The hormonal control of wheat leaf unrolling

Summary The unrolling of etiolated wheat leaf sections in the dark is stimulated by the application of gibberellic acid (GA₃). GA₃ is most effective if applied for a short time at the beginning of incubation. Kinetin also stimulated leaf unrolling in the dark. AMO1618 and CCC inhibit red light and kinetin-stimulated unrolling. Gibberellin-like substances extracted from red light-treated leaf tissue are effective in stimulating leaf unrolling.Ethylene production in leaf sections is stimulated by IAA, GA₃ and kinetin and inhibited by ABA. A brief exposure to red light decreases the ability of the tissue to produce ethylene. It is concluded that ethylene plays no important role in the control of leaf unrolling by red light or by the application of hormones.Holder of a Science Research Council Studentship.     

The effect of δ-aminolevulinic acid on red light-induced unrolling of dark-grown barley leaf sections

Unrolling of sections from dark grown barley leaves ( Hordeum vulgare L. cv. Klages) was stimulated by red irradiation. The unrolling started after a lag phase of 6-8 h and was completed after 24 h. The effect of several keto and amino acids on leaf unrolling was compared with their effect on coleoptile segment expansion growth. Of the substances tested δ-aminolevulinic acid had the most inhibitive effect on leaf unrolling and the least inhibitive effect on coleoptile segment expansion growth. Prolonged treatment caused a strong inhibition of the unrolling but with a short tretment, a stimulation of the unrolling occurred. The inhibitive effect of δ-aminolevulinic acid was only found if the treatment started within 6-8 h after the red irradiation. Kinetin and gibberellic acid could decrease the inhibitory effect of δ-aminolevulinic acid. A possible role for δ-aminolevulinic acid working as a triggering substance for leaf unrolling is discussed.     

Brassinosteroid leaf unrolling QTL mapping in durum wheat

Brassinosteroids are a newly reported class of plant growth phytohormones found in plants throughout the plant kingdom. Functioning at very low concentrations, they play an essential role in improving biomass yield and stress tolerance. There are no reports in the literature of the genetic variability of responsiveness of brassinosteroids in wheat; most studies on brassinosteroids have focused on the physiological effects of exogenous addition of brassinosteroids. Our aim was to study the genetic variation in the responsiveness of a doubled haploid durum wheat population to three brassinosteroid concentrations using the leaf unrolling test, which is a simple bioassay to test brassinosteroid activity. An F₁-derived doubled haploid population of 77 individuals from the cross Strongfield/Blackbird was used to construct a genetic map of 427 molecular marker loci. The leaf unrolling test was performed on the parents and doubled haploid genotypes of the population using 0.2, 2 and 20 nM brassinosteroid concentrations. The results indicated significant differences in leaf unrolling between the two parents, doubled haploid genotypes, treatments and genotype-by-treatment combinations. Transgressive segregation beyond Strongfield of leaf unrolling was observed for all concentrations, with the strongest response at 20 nM. Putative quantitative trait loci were revealed in the intervals Xgwm2–Xbarc45 on chromosome 3A and Xwmc643a–Xwmc625a on chromosome 3B. Additional quantitative trait loci were associated with markers Xwmc48a, Xwmc511, Xwmc89a and Xgwmc692 on chromosome 4B, and Xwmc17 on chromosome 7A. This work should enhance the understanding of the relationship between stress tolerance and productivity, and responsiveness to brassinosteroids.     

The light-induced unrolling of the grass leaf — A study of polarity, light-piping and stimulus transmission

Primary leaves of dark-grown barley ( Hordeum vulgare L. cv. Weibull's Ida) unroll in darkness when cut into sections shorter than about 2 cm; the shorter the more unrolling. The unrolling pattern of irradiated leaf sections longer than about 2 cm indicates a polarity in the leaves: the top end of the basal section of a divided leaf unrolls more than the base end of the distal section. Earlier reported findings of a stimulus transmission from irradiated to non-irradiated areas can be fully explained by light-piping and by mechanical, mutual influence between irradiated and non-irradiated areas.     

