Leaf Vacuoles Research Articles

Variation in foliar inorganic phosphorus concentrations in perennial ryegrass (Lolium perenne L.)

ABSTRACT Breeding ryegrass for reduced inorganic phosphorus (Pi) concentrations in leaf vacuoles could lower pasture P fertiliser requirements with economic and environmental benefits. We investigated foliar Pi% accumulation characteristics of a range of ryegrass cultivars and ecotypes in glasshouse experiments, and ryegrass cultivars from two field trials. Pi in ryegrass was also compared with Pi in cocksfoot, tall fescue, prairie grass and Phalaris in a glasshouse experiment. Pi expressed as a percentage of leaf total phosphorus was higher (average 82.5%) in the older ryegrass cultivars – Ruanui, and Nui and the New Zealand hill country ecotypes (85%) than in more recently bred cultivars like Aberdart and One50 (74.7%). In the other grass species, foliar Pi% was lowest in prairie grass, intermediate in tall fescue with the highest values in cocksfoot, Phalaris and ryegrass. The results suggest that including a lower Pi concentration in future ryegrass breeding programmes might improve ryegrass P use efficiency without compromising growth rates.

Differential gene expression patterns and physiological responses improve adaptation to high salinity concentration in pepper accessions.

High salinity decreases the productivity of crops worldwide. Pepper is particularly sensitive to high salt concentrations. Herein, we subjected three tolerant pepper accessions (C12, B14 and A25) to high sodium chloride concentration (70 mM NaCl). The aerial and root biomass, leaf and root osmotic potential (Ψπ ), Na+ , Cl- , K+ and proline concentrations and the relative expression of the putative genes CaSOS1, CaHKT1, three CaNHXs and CaP5CS were measured. Different salinity tolerance strategies depending on the pepper accession were identified. In C12, tolerance was attributed to the accumulation of Na+ in vacuoles and endosomes by the activation of vacuolar CaNHXs genes and the reduction in Ψπ ; additionally, the activation of CaHKT1 and CaSOS1 in leaves and roots moved and accumulated Na+ ions in the xylem and xylem parenchyma cells (XPC) as well as expulsed it out of the root cells. A25 accession, on the contrary, was specialized in compartmentalizing Na+ ions in root and leaf vacuoles and root XPC by the up-regulation of CaNHXs and CaHKT1, respectively, avoiding a toxic accumulation in leaves. Finally, B14 accession moved and accumulated Na+ in xylem and XPC, reducing its concentration in roots by the activation of CaSOS1 and CaHKT1. This study shade light on different tolerance mechanisms of pepper plants to overcome salt stress.

Citric acid and hydrogen sulfide cooperate to mitigate chromium stress in tomato plants by modulating the ascorbate-glutathione cycle, chromium sequestration, and subcellular allocation of chromium

The study aimed to investigate the role of hydrogen sulfide (H2S) in regulating chromium stress (Cr–S) tolerance of tomato plants treated with citric acid (CA). Prior to the Cr treatment, tomato plants were foliar-fed with CA (100 μM) daily for 3 days. Subsequently, the plants were grown for another ten days in a hydroponic system in a 50 μM Cr (VI) solution. Chromium treatment reduced photosynthetic pigments and plant biomass, but boosted the levels of hydrogen peroxide (H2O2) malondialdehyde (MDA), H2S, phytochelatins (PCs), and glutathione (GSH), electrolyte leakage (EL), and antioxidant enzyme activity in tomato plants. However, the foliar spray of CA mitigated the levels of H2O2, MDA, and EL, promoted plant growth and chlorophyll content, enhanced antioxidant enzymes’ activities, and increased H2S production in Cr–S-tomato plants. CA also increased the levels of GSH and PCs, potentially reducing the toxicity of Cr through regulated sequestration. Additionally, the application of sodium hydrogen sulfide (NaHS), a donor of H2S, improved CA-induced Cr stress tolerance. The addition of CA promoted Cr accumulation in root cell wall and leaf vacuoles to suppress its toxicity. To assess the involvement of H2S in CA-mediated Cr–S tolerance, 0.1 mM hypotaurine (HT), an H2S scavenger, was provided to the control and Cr–S-plants along with CA and CA + NaHS. HT reduced the beneficial effects of CA by decreasing H2S production in tomato plants. However, the NaHS addition with CA + HT inverted the adverse impacts of HT, indicating that H2S is required for CA-induced Cr–S tolerance in tomato plants.

