Signaling function of NH4+ in the activation of Fe-deficiency response in cucumber (Cucumis sativus L.).

MAIN CONCLUSION: NH4+ is necessary for full functionality of reduction-based Fe deficiency response in plants. Nitrogen (N) is present in soil mainly as nitrate (NO3-) or ammonium (NH4+). Although the significance of a balanced supply of NO3- and NH4+ for optimal growth has been generally accepted, its importance for iron (Fe) acquisition has not been sufficiently investigated. In this work, hydroponically grown cucumber (Cucumis sativus L. cv. Maximus) plants were supplied with NO3- as the sole N source under -Fe conditions. Upon the appearance of chlorosis, plants were supplemented with 2 mM NH4Cl by roots or leaves. The NH4+ treatment increased leaf SPAD and the HCl-extractable Fe concentration while decreased root apoplastic Fe. A concomitant increase in the root concentration of nitric oxide and activity of FRO and its abolishment by an ethylene action inhibitor, indicated activation of the components of Strategy I in NH4+-treated plants. Ammonium-pretreated plants showed higher utilization capacity of sparingly soluble Fe(OH)3 and higher root release of H+, phenolics, and organic acids. The expression of the master regulator of Fe deficiency response (FIT) and its downstream genes (AHA1, FRO2, and IRT1) along with EIN3 and STOP1 was increased by NH4+ application. Temporal analyses and the employment of a split-root system enabled us to suggest that a permanent presence of NH4+ at concentrations lower than 2 mM is adequate to produce an unknown signal and causes a sustained upregulation of Fe deficiency-related genes, thus augmenting the Fe-acquisition machinery. The results indicate that NH4+ appears to be a widespread and previously underappreciated component of plant reduction-based Fe deficiency response.

Diverse responses of halophyte and glycophyte Lepidium species to the salt-mediated amelioration of nickel toxicity and accumulation.

Tolerance mechanisms employed by plants under environmental stresses can protect them against other co-occurring stresses. In this study, the effect of pre-exposure and simultaneous salt treatment on nickel (Ni) toxicity tolerance in one halophyte (L. sativum) and one glycophyte (L. latifolium) Lepidium species in hydroponics was investigated. In order to compare the species independent from their salt and Ni tolerance level, the glycophyte was subjected to lower salt and Ni concentrations and for a shorter period of time than the halophyte. Salt (NaCl) was applied at 50 and 100 mM concentrations and Ni was provided at an equal free Ni2+ activity by adding 100 and 200 µM Ni as single stresses, but 130 and 300 µM Ni for the treatment of its combination with salt in the glycophyte and halophyte, respectively. Temporal analyses of signaling molecules revealed that the halophyte is characteristically different from the glycophyte in that it exhibits a higher constitutive level of nitric oxide and hydrogen peroxide, a longer duration of response to Ni, and its augmentation by salt. In addition to higher biomass and less Ni accumulation in salt-treated plants, the concentrations of free thiol groups, leaf pigments, proline, free and cell wall-bound phenolics contents, and the activity of phenolic metabolizing enzymes were higher in L. latifolium under the combined salt and Ni treatments than under the single Ni stress. In contrast, the biomass and most biochemical parameters of Ni-stressed L. sativum plants were not enhanced by salt treatment but rather decreased. Our findings shed light on cross-tolerance mechanisms in halophytes and uncovered halophyte survival strategies under multiple stresses.

The arbuscular mycorrhizal mycelium from barley differentially influences various defense parameters in the non-host sugar beet under co-cultivation.

