Calcium acetate‐induced reduction of cadmium accumulation in Oryza sativa: Expression of auto‐inhibited calcium‐ATPase and cadmium transporters

• Calcium (Ca) signalling has an essential role in regulating plant responses to various abiotic stresses.
• This study applied Ca in various forms (Ca acetate and CaCl₂) and concentrations to reduce cadmium (Cd) concentration in rice and propose a possible mechanism through which Ca acts to control the Cd concentration in rice.
• The results showed that supplementation of Cd‐contaminated soil with Ca acetate reduced the Cd concentration in rice after exposure for 7 days in both hydroponic and soil conditions. The possible involvement of the auto‐inhibited Ca²⁺‐ATPase gene (ACA) might act to control the primary signal of the Cd stress response. The messages from ACA3 and ACA13 tended to up‐regulate the low‐affinity cation transporter (OsLCT1) and down‐regulate Cd uptake and the Cd translocation transporter, including the genes, natural resistance‐associated macrophage protein 5 (Nramp5) and Zn/Cd‐transporting ATPase 2 (HMA2), which resulted in a reduction in the Cd concentration in rice. After cultivation for 120 days, the application of Ca acetate into Cd‐contaminated soil inhibited Cd uptake of rice.
• Increasing the Ca acetate concentration in the soil lowered the Cd concentration in rice shoots and grains. Moreover, Ca acetate maintained rice productivity and quality whereas both aspects decreased under Cd stress.

     

Involvement of soluble proteins in growth and metabolic adjustments of drought‐stressed Calligonum mongolicum seedlings under nitrogen addition

• The planting of seedlings is the most effective measure for vegetation restoration. However, this practice is challenging in desert ecosystems where water and nutrients are scarce. Calligonum mongolicum is a sand‐fixing pioneer shrub species, and its adaptive strategy for nitrogen (N) deposition and drought is poorly understood.
• Thus, in a pot experiment, we studied the impacts of four N levels (0, 3, 6, 9 gN·m^?2·year^?1) under drought or a well‐watered regime on multiple eco‐physiological responses of 1‐year‐old C. mongolicum seedlings.
• Compared to well‐watered conditions, drought considerably influenced seedling growth by impairing photosynthesis, osmolyte accumulation and activity of superoxide dismutase and enzymes related to N metabolism. Nitrogen addition improved the productivity of drought‐stressed seedlings, as revealed by increased water use efficiency, enhanced superoxide dismutase and nitrite reductase activity and elevated N and phosphorus (P) levels in seedlings. Nevertheless, the addition of moderate to high levels of N (6–9 gN·m^?2·year^?1) impaired net photosynthesis, osmolyte accumulation and nitrate reductase activity. N addition and water regimes did not markedly change the N:P ratios of aboveground parts; while more biomass and nutrients were allocated to fine roots to assimilate the insufficient resources. Soluble protein in assimilating shoots might play a vital role in adaptation to the desert environment.
• The response of C. mongolicum seedlings to N addtion and drought involved an interdependency between soluble protein and morphological, physiological and biochemical processes. These findings provide an important reference for vegetation restoration in arid lands under global change.

     

Physiological dissection revealed that both uptake and assimilation are the major components regulating different growth responses of two tobacco cultivars to nitrogen nutrition

• K326 and HD represent major tobacco cultivars in China, which required large N fertiliser input but at different application rates. To understand primary components affecting tobacco N use physiology, we adopted these two varieties as valuable genetic material to assess their growth response to N nutrition.
• We established a hydroponic culture system to grow plants supplied with different N regimes. Plant biomass, N, ammonium, nitrate, arginine, GS and NR activity, N transfer and use efficiency as well as root uptake were examined.
• Our data revealed the preference of K326 and HD to utilise nitrate or ammonium nitrate but not ammonium alone, with 2 mm N supply probably sufficient and economical to achieve good biomass production at the vegetative stage. Moreover, both varieties were very sensitive to ammonium, perhaps due to lack of or abnormal signalling related to nitrate and/or arginine rather than impairment of N acquisition and initial assimilation; this was supported by measurements of the plant content of N, ammonium and activities of GS and NR. Notably, short‐term ¹⁵N root influx studies identified differential uptake kinetics of K326 and HD, with distinct affinities and transport rates for ammonium and nitrate.
• The data suggest that the growth adaptation of K326 or HD to higher or lower N may be ascribed to different competences for effective N uptake/translocation and assimilation. Thus, our work provides valuable information to prompt deeper investigation of the molecular basis controlling plant N use efficiency.

