Maintenance and growth respiration in shoots and roots of sunflower plants grown at different root temperatures

Continuous measurements of CO₂-evolution and dry matter accumulation were carried out on shoots and roots separately of intact Helianthus annuus L. cv. Autumn Beauty plants grown in nutrient solution at different root temperatures. The data were used to distinguish between growth and maintenance components of respiration. The maintenance and growth coefficients were higher in the root system than in the shoots. The overall efficiency of assimilate utilization was within the range reported in the literature. An increase in root temperature increased the maintenance part of root respiration and, to a lesser degree, also shoot maintenance respiration. Neither root nor shoot growth respiration coefficients were affected by root temperature. It is concluded that the study of whole-plant respiration masks differences in energy utilization between shoots and roots.     

Leaf expansion as related to plant water availability in a wild and a cultivated sunflower

This study aimed to determine if two species of sunflower, Helianthus annus L. cv. Hysun 31 (cultivated, single-stemmed genotype) and Helianthus petiolaris Nuttall ssp. fallax (wild, many-hranched genotype) differed in the response of leaf growth to water deficits. Earlier published studies, concerned only with H. annuus, failed to reveal differences in the response of sunflowers to water stress. Plants of the two species were paired in large containers of soil and grown under high radiation in a glasshouse. One batch of plants was irrigated and the other allowed to dry so that predawn leaf water potentials declined at an average of 0.072 MPa day^?1. The dry batch was rewatered when predawn leaf water potentials reached ?0.85 MPa. The stress imposed was sufficient to curtail leaf growth so that plants in the dry treatment had only 60% of the leaf area of irrigated plants at the onset of rewatering. Both species were affected by stress to the same relative extent, though their leaf areas at this stage differed 7-fold. Both genotypes also recovered to the same degree in the long term, finally having leaf areas and gross dry matter distribution patterns which were indistinguishable from plants which were irrigated throughout. However, water stress resulted in different distribution patterns of leaf area: H. annuus produced larger leaves at the top of its single stem which compensated for the reduced area in lower leaves, whereas H. petiolaris compensated in the leaves on its branches. Leaves which emerged after the time of stress were most able to compensate in area subsequently. For example, those leaves of H. annuus which emerged one week after stress-relief were more than three times larger than comparable leaves on plants irrigated continuously. Leaf expansion rates were affected earlier in the stress cycle than leaf conductance in H. annuus, but not in H. petiolaris. But as with other plant responses to water stress, the differences between the two species were small.     

Effect of abscisic acid on exudation of sunflower roots as affected by nutrient status, glucose level and aeration

Three-week-old sunflower plants ( Helianthus annuus L. cv. Halcón) grown in nutrient solution at two K⁺ levels (0.3 and 2.5 m M ) were used to study the effect of 4 μ M abscisic acid (ABA) on the transport of K⁺ (Rb⁺) and water to the exuding stream of decapitated plants. Other conditions of the bathing medium of the roots were also assayed, such as presence of 10 m M glucose, aeration and time of ABA application. In the first 2 or 3 h after ABA application, ABA always promoted water and ion fluxes, even under the most unfavorable conditions such as low K⁺ roots without glucose or under anaerobiosis. The ABA-promoting effect on ion and water flow was higher with glucose in the medium. Under anaerobiosis the ABA effect disappeared after 3 h. With glucose and aeration the ABA-promoting effect appeared early and continued for several hours, although the effect decreased with time. If ABA was applied 24 h before excision, the effect was small or even negative. We suggest that ABA acts directly on membranes of certain root cells (endodermal or both endodermal and cortical cells) by increasing their permeability and thus releasing ions. This will decrease cell turgor pressure and, indirectly, the hydraulic conductivity of the whole root. Under conditions of higher hydraulic conductivity, the presence of ions and glucose in the root stimulates the transport of ions into the xylem. and thus increases the osmotic water flow.     