The interactions between arbuscular mycorrhizal fungi (AMF) and non-host species are poorly studied. Particularly scarce is information on members of the Amaranthaceae/Chenopodiaceae family. Sugar beet (Beta vulgaris) plants were co-cultivated with a host species (Hordeum vulgare) in the presence (+AMF) or absence of Rhizophagus intraradices to explore the hypothesis that the presence of an active, pre-established AMF mycelium induces defense responses in the non-host species. Biomass of sugar beet did not respond to the +AMF treatment, while its root exudation of organic acids and phenolic acids was drastically decreased upon co-cultivation with +AMF barley. The most conspicuous effect was observed on a wide range of potential defense parameters being differentially influenced by the +AMF treatment in this non-host species. Antioxidant defense enzymes were activated and the level of endogenous jasmonic acid was elevated accompanied by nitric oxide accumulation and lignin deposition in the roots after long-term +AMF treatment. In contrast, significant reductions in the levels of endogenous salicylic acid and tissue concentration and exudation of phenolic acids indicated that AM fungus hyphae in the substrate did not induce a hypersensitive-type response in the sugar beet roots and downregulated certain chemical defenses. Our results imply that the fitness of this non-host species is not reduced when grown in the presence of an AMF mycelium because of balanced defense costs. Further studies should address the question of whether or not such modulation of defense pattern influences the pest resistance of sugar beet plants under field conditions.

Salt tolerance mechanisms in three Irano-Turanian Brassicaceae halophytes relatives of Arabidopsis thaliana.

Salt tolerance mechanisms were studied in three Irano-Turanian halophytic species from the Brassicaceae (Lepidium latifolium, L. perfoliatum and Schrenkiella parvula) and compared with the glycophyte Arabidopsis thaliana. According to seed germination under salt stress, L. perfoliatum was the most tolerant species, while L. latifolium and S. parvula were rather susceptible. Contrastingly, based on biomass production L. perfoliatum was more salt sensitive than the other two species. In S. parvula biomass was increased up to 2.8-fold by 100 mM NaCl; no significant growth reduction was observed even when exposed to 400 mM NaCl. Stable activities of antioxidative defense enzymes, nil or negligible accumulation of superoxide anion and hydrogen peroxide, as well as stable membrane integrity in the three halophytes revealed that no oxidative stress occurred in these tolerant species under salt stress. Proline levels increased in response to salt treatment. However, it contributed only by 0.3‒2.0% to the total osmolyte concentration in the three halophytes (at 400 mM NaCl) and even less (0.04%) in the glycophyte, A. thaliana (at 100 mM NaCl). Soluble sugars in all three halophytes and free amino acids pool in S. parvula decreased under salt treatment in contrast to the glycophyte, A. thaliana. The contribution of organic osmolytes to the total osmolyte pool increased by salt treatment in the roots, while decreased in halophyte and glycophyte, A. thaliana leaves. Interestingly, this reduction was compensated by a higher relative contribution of K in the leaves of the halophytes, but of Na in A. thaliana. Taken together, biomass data and biochemical indicators show that S. parvula is more salt tolerant than the two Lepidium species. Our data indicate that L. latifolium, as a perennial halophyte with a large biomass, is highly suitable for both restoration of saline habitats and saline agriculture.

Silicon influences growth and mycorrhizal responsiveness in strawberry plants.

Effect of silicon (Si) on the response of strawberry (Fragaria × ananassa var. Parus) plants to arbuscular mycorrhizal fungus (AMF) was studied under growth chamber conditions. Plants were grown in perlite irrigated with nutrient solution without (- Si) or with (+ Si) 3 mmol L-1 Si (~ 84 mg L-1 Si as Na2SiO3) in the absence (- AMF) or presence (+ AMF) of fungus. Dry matter production, root colonization rate, photosynthesis rate and water relation parameters were all improved by both Si and AMF, and the highest amounts were achieved by + Si + AMF treatment. Mycorrhizal effectiveness increased by Si treatment associated with higher Si concentration in the + AMF plants. Leaf concentrations of total soluble and cell wall-bound phenolics were increased by Si accompanied by the enhanced activity of phenylalanine ammonia lyase, but not polyphenol oxidase. Profile of phenolics compound revealed that gallic acid, caffeic acid, epicatechin, chlorogenic acid, ellagic acid and kaempferol increased by both Si and AMF treatments, while p-coumaric acid decreased. In addition to vegetative growth, both treatments improved fruit yield and its quality parameters. Our results showed that Si and AMF acted in a synergistic manner and improved growth and biochemical parameters in strawberry plants. However, the mechanism for Si-mediated increase of mycorrhizal effectiveness is not known, thereby needing further elucidation.