Phytochrome-controlled Leaf Unrolling and Protein Synthesis

In the primary leaf sections of etiolated wheat (Triticum aestivum L.) seedlings, red light-induced unrolling is accompanied by an increase in incorporation of ¹⁴C-leucine into protein. By differential centrifugation, the unrolling response was found to be closely related to incorporation of the amino acid into the supernatant fraction (105,000g). Cycloheximide and chloramphenicol inhibit both leaf unrolling and synthesis of the supernatant protein, although chloramphenicol exerts its effect more strongly on the fraction which presumably contains the plastids. In a barley (Hordeum vulgare L.) albino mutant completely devoid of ribulose diphosphate carboxylase activity, only incorporation of ¹⁴C-leucine into the supernatant fraction is substantially promoted by red light. This mutant exhibits the photoresponse of leaf unrolling.     

Effects of vanadate on the plasma membrane ATPase of red beet and corn

The effect of vanadate on the plant plasma membrane ATPase were investigated in plasma membrane fractions derived from corn roots (Zea mays L.) and red beets (Beta vulgaris L.). The K_i for vanadate inhibition of the plasma membrane ATPase from corn roots and red beets was between 6 and 15 micromolar vanadate. In both membrane fractions, 80% to 90% of the total ATPase was inhibited at vanadate concentrations below 100 micromolar. Vanadate inhibition was optimal at pH 6.5, enhanced by the presence of K⁺, and was partially reversed by 1 millimolar EDTA. The Mg:ATP kinetics for the plasma membrane ATPase were hyperbolic in both the absence and presence of vanadate. Vanadate decreased both the K_m and V_max of the red beet plasma membrane ATPase, indicating that vanadate inhibits the ATPase uncompetitively. These results indicate many similarities with respect to vanadate inhibition between the plant plasma membrane ATPase and other major iontranslocating ATPases from fungal and animal cells. The high sensitivity to vanadate reported here, however, differs from other reports of vanadate inhibition of the plant plasma membrane ATPase from corn, beets, and in some instances oats.     

Effect of growth temperature and temperature shifts on spinach leaf morphology and photosynthesis

The growth kinetics of spinach plants (Spinacia oleracea L. cv Savoy) grown at 5°C or 16°C were determined to allow us to compare leaf tissues of the same developmental stage rather than chronological age. The second leaf pairs reached full expansion at a plant age of 32 and 92 days for the 16°C and 5°C plants, respectively. Growth at 5°C resulted in an increased leaf area, dry weight, dry weight per area, and leaf thickness. Despite these changes, pigment content and composition, room temperature in vivo fluorescence, and apparent quantum yield and light-saturated rates of CO₂ exchange or O₂ evolution were not affected by the growth temperature. Furthermore, 5°C expanded leaves were found to be more resistant to photoinhibition at 5°C than were 16°C expanded leaves. Thus, it is concluded that spinach grown at low temperature is not stressed. However, shifting spinach leaves from 5°C to 16°C or from 16°C to 5°C for 12 days after full leaf expansion had occurred resulted in a 20 to 25% reduction in apparent quantum yields and 50 to 60% reduction in light saturated rates of both CO₂ exchange and O₂ evolution. This was not accompanied by a change in the pigment content or composition or in the room temperature in vivo fluorescence. It appears that leaf aging during the temperature shift period can account for the reduction in photosynthesis. Comparison of cold-hardened and non-hardened winter rye (Secale cereale L. cv Muskateer) with spinach by in vivo fluorescence indicated that rye is more sensitive to both short term and longer duration temperature shifts than is spinach. Thus, susceptibility to an abrupt temperature shift appears to be species dependent.     

Deposition pattern of hydrogen peroxide in the leaf sheaths of rice under salt stress

Deposition pattern of hydrogen peroxide (H₂O₂) under salt stress (100 mM NaCl) was examined cytochemically in rice (Oryza sativa L. cv. Pokkali) through the reaction of H₂O₂ with cerium chloride (CeCl₃) to produce electron dense precipitates of cerium perhydroxide. The distribution pattern of cerium perhydroxide precipitates in leaf sheath was considerably different from other parts of rice under salinity stress. Cerium perhydroxide precipitates were mainly accumulated on the tonoplast of leaf sheath under salinity, although they were localized on the cell wall and plasma membrane in all other tissues such as leaf blade and root.     