Induction of phenylalanine ammonia-lyase in hypocotyls of sunflower seedlings by light, excision and sucrose

Light, excision and sucrose increased extractable phenylalanine ammonia-lyase (PAL) activity from hypocotyl tissue of sunflower ( Helianthus annuus L. cv. Peredovik) to 2–6 times the basal level. Intact sunflower seedlings or whole hypocotyls incubated in water or 0.1 M sucrose exhibited, in continuous light, a pattern in which PAL peaked 4 and 28 h after the beginning of the illumination. When 0.5 cm long hypocotyl segments were incubated in water or 0.1 M sucrose, they exhibited, both in continuous light and in the dark, a pattern in which PAL rose during an initial period of 10 h (assay in sucrose and light) to 48 h (assay in water and dark) and then remained nearly constant at a high value for at least the next 10 h. When whole hypocotyls were incubated in 0.1 M sucrose, a third pattern in PAL activity was found in which PAL peaked after 28 h and subsequently declined. In all the above systems the increase in PAL activity was significantly reduced by cycloheximide. Furthermore, the subsequent decay of PAL activity following illumination was prevented by delayed transfer to cycloheximide. It is suggested that the results can be explained on the basis of a turnover mechanism involving continued de novo enzyme synthesis and subsequent synthesis of a PAL-inactivating system.     

Cloning and characterization of a root sunflower peroxidase gene putatively involved in cell elongation

A cDNA-encoding a peroxidase (Helianthus annuus POX (HaPOX)1) was isolated and characterized from the roots of sunflower seedlings. This gene exhibited homology with other peroxidases from several higher-plant species, and its expression in the root growth was particularly abundant during cell expansion. To elucidate the precise functions of HaPOX1 in sunflower root, we examined its expression pattern in response to several plant growth regulators. Expression of HaPOX1 is down-regulated by abscisic acid (ABA), whereas indole-3-acetic acid (IAA) induced its expression. These results suggest that HaPOX1 expression is differentially regulated by phytohormonal components of signaling cascades. Since IAA appears to participate in the regulation of HaPOX1 expression, we postulate that the peroxidase encoded by HaPOX1 may be involved in the reactions that promote cell elongation during the early stage of root growth.     

Glyphosate inhibition of ferric reductase activity in iron deficient sunflower roots

Iron (Fe) deficiency is increasingly being observed in cropping systems with frequent glyphosate applications. A likely reason for this is that glyphosate interferes with root uptake of Fe by inhibiting ferric reductase in roots required for Fe acquisition by dicot and nongrass species. This study investigated the role of drift rates of glyphosate (0.32, 0.95 or 1.89 mm glyphosate corresponding to 1, 3 and 6% of the recommended herbicidal dose, respectively) on ferric reductase activity of sunflower (Helianthus annuus) roots grown under Fe deficiency conditions. Application of 1.89 mm glyphosate resulted in almost 50% inhibition of ferric reductase within 6 h and complete inhibition 24 h after the treatment. Even at lower rates of glyphosate (e.g. 0.32 mm and 0.95 mm), ferric reductase was inhibited. Soluble sugar concentration and the NAD(P)H oxidizing capacity of apical roots were not decreased by the glyphosate applications. To our knowledge, this is the first study reporting the effects of glyphosate on ferric reductase activity. The nature of the inhibitory effect of glyphosate on ferric reductase could not be identified. Impaired ferric reductase could be a major reason for the increasingly observed Fe deficiency in cropping systems associated with widespread glyphosate usage.     

Iron deficiency stress induced morphological and physiological changes in root tips of sunflower

Typical morphological and physiological changes were observed in iron deficient sunflower ( Helianthus annuus L. cv. Sobrid) roots. These changes, or so-called iron stress reactions, are exclusively confined to the root tips. Typical morphological changes included additional cell division in the rhizodermis layer and enhanced formation of root hairs, leading to an increase in root diameter ("swollen root tips"). These morphological changes were correlated with physiological changes such as increased release of protons, accumulation of phenols in the rhizodermis, and an increased ability of the roots to reduce iron-III compounds ("reducing capacity"). A marked increase in ability of the root tips to take up and translocate iron occurred simultaneously with these changes. There is good evidence that these morphological and physiological changes are reflections of an effective regulatory mechanism for enhanced mobilization of sparingly soluble iron-III compounds in the rhizosphere and for iron uptake by sunflower plants.     