Phenolics metabolism in boron-deficient tea [Camellia sinensis (L.) O. Kuntze] plants.

Modification in the metabolism of phenolic compounds under boron (B) deficiency conditions was studied in tea plants. Plants were grown from seed, treated with low B in hydroponic medium under environmentally controlled conditions for six weeks. Dry matter production and B content of plants were significantly declined under B deficiency conditions. Boron starvation resulted in rising phenylalanine ammonia lyase activity in the young leaves and declining polyphenol oxidase activity in the roots. Soluble phenolics fraction was increased up to 3.4-fold in the young leaves while did not influence by B nutrition in the old leaves and roots. Cell wall (CW) bound phenolics and lignin content was lower in B-deficient plants compared with B-sufficient ones. Boron deficiency increased significantly activity of soluble peroxidase (POD) only in the leaves. Activity of ionically bound POD was decreased in the old leaf and roots while it increased in the young leaves upon B deprivation. Activity of covalently bound POD decreased in the roots and leaves of different age in low B plants. Our results suggested that tea plant is highly tolerant species to B deficiency and CW tightening and accumulation of oxidized phenolics are not mechanisms for growth inhibition under B deficiency conditions.

Weed Species from Tea Gardens as a Source of Novel Aluminum Hyperaccumulators.

Increased availability of toxic Al3+ is the main constraint limiting plant growth on acid soils. Plants adapted to acid soils, however, tolerate toxic Al3+, and some can accumulate Al in their aerial parts to a significant degree. Studies on Al-tolerant and Al-accumulating species have mainly focused on the vegetation of acid soils distributed as two global belts in the northern and southern hemispheres, while acid soils formed outside these regions have been largely neglected. The acid soils (pH 3.4-4.2) of the tea plantations in the south Caspian region of Northern Iran were surveyed over three seasons at two main locations. Aluminum and other mineral elements (including nutrients) were measured in 499 plant specimens representing 86 species from 43 families. Al accumulation exceeding the criterion for accumulator species (>1000 µg g-1 DW) was found in 36 species belonging to 23 families of herbaceous annual or perennial angiosperms, in addition to three bryophyte species. Besides Al, Fe accumulation (1026-5155 µg g-1 DW) was also observed in the accumulator species that exceeded the critical toxicity concentration, whereas no such accumulation was observed for Mn. The majority of analyzed accumulator plants (64%) were cosmopolitan or pluriregional species, with a considerable rate of Euro-Siberian elements (37%). Our findings, which may contribute to phylogenetic studies of Al accumulators, also suggest suitable accumulator and excluder species for the rehabilitation of acid-eroded soils and introduce new model species for investigating Al accumulation and exclusion mechanisms.

Phytoremediation of nitrate contamination using two halophytic species, Portulaca oleracea and Salicornia europaea.

Nitrate is a common form of nitrogen fertilizer, and its excess application combined with easy leaching from agricultural fields causes water and soil contamination, hazards on human health, and eutrophication of aquatic ecosystems. Compared to other pollutants, the application of phytoremediation technology for nitrate-contaminated sites has received less attention. Nitrophilous halophyte species are suitable candidates for this purpose particularly by application of additional treatments for assisting nitrate accumulation. In this work, two annual halophyte species, Portulaca oleracea and Salicornia europaea were studied for their phytoremediation capacity of nitrate-contaminated water and soils. Plants were treated with three nitrate levels (2, 14, and 50 mM) combined with either selenium (10 µM as Na2SeO4) or salt (100 mM NaCl) in the hydroponics and sand culture medium, respectively. A fast growth and production of higher biomass enables P. oleracea for higher nitrate removal compared with S. europaea in both experiments. In S. europaea, both selenium and salt treatments enhanced nitrate removal competence through increasing the biomass and nitrate uptake or assimilation capacity. Salt treatment, however, reduced these parameters in P. oleracea. Based on data, selenium-assisted phytoremediation of nitrate contamination is a feasible strategy for both species and S. europaea is better suited to nitrate-contaminated saline water and soils. Nitrate accumulation in both species, however, exceeds that of the permitted nitrate level in the forage crops suggesting that the phytoremediation byproducts could not be consumed and other management strategies should be applied to the residual biomass.

