Seed priming to optimize germination in Arthrocnemum Moq.

Background: Seed germination and seedling growth constitute the first stage of a plant's life cycle for crop establishment. Arthrocnemum Moq. is a halophyte of the subfamily Salicornioideae (Amaranthaceae), which could be recognized in the foreseeable future as an emerging candidate in applied biosaline agricultural programs, mainly due to the large biomass it represents in coastal and inland saltmarshes, in addition to its interesting nutritional and pharmacological properties. However, to ensure their subsequent use as a crop, it is necessary to optimize their germination through appropriate seed priming treatments. The main goal of this work was to seek the optimization of Arthrocnemum germination process using different pretreatments: exposure to sodium chloride (100 to 1200 mM) in the dark and its subsequent transferred to distilled water separately and together with the combination of pH (5, 7, 9), salinity (0, 100, 200 mM NaCl), and iron conditions (0, 200, 400 µM FeSO4). The experiments were tested on six samples of two different species: A. meridionale (from Tunisia) and A. macrostachyum (from Spain). Results: Salinity priming of seeds for 15 days in darkness improved germination percentages by almost 25% at 600 mM NaCl, in both Tunisian and Spanish species. However, keeping seeds at different salt concentrations for 30 days produced higher improvement percentages at lower concentrations in A. meridionale (100–200 mM NaCl), while in A. macrostachyum the highest improvement percentages were obtained at 600 mM NaCl (percentage improvement of 47%). When the dark time period is reduced to 5 days at higher salt concentrations, the greater germination percentages were reached in all the samples at the concentration of 800 mM NaCl, increasing the improvement of germination between 17 and 50%. Finally, the conditions of pH = 7, pretreatment in darkness at 800 mM NaCl and 400 µM or iron, turned out to be an effective medium for seed germination. Conclusions: Therefore, before using Arthrocnemum seeds in applied biotechnological programs, a seed priming treatment based on prior exposure to high salt concentrations (600–1000 mM NaCl) is recommended in order to maximize germination percentage

Identification and measurements of sub-individual biomarkers in date palm (Phoenix dactylifera) during metal contamination : prediction of metabolic pathways and description of metal detoxification mechanisms involved

Les industries de traitement du phosphate dans le but de produire des engrais phosphatés, très présentes dans la zone côtière sud du Grand Sfax en Tunisie, sont à l’origine d’émissions atmosphériques et de rejets de déchets ; en l’occurrence du phosphogypse chargés de contaminants métalliques. La contamination des sols qui en résulte est une contamination persistante. Dans le but de mieux comprendre les effets de la contamination par les métaux engendrés par cette activité industrielle, une attention particulière a été portée sur une espèce d’importance économique, le palmier dattier Phoenix dactylifera. Le présent travail avait pour objectifs d'étudier la germination des graines et la variété Deglet Nour dans différents contextes de contamination métallique aux moyens de techniques de biotechnologie végétale impliquant la recherche in silico et la culture in vitro de vitroplant et à l’aide de deux approches. Une approche individuelle intégrative qui s’est servie de nombreuses mesures de biomarqueurs morphologiques et biochimiques chez l’espèce étudiée exposée à divers stress métalliques. Une deuxième approche, plus moléculaire et mécanistique avait pour but d’identifier des gènes qui répondent à une exposition au Cd, ou au Cu, ou au Cr capables de prédire les voies métaboliques impliquées. Cette seconde approche, basée sur l’exploitation d’une banque de cDNA de la variété Deglet Nour a permis l’identification de gènes de chélateurs et transporteurs de métaux. Le suivi des niveaux d’expression de ces gènes chez des plantes placées dans des conditions d’expositions métalliques variées a permis de mieux comprendre les mécanismes de détoxication des métaux mis en œuvre.The phosphate processing industries for the production of phosphate fertilizers, which are present in the southern coastal zone of the Grand Sfax in Tunisia, caused atmospheric emissions and waste discharges i.e., phosphogypsum loaded with metal contaminants. The resulting contamination of soils is a persistent contamination. In order to better understand the effects of metal contamination caused by this industrial activity and ultimately to propose measures for the rehabilitation and/or ecological restoration of the sites but also because of its economic importance, particular attention has been paid on the palm date (Phoenix dactylifera). The aim of this work was to study seed germination and growth of the Deglet Nour variety in various metal contamination contexts by means of plant biotechnology techniques involving in silico research and in vitro culture of vitroplant. Two approaches have been performed. An individual integrative approach that used numerous measurements of morphological and biochemical biomarkers in date palm exposed to various metal stresses. A second approach, more molecular and mechanistic, was performed to identify genes that respond when plants are exposed to Cd, Cu, or Cr which help for the prediction of the metabolic pathways that are affected by contaminants or involved in detoxification processes. This second approach, based on the exploitation of a cDNA library of the Deglet Nour variety, allowed the identification of genes coding for metal chelators and transporters. Monitoring of the levels of expression of these genes made it possible to better understand the detoxification mechanisms of metals in the palm date

In-Site and Ex-Site Date Palm Exposure to Heavy Metals Involved Infra-Individual Biomarkers Upregulation