Effects of nitrate on influx, efflux and translocation of potassium in young sunflower plants

Influx, efflux and translocation of K⁺(⁸⁶Rb) were studied in the roots of sunflower seedlings ( Helianthus annuus L. cv. Uniflorus) treated with 0–4.0 m M NO₃⁻ during a 9 day growth period or a 24 h pretreatment period. Roots treated with high levels of NO₃⁻ absorbed and translocated more K⁺(⁸⁶Rb) than seedlings treated with low levels of NO₃⁻. The content of K⁺ in the shoots was, however, higher in seedlings treated with low levels of NO₃⁻, indicating a low rate of retranslocation of K⁺ in those plants. K⁺(⁸⁶Rb) efflux was highest into the low-NO₃⁻ solutions. All effects on K⁺(⁸⁶Rb)-fluxes were more obvious in high-K plants than in low-K plants. The results are discussed in relation to the Dijkshoorn-Ben Zioni hypothesis for K⁺+ NO₃⁻-uptake and translocation in plants.     

Early degradation of photosynthetic membranes in carob and sunflower cotyledons

The assimilatory activity of cotyledons can play an essential role in the survival of seedlings with a slow and delayed development of primary leaves. Changes in the photosynthetic activity of the cotyledon, from the onset of greening through senescence, were studied in two such plants, carob and sunflower, in order to determine its efficiency and duration, also in connection with the achievement of assimilatory autonomy by the plantlet. Chlorophyll analyses showed that the cotyledon's chloroplasts reached maximal greening in plantlets with a pair of expanded leaves. In contrast, the cotyledon's photosynthetic activity, measured as the rate of oxygen release, started to decrease early, before expansion of primary leaves. The decrease was due to the inactivation of a number of photosystem II (PSII) units, as revealed by immunodetection of breackdown products of the reaction centre's D1 and D2 thylakoid proteins. No signals of PSII alteration were noticed in the primary leaf chloroplasts that differentiated under the same environmental conditions. The damage to the cotyledon PSII, occurring in a non-photoinhibitory situation, might be due to a slower rate of turnover of D1 polypeptide than in the leaf thylakoids. The differential turnover of this protein in cotyledons and in leaves might represent an organ-specific regulation of the photosynthetic activity. The peculiarity of the cotyledon thylakoids make these organs useful objects for studying the metabolic cycle of both D1 and D2 proteins in vivo, under non-photoinhibiting conditions.     

Lipid composition of plasma membranes isolated from sunflower seedlings grown under water-stress

Plasma membranes were isolated by aqueous two-phase-partitioning from sunflower ( Helianthus annuus cv. Isabel) seedlings grown both under field irrigation and dryland conditions. Water-stressed plants showed a decrease in the leaf water potential and in the osmotic potential at full turgor, with the turgor pressure remaining at positive values. Dryland conditions also induced a reduction in the bulk modulus of elasticity. Plasma membranes of irrigated plants were characterized by high contents of phospholipids (68% of total lipids), free sterols (15. 7%) and glycolipids (9. 1%), mainly glycosphingolipids and steryl glycosides. Diacylglycerols, triacylglycerols and free fatty acids were also present. The major phospholipids were phosphatidylcholine and phosphatidylethanolamine with smaller amounts of phosphatidylinositol and phosphatidylglycerol. Following water stress, the plasma membranes showed a reduction of about 24 and 31% in total lipids and phospholipids, respectively. Also the amounts of glycolipids and diacylglycerols decreased significantly upon water stress. There was no change in free fatty acids, however, and triacylglycerols and free sterols increased. As a consequence, the free sterol to phospholipid molar ratio increased from 0. 4 to 0. 7 under water deficit conditions. The ratio of phosphatidylcholine to phosphatidylethanolamine increased from 1. 1 (control plants) to 1. 6 (water-stressed plants), while phosphatidic acid rose to 4% of total phospholipids. Dehydration did not result in any substantial change in the unsaturation level of the individual lipid classes, however. The results show that dryland conditions resulted in a marked alteration in the lipid composition of the sunflower leaf plasma membrane     

