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The emergence of microplastics (MPs) as pollutants in agricultural soils is increasingly alarming, presenting significant threats to soil ecosystems. Given the widespread contamination of ecosystems by various types of MPs, including polystyrene (PS), polyvinyl chloride (PVC), and polyethylene (PE), it is crucial to understand their effects on agricultural productivity. The present study was conducted to investigate the effects of different types of MPs (PS, PVC, and PE) on various aspects of sunflower (Helianthus annuus L.) growth with the addition of rice straw biochar (RSB). This study aimed to examine plant growth and biomass, photosynthetic pigments and gas exchange characteristics, oxidative stress indicators, and the response of various antioxidants (enzymatic and non-enzymatic) and their specific gene expression, proline metabolism, the AsA-GSH cycle, cellular fractionation in the plants and post-harvest soil properties. The research outcomes indicated that elevated levels of different types of MPs in the soil notably reduced plant growth and biomass, photosynthetic pigments, and gas exchange attributes. Different types of MPs also induced oxidative stress, which caused an increase in various enzymatic and non-enzymatic antioxidant compounds, gene expression and sugar content; notably, a significant increase in proline metabolism, AsA-GSH cycle, and pigmentation of cellular components was also observed. Favorably, the addition of RSB significantly increased plant growth and biomass, gas exchange characteristics, enzymatic and non-enzymatic compounds, and relevant gene expression while decreasing oxidative stress. In addition, RSB amendment decreased proline metabolism and AsA-GSH cycle in H. annuus plants, thereby enhancing cellular fractionation and improving post-harvest soil properties. These results open new avenues for sustainable agriculture practices and show great potential for resolving the urgent issues caused by microplastic contamination in agricultural soils., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Drought is the most critical climatic factor instigating severe threats to crop production worldwide. As stress ameliorants, exogenous sodium nitroprusside (SNP) or spermidine (Spd) supply has positive responses in alleviating the drought adversities in crops, however, reports regarding their combined effects is still elusive. Here, the protective role of SNP and Spd to confer drought resistance in sunflower (Helianthus annuus L.) through up-regulation of physiological and metabolic processes was investigated. Plants were foliar sprayed with individual or combined SNP (100 μM) or Spd (100 μM). Drought was induced by keeping the soil at 100% (normal) and 60% (drought stress) field capacity levels. Drought exposure caused a marked decline in relative water content (RWC), excised leaf water retention (ELWR), net photosynthesis (P N ), transpiration rate (E), stomatal conductance (gs), and sub-stomatal conductance (Ci) with substantial increase in catalase (CAT), superoxide dismutase (SOD), and peroxidase (POX). SNP plus Spd exhibited a considerable increase in CAT, SOD, and POX activities under drought, and helped the plants to retain optimum water status and gas exchange attributes. Similarly, hydrogen peroxide (H 2 O 2 ) and malondialdehyde (MDA) contents were increased significantly to drought; however, a notable decline was recorded in drought prone plants treated with exogenous SNP plus Spd. Moreover, addition of SNP plus Spd under drought caused a remarkable increase in chlorophyll a (Chl a), chlorophyll b (Chl b), chlorophyll total (Chl t), carotenoids (Car), and growth traits like shoot length (SL), root length (RL), shoot fresh weight (SFW), shoot dry weight (SDW), root dry weight (RDW). Combined SNP and Spd application could potentially alleviate the drought-induced damages in sunflower through increased water status (8-10%), antioxidant enzymes (17-28%), chlorophyll pigments (14-21%), and growth performance (12-22%) under drought stress., Competing Interests: Declaration of competing interest The authors describe no known competing financial interest in connection with this publication., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

A rhizobacterium, Pantoea conspicua, was examined against sunflower seedlings' growth under arsenate stress. Sunflower upon exposure to arsenate resulted in compromised growth that might be due to the accumulation of higher concentrations of arsenate and reactive oxygen species (ROS) in seedlings' tissues. The deposited arsenate led to oxidative damage and electrolyte leakage, making the sunflower seedlings vulnerable to compromise its growth and development. However, inoculation of sunflower seedlings with P. conspicua alleviated arsenate stress in host by initiating a multilayered defence mechanism. In fact, P. conspicua filtered out 75.1% of the arsenate from growth medium that were available to the plant roots in the absence of the said strain. To accomplish such