Modulation of inflammatory responses by diterpene acids from Helianthus annuus L

Fractionation of a petroleum ether extract of Helianthus annuus L. led to the isolation of three diterpene acids: grandiflorolic, kaurenoic and trachylobanoic acids. These compounds were studied for potential anti-inflammatory activity on the generation of inflammatory mediators in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. At non-toxic concentrations, these compounds reduced, in a concentration-dependent manner nitric oxide (NO), prostaglandin E₂ (PGE₂) and tumor necrosis factor (TNF-α) production, as well as expression of inducible nitric oxide synthase (NOS-2) and cyclooxygenase-2 (COX-2).All diterpenoids displayed significant in vivo anti-inflammatory activity and suppressed the 12-O-tetradecanoylphorbol-13-acetate (TPA)-mouse ear edema. In addition, inhibition of myeloperoxidase (MPO) activity, an index of cellular infiltration, was observed.In summary, our results suggest that the inhibition of the expression of NOS-2, COX-2 and the release of inflammatory cytokines, is responsible for the anti-inflammatory effects of the diterpenoids isolated from H. annuus L. which likely contributes to the pharmacological action of sunflower.     

Induction of transfer-cell formation by iron deficiency in the root epidermis of Helianthus annuus L.

Helianthus annuus L. responds to iron deficiency by forming a thickened cortex and abundant root hairs in a zone near the root apex that corresponds to the primary developmental stage. Cytological investigations revealed that within 24 to 48 h of iron deficiency most of the peripheral cells differentiate into transfer cells. The wall labyrinth is always situated on the peripheral walls that face the external medium. The cytoplasm of these cells is characterized by numerous mitochondria, extensive rough endoplasmic reticulum, and large leucoplasts containing protein bodies. These observations are discussed in relation to the fact that Helianthus, as an iron efficient plant, responds physiologically to iron deficiency by extrusion of H⁺, production of reducing substances, and a steep increase in the uptake efficiency of Fe.     

Characteristics of action potentials in Helianthus annuus

The action potentials induced by nondamaging electrical stimuli in 16- to 22-day-old plants of Helianthus annuus were examined. Typical recordings are presented. Mean values of their amplitudes and conduction velocities in the stem, the strength-duration relation, the 'all-or-none' law and the refractory periods have been determined. The amplitude and velocity of propagation were essentially identical in the upward and downward direction, but greater in the upper than in the lower half. In 'electrically active' plants, the rheobase value is 2 V, the minimum period for stimulation is 1.8 s. and the chronaxie 2.3 s. It is noted that the excitability level between similar plants on the same day and in the same plant on different days is highly variable and undergoes periodic changes.     

A comparison of the water relations characteristics of Helianthus annuus and Helianthus petiolaris when subjected to water deficits

The effect of water deficits on the water relations and stomatal responses of Helianthus annuus and Helianthus petiolaris were compared in plants growing in the glasshouse under controlled conditions. Unirrigated plants of both genotypes were subjected to two different stress rates in which predawn leaf water potentials declined steadily at either 0.15 MPa day^?1 or 0.50 MPa day^?1. In both genotypes water stress induced a gradual and similar decrease in leaf conductance from 1.6 to 0.3 cm s^?1 as water potential decreased from-0.5 to-2.0 MPa. The relationship between leaf conductance and leaf water potential was not affected by the rate of stress development. Development of predawn leaf water potentials of-1.3 MPa had no significant effect on the relative water content at zero turgor, the apoplastic water content or the volumetric elastic modulus of whole leaves in either species, but decreased the osmotic potential at full turgor and zero turgor by 0.22 MPa and decreased the turgid weight: dry weight ratio from 10.6 to 8.4 in H. annuus, but not in H. petiolaris. In H. annuus leaves expanded during stress development, changes in the osmotic potential at full turgor induced by water deficits did not disappear on rewatering.     

Hairy roots of Helianthus annuus: a model system to study phytoremediation of tetracycline and oxytetracycline

The release of antibiotics to the environment has to be controlled because of serious threats to human health. Hairy root cultures of Helianthus annuus (sunflower), along with their inherent rhizospheric activity, provide a fast growing, microbe-free environment for understanding plant-pollutant interactions. The root system catalyzes rapid disappearance of tetracycline (TC) and oxytetracycline (OTC) from aqueous media, which suggests roots have potential for phytoremediation of the two antibiotics in vivo. In addition, in vitro modifications of the two antibiotics by filtered, cell- and microbe-free root exudates suggest involvement of root-secreted compounds. The modification is confirmed from changes observed in UV spectra of exudate-treated OTC. Modification appears to be more dominant at the BCD chromophore of the antibiotic molecule. Kinetic analyses dismiss direct enzyme catalysis; the modification rates decrease with increasing OTC concentrations. The rates increase with increasing age of cultures from which root exudates are prepared. The decrease in modification rates upon addition of the antioxidant ascorbic acid (AA) suggests involvement of reactive oxygen species (ROS) in the antibiotic modification process.     