Influence of the g-force on the circumnutations of sunflower hypocotyls

Circumnutations of hypocotyls of sunflower ( Helianthus annuus L. cv. Californicus) were studied under 1 g and 3 g conditions. Root mean square values of the hypocotyl deviation from the plumbline and period of the movements were determined from Calculations of the autocorrelation functions of the movements. The amplitude and the period of the circumnutations increased under 3 g as compared to 1 g . A transition from 3 to 1 g or vice versa also caused changes in period and amplitude of the movoments. The results are interpreted as a support for the idea that gravity influences the circumnutation parameters in this sunflower variety. A comparison is made with published results on the dwarf sunflower ev. Teddy Bear where the force influence is very small or negligible. Simulations of a model for circumnutations show movements which are in qualitative agreement with the experimental results, provided adaptation to g -levels is included in the model. Finally, the results are discussed with the recent Spacelab-experiment (SLI) as a background.     

Relationship between virulence and aggressiveness in Plasmopara halstedii (sunflower downy mildew)

Relationship between virulence and aggressiveness was studied in seven Plasmopara halstedii (sunflower downy mildew) pathotypes including five progeny pathotypes of races 300, 304, 314, 704 and 714 arising from two parental pathotypes of races 100 and 710. Aggressiveness criteria including percentage infection, latent period, sporulation density and reduction of hypocotyl length were analysed in one sunflower inbred line showing a high level of quantitative resistance. There were significant differences between P. halstedii pathotypes for all aggressiveness criteria. Pathogenicity of progeny pathotypes as compared with parental ones (relationship between virulence and aggressiveness) seems to be positive, negative or uncorrelated. Hypothesis explaining these cases are discussed.     

Measurements of pH in the apoplast of sunflower leaves by means of fluorescence

A method was elaborated by which the pH in leaf apoplast can be measured. The technique is based on the pH dependent fluorescence of 5-carboxyfluorescein (5-CF) or fluorescein isothiocyanate (FITC). The fluorescein isothiocyanate is coupled with a macromolecular dextran molecule (FITC-dextran). For eliminating the effect of the absolute dye concentration the dual excitation technique was applied. It was shown that the ratio of fluorescence excited by light of 491 nm and 463 nm was virtually independent of the concentration of 5-CF and that this fluorescence ratio was related to the pH. The plasmalemma is practically impermeable to FITC-dextran and in the test we carried out over a period of 6 h not the slightest indication was found that it may penetrate the plasma membrane. For 5-CF this cannot be ruled out completely. It is possible that at pH values below 4.5 it may penetrate biological membranes at low rates.
Experiments with leaves of sunflower ( Helianthus animus cv. Erika) perfused with 5-carboxyfluorescein and supplied with different nitrogen forms showed that NH⁺₄ application resulted in a decrease and NO⁺₃ application in an increase of the leaf apoplast pH. Leaf spraying with fasicoccin was followed by a pH decrease, while leaf spraying with the protonophores p -trifluoromethoxy carbonytcyanide phenylhydra-zon (FCCP) or nigericin resulted in neutral apoplastic pH. These results provide evidence that the method is well suited for measuring the response of the leaf apoplast pH to changing physiological conditions.     

Genetic distance as a predictor of heterosis and hybrid performance within and between heterotic groups in sunflower