Ozone-induced ethylene release from leaf surfaces

Ozone-induced stress ethylene emissions from the adaxial and abaxial leaf surfaces of four plant species (Glycine max [L.] Merr. cv. Dare, Lycopersicon esculentum Mill cv. Roma VF, Eucalyptus globulus Labill. and Hedera helix L.) were studied to determine if the stress ethylene diffused through the stomata or cuticle. In plants not exposed to ozone, basal ethylene was detected above both the adaxial and abaxial leaf surfaces of all the plant species examined, indicating that some ethylene can diffuse across the leaf cuticle. Ozone induced stress ethylene production in all species examined. Significant ozone-induced ethylene concentrations were detected above both surfaces of amphistomatous soybean (Glycine) and tomato (Lycopersicon) leaves. In contrast, ozone-induced ethylene production was associated only with the leaf surface (abaxial) that contained stomata for hypostomatous blue gum eucalyptus and English ivy (Hedera) leaves; the leaf surface (adaxial) of the eucalyptus and ivy leaves which did not contain stomata did not release significant amounts of stress ethylene. These data indicate that ozone-induced stress ethylene primarily diffuses from the leaf via the stomata.     

Purification and characterization of a soybean leaf storage glycoprotein

Removing the pods from soybean (Glycine max [L.] Merr. cv Wye) plants induces a change in leaf function which is characterized by a change in the leaf soluble protein pattern. The synthesis of at least four polypeptides (~27, 29, 54, and 80 kilodaltons) is enhanced, and these polypeptides accumulate to levels comprising over 50% of the soluble protein. Heat girdling the petiole also causes the accumulation of these polypeptides, suggesting that the signal for changing leaf function may be associated with inhibition of phloem transport. The 27 and 29 kilodalton polypeptides are glycosylated and have been purified to greater than 90% by (NH₄)₂SO₄ fractionation, concanavilin A affinity, and gel filtration chromatography. These peptides appear to comprise a single protein. Mouse antiserum has been prepared against this glycoprotein and has been used to check for cross-reactivity with seed proteins and to quantitate changes with leaf development. No cross-reactivity was observed with seed soluble proteins from several stages of development. Quantitation showed the highest content in podded plants at, and shortly following, flowering, with levels subsequently declining in conjunction with seed growth. In depodded plants, the level of glycoprotein continued to increase following flowering and accounted for 45% of the soluble leaf protein by 4 weeks after depodding.     

Effects of fully open-air [CO2] elevation on leaf ultrastructure, photosynthesis, and yield of two soybean cultivars

The objective of this study was to investigate the effect of elevated (550 ± 17 ??mol mol^?1) CO₂ concentration ([CO₂]) on leaf ultrastructure, leaf photosynthesis and seed yield of two soybean cultivars [Glycine max (L.) Merr. cv. Zhonghuang 13 and cv. Zhonghuang 35] at the Free-Air Carbon dioxide Enrichment (FACE) experimental facility in North China. Photosynthetic acclimation occurred in soybean plants exposed to long-term elevated [CO₂] and varied with cultivars and developmental stages. Photosynthetic acclimation occurred at the beginning bloom (R1) stage for both cultivars, but at the beginning seed (R5) stage only for Zhonghuang 13. No photosynthetic acclimation occurred at the beginning pod (R3) stage for either cultivar. Elevated [CO₂] increased the number and size of starch grains in chloroplasts of the two cultivars. Soybean leaf senescence was accelerated under elevated [CO₂], determined by unclear chloroplast membrane and blurred grana layer at the beginning bloom (R1) stage. The different photosynthesis response to elevated [CO₂] between cultivars at the beginning seed (R5) contributed to the yield difference under elevated [CO₂]. Elevated [CO₂] significantly increased the yield of Zhonghuang 35 by 26% with the increased pod number of 31%, but not for Zhonghuang 13 without changes of pod number. We conclude that the occurrence of photosynthetic acclimation at the beginning seed (R5) stage for Zhonghuang 13 restricted the development of extra C sink under elevated [CO₂], thereby limiting the response to elevated [CO₂] for the seed yield of this cultivar.     