Localization of aluminium in tea (Camellia sinensis) leaves using low energy X-ray fluorescence spectro-microscopy.

Information on localization of Al in tea leaf tissues is required in order to better understand Al tolerance mechanism in this Al-accumulating plant species. Here, we have used low-energy X-ray fluorescence spectro-microscopy (LEXRF) to study localization of Al and other low Z-elements, namely C, O, Mg, Si and P, in fully developed leaves of the tea plant [Camellia sinensis (L.) O. Kuntze]. Plants were grown from seeds for 3 months in a hydroponic solution, and then exposed to 200 microM AlCl(3) for 2 weeks. Epidermal-mesophyll and xylem phloem regions of 20 microm thick cryo-fixed freeze-dried tea-leaf cross-sections were raster scanned with 1.7 and 2.2 keV excitation energies to reach the Al-K and P-K absorption edges. Al was mainly localized in the cell walls of the leaf epidermal cells, while almost no Al signal was obtained from the leaf symplast. The results suggest that the retention of Al in epidermal leaf apoplast represent the main tolerance mechanism to Al in tea plants. In addition LEXRF proved to be a powerful tool for localization of Al in plant tissues, which can help in our understanding of the processes of Al uptake, transport and tolerance in plants.

Aluminium alters mineral composition and polyphenol metabolism in leaves of tea plants (Camellia sinensis).

Tea plants (Camellia sinensis) can hyperaccumulate and tolerate high leaf concentrations of aluminium (Al). The quality of tealeaves and the positive health effects of their infusion depend on the leaf concentrations of both polyphenolic substances and mineral elements. This study explored the influence of Al supply on these leaf components under low and optimal phosphorus (P) availability. After 8 weeks exposure in hydroponics, multifactorial analysis revealed a negative influence of leaf Al on magnesium (Mg), P, boron (B), and manganese (Mn) leaf concentrations. Contrastingly, these essential mineral nutrients were positively related to leaf epigallocatechin. Galloylated catechins were positively related to leaf iron (Fe). After short-term exposure (24 and 96 h), RT-qPCR (Reverse Transcription-quantitative Polymerase Chain Reaction) analysis revealed upregulation of galloylation-related genes by substrate acidification both in old and young leaves. Only the extremely high Al accumulation in old leaves activated genes involved in biosynthesis of galloylated catechins, while in young leaves the lower Al leaf concentrations activated genes involved in anthocyanin accumulation. In conclusion, low pH and enhanced Al availability to tea plants have a strong influence on the polyphenolic pattern of tealeaves and therefore may alter both the leaves' antioxidant properties and their ability to bind Al and Fe in non-toxic form.

Contrasting allocation of magnesium, calcium and manganese in leaves of tea (Camellia sinensis (L.) Kuntze) plants may explain their different extraction efficiency into tea.

During tea preparation mineral elements are extracted from the dried leaves of tea (Camellia sinensis (L.) Kuntze) plants into the solution. Micro-particle induced X-ray emission was employed to investigate the spatial distribution of magnesium (Mg), calcium (Ca) and manganese (Mn) in the young and old leaves of tea plants grown in the absence and presence of aluminium (Al) in the substrate. Results revealed that in tea leaves the largest concentrations of Mg occurred in the epidermis, of Ca in oxalate crystals and of Mn in epidermis and oxalate crystals; there was a leaf-age effect on tissue-specific concentrations of Mg, Ca and Mn with all tissues of old leaves containing larger concentrations of Mg, Ca and Mn than young leaves; supplementation of substrate with Al reduced concentrations of Mg, Ca and Mn in the old leaves, and a link between the distribution of Mg, Ca and Mn in the tea leaves with the extraction efficiencies of these elements into the tea was possible. We conclude that old leaves of tea plants cultivated under conditions of low Al availability will have the largest concentrations of Mg, Ca and Mn and may represent most acceptable ingredient for the preparation of tea.