Photon costs of shoot and root NO3-, and root NH4+, assimilation in terrestrial vascular plants considering associated pH regulation, osmotic and ontogenetic effects.

The photon costs of photoreduction/assimilation of nitrate (NO3-) into organic nitrogen in shoots and respiratory driven NO3- and NH4+ assimilation in roots are compared for terrestrial vascular plants, considering associated pH regulation, osmotic and ontogenetic effects. Different mechanisms of neutralisation of the hydroxyl (OH-) ion necessarily generated in shoot NO3- assimilation are considered. Photoreduction/assimilation of NO3- in shoots with malic acid synthesis and either accumulation of malate in leaf vacuoles or transport of malate to roots and catabolism there have a similar cost which is around 35% less than that for root NO3- assimilation and around 20% less than that for photoreduction/assimilation of NO3-, oxalate production and storage of Ca oxalate in leaf vacuoles. The photon cost of root NH4+ assimilation with H+ efflux to the root medium is around 70% less than that of root NO3- assimilation. These differences in photon cost must be considered in the context of the use of a combination of locations of NO3- assimilation and mechanisms of acid-base regulation, and a maximum of 4.9-9.1% of total photon absorption needed for growth and maintenance that is devoted to NO3- assimilation and acid-base regulation.

Subcellular distribution, chemical forms of cadmium and rhizosphere microbial community in the process of cadmium hyperaccumulation in duckweed

Duckweed is a newly reported Cd hyperaccumulator that grow rapidly; however, little is known about its tolerance and detoxification mechanisms. In this study, we investigated the tissue, subcellular, and chemical form distribution of the Cd in duckweed and studied the influences of Cd on duckweed growth, ultrastructure, and rhizosphere microbial community. The results showed that Cd could negatively affect the growth of duckweed and shorten the root length. More Cd accumulated in the roots than in the leaves, and Cd was transferred from the roots to the leaves with time. During 12–24 h, Cd mainly existed in the cell wall fraction (2.05 %–95.52 %) and the organelle fraction (5.03 %–97.80 %), followed the soluble fraction (0.14 %–16.98 %). Over time, the proportion of Cd in the organelles increased (46.64 %–92.83 %), exceeding that in the cell wall (6.79 %–66.23 %), which indicated that duckweed detoxification mechanism may be related to the retention of cell wall and vacuole. The main chemical form of Cd was the NaCl-extracted state (30.15 %–88.66 %), which was integrated with pectate and protein. With increasing stress concentration and time, the proportion of the HCl-extracted state and HAc-extracted state increased, and they were low-toxic Cd oxalate and Cd phosphate, respectively. Cd damaged the ultrastructure of cells such as chloroplasts and mitochondria and inhibited the diversity of microbial communities in the duckweed rhizosphere; however, the dominant populations that could tolerate heavy metals increased. It was speculated that duckweed distributed Cd in a less toxic chemical form in a less active location, mainly through retention in the root cell wall and sequestration in the leaf vacuoles, and is dynamically adjusted. The rhizosphere microbial communities tolerate heavy metals may also be one of the mechanisms by which duckweed can tolerate Cd. This study revealed the mechanism of duckweed tolerance and detoxification of Cd at the molecular level and provides a theoretical basis for further development of duckweed.

The vacuolar transporter LaMTP8.1 detoxifies manganese in leaves of Lupinus albus.