Response of three broccoli cultivars to salt stress, in relation to water status and expression of two leaf aquaporins

The aim of this study was to compare differences in water relations in the leaves of three broccoli cultivars and differential induction of the expression of PIP2 aquaporin isoforms under salt stress. Although broccoli is known to be moderately tolerant to salinity, scarce information exists about the involvement of leaf aquaporins in its adaptation to salinity. Thus, leaf water relations, leaf cell hydraulic conductivity (Lp_c), gas exchange parameters and the PIP2 expression pattern were determined for short- (15 h) and long- (15 days) term NaCl treatments. In the long term, the lower half-time of water exchange in the cells of cv. Naxos, compared with Parthenon and Chronos, and its increased PIP2 abundance may have contributed to its Lp_c maintenance. This unmodified Lp_c in cv. Naxos under prolonged salinity may have diluted NaCl in the leaves, as suggested by lower Na⁺ concentrations in the leaf sap. By contrast, the increase in the half-time of water exchange and the lower PIP2 abundance in cvs. Chronos and Parthenon would have contributed to the reduced Lp_c values. In cv. Parthenon, there were no differences between the ε values of control and salt-stressed plants; in consequence, cell turgor was enhanced. Also, the increases in BoPIP2;2 and BoPIP2;3 expression in cv. Chronos for the short-term NaCl treatment suggest that these isoforms are involved in osmotic regulation as downstream factors in this cultivar, in fact, in the short-term, Chronos had a significantly reduced osmotic potential and higher PIP2 isoforms expression.     

Effect of different leaf age on the relationship between the CO2 uptake and water vapour efflux in tobacco plants

CO₂ absorption (PAT) and transpiration (E) rates, and leaf diffusion resistance (r_i) were individually studied in all leaves of tobacco plants (Nicotiana tabacum L. cv. Wisconsin 38) before flowering. Differences between old, middle age and young leaves were in all characteristics studied and found statistically significant. In all three leaf age groups E was closely correlated to r_i. No similar correlation was discovered between P_N and r_i. The highest ratiosP _N /E in young and middle age leaves indicate that the increase of the internal resistance to photosynthesis with leaf age was more rapid than that of r_i.     

An analysis of the light-induced unrolling of the grass leaf

The light-induced unrolling of primary leaves of dark-grown wheat ( Triticum aestivum L. cv. Weibull's Folke) and barley ( Hordeum vulgare L. cv. Weibull's Ida) does not follow the reciprocity law but shows maxima and minima like the phototropic dose-response curve. These fluctuations of the unrolling can be explained by differences and non-synchronizations of the growth in width of the two sides of the leaves. As the thickness of the leaves is small, even minute differences between the widths of the two sides will give rise to great changes in the unrolling and thus in the projection values.     

Carbon dioxide enhances the development of the ethylene forming enzyme in tobacco leaf discs

Since CO₂ is known to stimulate ethylene production by promoting the conversion of 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene, the effect of CO₂ on the activity and the development of the ethylene forming enzyme (EFE) was studied in tobacco (Nicotiana tabacum L. cv Havana 425 and Xanthi) leaf discs. In addition to previous observations that EFE activity is dependent on CO₂ concentration and is saturable with 2% CO₂, present data show two saturation curves at 2% and 10% CO₂. Promotion of EFE development was dependent also on CO₂ concentration (saturated at 2% CO₂) and duration (maximum at 24 in the dark), and was abolished by 20 micromolar cycloheximide. Application of exogenous ethylene (20 microliters per liter) or light treatment further increased the CO₂-enhanced development of EFE, implying that these two factors can also affect EFE development via interaction with CO₂. The results suggest that CO₂ exerts its stimulatory effect on the conversion of ACC to ethylene by enhancing not only the activity but also the synthesis of EFE in leaf discs.     

Accelerated leaf senescence takes part in enhanced resistance in cucumber mosaic virus inoculated pepper leaves

Altered photosynthetic reactions in cucumber mosaic virus (CMV) inoculated leaves of virus resistant lines L113 and L57 and susceptible pepper (Capsicum annuum L.) plants cv. Albena grown in controlled environment and in the field were investigated. The CMV inoculated leaves of virus resistant lines developed different symptoms—necrotic local lesions on L113 and chlorotic spots on L57 while the same leaves of susceptible cv. Albena were symptomless. The changes in Photosystem II (PSII) and PSI electron transport were evaluated by chlorophyll fluorescence, and far-red (FR) light induced leaf absorbance A _810–860. CMV infection caused a decrease in maximal PSII quantum yield, F _v/F _m, in susceptible leaves. Increased non-photochemical fluorescence quenching in CMV-inoculated leaves of both resistant lines were observed. In CMV-inoculated leaves of all tested plants FR light induced P700 oxidation was decreased. In the present study, the viral-infected pepper plants grown in controlled environment to avoid the effects of abiotic factors were used as model system that allow us to investigate the differences in leaf senescence in CMV-inoculated leaves of susceptible and resistant pepper lines expressing different symptoms. Earlier leaf falls of inoculated leaves as a result of accelerated leaf senescence is important for building successful secondary virus resistance strategy following fast responses such as hypersensitive reaction.     