activity, P. conspicua secreted exopolysaccharides as well as altered lignification in host roots. The arsenate (24.9%) that made its way to plant tissues was countered by helping the host seedlings to produce higher levels of indole acetic acid, non enzymatic antioxidants (phenolics and flavonoids) and antioxidant enzymes (catalase, ascorbte peroxidase, peroxidase, superoxide dismutase). As a result, ROS accumulation and electrolyte leakage were brought back to normal levels as observed in control seedlings. Hence, the rhizobacterium associated host seedlings achieved higher net assimilation (127.7%) and relative growth rate (113.5%) under 100 ppm of arsenate stress. The work concluded that P. conspicua alleviated arsenate stress in the host plants by imposing physical barrier as well as improving host seedlings' physiology and biochemistry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Chromium (Cr) in the soil is one of the major pollutants for agricultural production. This study examined the efficiency of sunflower plants to remediate Cr-contaminated soils using a plant growth regulator, 5-aminolevolinic acid (ALA). At six leaf stage, sunflower plants were exposed to soil-applied Cr (0.15 g kg -1 ), manganese (Mn, 0.3 g kg -1 ) and trisodium (S,S)-ethylenediamine-N,N'-disuccinic acid (EDDS, 2.5 mmol kg -1 ), ALA (10 mg L -1 ) was sprayed. After ALA treatment, the plants were harvested for further biochemical analyses. Results showed that EDDS and Mn improved the Cr accumulation but restrained plant growth. Conversely, ALA improved the growth of Cr-stressed plants by promoting chlorophyll concentration in the top fully expanded leaves. The bioaccumulation quantity and removal efficiency of sunflowers treated by Cr + EDDS + ALA was improved by 47.92% and 47.94%, respectively, as compared to the Cr treatment. This was further supported by qRT-PCR analysis, where the expression of heavy metal transport genes such as ZIP6 and NRAMP6 and subsequently Cr accumulation in sunflower tissues increased by EDDS, Mn, and ALA application. However, compared with other treatments, ALA ameliorated cellular injury from Cr-stress by uptake or movement of Cr prevention, modulation of antioxidant enzymes, and elimination of reactive oxygen species. Our study suggested that ALA as an ideal option for the phytoremediation of Cr-contaminated soils., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Fatty acids play many roles in plants, but the function of some key genes involved in fatty acid biosynthesis in plant development are not yet properly understood. Here, we clone two β-ketoacyl-[ACP] reductase (KAR) genes from sunflower, HaKAR1 and HaKAR2, and characterize their functional roles. The enzymes cloned were the only two copies present in the sunflower genome. Both displayed a high degree of similarity, but their promoters infer different regulation. The two sunflower KAR genes were constitutively expressed in all tissues examined, being maximum in developing cotyledons at the start of oil synthesis. Over-expression of HaKAR1 in E. coli changed the fatty acid composition by promoting the elongation of C16:0 to C18:0 fatty acids. The enzymatic characterization of HaKAR1 revealed similar kinetic parameters to homologues from other oil accumulating species. The results point to a partially functional redundancy between HaKAR1 and HaKAR2. This study clearly revealed that these genes play a prominent role in de novo fatty acids synthesis in sunflower seeds., (Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Iron (Fe)-deficiency causes chlorosis and growth inhibition in sunflower, an important commercial crop. This study examines whether and how arbuscular mycorrhizal fungi (AMF) ameliorate Fe-deficiency symptoms in Fe-deficiency sensitive sunflower plants. AMF supplementation showed a significant improvement in plant biomass, chlorophyll score, Fv/Fm (quantum efficiency of photosystem II), and Pi_ABS (photosynthesis performance index), suggesting its beneficial effect under Fe deficiency. This AM-driven amelioration of Fe deficiency was further supported by the improvement of biochemical stress indicators, such as cell death, electrolyte leakage, superoxide anion, and hydrogen peroxide. In this study, the AMF supplementations resulted in significant improvement in Fe as well as Zn concentrations in root and shoot of sunflower under Fe deficiency. One of the primary Strategy-I responses, ferric reductase activity along with the expression of its respective gene (HaFRO1), significantly increased in roots due to AMF ensuring Fe availability in the rhizosphere under Fe deficiency. Our qPCR analysis also showed a significant upregulation of HaIRT1, HaNramp1, and HaZIP1 in roots of sunflower in the presence of AMF, suggesting that Fe and Zn transporters are concurrently involved with AMF-mediated alleviation of Fe deficiency. Further, AMF accelerates the activities of CAT and SOD, predominantly in roots to protect sunflower plants from Fe-deficiency reactive oxygen species (ROS). This study unveils the mechanistic basis