Senescence is delayed by selenium in oilseed rape plants.

Leaf senescence is a genetically programmed process that can also be induced by nitrogen (N) deficiency. Although selenium (Se) delays leaf senescence, the underlying mechanisms are still unknown. To explore the mechanisms of Se-mediated delay of leaf senescence, we studied the biochemical and molecular events that occur during developmental and N deficiency-induced senescence. Oilseed rape (Brassica napus L.) plants were grown under adequate N (AN, 16 mM) or low N (LN, 4 mM) conditions during the rosette growth stage and treated with Se (15 µg plant-1 as Na2SeO4) either through roots or leaves for four weeks. Shoot dry matter production was not influenced, while the photosynthetic parameters were improved by Se application in both young and old leaves under both AN and LN conditions. The Se treatment rarely influenced the concentrations of reactive oxygen species (ROS), while it increased the nitric oxide (NO) levels in young and old leaves under both AN and LN conditions. The positive correlation between the NO level and leaf photosynthetic parameters in old leaves of LN plants suggested a role for NO boosting, mediated by Se, in the protection of aging leaves from LN-induced accelerated senescence. This implication was further supported by the clear down-regulation of SAG12-1 and up-regulation of Cab, particularly by root application of Se in old leaves of LN plants. Our results provide the first evidence that Se influences the expression of senescence-associated genes and delays senescence through NO signalling but is independent of the ROS defence system.

Presence of Belowground Neighbors Activates Defense Pathways at the Expense of Growth in Tobacco Plants.

Plants can detect the presence of their neighbors belowground, often responding with changes in root growth for resource competition. Recent evidence also implies that perception of neighbors may also elicit defense responses, however, the associated metabolic activities are unclear. We investigated primary and defense-related secondary metabolisms and hormone expressions in tobaccos (Nicotiana rustica) grown either with own roots or roots of another conspecifics in hydroponic condition. The results showed that non-self root interaction significantly reduced photosynthetic activity and assimilate production, leading to a reduction of growth. Non-self interaction also modified plant phenylpropanoids metabolism, yielding higher lignin content (i.e., structural resistance) at whole plant level and higher phenolics accumulation (i.e., chemical defense) in roots. All these metabolic responses were associated with enhanced expressions of phytohormones, particularly jasmonic acid, salicylic acid and cytokinin in roots and abscisic acid in leaves, at the early stage of non-self interaction. Since the presence of neighbors often increase the probability of attacks from, e.g., pathogens and pests, this defense activation may act as an adaptation of plants to these possible upcoming attacks.

Amelioration of iron toxicity: A mechanism for aluminum-induced growth stimulation in tea plants.

Tea plants (Camellia sinensis) are well adapted to acid soils with high Al availability. These plants not only accumulate high leaf Al concentrations, but also respond to Al with growth stimulation. Decreased oxidative stress has been associated with this effect. Why tea plants not exposed to Al suffer from oxidative stress has not been clarified. In this study, hydroponically grown tea plants treated with 0 to 300 µM Al were analyzed for growth, Al and Fe accumulation, and Al distribution by means of morin and hematoxylin staining. Roots of control plants stained black with hematoxylin. This indicates the formation of a Fe-hematoxylin complex. Young leaves of controls accumulated more than 1000 mg Fe kg(-1) dry weight. This concentration is above the Fe-toxicity threshold in most species. Supply of Al stimulated growth and reduced Fe uptake and transport. These results indicate that Al-induced growth stimulation might be due to alleviation of a latent Fe toxicity occurring in tea plants without Al supply.