Behaviour of antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells

A relationship between the antioxidant defense system and salt tolerance in two types of sunflower calli differing in salt sensitivity was studied. No reduction in growth occurred in the NaCl-salt-adapted cell line (T) when grown on 175 mM NaCl but growth of the salt-stressed cell line (S) was reduced by 83%. Lipid peroxidation and protein oxidation increased during acute stress of salt stressed cells at 14 and 28 d of the experiment, while salt-adapted calli (T) remained similar to non-shocked (C) values. The antioxidant defense system of callus adapted to growth under NaCl responded differently to 175 mM of salt compared with the corresponding controls under shock treatment. Salt-adapted and salt-stressed calli showed a similar pattern in GSH content at day 14 but at day 28 in S calli, GSH content was increased 100% over the non-shocked calli, while T calli returned to the initial values. In the salt-stressed calli, a general decrease in all the antioxidant enzymes studied (except for glutathione reductase and dehydroascorbate reductase activities) was observed at day 28. Except for catalase, the antioxidant enzymes were elevated constitutively in adapted calli as compared to stressed cells, when both were grown in the absence of NaCl (time 0), and remained unaltered until 28 d after the beginning of the experiment. These results suggest the involvement of an enzymatic antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells.     

Basal leaf senescence in a sunflower (Helianthus annuus) canopy: responses to increased R/FR ratio

Senescence of lower leaves (LS) begins before anthesis in sunflower crop canopies. Using isolated field-grown sunflower plants, it has previously been shown that pre-anthesis LS is dependent on photosynthetic photon flux density (PPFD) and is hastened by increases in far-red light. We tested the hypothesis that increasing the red/far-red ratio (R/FR) perceived by basal leaves within canopies delays LS. To do this, light impinging on the lower surface of north-oriented 8th leaves (cotyledons=0) of crops with maximum leaf area indexes of 3.3 (Experiment 1) and 2.4 (Experiment 2) was enriched (+8.33 μmol m⁻² s⁻¹) with red light using light emitting diode (LED) panels. LED panels constructed with unlit LED or with green LED (PPFD slightly greater than the red LED panels, to compensate for lower efficiency) were used as controls. Compared with controls, additional R significantly ( P <0.05) increased R/FR perceived by the lower surface and significantly ( P <0.01) delayed LS. On average, leaf duration, as time between full expansion and a 70% diminution of chlorophyll content, was 5 days greater for leaves receiving extra red light (maximum observed LD=27 days). We conclude that an increase in the R/FR ratio can delay LS in crop canopies.     

Relationships between root permeability to water,leaf conductance and transpiration rate in sunflower (Helianthus annuus L.) cultivars

Summary The relations between leaf conductance (g_l) transpiration rate and root permeability to water (R_p) of three sunflower (Helianthus annuus L.) cultivars grown in a controlled environment cabinet are described.No differences in transpiration rates were found but it was shown that plants with low values of R_p have active stomatal closure with favourable consequences for water use efficiency under water limiting conditions.R_p was estimated by applying hydrostatic pressure on the root system. Values of R_p per unit root volume ranged from 0.34×10^–5 to 16.75×10^–5 (s MPa^–1). There were significant inter-cultivar differences (P<0.05) in R_p and g_l and an inverse correlation between R_p and the maximum values cf g_l within cultivars.Pressure applied on the root system is proposed as a useful tool for the determination of differences in the root permeability to water amongst sunflower cultivars.     

Transport of newly synthesized ribosome precedes initiation of S phase in root meristem cells in Helianthus annuus L

According to earlier results, cold treatment blocks most of cells in G1 phase; after 3 h of postincubation at 20 degrees C these cells initiate S phase. Simultaneous cytophotometric (DNA stained with Methyl Green) and autoradiographic (3H-uridine, 3H-arginine) analyses of cold pretreated cells have shown that transport of 3H-RNA into cytoplasm is faster in G2 than in G1 cells: radioactivity of cytoplasm is faster in G2 cells becomes higher than that of nucleoli as early as between 2 h and 2.5 h postincubation, while in G1 cells--not until between 2.5. and 3.0 h. Simultaneous cytophotometric measurements of DNA (Feulgen) and protein (Naphthol Yellow S) contents demonstrate the considerable increase in cytoplasmic protein contents during the 2nd h of postincubation at 20 degrees C; therefore it precedes the export of more than 50% of 3H-RNA synthesized after cold treatment. The results of these experiments indicate two separate events in G1 cells: increase in cytoplasmic protein amounts (up to 2nd h of postincubation) and enhanced transport of newly-synthesized ribosome (2.5 h of postincubation), the later event immediately precede the entry of cells into S phase.     