     

Differences in feeding behaviour among Chironomus species revealed by measurements of sulphur stable isotopes and cadmium in larvae

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Atmospheric nitrogen deposition on petals enhances seed quality of the forest herb Anemone nemorosa

• Elevated atmospheric input of nitrogen (N) is currently affecting plant biodiversity and ecosystem functioning. The growth and survival of numerous plant species is known to respond strongly to N fertilisation. Yet, few studies have assessed the effects of N deposition on seed quality and reproductive performance, which is an important life‐history stage of plants.
• Here we address this knowledge gap by assessing the effects of atmospheric N deposition on seed quality of the ancient forest herb Anemone nemorosa using two complementary approaches.
• By taking advantage of the wide spatiotemporal variation in N deposition rates in pan‐European temperate and boreal forests over 2 years, we detected positive effects of N deposition on the N concentration (percentage N per unit seed mass, increased from 2.8% to 4.1%) and N content (total N mass per seed more than doubled) of A. nemorosa seeds. In a complementary experiment, we applied ammonium nitrate to aboveground plant tissues and the soil surface to determine whether dissolved N sources in precipitation could be incorporated into seeds. Although the addition of N to leaves and the soil surface had no effect, a concentrated N solution applied to petals during anthesis resulted in increased seed mass, seed N concentration and N content.
• Our results demonstrate that N deposition on the petals enhances bioaccumulation of N in the seeds of A. nemorosa. Enhanced atmospheric inputs of N can thus not only affect growth and population dynamics via root or canopy uptake, but can also influence seed quality and reproduction via intake through the inflorescences.

     

pH affects ammonium,nitrate and proton fluxes in the apical region of conifer and soybean roots

The effect of pH on nitrate and ammonium uptake in the high‐affinity transport system and low‐affinity transport system ranges was compared in two conifers and one crop species. Many conifers grow on acidic soils, thus their preference for ammonium vs nitrate uptake can differ from that of crop plants, and the effect of pH on nitrogen (N) uptake may differ. Proton, ammonium and nitrate net fluxes were measured at seedling root tips and 5, 10, 20 and 30 mm from the tips using a non‐invasive microelectrode ion flux measurement system in solutions of 50 or 1500 µM NH₄NO₃ at pH 4 and 7. In Glycine max and Pinus contorta, efflux of protons was observed at pH 7 while pH 4 resulted in net proton uptake in some root regions. Pseudotsuga menziesii roots consistently showed proton efflux behind the root tip, and thus appear better adapted to maintain proton efflux in acid soils. P. menziesii's ability to maintain ammonium uptake at low pH may relate to its ability to maintain proton efflux. In all three species, net nitrate uptake was greatest at neutral pH. Net ammonium uptake in G. max and net nitrate uptake in P. menziesii were greatly reduced at pH 4, particularly at high N concentration, thus N concentration should be considered when determining optimum pH for N uptake. In P. menziesii and G. max, net N uptake was greater in 1500 than 50 µM NH₄NO₃ solution, but flux profiles of all ions varied among species.     

The expression of heterologous Fe (III) phytosiderophore transporter HvYS1 in rice increases Fe uptake,translocation and seed loading and excludes heavy metals by selective Fe transport

Many metal transporters in plants are promiscuous, accommodating multiple divalent cations including some which are toxic to humans. Previous attempts to increase the iron (Fe) and zinc (Zn) content of rice endosperm by overexpressing different metal transporters have therefore led unintentionally to the accumulation of copper (Cu), manganese (Mn) and cadmium (Cd). Unlike other metal transporters, barley Yellow Stripe 1 (HvYS1) is specific for Fe. We investigated the mechanistic basis of this preference by constitutively expressing HvYS1 in rice under the control of the maize ubiquitin1 promoter and comparing the mobilization and loading of different metals. Plants expressing HvYS1 showed modest increases in Fe uptake, root‐to‐shoot translocation, seed accumulation and endosperm loading, but without any change in the uptake and root‐to‐shoot translocation of Zn, Mn or Cu, confirming the selective transport of Fe. The concentrations of Zn and Mn in the endosperm did not differ significantly between the wild‐type and HvYS1 lines, but the transgenic endosperm contained significantly lower concentrations of Cu. Furthermore, the transgenic lines showed a significantly reduced Cd uptake, root‐to‐shoot translocation and accumulation in the seeds. The underlying mechanism of metal uptake and translocation reflects the down‐regulation of promiscuous endogenous metal transporters revealing an internal feedback mechanism that limits seed loading with Fe. This promotes the preferential mobilization and loading of Fe, therefore displacing Cu and Cd in the seed.     