of AMF to limit Fe deficiency retardation in sunflower., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Salicylic acid (SA) is involved in several responses associated with plant development and defence against biotic and abiotic stress, but its role on photosynthetic regulation is still under debate. This work investigated energy conversion processes and related gene expression in the brachytic mutant of sunflower lingering hope (linho). This mutant was characterized by a higher ratio between the free SA form and its conjugate form SA O-β-D-glucoside (SAG) compared to wild type (WT), without significant changes in the endogenous level of abscisic acid and hydrogen peroxide. The mutant showed an inhibition of photosynthesis due to a combination of both stomatal and non-stomatal limitations, although the latter seemed to play a major role. The reduced carboxylation efficiency was associated with a down-regulation of the gene expression for both the large and small subunits of Rubisco and the Rubisco activase enzyme. Moreover, linho showed an alteration of photosystem II (PSII) functionality, with reduced PSII photochemistry, increased PSII excitation pressure and decreased thermal energy dissipation of excessive light energy. These responses were associated with a lower photosynthetic pigments concentration and a reduced expression of genes encoding for light-harvesting chlorophyll a/b binding proteins (i.e. HaLhcA), chlorophyll binding subunits of PSII proteins (i.e. HaPsbS and HaPsbX), phytoene synthase enzyme and a different expression level for genes related to PSII repair cycle, such as HaPsbA and HaPsbD. The concomitant stimulation of respiratory metabolism, suggests that linho activated a coordinate modulation of chloroplast and mitochondria activities to compensate the energy imbalance and regulate energy conversion processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Photosynthetic processes in the chloroplast depend on the abundance of magnesium (Mg) in relatively high amounts; hence chloroplasts might react more sensitive to Mg-deficiency than other physiological processes within other organelles. Most authors suggest a critical Mg concentration to be 1.5 mg g -1 DM for biomass and yield formation. However, it is not yet elucidated whether this value also applies to photosynthetic processes. The present study focused on the response of photosynthetic processes to different Mg tissue concentrations. Wheat (Triticum aestivum) and sunflower (Helianthus annuus) plants were grown hydroponically for 10 days with 8 different levels of Mg supply (1.0, 0.5, 0.25, 0.1, 0.075, 0.05, 0.025, 0.01 mM Mg). Specific leaf mass, SPAD values, assimilation rate, F v /F m , electron transport rate and photochemical and non-photochemical quenching parameters were determined on youngest mature leaves. Tissue Mg concentrations decreased with lowering Mg supply to lowest concentrations of 0.7 mg g -1 DM in wheat leaves, but photosynthetic capacity was not affected. In sunflower leaves, lowest Mg concentrations of 0.56 mg g -1 DM were achieved and a diminished photosynthetic capacity was observed. The study shows that a Mg tissue concentration of 1.5 mg g -1 DM did not induce a negative effect on the photosynthetic capacity of wheat and sunflower leaves under our experimental conditions and hence, the critical Mg concentration for photosynthetic processes might be lower than for biomass and yield formation., (Copyright © 2019 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Salt stress is known to negatively affect the fundamental processes in plants, reducing their growth and yield. The role of Silicon (Si) to protect the sorghum and sunflower plants against salinity stress was assessed. The objective of this study was to evaluate the effects of different forms of Si application on the uptake and use efficiency of macronutrients and micronutrients in sorghum and sunflower plants under salinity stress under greenhouse conditions. Two experiments were conducted using sorghum and sunflower under greenhouse conditions. Four Si levels were applied to plants: no Si application; foliar application of 28.6 mmol.L -1 ; root application of 2.0 mmol.L -1 ; and combined Si application with both via nutrient solution and foliar spraying. Each Si treatment was applied in the absence and presence of NaCl (100 mM). Thirty days after treatments, sodium (Na + ) and Si accumulation, nutrient uptake and use efficiency, and the roots and total plant dry weight were measured. Salinity stress induced nutritional imbalance and decreased dry weight production in both plant species. Our results showed that Si application alleviated the salinity stress by decreased Na + uptake and increased nutritional efficiency, thereby favoring the total plant dry weight in sorghum by 27% and sunflower by 41%. This occurred when Si was applied either via root or in combination via root and foliar application, respectively. Collectively, our findings indicate that Si application can attenuate the deleterious effects of salt stress and