Manganese (Mn) is an essential microelement, but overaccumulation is harmful to many plant species. Most plants have similar minimal Mn requirements, but the tolerance to elevated Mn varies considerably. Mobilization of phosphate (P) by plant roots leads to increased Mn uptake, and shoot Mn levels have been reported to serve as an indicator for P mobilization efficiency in the presence of P deficiency. White lupin (Lupinus albus L.) mobilizes P and Mn with outstanding efficiency due to the formation of determinate cluster roots that release carboxylates. The high Mn tolerance of L. albus goes along with shoot Mn accumulation, but the molecular basis of this detoxification mechanism has been unknown. In this study, we identify LaMTP8.1 as the transporter mediating vacuolar sequestration of Mn in the shoot of white lupin. The function of Mn transport was demonstrated by yeast complementation analysis, in which LaMTP8.1 detoxified Mn in pmr1∆ mutant cells upon elevated Mn supply. In addition, LaMTP8.1 also functioned as an iron (Fe) transporter in yeast assays. The expression of LaMTP8.1 was particularly high in old leaves under high Mn stress. However, low P availability per se did not result in transcriptional upregulation of LaMTP8.1. Moreover, LaMTP8.1 expression was strongly upregulated under Fe deficiency, where it was accompanied by Mn accumulation, indicating a role in the interaction of these micronutrients in L. albus. In conclusion, the tonoplast-localized Mn transporter LaMTP8.1 mediates Mn detoxification in leaf vacuoles, providing a mechanistic explanation for the high Mn accumulation and Mn tolerance in this species.

Metal tolerance protein family members are involved in Mn homeostasis through internal compartmentation and exocytosis in Brassica napus

Mineral elements, including manganese (Mn), are indispensable for plant growth and development. However, plants cultivated on acidic soils are vulnerable to Mn2+ toxicity due to a prevalently phytotoxic level of Mn2+. Metal tolerance proteins (MTPs) were demonstrated to be essential for metal homeostasis and tolerance in different plant species. In this study, we present the functional characteristics of BnMTP8 and BnMTP9 from Brassica napus. The expression level of BnMTP8 was most prominent in rapeseed roots and was only markedly up-regulated in shoots by excess Mn2+. In contrast, BnMTP9 was most abundant in shoots. Expression of BnMTP8 and BnMTP9 in Saccharomyces cerevisiae compensated the Mn2+-hypersensitivity of Δpmr1 by increasing metal sequestration and efflux, respectively. In addition, heteroexpression of BnMTP8 restored Mn2+ tolerance of Arabidopsis mtp8 mutants and increased Mn2+ accumulation in tissues and leaf vacuoles. Expression of BnMTP9 in Arabidopsis mtp11 mutants restored their growth and reduced Mn2+ concentrations in mesophyll protoplasts and tissues upon elevated levels of Mn2+. Transient expression of GFP-fusion constructs in protoplasts showed that BnMTP8 and BnMTP9 localize to pre-vacuolar compartments and the trans-Golgi network, respectively. However, high extracellular Mn2+ in Arabidopsis root cells triggered BnMTP8-GFP relocation to the tonoplast and BnMTP9-GFP relocation to the plasma membrane, suggesting that these two proteins are involved in vacuolar Mn2+ sequestration and cytosolic Mn2+ export, respectively. Taken together, these findings indicate that BnMTP8 and BnMTP9 load Mn2+ into vesicles for subsequent delivery to the vacuole or secretion into extracellular spaces, respectively, thereby cooperating to maintain Mn2+ homeostasis in the roots and shoots of rapeseed.

Effect of arbuscular mycorrhizal symbiosis on ion homeostasis and salt tolerance-related gene expression in halophyte Suaeda salsa under salt treatments