Involvement of an antioxidant defence system in the adaptive response to heavy metal ions in Helianthus annuus L. cells

A relationship between the antioxidant defence system and metal iontolerance in two types of sunflower callus differing in metal ion sensitivitywas studied. The antioxidant defence system of callus subjected to anadaptationtreatment of Cd(II), Al(III) and Cr(III) responded differently to 150M of each metal compared with the corresponding controls undershock treatment. The GSH/GSSG ratio remained similar to control values for thethree metal-acclimated calli and in the chromium shock treatment, decreasingmoderately in the acute treatment with cadmium and aluminum. In contrast, theAs/DAs ratio was decreased in the two different treatments for the three metalsions, but the decrease was greater with acute stress. The antioxidant enzymesresponded differently according to the metal and treatment used. In chromiumadapted callus, all antioxidant enzymes increased except for glutathionereductase. However, in the shock treatment ascorbate peroxidase activity wasdiminished with each metal ion assayed. Guaiacol peroxidase was decreased bycadmium and chromium and remained similar to control values with aluminum.Glutathione reductase was only decreased by cadmium, and superoxide dismutaseand catalase activities were less increased than in tolerant cells. Theseresults suggest the involvement of an antioxidant defence system in theadaptiveresponse to heavy metal ions in Helianthus annuus L.cells.     

向日葵单子叶和三子叶突变体的形态特征

Phyllotaxis and morphological characteristics of monocotyledonous and tricotyledonous mutants ( found during cultivation process) and dicotyledonous plants of Helianthus annuus Linn. were observed. The results show that numbers of monocotyledonous, dicotyledonous and tricotyledonous plants are 8, 339 and 3, respectively in 350 cultivated plants of H. annuus, accounting for 2. 3%, 96. 9% and 0. 9% of total individual number, respectively, and their corresponding phyllotaxis is alternate, opposite and whorled types, respectively. Monocotyledonous plant has the fewest leaf number, the most leaf layer number, and the largest flower diameter; tricotyledonous plant has the most leaf number, the fewest leaf layer number, and the smallest flower diameter. There is no obvious difference in plant height of individuals with different cotyledon types. It is suggested that cotyledon number of H. annuus is obviously correlated with phyllotaxis type.     

Response of sunflower (Helianthus annuus L) genotypes to PEG-mediated water stress

Drought tolerance of two sunflower (Helianthus annuus L.) genotypes, cultivated cultivar 1114 and interspecific line H. annuus × H. mollis, was studied under laboratory conditions using PEG-6000. Four levels of osmotic stress (?0.4, ?0.6, ?0.8 and ?1.0 MPa) were created and performances were monitored against a control. Physiological and biochemical stress determining parameters such as malondialdechyde (MDA), proline content, and hydrogen peroxide (H₂O₂) were compared between seedlings of both genotypes. The results indicated that both genotypes have similar responses at four osmotic potentials for all traits studied. All seedling growth parameters such as germination percentage, root length, shoot length, root and shoot dry weight decreased with increasing osmotic stress. MDA, proline, and H₂O₂ were found to be increased at different osmotic gradients in comparison to control. Cultivar 1114 was less affected than the interspecific line under these stress conditions. The data observed in the experiments revealed that perennial wild H. mollis can hardly be considered to be an excellent candidate of drought tolerance genes.     

The domestication of Helianthus annuus L. (sunflower)

All modern domesticated sunflowers can be traced to a single center of domestication in the interior mid-latitudes of eastern North America. The sunflower achenes and kernels recovered from six eastern North American sites predating 3000 b.p. that document the early history of this important crop plant are reanalyzed, and two major difficulties in the interpretation of archaeological sunflower specimens are addressed. First, achenes and kernels obtained from a modern wild sunflower population included in a prior genetic study because of its minimal likelihood for crop-wild gene flow, and its close genetic relationship to domesticated sunflowers, provide a new and more tightly drawn basis of comparison for distinguishing between wild and domesticated achene and kernel specimens recovered from archaeological contexts. Second, achenes and kernels from this modern wild baseline population were carbonized, allowing a direct comparison between carbonized archaeological specimens and a carbonized modern wild reference class, thereby avoiding the need for the various problematic shrinkage correction conversion formulas that have been employed over the past half century. The need for further research on museum collections is underscored, and new research directions are identified.     

Composition of lipids from sunflower pollen (Helianthus annuus)

The contents of the pollen lipids of the sunflower Helianthus annuus are described. The major component is the seco-triterpene helianyl octanoate, followed by new beta-diketones as second major group of compounds. They exhibit a shorter chain length and often other positions of the functional group compared to already known beta-diketones. Of particular note are the 1-phenyl-beta-diketones, not previously reported from nature. Further lipid classes present are related hydroxyketones and diols. Interestingly, new beta-dioxoalkanoic acids are present in the extracts, which most likely are biogenetic precursors of the diketones. Additionally, we investigated the composition of the pollen coat which resembles the total extract, but lacks the dioxoalkanoic acids and certain estolides.     