increase yield in sorghum and sunflower plants in a sustainable manner., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Fumarase (EC 4.2.1.2) is encoded in sunflower (Helianthus annuus L.) by two genes (FUM1 and FUM2) expressing correspondingly the mitochondrial and the cytosolic form. Both forms have been purified from sunflower cotyledons and characterized. Three quarters of fumarase activity is located in the mitochondrial and one quarter in the cytosolic fraction. The cytosolic form has lower pH optimum than the mitochondrial form, it possesses higher affinity to malate, activated by Mn 2+ and less efficiently by Mg 2+ while the mitochondrial form is activated only by Mg 2+ . It is proposed that the mitochondrial form is involved in the respiratory processes linked to the tricarboxylic acid cycle and the cytosolic form participates in the utilization of succinate produced in the glyoxylate cycle providing the flux to gluconeogenesis in germinating sunflower seeds., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Magnesium (Mg) often leaches down in sandy soils due to high mobility and its foliar application proves to be beneficial. Organic biostimulants also prove to be helpful to affect plant physio-biochemistry and antioxidative defense system. The present study evaluated the beneficial effects of seed soaking in maize grain extract (MGE; 3%) in integration with or without foliar Mg (1 mM) in comparison to control (no treatment) on growth, yield performance, seed oil and fatty acid profile including physiological and biochemical basis of Hysun-336 sunflower hybrid grown on a sandy soil under greenhouse conditions. The integrative treatment (seed soaking in MGE + foliar spray with Mg) elevated growth traits, plant water status and membrane stability index, and reduced electrolyte leakage. Improved leaf contents of chlorophylls, carotenoids, total soluble sugars and proline, activities of non-enzymatic and enzymatic antioxidants were also observed. In addition, enhanced uptake of N, P, K including Mg and endogenous levels of plant hormones IAA, GA 3 and zeatin were recorded with the integrative treatment. Seed yield and oil contents including oleic (mono-unsaturated) and linoleic (poly-unsaturated) fatty acids also increased; however, a decrease in other saturated, mono-unsaturated and poly-unsaturated fatty acids was noticed. Improved seed and seedling vigor traits were also observed in progeny of sunflower that harvested from the integrative treatment. In summary, improved plant performance by the integrative treatment may be attributed to improved activities of antioxidants contributing to improved plant water content, nutrient uptake and endogenous hormonal levels in sunflower plants grown under sandy soil conditions., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Jerusalem artichoke (Helianthus tuberosus L.) is an important energy crop for utilizing coastal marginal land. This study was to investigate waterlogging tolerance of Jerusalem artichoke through photosynthetic diagnose with emphasis on photosystem II (PSII) and photosystem I (PSI) performance. Potted plants were subjected to severe (liquid level 5 cm above vermiculite surface) and moderate (liquid level 5 cm below vermiculite surface) waterlogging for 9 days. Large decreased photosynthetic rate suggested photosynthesis vulnerability upon waterlogging. After 7 days of severe waterlogging, PSII and PSI photoinhibition arose, indicated by significant decrease in the maximal photochemical efficiency of PSII (Fv/Fm) and PSI (△MR/MR 0 ), and PSI seemed more vulnerable because of greater decrease in △MR/MR 0 than Fv/Fm. In line with decreased △MR/MR 0 and unchanged Fv/Fm after 9 days of moderate waterlogging, the amount of PSI reaction center protein rather than PSII reaction center protein was lowered, confirming greater PSI vulnerability. According to positive correlation between △MR/MR 0 and efficiency that an electron moves beyond primary quinone and negative correlation between △MR/MR 0 and PSII excitation pressure, PSI inactivation elevated PSII excitation pressure by depressing electron transport at PSII acceptor side. Thus, PSI vulnerability induced PSII photoinhibition and endangered the stability of whole photosynthetic apparatus under waterlogging. In agreement with photosystems photoinhibition, elevated H 2 O 2 concentration and lipid peroxidation in the leaves corroborated waterlogging-induced oxidative stress. In conclusion, Jerusalem artichoke is a waterlogging sensitive species in terms of photosynthesis and PSI vulnerability. Consistently, tuber yield was tremendously reduced by waterlogging, confirming waterlogging sensitivity of Jerusalem artichoke., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