Involvement of LeMRP,an ATP‐binding cassette transporter,in shikonin transport and biosynthesis in Lithospermum erythrorhizon

• Shikonin and its derivatives are important medicinal secondary metabolites accumulating in roots of Lithospermum erythrorhizon. Although some membrane proteins have been identified as transporters of secondary metabolites, the mechanisms underlying shikonin transport and accumulation in L. erythrorhizon cells still remain largely unknown.
• In this study, we isolated a cDNA encoding LeMRP, an ATP‐binding cassette transporter from L. erythrorhizon, and further investigated its functions in the transport and biosynthesis of shikonin using the yeast transformation and transgenic hairy root methods, respectively. Real‐time PCR was applied for expression analyses of LeMRP and shikonin biosynthetic enzyme genes.
• Functional analysis of LeMRP using the heterologous yeast cell expression system showed that LeMRP could be involved in shikonin transport. Transgenic hairy roots of L. erythrorhizon demonstrated that LeMRP overexpressing hairy roots produced more shikonin than the empty vector (EV) control. Real‐time PCR results revealed that the enhanced shikonin biosynthesis in the overexpression lines was mainly caused by highly up‐regulated expression of genes coding key enzymes (LePAL, HMGR, Le4CL and LePGT) involved in shikonin biosynthesis. Conversely, LeMRP RNAi decreased the accumulation of shikonin and effectively down‐regulated expression level of the above genes. Typical inhibitors of ABC proteins, such as azide and buthionine sulphoximine, dramatically inhibited accumulation of shikonin in hairy roots.
• Our findings provide evidence for the important direct or indirect role of LeMRP in transmembrane transport and biosynthesis of shikonin.

     

Melatonin mitigates cadmium and aluminium toxicity through modulation of antioxidant potential in Brassica napus L

• Melatonin has emerged as an essential molecule in plants, due to its role in defence against metal toxicity. Aluminium (Al) and cadmium (Cd) toxicity inhibit rapeseed seedling growth.
• In this study, we applied different doses of melatonin (50 and 100 µm ) to alleviate Al (25 µm ) and Cd (25 µm ) stress in rapeseed seedlings. Results show that Al and Cd caused toxicity in rapeseed seedling, as evidenced by a decrease in height, biomass and antioxidant enzyme activity.
• Melatonin increased the expression of melatonin biosynthesis‐related Brassica napus genes for caffeic acid O‐methyl transferase (BnCOMT) under Al and Cd stress. The genes BnCOMT‐1, BnCOMT‐5 and BnCOMT‐8 showed up‐regulated expression, while BnCOMT‐4 and BnCOMT‐6 were down‐regulated during incubation in water. Melatonin application increased the germination rate, shoot length, root length, fresh and dry weight of seedlings. Melatonin supplementation under Al and Cd stress increased superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, proline, chlorophyll and anthocyanin content, as well as photosynthesis rate. Both Cd and Al treatments significantly increased hydrogen peroxide and malondialdehyde levels in rapeseed seedlings, which were strictly counterbalanced by melatonin. Analysis of Cd and Al in different subcellular compartments showed that melatonin enhanced cell wall and soluble fractions, but reduced the vacuolar and organelle fractions in Al‐ and Cd‐treated seedlings.
• These results suggest that melatonin‐induced improvements in antioxidant potential, biomass, photosynthesis rate and successive Cd and Al sequestration play a pivotal role in plant tolerance to Al and Cd stress. This mechanism may have potential implications in safe food production.

     

Cadmium uptake and translocation in seedlings of near isogenic lines of durum wheat that differ in grain cadmium accumulation

Background  

Cadmium (Cd) concentrations in durum wheat (Triticum turgidum L. var durum) grain grown in North American prairie soils often exceed proposed international trade standards. To understand the physiological processes responsible for elevated Cd accumulation in shoots and grain, Cd uptake and translocation were studied in seedlings of a pair of near-isogenic durum wheat lines, high and low for Cd accumulation in grain.     