Halophytes can remove large quantities of salts from saline soils, so their importance in ecology has received increasing attention. Preliminary studies have shown that arbuscular mycorrhizal (AM) fungi can improve the salt tolerance of halophytes. However, few studies have focused on the molecular mechanisms and effects of AM fungi in halophytes under different salt conditions. A pot experiment was carried out to investigate the effects of Funneliformis mosseae inoculation on growth, nutrient uptake, ion homeostasis and the expression of salt tolerance-related genes in Suaeda salsa under 0, 100, 200 and 400 mM NaCl. The results showed that F. mosseae promoted the growth of S. salsa and increased the shoot Ca2+ and Mg2+ concentrations under no-salt condition and high-salt condition. In addition, AM fungi increased the K+ concentration and maintained a high K+/Na+ ratio at 400 mM NaCl, while AM fungi decreased the K+ concentration and reduced the K+/Na+ ratio at 0 mM NaCl. AM fungi downregulated the expression of SsNHX1 in shoots and the expression of SsSOS1 in roots at 400 mM NaCl. These effects may decrease the compartmentation of Na+ into leaf vacuoles and restrict Na+ transport from roots to shoots, leading to an increase in root Na+ concentration. AM symbiosis upregulated the expression of SsSOS1 in shoots and downregulated the expression of SsSOS1 and SsNHX1 in roots at 100 mM NaCl. However, regulation of the genes (SsNHX1, SsSOS, SsVHA-B and SsPIP) was not significantly different with AM symbiosis at 0 mM or 200 mM NaCl. The results revealed that AM symbiosis might induce diverse modulation strategies in S. salsa, depending on external Na+ concentrations. These findings suggest that AM fungi may play significant ecological roles in the phytoremediation of salinized ecosystems.

Responses to Increased Salinity and Severe Drought in the Eastern Iberian Endemic Species Thalictrum maritimum (Ranunculaceae), Threatened by Climate Change

Thalictrum maritimum is an endangered, endemic species in East Spain, growing in areas of relatively low salinity in littoral salt marshes. A regression of its populations and the number of individuals has been registered in the last decade. This study aimed at establishing the causes of this reduction using a multidisciplinary approach, including climatic, ecological, physiological and biochemical analyses. The climatic data indicated that there was a direct negative correlation between increased drought, especially during autumn, and the number of individuals censused in the area of study. The susceptibility of this species to water deficit was confirmed by the analysis of growth parameters upon a water deficit treatment applied under controlled greenhouse conditions, with the plants withstanding only 23 days of complete absence of irrigation. On the other hand, increased salinity does not seem to be a risk factor for this species, which behaves as a halophyte, tolerating in controlled treatments salinities much higher than those registered in its natural habitat. The most relevant mechanisms of salt tolerance in T. maritimum appear to be based on the control of ion transport, by (i) the active transport of toxic ions to the aerial parts of the plants at high external salinity—where they are presumably stored in the leaf vacuoles to avoid their deleterious effects in the cytosol, (ii) the maintenance of K+ concentrations in belowground and aboveground organs, despite the increase of Na+ levels, and (iii) the salt-induced accumulation of Ca2+, particularly in stems and leaves. This study provides useful information for the management of the conservation plans of this rare and endangered species.

CmCLCa Plays a Key Role in the Storage of Nitrate in Chrysanthemum Leaf Vacuoles

Nitrogen (N) is very important for chrysanthemum yield and quality. Nitrate (NO3−) is the main form of N absorbed by chrysanthemum. The NO3− accumulated in the leaf vacuoles can be released for redistribution and reutilization within the plant when the external N source is insufficient. Therefore, understanding the storage mechanism of NO3− in the vacuoles of leaves is essential for the cultivation of chrysanthemum varieties with high N-use efficiency and high N-starvation tolerance. In this study, we found that the transcript level of CmCLCa was significantly increased by exogenous NO3−. We generated CmCLCa-RNA interference transgenic lines with knocked-down CmCLCa expression. In 5 mM NO3− hydroponic culture, the vacuolar H+-ATPase (V-ATPase) and vacuolar H+-pyrophosphatase (V-PPase) activities and vacuolar NO3− content were decreased in CmCLCa-RNAi transgenic lines compared with wild-type (WT) chrysanthemum. Next, WT and CmCLCa-RNAi transgenic plants were grown in hydroponic culture for 28 days with 5 mM NO3− and then subjected to a 7-day N-starvation treatment. The CmCLCa-RNAi transgenic lines were more sensitive to N-starvation than were WT plants and showed significantly decreased vacuolar NO3− content in the leaves. Our results indicate that inhibition of CmCLCa results in reduced NO3− accumulation in the leaf vacuole and lower tolerance to N-starvation. Our findings show that CmCLCa plays a key role in NO3− storage in leaf vacuoles and is a candidate gene for improving the N-starvation tolerance of chrysanthemum.