In addition to their role as reserve carbohydrates, fructans have been recognized as compounds that are protective against adverse environments. The aim of this study was to identify changes in the content and the degree of polymerization (DP) of fructan in sprouting tubers of Jerusalem artichoke under salt stress. Fructan was extracted from tubers at 1, 3, 5, and 7 days after planting in sandy loam soil irrigated with NaCl solution. Fructan accumulation and polymerization and the expression of genes encoding enzymes for fructan synthesis and degradation were evaluated. No significant differences between the control and treatment groups were observed until 5 days after sowing. The highest level of salinity (250 mM) not only inhibited sprouting and root growth but also decreased the level of fructan in the tubers. The proportion of fructan at DP 2-5 rapidly increased one day after sowing and then decreased over time. Under various NaCl treatments, at 7 days after sowing, all fructans except fructan at DP 6-10 were present in proportions less than or equal to the control. The variation in the DP of fructan was related to the transcription level of fructan metabolism genes. Fructan may support sprouting or resistance to salt stress by changing the DP of fructan molecules through hydrolysis without changing the total amount of fructan. The low-molecular-weight oligosaccharides (DP < 5) may be the major carbohydrates that support tuber sprouting or that are involved in protection from salt stress., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Salinity results in significant reduction in sunflower (Helianthus annuus L.) seedling growth and excessive generation of reactive oxygen species (ROS). Present work highlights the possible role of melatonin as an antioxidant through its interaction with nitric oxide (NO), and as an early and long distance NaCl-stress sensing signaling molecule in seedling cotyledons. Exogenous melatonin (15µM)±NaCl (120mM) inhibit seedling growth, which is also correlated with NO availability, accumulation of potential superoxide anion (O 2 •- ) and peroxynitrite anion (ONOO - ), extent of tyrosine-nitration of proteins, spatial localization and activity of superoxide dismutase (SOD) isoforms. NO acts as a positive modulator of melatonin accumulation in seedling cotyledons as a long-distance signaling response. Modulation of superoxide anion and peroxynitrite anion content by melatonin highlights its crucial role in combating deleterious effects of ROS and reactive nitrogen species (RNS). Present findings provide evidence for an interaction between melatonin and NO in their effect on seedling growth under salt stress accompanying differential modulation of two SOD isoforms, i.e. Cu/Zn SOD and Mn SOD., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Seed germination and dormancy are tightly regulated by hormone metabolism and signaling pathway. We investigated the endogenous content of abscisic acid (ABA), its catabolites, and gibberellins (GAs), as well as the expression level of certain ABA and GAs metabolic and signaling genes in embryo of dry and imbibed cypselas of inbred sunflower (Helianthus annuus L., Asteraceae) lines: B123 (dormant) and B91 (non-dormant). Under our experimental conditions, the expression of RGL2 gene might be related to the ABA peak in B123 line at 3 h of imbibition. Indeed, RGL2 transcripts are absent in dry and early embedded cypselas of the non-dormant line B91. ABA increase was accompanied by a significant ABA-Glucosyl ester (ABA-GE) and phaseic acid (PA) (two ABA catabolites) decrease in B123 line (3 h) which indicates that ABA metabolism seems to be more active in this line, and that it would be involved in the imposition and maintenance of sunflower seed dormancy, as it has been reported for many species. Finally, an increase of bioactive GAs (GA1 and GA3) occurs at 12 h of imbibition in both lines after a decrease in ABA content. This study shows the first report about the RGL2 tissue-specific gene expression in sunflower inbred lines with contrasting dormancy level. Furthermore, our results provide evidence that ABA and GAs content and differential expression of metabolism and signaling genes would be interacting in seed dormancy regulation through a mechanism of action related to embryo itself., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Acyl-CoA-binding proteins (ACBPs) bind to acyl-CoA esters and promote their interaction with other proteins, lipids and cell structures. Small class I ACBPs have been identified in different plants, such as Arabidopsis thaliana (AtACBP6), Brassica napus (BnACBP) and Oryza sativa (OsACBP1, OsACBP2, OsACBP3), and they are capable of binding to different acyl-CoA esters and phospholipids. Here we characterize HaACBP6, a class I ACBP expressed in sunflower (Helianthus annuus) tissues, studying the specificity of its corresponding recombinant HaACBP6 protein towards various acyl-CoA esters and phospholipids in vitro, particularly using isothermal titration calorimetry and protein phospholipid binding assays. This protein binds with high affinity to de novo synthetized derivatives palmitoly-CoA, stearoyl-CoA and oleoyl-CoA (Kd 0.29, 0.14 and 0.15 μM respectively). On the contrary, it showed lower affinity towards linoleoyl-CoA (Kd 5.6 μM). Moreover, rHaACBP6 binds to different phosphatidylcholine species (dipalmitoyl-PC, dioleoyl-PC and dilinoleoyl-PC), yet it displays no affinity towards other phospholipids like lyso-PC, phosphatidic acid and lysophosphatidic acid derivatives. In the light of these results, the possible involvement of this protein in sunflower oil synthesis is considered., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