滨海盐渍土壤中不同类型盐生植物富集镉的效应

为了利用被镉污染的滨海盐渍土壤,通过实验对比分析3种不同类型盐生植物对盐渍土中镉的富集效应,以期初步探明不同类型盐生植物在镉污染盐渍土壤修复中的效果。选择的3种盐生植物类型是:聚盐盐生植物,泌盐盐生植物和避盐盐生植物。通过温室盆栽实验,将植物在不同镉含量的盐渍土壤中种植培养60 d,测定和分析不同类型盐生植物对镉的生物浓缩因子、转移系数以及植株内地上部分和根部生物量和镉含量的变化。结果表明,不同镉含量的土壤对碱蓬和芦苇的生长影响较小,对二色补血草的生长影响较大。不同镉含量的土壤中,芦苇地上部分镉的生物浓缩因子变化差异不显著,并且其地上部分吸收镉的百分率较高。碱蓬和芦苇的转移系数大于二色补血草的转移系数,并且碱蓬的转移系数在不同镉含量的土壤中变化不显著;二色补血草的转移系数随着土壤中镉含量的增加而显著增大。3种盐生植物中,碱蓬最具修复镉污染盐渍土壤的潜力,这可能和它是聚盐盐生植物的生理类型有关。芦苇整个植株的地上部分富集镉的总量在3种植物中是最高的,因此,芦苇在镉含量较低时也可以做为镉污染盐渍土壤的修复材料。     

Ultrastructural and metabolic disorders induced by short‐term cadmium exposure in Avicennia schaueriana plants and its excretion through leaf salt glands

• Environmental cadmium (Cd) sources have increased in mangrove sediments in recent decades, inducing cellular damage to many plants. Avicennia schaueriana is abundant in mangrove sites and has been subject to Cd contamination. The possible effects of Cd toxicity and the structural and physiological disturbances to this plant were studied. Can this plant express early cellular tolerance mechanisms to such metal contamination?
• Seedlings of A. schaueriana were collected from sites of their natural occurrence, placed in plastic pots containing nutrient solution for 60 days, and subsequently exposed to increasing Cd concentrations for 5 days under experimental conditions. The anatomical, ultrastructural and physiological changes induced by Cd were analysed.
• Cd accumulated mainly in the root system and in pneumatophores, stems and leaves, induced differential accumulation of mineral nutrients, but did not induce necrosis or changes in leaf anatomy. However, there was a decrease in starch grains and an increase in deposited electron‐dense material in the cortex and vascular bundles. Cd induced both increases in calcium (Ca) content in shoots and Ca oxalate crystal precipitation in leaf mesophyll and was detected in crystals and in the secretion of salt glands.
• Our observations and experimental results provide evidence of Cd tolerance in A. schaueriana. As a new feature, despite the clear cellular physiological disorders, this plant is able to eliminate Cd through leaf salt glands and immobilise it in Ca crystals, representing fast mechanisms for Cd exclusion and complexation in leaves in heavy metal coastal polluted marine ecosystems.

     

Comparative growth and metabolism of gelatinous colonies of three cyanobacteria,Nostoc commune,Nostoc pruniforme and Nostoc zetterstedtii,at different temperatures

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Seasonal nitrogen uptake strategies in a temperate desert ecosystem depends on N form and plant species

• Symbiotic plants might be able to regulate a limited nitrogen (N) pool, thus avoiding and reducing competition for resources, through the uptake of different chemical N forms. Our aim was to see whether coexisting herbs showed preference for different forms of N in a temperate desert.
• We conducted a situ experiment using the ¹⁵N labeling method in the Gurbantunggut Desert of Northwestern China dominated by Erodium oxyrrhynchum, Hyalea pulchella, Nonea caspica and Lactuca undulata during their growing period (April and May).
• Four desert herb species preferentially relied on ¹⁵N‐NO₃ for their N nutrition. Multi‐factor analysis of variance (ANOVA) analysis results showed that species, N forms, months, and soil depths strongly affected N uptake rate. The uptake rate by herbs was higher in May than in April, and higher at 0–5 cm than at 5–15 cm soil layers. Erodium oxyrrhynchum, N. caspica and L. undulata showed different preference on N form over months. Erodium oxyrrhynchum and L. undulata changed their uptake preference from more ¹⁵N‐Glycine in April to more ¹⁵N‐NH4 in May.
• Although the N uptake rate of four desert herbs varied across different soil depths and months, all species absorbed more inorganic N compared with organic N. The higher preference for ¹⁵N‐NO₃ and ¹⁵N‐NH4 over ¹⁵N‐Gly possibly reflects adaptation to different N forms in temperate desert.