Glutamate synthase isoforms in rice: immunological studies of enzymes in green leaf, etiolated leaf, and root tissues

Rabbit antiserum was raised against ferredoxin-dependent glutamate synthase (EC 1.4.7.1) purified from green leaves of Oryza sativa L. cv Delta. Ferredoxin-dependent glutamate synthase, detected in green leaf, etiolated leaf, and root tissues cross-reacted completely with the antiferredoxin glutamate synthase immunoglobulin G. In contrast, the immunoglobulin G did not cross-react with NADH-dependent (EC 1.4.1.14) and NADPH-dependent (EC 1.4.1.13) glutamate synthases found in nonphotosynthetic etiolated leaf and root tissues. In addition, ferredoxin-dependent glutamate synthase was separated and distinguished by its affinity to ferredoxin from NAD(P)H-dependent glutamate synthase on ferredoxin-Sepharose affinity chromatography. Based on the immunological studies, it is suggested that ferredoxin-dependent glutamate synthases in green leaf and etiolated leaf tissues are closely related proteins; in contrast, ferredoxin-dependent glutamate synthase in root tissue is a distinct protein from the leaf enzymes.     

H-pumping driven by the vanadate-sensitive ATPase in membrane vesicles from corn roots

The initial rate of quenching of quinacrine fluorescence was used to monitor Mg:ATP-dependent H⁺-pumping in membrane vesicles from corn (Zea mays L. cv WF9 × MO17) roots and obtain a preparation in which vanadate-sensitive H⁺-pumping could be observed. Separation of membranes on a linear sucrose density gradient resulted in two distinct peaks of H⁺-pumping activity: a major one, at density 1.11 grams per cubic centimeter, was sensitive to NO₃⁻ and resistant to vanadate, while a minor one, at density 1.17 grams per cubic centimeter, was substantially resistant to NO₃⁻ and sensitive to vanadate. A membrane fraction enriched in the vanadate-sensitive H⁺-pump could be obtained by washing microsomes prepared in the presence of 10% glycerol with 0.25 molar KI. The kinetics of inhibition of H⁺-pumping by vanadate in this membrane preparation indicated that most of the H⁺-pumping activity in this fraction is sensitive to inhibition by vanadate, 50% inhibition being reached at about 60 micromolar vanadate. This value is fairly close to that observed for inhibition by vanadate of the ATPase activity in similar experimental conditions (40 micromolar). The inhibitor sensitivity, divalent cation dependence, pH optimum (6.5), and K_m for ATP (0.7 millimolar) of the H⁺-pumping activity match quite closely those reported for the plasma membrane ATPase of corn roots and other plant materials.     

Photosynthetic capacity,irradiance and sequential senescence of sugar beet leaves

In field-grown sugar beet plants (Beta vulgaris L. cv. Dobrovická A), each of66 successive leaves produoed in the course of the vegetation period was different with respect to its photosynthetic capaoity (P_c), life span, duration of leaf area expansion, and longevity after its maximum leaf area (A_max) has developed. The proportionality between the seasonal changes in these characteristics was not the same if the sequential senescence of leaves was taken into account. With aging of individual leaves, P_c increased with the leaf area expansion having attained the peak value between 75% to 100% of A_max The rate of ontogenetic changes in P_c of each leaf was specified by the rate of its growth and development so that even at comparable ages the successive leaves constituted a series of different physiological units. The seasonal changes in quantum irradiance (PAR) were found to be responsible for differences in the growth characteristics between the successive leaves: Leaf expansion period was related with daily integrals of the incoming PAR (Io), while leaf longevity, after the A_max had been attained, was closely linked with PAR intercepted by the canopy (I). P_c expressed per the total leaf area of the plant was significantly correlated withI, while P_c calculated per unit leaf area of the plant was related toI _o Leaf potential to adapt P_c correspondingly to changes in PAR was greatest during leaf blade expansion; after the leaf had ceased to expand, changes in P_c were independent of differences in leaf irradiance. The results stress, at least for field conditions, the inadmissibility of the extrapolation of attributes from one leaf to the other ones sequentially senescing on the plant.