There is increasing evidence showing that low molecular weight organic acids (LMWOA) are involved in heavy metal resistance mechanisms in plants. The aim of this study was to investigate the effects of exogenous malic (MA) or acetic (AA) acids on the toxicity and accumulation of cadmium (Cd) in sunflower (Helianthus annuus L.). For this purpose, plants were grown in hydroponics under controlled conditions. Single Cd stress (5 μM Cd for 14 days) induced strong phytotoxic effects, as indicated by a decrease in all growth parameters, concentration of photosynthetic pigments, and root activity, as well as a high level of hydrogen peroxide (H2O2) accumulation. Exogenous MA or AA (250 or 500 μM) applied to the Cd-containing medium enhanced the accumulation of Cd by the roots and limited Cd translocation to the shoots. Moreover, the MA or AA applied more or less reduced Cd phytotoxicity by increasing the growth parameters, photosynthetic pigment concentrations, decreasing accumulation of H2O2, and improving the root activity. Of the studied organic acids, MA was much more efficient in mitigation of Cd toxicity than AA, probably by its antioxidant effects, which were stronger than those of AA. Plant response to Cd involved decreased production of endogenous LMWOA, probably as a consequence of severe Cd toxicity. The addition of MA or AA to the medium increased endogenous accumulation of LMWOA, especially in the roots, which could be beneficial for plant metabolism. These results imply that especially MA may be involved in the processes of Cd uptake, translocation, and tolerance in plants., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Thioredoxins are believed to mediate starch and protein mobilization in germinating cereals and dicotyledons. Nothing is known about redox regulation of lipid mobilization in plants. The possible redox regulation by thioredoxin h (Trx h) of a thiol-protease which degrades the oleosin coat of the oil body and its impacts on lipid mobilization was investigated in sunflower (Helianthus annuus L.) seedlings. An alkaline proteolytic activity stimulated by light was detected in seedlings. In vitro, the activity of this alkaline protease increased after reduction by NADPH-thiordoxin reductase system (NTS). The expression pattern of an alkaline 65 kDa thiol protease detected by gelatin SDS-PAGE technique, corresponded to the activity profile of the NTS-activated protease. The thiol-specific fuorochrome monobromobimane (mBBr) showed that a 65 kDa protein was also in a reduced state in vivo and becomes reduced in vitro by NTS. Except for 17-20 kDa oleosins, other oil body associated mBBr-labeled proteins were disappeared within three days following germination. Treatments of sunflower oil bodies by the NTS-activated alkaline protease made them more susceptible to maize lipase action. Ascorbate application enhanced lipid mobilization of seedlings. A model for seedling oil body mobilization was proposed according to which Trx h or other Trx isoforms, reductively activates an oleosin degrading thiol-protease and some oil body proteins, thus renders the organelle more susceptible to subsequent lipolytic actions. For the first time the potential role of Trx in the mobilization of lipid reserves in plants has been shown., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Sunflower (Helianthus annuus L.) is a commercially important oilseed crop. Previous studies proved that this crop is a promising plant species for phytoextraction of excess soil phosphorus (P) because of its superior P accumulating characteristics. Suppression subtractive hybridization (SSH) strategy was employed to isolate and characterize genes that are induced in response to high P in this crop. SSH library was prepared using cDNA generated from plants treated with high P as the 'tester'. Based on the results of dot blot analysis, 360 positive cDNA clones were selected from the SSH library for sequencing. A total of 89 non-redundant expressed sequence tags (ESTs) were identified as high P-responsive genes and they were classified into 6 functional groups. Several genes involved in metabolism showed markedly preferential expression in the library. For further confirmation, thirteen of the representative ESTs were selected from all categories for RT-PCR analysis and the results showed up-regulation of these genes in response to high P-treatment. The gene expression data derived from this study suggested that several of the up-regulated genes identified under high P-treatment might be involved in P-accumulation and tolerance in this plant., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)

