concomitantly, increasing leaf turgor pressure and chlorophyll concentrations resulting in an enhancement of overall plant photosynthetic activity (i.e., photosynthetic rate and water use efficiency). Furthermore, NaCl (50 mM) could alleviate the inhibitory effect of water deficit on the activity of photosystem II in Z. xanthoxylum. The contribution of Na(+) to the total osmotic potential varied from 8% in the control to 13% in plants subjected to water deficit and, surprisingly, to 28% in plants grown in the presence of 50 mM NaCl under water deficit; however, the contribution of K(+) significantly decreased from 13 to 8%. These findings suggest that, under arid environments, Z. xanthoxylum is able to accumulate a high concentration of Na(+) in its leaves and use it directly for osmotic adjustment, which was coupled with an improvement in leaf hydration and photosynthetic activity.

Impact of cadmium and zinc on growth and water status of Zygophyllum fabago in two contrasting metallicolous populations from SE Spain: comparison at whole plant and tissue level.

Cadmium and zinc accumulation and toxicity were assessed in whole plants and callus culture of two Zygophyllum fabago populations originating from two metallicolous sites in Murcia (southeast Spain), La Peña and Mazarrón, the first containing 2.8-times more Cd and five-times more Zn than the second. Seedlings from both ecotypes were exposed for 3 weeks to 1 or 10 µm Cd, and to 10 or 100 µm Zn in nutrient solution in a controlled environment. Calli from both ecotypes were exposed to 0.01, 0.1 or 1 mm Cd, and to 0.1, 1 or 5 mm Zn. Plants from both populations exhibited similar tolerance to Zn, while tolerance to Cd appeared more important in plants from La Peña than those from Mazarrón. Only minor differences were recorded in final Cd accumulation, with higher Cd retention in roots and stems of plants from La Peña. In both populations, transient decreases in the rate of Zn intake and translocation from root to shoot were recorded. This reduction in ion uptake was not more efficient for the population from the most contaminated area compared to the less contaminated area. Similar concentrations of Cd were found in cotyledon-derived calli from the two populations, but absorbed Cd induced conspicuous water stress in calli issues from Mazarrón but not in those from La Peña. It is concluded that, beside comparable levels of heavy metal concentration in tissues, the physiological strategy of tolerance may differ according to the metal and according to the considered population.

[Water parameters of desert xeric shrubs in west Erdos region].

By using PV technique, this paper studied the turgor pressure (psi P), cell elastic modulus (epsilon), and relative cell volume (RCV) of super xerophytes Potaninia mongolica, Reaumuria soongorica, Tetraena mongolica and Zygophyllum xanthoxylon in west Alashan, with the relationships among the parameters analyzed. The results showed that R. soongorica had the strongest ability to maintain maximum turgor pressure (a = 2.4593). The four plants maintained their turgor pressure by different ways, i.e., P. mongolica maintained it by elastic adjustment (epsilon max = 8.4005 MPa), R. soongorica by osmotic adjustment (psi pi100 = -3.1302 MPa; psi0 = -3.5074 MPa), T. mongolica by both osmotic and elastic adjustment, and Z. xanthoxylon by osmotic adjustment, which had weak adjustment ability. The cell wall of P. mongolica was soft and highly elastic, benefiting to the water absorption by root and stem and to the fast water transmission. T. mongolica also had relatively soft and high elastic cell wall, and its psi P, and epsilon changed slowly with decreasing RCV, suggesting that this plant had strong ability of holding water and resisting dehydration.

[Characteristics of natural radionuclide distribution in different organs of plants growing on territory with increased radiation background].

We investigated wild-growing grassy plants such as Alhagi pseudalhagi, Zygophyllum, Juncus acutus and Argusia sibirica with the purpose of studying the mechanism of carry of radionuclides from ground in plants and an establishment most the common laws of distribution of radionuclides between different organs of plants. The results of the research show that the researched plants are mainly accumulated 40K. On a degree of accumulation, 226Ra occupies the second place, but 232Th doesn't participate almost in processes of carry from ground in plants. It is established that root systems of plants possesses unequal ability to absorb same radionuclides from the same ground. For example, the root system of Argusia sibirica is characterized by smaller ability to absorb 226Ra, but 40K is more accessible. It is shown that biological availability radionuclides in the given ground depends both on a kind of plants and on radionuclides. For example, Argusia sibirica, Zygophyllum show higher accumulating ability to 40K and in relation to 226Ra, Alhagi pseudalhagi and Juncus acutus are more sensitive. It is established that accumulating ability of stalks of different plants in relation to 40K are appreciably different. Distribution of radionuclides in seeds of plants has other character. Thus Alhagi pseudalhagi in the seeds had also certain amount of 232Th, and leaves Argusia sibirica in comparison with Zygophyllum have a high degree of accumulation as for 40K, so for 226Ra.

Heavy metal accumulation and tolerance in plants from mine tailings of the semiarid Cartagena-La Unión mining district (SE Spain).

Mine tailings are a characteristic of landscapes where mineral extraction has occurred. These tailings usually contain high heavy metal concentrations and have low fertility. In arid and semiarid zones, erosion may be an additional problem. The removal of these tailings is often impractical due to their large volumes. Therefore, a need exists to develop in situ low cost technologies to effect surface stabilization. The use of vegetation can be an attractive option, since there are some native plant species that can colonize parts of these polluted sites unaided. Some edaphic factors were investigated, including heavy metal concentrations, in three mine tailings from a semiarid mining zone in Southeast Spain. High total metal concentrations were found: 5000-8000 mg/kg for lead and 7600-12300 mg/kg for zinc. Two of these mine tailings had pH values between 6 and 7, while the other was acidic, with a pH of 2.5. Metal solubility was pH dependent, with more than 10% of the total zinc soluble in the acid substrate and less than 1% in the neutral substrates. The metal concentrations (copper, lead and zinc) in shoots of native vegetation colonizing in these sites were studied. No species of hyperaccumulators were found. The highest concentrations were found in Zygophyllum fabago, with 530 mg/kg zinc, Helichrysum decumbens with 390 mg/kg lead, and Tamarix sp. with 11 mg/kg copper. An analysis of the rhizospheric soil of these plants indicated that Lygeum spartum grew in pH 3 soil and had low metal concentration in shoots (40 mg/kg zinc and 41 mg/kg lead).