SsHKT1;1 is coordinated with SsSOS1 and SsNHX1 to regulate Na+ homeostasis in Suaeda salsa under saline conditions

Under saline conditions, Suaeda salsa, as a typical halophyte, accumulates large amounts of Na+ in its leaves during optimal growth. Key transporters involved in Na+ accumulation in plants are HKT-type protein, the plasma membrane Na+/H+ transporter SOS1, and the tonoplast Na+/H+ antiporter NHX1. In this study, the function of SsHKT1;1 and its coordinate expression with SsSOS1 and SsNHX1 to regulate Na+ homeostasis in S. salsa was investigated. We showed, by yeast complementation assays, that SsHKT1;1 encoded a Na+-selective transporter, which located to the plasma membrane and was preferentially expressed within the stele, and was particularly abundant in xylem parenchyma and pericycle cells. When compared with a treatment of 25 mM NaCl, 150 mM NaCl greatly decreased the transcripts of SsHKT1;1, but maintained a relatively constant level of the expression of SsSOS1 in roots. Consequently, the synergistic effect of SsHKT1;1 and SsSOS1 would result in greater Na+ loading into the xylem under 150 mM NaCl than 25 mM NaCl. In leaves, 150 mM NaCl up-regulated the abundance of SsNHX1 compared with levels in 25 mM NaCl. This enabled the permanent sequestering of Na+ into leaf vacuoles. Overall, SsHKT1;1 functioned in reducing Na+ retrieval from the root xylem, and played an important role in coordinating with SsSOS1 and SsNHX1 to maintain Na+ accumulation in S. salsa under saline conditions.

Transcriptome profiling of Fagopyrum tataricum leaves in response to lead stress

BackgroundLead (Pb) pollution is a widespread environmental problem that is harmful to living organisms. Tartary buckwheat (Fagopyrum tataricum), a member of the family Polygonaceae, exhibits short growth cycles and abundant biomass production, could be an ideal plant for phytoremediation due to its high Pb tolerance. Here, we aimed to explore the molecular basis underlying the responses of this plant to Pb stress.ResultsIn our study, ultrastructural localization assays revealed that Pb ions primarily accumulate in leaf vacuoles. RNA deep sequencing (RNA-Seq) of tartary buckwheat leaves was performed on two Pb-treated samples, named Pb1 (2000 mg/kg Pb (NO3)2) and Pb2 (10,000 mg/kg Pb (NO3)2), and a control (CK). A total of 88,977 assembled unigenes with 125,203,555 bases were obtained. In total, 2400 up-regulated and 3413 down-regulated differentially expressed genes (DEGs) were identified between CK and Pb1, and 2948 up-regulated DEGs and 3834 down-regulated DEGs were generated between CK and Pb2, respectively. Gene Ontology (GO) and pathway enrichment analyses showed that these DEGs were primarily associated with ‘cell wall’, ‘binding’, ‘transport’, and ‘lipid and energy’ metabolism. The results of quantitative real-time PCR (qRT-PCR) analyses of 15 randomly selected candidate DEGs and 6 regulated genes were consistent with the results of the transcriptome analysis. Heterologous expression assays in the yeast strain Δycf1 indicated that overexpressing CCCH-type zinc finger protein 14 (ZFP14) enhanced sensitivity to Pb2+, while 5 other genes, namely, metal transporter protein C2 (MTPC2), phytochelatin synthetase-like family protein (PCSL), vacuolar cation/proton exchanger 1a (VCE1a), natural resistance-associated macrophage protein 3 (Nramp3), and phytochelatin synthetase (PCS), enhanced the Pb tolerance of the mutant strain.ConclusionCombining our findings with those of previous studies, we generated a schematic model that shows the metabolic processes of tartary buckwheat under Pb stress. This study provides important data for further genomic analyses of the biological and molecular mechanisms of Pb tolerance and accumulation in tartary buckwheat.

Disentangling the complexity and diversity of crosstalk between sulfur and other mineral nutrients in cultivated plants.