Fourteen monocyclic analogues of trypsin inhibitor SFTI-1 isolated from sunflower seeds were synthesized by the solid-phase method. The purpose of this work was to establish the role of a disulfide bridge present in inhibitor's side chains of Cys3 and Cys11 in association with serine proteinases. This cyclic fragment was replaced by the disulfide bridges formed by l-pencillamine (Pen), homo-l-cysteine (Hcy), N-sulfanylethylglycine (Nhcy) or combination of the three with Cys. As in the substrate specificity the P(1) position of the synthesized analogues Lys, Nlys [N-(4-aminobutyl)glycine], Phe or Nphe (N-benzylglycine) were present, and they were checked for trypsin and chymotrypsin inhibitory activity. The results clearly indicated that Pen and Nhcy were not acceptable at the position 3, yielding inactive analogues, whereas another residue (Cys11) could be substituted without any significant impact on the affinity towards proteinase. On the other hand, elongation of the Cys3 side chain by introduction of Hcy did not affect inhibitory activity, and an analogue with the Hcy-Hcy disulfide bridge was more than twice as effective as the reference compound ([Phe⁵] SFTI-1) in inhibition of bovine α-chymotrypsin., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

As opposed to other oilseeds, developing sunflower seeds do not accumulate starch initially. They rely on the sucrose that comes from the mother plant to synthesise lipid precursors. Glycolysis is the principal source of carbon skeletons and reducing power for lipid biosynthesis. In this work, glycolytic initial metabolites and enzyme activities from developing seed of two different sunflower lines, of high and low oil content, were compared during storage lipid synthesis. These two lines showed different kinetic lipid accumulation in the developing embryos. Fatty acids levels during the initial and final stage of lipid synthesis were higher in CAS-6 than in ZEN-8. The analysis of the photosynthate and sugars content suggests that, although the hexoses levels were quite similar in both lines, the amount of sucrose produced by the mother plant and available for lipid synthesis was higher in CAS-6. Although, a smaller amount of sucrose is available in the ZEN-8 line, its seeds maintain the levels of intermediate sugars in the initial steps of glycolysis due to an increase in the levels of the invertase, hexokinase and phosphoglucose isomerase activities in ZEN-8, with respect to CAS-6. Also, a readjustment in the final part of this metabolic route took place, with the activities of phosphoglycerate kinase and enolase in CAS-6 being higher, allowing increased synthesis of phosphoenolpiruvate, the intermediate carbon donor for fatty acid synthesis. In addition, recently, it has been shown that Arabidopsis mutants with a lower fat content in their seeds have a higher amount of sucrose. These data together point to these last two enzymatic activities, phosphoglycerate kinase and enolase, as being responsible for the lower fat content in the ZEN-8 line., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

The study deals with the quantitative expression pattern of genes involved in signaling transduction pathways in response to water stress in leaves and embryos of a water stress tolerant genotype compared to a non-tolerant genotype using real-time quantitative PCR. The experiment was conducted in the field. The results showed a high quantitative up-regulation of genes belonging to protein kinase, phosphatase and transcription factor pathways (from two to 70 fold) only in leaves of the tolerant genotype compared to the non-tolerant genotype. Moreover, genes related to the protein kinase pathway were down-regulated in leaves of the non-tolerant genotype. On the contrary, in seeds, our study showed that the positive regulation of genes related to the signal transduction pathway observed in leaves of the tolerant genotype is turned off, suggesting different transcriptional control of signaling water stress in reproductive organs compared to vegetative organs.

Treatment of 14-day-old sunflower seedlings with a toxic amount of copper (50 microM of CuSO(4)) during 5days caused significant increase in peroxidase activity in roots. Qualitative analysis of soluble proteins using native anionic PAGE followed by detection of peroxidase activity with guaïacol as electron donor in the presence of H(2)O(2) revealed five stimulated peroxidases, named A1, A2, A3, A4, and A5. These peroxidases had differential behavior during the period of treatment. A1, A2, A3 and A4 were stimulated in the first period of stress, but rapidly suppressed at 72h. A5 showed a progressive stimulation which was even increased at 120h. A1 was partially purified, identified using liquid chromatography coupled to mass spectrometry (LC-MS/MS), and characterized. Effects of pH and temperature on its activity were determined with guaïacol as electron donor. Optima were obtained at pH 8 and at 40 degrees C. Analysis of substrate specificity showed that A1 was active on coniferyl alcohol but not on IAA. Enzymatic activity was inhibited by a high concentration of H(2)O(2).