Heterosis is significant for seed yield and is one of the driving forces behind the hybrid seed industry in cultivated sunflower (Helianthus annuus L). Heterotic groups in sunflower, if any other than the female and male inbred-line groups exist, have not been well studied or described. The primary aims of this study were to assess the utility and validity of a series of proposed heterotic groups and estimate correlations between genetic distance, heterosis, and hybrid performance for seed yield in sunflower. Fortytwo female by male heterotic group (A × R) and 81 female by female heterotic group (A × B) single-cross hybrids were grown in Corvallis, Ore., and Casselton, N.D., in 1996 and 1997. Heterosis was significant for seed yield and plant height but not for seed oil concentration and days to flowering. Genetic distances were significantly correlated with hybrid seed yield when estimated from AFLP fingerprints (G _D) (r = 0.63 for A × R and 0.79 for A × B hybrids), but not from coancestries (G _C) (r = -0.02 for A × R and 0.54 for A × B hybrids). G _D (R ² = 0.4) was a poor predictor of hybrid seed yield. The proposed heterotic groups in sunflower seem to have utility, but do not seem to be as strongly differentiated as those in corn (Zea mays L.). The highest-yielding hybrids were from the B_C× R_B heterotic pattern; however, several B_C× B_C hybrids (within-group hybrids) were among the top-yielding hybrids. The outstanding performance of certain B_C× B_C hybrids casts some doubt on the validity of the B_C group. Substantial genetic diversity seems to be present within and between heterotic groups in sunflower. Received: 1 September 1998 / Accepted: 14 September 1999     

Aggressiveness variation in Plasmopara halstedii and its alternation with non-race-specific resistance in sunflower

Aggressiveness variation and its alternation with non-race specific resistance in sunflower were studied in 19 Plasmopara halstedii isolates belonging to several races. Regarding aggressiveness criteria, percentage infection, latent period, sporulation density and dwarfing, on two sunflower inbred lines showing different levels of non-race specific resistance resistance FU and BT, there were significant differences in aggressiveness for P. halstedii isolates. The index of aggressiveness varied between 9.4 and 31.4. The inbred line BT, rather susceptible in the field, showed a higher percentage infection, a higher sporulation density, a shorter latent period and less reduced hypocotyl length than inbred line FU, which showed a greater resistance in the field. Percentage infection on FU was 1.4% less than BT, latent period on BT was 12.4% less than FU, sporulation density on FU was 22.3% less than BT and reduced hypocotyl length on BT was 15.3% less than FU. Consequently, it seems that the criteria as latent period, sporulation density and reduction of hypocotyl length could be used to measure non-race specific resistance in sunflower to P. halstedii under controlled conditions.     

Chitinase and peroxidase activities in sunflower hypocotyls: Effects of BTH and inoculation with <Emphasis Type="Italic">Plasmopara halstedii</Emphasis>

Systemic acquired resistance (SAR) can be induced in plants by incompatible pathogens, pathogen derived extracts, or certain chemicals as benzothiadiazole (BTH). The aim of this work was to compare changes in peroxidase and chitinase activities, enzymes considered as PR-proteins, caused by BTH and the pathogen Plasmopara halstedii. Hypocotyls from susceptible and resistant BTH-treated sunflower seedlings showed increased peroxidase and chitinase activities. Inoculation with P. halstedii increased chitinase and peroxidase activities in inoculated hypocotyls from susceptible but not from resistant sunflower seedlings.     

Trichoderma harzianum antagonistic activity and competition for seed colonization against seedborne pathogenic fungi of sunflower

This study investigated the antagonistic effects of Trichoderma harzianum isolate (TRIC8) on mycelial growth, hyphal alteration, conidial germination, germ tube length and seed colonization by the seedborne fungal pathogens Alternaria alternata, Bipolaris cynodontis, Fusarium culmorum and F. oxysporum, the causes of seedling rot in over 30% of sunflowers. The antagonistic effect of TRIC8 on mycelial growth of pathogens was evaluated on dual culture that included two inoculation assays: inoculation of antagonist at 48 h before pathogen (deferred inoculation) and inoculation at the same time with pathogen (simultaneous inoculation). TRIC8 inhibited mycelial growth of the fungal pathogens between 70·67 and 76·87% with the strongest inhibition seen with deferred inoculation. Alterations in hyphae were observed in all pathogens. Conidial germination of F. culmorum was inhibited by most of the fungal pathogens (38·28%) by TRIC8. Inhibition of germ tube length by the antagonist varied from 31·83 to 37·67%. In seed colonization experiments, TRIC8 was applied in combination with each pathogen to seeds of a sunflower genotype that is highly tolerant to downy mildew. Seed death was inhibited by TRIC8 and the antagonist did not allow growth of A. alternata, B. cynodontis and F. culmorum on seeds and inhibited the growth of F. oxysporum at the rate of 58·32%.