Stomatal response to drying soil in relation to changes in the xylem sap composition of Helianthus annuus. II. Stomatal sensitivity to abscisic acid imported from the xylem sap

Sunflower plants [Helianthus annuus L.) were subjected to soil drought. Leaf conductance declined with soil water content even when the shoot was kept turgid throughout the drying period. The concentration of abscisic acid in the xylem sap increased with decreasing soil water content. No general relation could be established between abscisic acid concentration in the xylem sap and leaf conductance due to marked differences in the sensitivity of leaf conductance of individual plants to abscisic acid from the xylem sap. The combination of these results with data from Gollan, Schurr & Schulze (1992, see pp. 551–559, this issue) reveals close connection of the effectiveness of abscisic acid as a root to shoot signal to the nutritional status of the plant.     

Influence of cytokinins on the methyl jasmonate-promoted senescence in Helianthus annuus cotyledons

Cotyledons of seven-day-old sunflower seedlings were pretreated withcytokinins prior to exposure to methyl jasmonate (MeJA). The rate of senescencewas then followed by measuring chlorophyll loss, electrolyte leakage, ethyleneproduction, and 1-aminocyclopropane-1-carboxylase (ACC) oxidase activity. MeJApromoted all these senescence parameters and the MeJA effects were partiallyblocked by cytokinin pretreatment. 8-azaguanine, which has been reported tohaveanticytokinin activity, blocked the ability of benzyl adenine (BA) to reversethe effect of MeJA on senescence. MeJA also increased SOD and catalaseactivity,decreased protein content. However, while the cytokinin BA more than overcamethe MeJA effect on protein content and SOD activity it did not antagonize theeffect of MeJA on catalase.     

Ent-Kaurene Biosynthesis in Extracts of Helianthus annuus L. Seedlings

Kaurene synthetase B activity (conversion of copalyl pyrophosphate to ent-kaurene) is readily detectable in crude cell-free extracts of 3- to 4-day old dark-grown sunflower (Helianthus annuus cv. Mammoth) seedlings, whereas little or no kaurene synthetase AB activity (conversion of geranylgeranyl pyrophosphate to ent-kaurene) can be found in these extracts under comparable assay conditions. A low amount of AB activity is evident only if an extensively dialyzed extract is used in low concentrations as the enzyme source. One factor which may contribute to the low apparent levels of AB activity is the presence of inhibitory factors in the crude sunflower extract since these extracts can be shown to act as a potent inhibitor of Marah macrocarpus endosperm kaurene synthetase AB activity. Heat treatment (100°C) or dialysis of the sunflower extract reduces the amount of its inhibitory activity. Also, it was observed that low concentrations of extensively dialyzed sunflower extracts act to stimulate M. macrocarpus AB activity. There is no evidence for the presence of an inhibitory factor for M. macrocarpus kaurene synthetase B activity in sunflower extracts. However, there does appear to be present in the crude preparation of sunflower extract a dialyzable factor(s) that impedes its own B activity. There is little information to date on the nature of these inhibitory and stimulatory factors for kaurene synthetase activity or their possible roles in physiological regulation. The possible presence of such factors should be considered, however, when attempting to evaluate kaurene synthetase activities in extracts of vegetative plants.     

Longitudinal variation in cadmium influx in sunflower (Helianthus annuus L.) roots as depending on the growth substrate,root age and root order

Aims

In a previous work, we observed a longitudinal decrease in Cd²⁺ influx starting from the root tip in first order lateral roots of sunflower (Helianthus annuus L.) grown in hydroponics. This variable influx was expected to impact the total Cd²⁺ uptake depending on the root system architecture and on how steep was the decrease of the influx. Here, we examined the influence of the culture substrate, of the age and order of lateral roots on the longitudinal variation of Cd²⁺ influx.

Methods

By using short-term exposures to ¹⁰⁹Cd-labelled solution (5 to 200 nM), we compared the longitudinal variations in Cd²⁺ roots influx depending on the growth substrate (hydroponics or sand), on the root age and order.

Results

In second order laterals, Cd²⁺ influx decreased from the apex to the root base, as for first order laterals. For sand cultures compared to hydroponics, the mean Cd²⁺ influx was lower and decreased more steeply with the distance from the apex. The influx also decreased with increasing root age and order, markedly in hydroponics but less for sand cultures.

Conclusion

Results suggested that for a given root surface area, the Cd²⁺ uptake by a root system should increase with increasing number of root tips and decreasing individual root length.