In this study, we have biochemically characterized the high-stearic sunflower (Helianthus annuus L.) mutant CAS-14. This mutant displays an abnormal fatty acid composition along the length of the seed when grown at high temperatures. Thus, at the expense of oleate and linoleate, CAS-14 seeds present an increasing axial gradient of stearic acid from the embryo to the distal extreme of the seed. The accumulation of oil in this mutant was initially characterized by analysing the incorporation of radiolabelled acetate, a fatty acid synthetic precursor, into the developing seed tissues in vivo. These experiments indicated that the mutant phenotype was associated with a decrease in the soluble stearoyl-acyl carrier protein desaturase (SAD) activity, as later confirmed when assessing this activity in cell-free extracts from developing sunflower kernels. Furthermore, SAD enzyme gene transcription was also examined in this tissue, identifying the coordinated decrease in the transcription of the sad17 and sad6 genes as underlying the decrease in enzyme activity. On the basis of these results and those previously obtained on the inheritance of the CAS-14 trait, we discuss the possible regulatory mechanisms acting on plant soluble desaturases.

One of the main antioxidant enzymes, catalase (CAT, EC 1.11.1.6), is capable of catalyzing the dismutation of H(2)O(2). This enzyme is involved in signal transduction pathway in plants, controlling the cellular level of this reactive oxygen species. Four different genes, CATA1-CATA4, were identified in Helianthus annuus L. cotyledons. Incubation of sunflower leaf discs with 300 and 500 microM CdCl(2) under light conditions increased CATA3 transcript level. However, it was not induced by Cd(2+) in etiolated plants. This Cd(2+)-induced increase was reverted by adding 10mM ascorbate. Treatments with 0.4 and 10 microM rose bengal (a generator of (1)O(2)) did not activate CATA3, but 10 microM methyl viologen (an enhancer of O(2)(-) production) and 10 mM H(2)O(2) increased its expression. In isolated chloroplasts, Cd(2+) and methyl viologen produced oxidation of the probe 2',7'-dichlorofluorescein diacetate indicating ROS formation. Besides, Cd(2+) treatment of leaf discs under light decreased CAT activity and increased carbonyl groups content, thus suggesting that enzyme inactivation could be due - in part - to a protein oxidation. These results indicate that light is involved in Cd(2+)-induced CATA3 enhancement, which leads to the synthesis of CAT isoforms less sensible to oxidation, and that chloroplast might be the main source of ROS responsible for this process.

Using the perspective of full scale application of phytoremediation techniques, research is focusing on the optimization of agronomic practices. Two annual high biomass yield crops, Sorghum bicolor and Helianthus annuus, were grown in a polymetallic soil. The experimental site, polluted by pyrite cinders, is located in an industrial site that has been listed in the clean-up national priority list since 2001. Specific aims of this work were to observe the concentration of metals in plants during the crop cycle and to establish the amount of metal removed by the crops. The field trial, arranged in a randomized block design, started in 2005. The concentrations of heavy metals in the soil were: As 309, Cd 4.29, Co 50.9, Cu 1527 and Zn 980mg kg(-1). The crops grown on the polluted soil received mineral fertilization (Fert) and organic amendment (Org), while plants in control soil (Ctrl) did not receive anything. The plots were watered during the crop cycle during two drought periods, using a sprinkler irrigation system. The phytoextraction potential of crops was estimated during the whole growth cycle and the plant biomass that was collected in each sampling date was ICP-analyzed. Plant-biomass growth curves were obtained. The concentrations of the metals in the shoots and in the total plant biomass were recorded. Finally, the metal removal was calculated for the harvestable parts of the crops. The amelioration of the nutritive status of the substrate that resulted, was highly effective for the biomass yield. However, fertilization and soil amendment did not heighten the concentration of metals in the harvestable tissue of the plants during the crop cycle. In some cases, organic matter appeared to bind the elements making them less available for the plants. The evaluation of the potential of phytoremediation of our plants compared to other crops in terms of metal removal was positive. Our results of metal removal are consistent with the results from other in situ experiments. The Zn removal by S. bicolor and H. annuus reached about 2000g ha(-1) and 1000g ha(-1), respectively.