A complete understanding of ionome homeostasis requires a thorough investigation of the dynamics of the nutrient networks in plants. This review focuses on the complexity of interactions occurring between S and other nutrients, and these are addressed at the level of the whole plant, the individual tissues, and the cellular compartments. With regards to macronutrients, S deficiency mainly acts by reducing plant growth, which in turn restricts the root uptake of, for example, N, K, and Mg. Conversely, deficiencies in N, K, or Mg reduce uptake of S. TOR (target of rapamycin) protein kinase, whose involvement in the co-regulation of C/N and S metabolism has recently been unravelled, provides a clue to understanding the links between S and plant growth. In legumes, the original crosstalk between N and S can be found at the level of nodules, which show high requirements for S, and hence specifically express a number of sulfate transporters. With regards to micronutrients, except for Fe, their uptake can be increased under S deficiency through various mechanisms. One of these results from the broad specificity of root sulfate transporters that are up-regulated during S deficiency, which can also take up some molybdate and selenate. A second mechanism is linked to the large accumulation of sulfate in the leaf vacuoles, with its reduced osmotic contribution under S deficiency being compensated for by an increase in Cl uptake and accumulation. A third group of broader mechanisms that can explain at least some of the interactions between S and micronutrients concerns metabolic networks where several nutrients are essential, such as the synthesis of the Mo co-factor needed by some essential enzymes, which requires S, Fe, Zn and Cu for its synthesis, and the synthesis and regulation of Fe-S clusters. Finally, we briefly review recent developments in the modelling of S responses in crops (allocation amongst plant parts and distribution of mineral versus organic forms) in order to provide perspectives on prediction-based approaches that take into account the interactions with other minerals such as N.

Nicotine Biosynthesis inNicotiana: A Metabolic Overview

Alkaloids are important compounds found in Nicotiana plants, essential in plant defense against herbivores. The main alkaloid of Nicotiana tabacum, nicotine, is produced in roots and translocated to the leaves. Nicotine is formed by a pyrrolidine and a pyridine ring in a process involving several enzymes. The pyridine ring of nicotine is derived from nicotinic acid, whereas the pyrrolidine ring originates from polyamine putrescine metabolism. After synthesis in root cortical cells, a set of transporters is known to transport nicotine upward to the aerial part and store it in leaf vacuoles. Moreover, nicotine can be metabolized in leaves, giving rise to nornicotine through the N-demethylation process. Some Nicotiana wild species produce acyltransferase enzymes, which allow the plant to make N-acyl-nornicotine, an alkaloid with more potent insecticidal properties than nicotine. However, although we can find a wealth of information about the alkaloid production in Nicotiana spp., our understanding about nicotine biosynthesis, transport, and metabolism is still incomplete. This review will summarize these pathways on the basis on recent literature, as well as highlighting questions that need further investigation.

Differential physiological responses of Tunisian wild grapevines (Vitis vinifera L. subsp. sylvestris) to NaCl salt stress

The effects of salt stress on growth, organic and inorganic solute accumulation and chlorophyll florescence were studied on 3-month-old plants of six Tunisian wild grapevine accessions in order to identify salt tolerance mechanisms and select tolerant genotypes. Potted plants were grown under controlled conditions and irrigated for 14 days with 0, 100 and 150 mM NaCl Long Ashton nutrient solution. Salt begins to adversely affect plant growth and plant nutrition at 100 mM NaCl. Compared to control, shoot growth rates were 21.5% less for Khedhayria, Tebaba and Ouchteta, 33% for Djebba and Zouarâa and 49% for Houamdia. They were assigned to stomatal and non-stomatal factors. Stomatal conductance decreased after 1 day at 150 mM NaCl in all accessions in response to reduced leaf water potential. Leaves in tolerant accessions were well hydrated through efficient osmotic adjustment, sufficient potassium flux and selectivity of K+ versus Na+. In addition, salt tolerance of wild grapes was related to limiting Na+ transport to lamina and to compartmentalization of Cl− on root and leaf vacuoles, improved in Tebaba and Khedhayria by the uptake of K+. At the same time, disturbances of the PSII have been noted as non-stomatal factors, and the most important photoprotective mechanism against photosynthetic damages was non-photochemical energy dissipation. However, chlorophyll fluorescence parameters were more stable in Tebaba compared to Khedhayria, thus showing better salt tolerance. Based on our results, wild grapevine accessions could be classified from most tolerant to most sensitive as follows: Tebaba > Khedhayria > Ouchteta > Zouarâa > Djebba > Houamdia.