As a tree of considerable importance in arid regions—date palm, Phoenix dactylifera L. survival in contaminated areas of Sfax city has drawn our attention. Leaf samples of the plants grown in the study area showed high levels of cadmium (Cd), copper (Cu), and chromium (Cr). On the basis of this finding, the cellular mechanisms that explain these metal accumulations were investigated in controlled conditions. After four months of exposure to Cd, Cr, or Cu, high bioconcentration and translocation factor (TF > 1) have been shown for date palm plantlets exposed to Cd and low TF values were obtained for plantlets treated with Cr and Cu. Moreover, accumulation of oxidants and antioxidant enzyme activities occurred in exposed roots to Cu and Cd. Secondary metabolites, such as polyphenols and flavonoids, were enhanced in plants exposed at low metal concentrations and declined thereafter. Accumulation of flavonoids in cells may be correlated with the expression of the gene encoding Pdmate5, responsible for the transport of secondary metabolites, especially flavonoids. Other transporter genes responded positively to metal incorporation, especially Pdhma2, but also Pdabcc and Pdnramp6. The latter would be a new candidate gene sensitive to metallic stress in plants. Expressions of gene coding metal chelators were also investigated. Pdpcs1 and Pdmt3 exhibited a strong induction in plants exposed to Cr. These modifications of the expression of some biochemical and molecular based-markers in date palm helped to better understand the ability of the plant to tolerate metals. They could be useful in assessing heavy metal contaminations in polluted soils and may improve accumulation capacity of other plants

Copper toxicity and date palm (Phoenix dactylifera) seedling tolerance: Monitoring of related biomarkers

ACLDate palm (Phoenix dactylifera) seeds were exposed to different copper (Cu) solutions to examine plant stress responses. Low Cu concentrations (0.02 and 0.2 mM) caused an increase of seed germination, whereas higher Cu amounts (2 mM) significantly inhibited seed germination, delayed hypocotyl elongation, increased seedling mortality, and reduced the germination index by more than 90%. Metal-related toxicity symptoms appeared after 15 d of 2 mM of Cu exposure. Biochemical activities such as amylase activity and redox balance elements were examined to study the relationship between external Cu amount and internal plant response. The present study showed that amylolytic activity was dose- and time-dependent. Likewise, H2 O2 production increased after exposure to Cu, which was correlated with thiobarbituric acid reactive substance (TBARS) accumulation. Furthermore at low Cu concentrations, superoxide dismutase (SOD) and catalase (CAT) activities increased, suggesting that date palm seed stimulated its metal homeostasis networks. However, the highest cupric ion amounts increased cell oxidant accumulation and reduced enzyme production. Gene expression level measures of P. dactylifera phytochelatin synthase (Pdpcs) and P. dactylifera metallothionein (Pdmt) encoding genes have been carried out to investigate the implication of PdPCS and PdMT proteins in Cu homeostasis and/or its sequestration. Phoenix dactylifera metallothionein induction reached a peak after 30 d of exposure to 0.2 mM of Cu. However, it was down-regulated in plants exposed to higher Cu concentrations. In the same conditions, Pdpcs was overexpressed during 1 mo of exposure before it decreased thereafter. These observations provide a new insight into date palm cell response to Cu, a metal that can be toxic but that is also an essential element

In-Site and Ex-Site Date Palm Exposure to Heavy Metals Involved Infra-Individual Biomarkers Upregulation

As a tree of considerable importance in arid regions—date palm, Phoenix dactylifera L. survival in contaminated areas of Sfax city has drawn our attention. Leaf samples of the plants grown in the study area showed high levels of cadmium (Cd), copper (Cu), and chromium (Cr). On the basis of this finding, the cellular mechanisms that explain these metal accumulations were investigated in controlled conditions. After four months of exposure to Cd, Cr, or Cu, high bioconcentration and translocation factor (TF > 1) have been shown for date palm plantlets exposed to Cd and low TF values were obtained for plantlets treated with Cr and Cu. Moreover, accumulation of oxidants and antioxidant enzyme activities occurred in exposed roots to Cu and Cd. Secondary metabolites, such as polyphenols and flavonoids, were enhanced in plants exposed at low metal concentrations and declined thereafter. Accumulation of flavonoids in cells may be correlated with the expression of the gene encoding Pdmate5, responsible for the transport of secondary metabolites, especially flavonoids. Other transporter genes responded positively to metal incorporation, especially Pdhma2, but also Pdabcc and Pdnramp6. The latter would be a new candidate gene sensitive to metallic stress in plants. Expressions of gene coding metal chelators were also investigated. Pdpcs1 and Pdmt3 exhibited a strong induction in plants exposed to Cr. These modifications of the expression of some biochemical and molecular based-markers in date palm helped to better understand the ability of the plant to tolerate metals. They could be useful in assessing heavy metal contaminations in polluted soils and may improve accumulation capacity of other plants

Insight into the expression variation of metal-responsive genes in the seedling of date palm (Phoenix dactylifera)

ACLPhytochelatin synthase and metallothionein gene expressions were monitored via qPCR in order to investigate the molecular mechanisms involved in Cd and Cr detoxification in date palm (Phoenix dactylifera). A specific reference gene validation procedure using BestKeeper, NormFinder and geNorm programs allowed selection of the three most stable reference genes in a context of Cd or Cr contamination among six reference gene candidates, namely elongation factor α1, actin, aldehyde dehydrogenase, SAND family, tubulin 6 and TaTa box binding protein. Phytochelatin synthase (pcs) and metallothionein (mt) encoding gene expression were induced from the first days of exposure. At low Cd stress (0.02mM), genes were still up- regulated until 60th day of exposure. At the highest metal concentrations, however, pcs and mt gene expressions decreased. pcs encoding gene was significantly up-regulated under Cr exposure, and was more responsive to increasing Cr concentration than mt encoding gene. Moreover, exposure to Cd or Cr influenced clearly seed germination and hypocotyls elongation. Thus, the results have proved that both analyzed genes participate in metal detoxification and their expression is regulated at transcriptional level in date palm subjected to Cr and Cd stress. Consequently, variations of expression of mt and pcs genes may serve as early-warning biomarkers of metal stress in this species