Subcellular distribution and chemical forms of lithium in Li-accumulator Apocynum venetum

Apocynum venetum is a promising species to remediate an emerging environmental contaminant lithium (Li). However, no research has been conducted so far relating Li tolerance mechanism. In order to improve the understanding of Li transportation and detoxification, subcellular accumulation and distribution of different chemical forms of Li was studied in Apocynum venetum. Subcellular Li compartmentalization analysis showed that majority of Li was located in vacuole (45.52–72.65%) and cell wall (14.84–29.02%) under Li treatment. Furthermore, water soluble and ethonal extracted Li (inorganic Li) are the main chemical forms of Li taken up by A. venetum. With the increase of Li concentration in the medium, Li content in all subcellular fractions and proportion of F-ethanol form with high mobility increased. The greatest amount of Li was found in soluble fraction in leaves at 25 mg L−1 Li treatment, followed by soluble fraction in leaves at 2.5 mg L−1. These results suggest that Li compartmentation in leaf vacuoles is important in Li detoxification and Li accumulation of A. venetum.

Transcriptome sequencing and comparative analysis of differentially-expressed isoforms in the roots of Halogeton glomeratus under salt stress

Although Halogeton glomeratus (H. glomeratus) has been confirmed to have a unique mechanism to regulate Na+ efflux from the cytoplasm and compartmentalize Na+ into leaf vacuoles, little is known about the salt tolerance mechanisms of roots under salinity stress. In the present study, transcripts were sequenced using the BGISEQ-500 sequencing platform (BGI, Wuhan, China). After quality control, approximately 24.08 million clean reads were obtained and the average mapping ratio to the reference gene was 70.00%. When comparing salt-treated samples with the control, a total of 550, 590, 1411 and 2063 DEIs were identified at 2, 6, 24 and 72h, respectively. Numerous differentially-expressed isoforms that play important roles in response and adaptation to salt condition are related to metabolic processes, cellular processes, single-organism processes, localization, biological regulation, responses to stimulus, binding, catalytic activity and transporter activity. Fifty-eight salt-induced isoforms were common to different stages of salt stress; most of these DEIs were related to signal transduction and transporters, which maybe the core isoforms regulating Na+ uptake and transport in the roots of H. glomeratus. The expression patterns of 18 DEIs that were detected by quantitative real-time polymerase chain reaction were consistent with their respective changes in transcript abundance as identified by RNA-Seq technology. The present study thoroughly explored potential isoforms involved in salt tolerance on H. glomeratus roots at five time points. Our results may serve as an important resource for the H. glomeratus research community, improving our understanding of salt tolerance in halophyte survival under high salinity stress.

Influence of Se concentrations and species in hydroponic cultures on Se uptake, translocation and assimilation in non-accumulator ryegrass

The success of biofortification and phytoremediation practices, addressing Se deficiency and Se pollution issues, hinges crucially on the fate of selenium in the plant media in response to uptake, translocation and assimilation processes. We investigate the fate of selenium in root and shoot compartments after 3 and 6 weeks of experiment using a total of 128 plants grown in hydroponic solution supplied with 0.2, 2, 5, 20 and 100 mg L−1 of selenium in the form of selenite, selenate and a mixture of both species. Selenate-treated plants exhibited higher root-to-shoot Se translocation and total Se uptake than selenite-treated plants. Plants took advantage of the selenate mobility and presumably of the storage capacity of leaf vacuoles to circumvent selenium toxicity within the plant. Surprisingly, 28% of selenate was found in shoots of selenite-treated plants, questioning the ability of plants to oxidize selenite into selenate. Selenomethionine and methylated organo-selenium amounted to 30% and 8% respectively in shoots and 35% and 9% in roots of the identified Se, suggesting that selenium metabolization occurred concomitantly in root and shoot plant compartments and demonstrating that non-accumulator plants can synthesize notable quantities of precursor compound for volatilization. The present study demonstrated that non-accumulator plants can develop the same strategies as hyper-accumulator plants to limit selenium toxicity. When both selenate and selenite were supplied together, plants used selenate in a storage pathway and selenite in an assimilation pathway. Plants might thereby benefit from mixed supplies of selenite and selenate by saving enzymes and energy required for selenate reduction.

Comparison of subcellular distribution and chemical forms of cadmium among four soybean cultivars at young seedlings.

The hydroponic experiment was carried out to investigate the Cd subcellular distribution and chemical forms in roots and shoots among four soybean seedling cultivars with two Cd treatments. HX3 and GC8, two tolerant and low-grain-Cd-accumulating cultivars, had the lowest Cd concentration in roots and high Cd concentration in shoots, while BX10 and ZH24, two sensitive and high-grain-Cd-accumulating cultivars, had the highest Cd concentration in roots and the lowest Cd concentration in shoots at young seedling stage. Furthermore, the sequence of Cd subcellular distribution in roots at two Cd levels was cell wall (53.4-75.5 %) > soluble fraction (15.8-40.4 %) > organelle fraction (2.0-14.7 %), but in shoots, was soluble fraction (39.3-74.8 %) > cell wall (16.0-52.0 %) > organelle (4.8-19.5 %). BX10 and ZH24 had higher Cd concentration in all subcellular fractions in roots, but HX3 and GC8 had higher Cd concentration of soluble fraction in shoots. The sequence of Cd chemical forms in roots was FNacl (64.1-79.5 %) > FHAC (3.4-21.5 %) > Fd-H2O (3.6-13.0 %) > Fethanol (1.4-21.8) > FHCl (0.3-1.6 %) > Fother (0.2-1.4 %) at two Cd levels but, in shoots, was FNacl (19.7-51.4 %) ≥ FHAC (10.2-31.4 %) ≥ Fd-H2O (8.8-28.2 %) ≥ Fethanol (8.9-38.6 %) > FHCl (0.2-9.6 %) > Fother (2.5-11.2 %). BX10 and ZH24 had higher Cd concentrations in each extracted solutions from roots, but from shoots for GC8 and HX3. Taken together, the results uncover that root cell walls and leaf vacuoles might play important roles in Cd detoxification and limiting the symplastic movement of Cd.

A Prominent Role for Leaf Calcium as a Yield and Quality Determinant in Upland Cotton (Gossypium hirsutum L.) Varieties Grown Under Irrigated Mediterranean Conditions

Seven cotton (Gossypium hirsutum L.) accessions were tested over 2 years under irrigated Mediterranean conditions on a loamy soil with nitrogen (N) as the only nutrient input. The study aimed to identify the critical nutritional and physiological factors determining seedcotton yield and fibre quality. A suite of leaf physiological traits [chlorophyll content (assessed by SPAD), carbon isotope discrimination (Δ), 15N natural abundance (δ15N), leaf water potential, N and C concentrations, C/N ratio, K, Na, Ca and Mg concentrations, their sum and ratios] was assessed, and their interrelationships then analysed. It was found that physiological indices such as SPAD, Δ and δ15N failed to discern genotypes for yield and did not relate with fibre quality traits. At the same time, leaf Ca concentration was the trait that showed the strongest correlation with both seedcotton (SY) and lint yield (LY). An increase of K/Na ratio up to 5.74 was beneficial for SY but higher ratios impacted yield adversely. In this line, exclusion of K in favour of Ca (lower K/Ca ratios) increased both SY and LY. The above results could be explained by Ca2+ control over activity of tonoplast and plasma membrane cation channels, resulting in redistribution of K+ between cell compartments. It is suggested that Ca2+-rich plants are more efficient in sequestering higher K+ quantities in leaf vacuoles, at the expense of cytosolic K+. Under K+-limiting conditions, such redistribution may trigger programmed cell death and enhance leaf senescence. This would remobilize and translocate nutrients (e.g. N) and organic substances to sinks (seedcotton), contributing to higher yields reported in the present work.