Operation of the xanthophyll cycle in higher plants in response to diurnal changes in incident sunlight

Changes in the carotenoid composition of leaves in response to diurnal changes in sunlight were determined in the crop species Helianthus annuus L. (sunflower), Cucurbita pepo L. (pumpkin), and Cucumls sativus L. (cucumber), in the diaheliotropic mesophyte Malva neglecta Wallr., and in the perennial shrub Euonymus kiautschovicus Loesner. Large daily changes were observed in the relative proportions of the components of the xanthophyll cycle, violaxanthin (V), antheraxanthin (A), and zeaxanthin (Z) in plants grown in full sunlight. In all leaves large amounts of Z were formed at peak irradiance, with the changes in Z content closely following changes in incident photon flux density (PFD) over the course of the day. All leaves also contained large total pools of the three xanthophyll-cycle components. However, the extent to which the V pool present at dawn became de-epoxidized during the day varied widely among leaves, from a 27% decrease in M. neglecta to a 90% decrease in E. kiautschovicus. The largest amounts of Z and the lowest amounts of V at peak irradiance (full sunlight) were observed in the species with the lower rates of photosynthesis (particularly in E. kiautschovicus and pumpkin), and smaller amounts of Z and a lesser decrease in V content were found at peak irradiance in those species with the higher rates of photosynthesis (particularly in M. neglecta and sunflower). In all species some Z was present in the leaves prior to sunrise. Furthermore, in individuals of sunflower, pumpkin, and cucumber grown at 85% of full sunlight and transferred to full sunlight, a further increase in the already large pool of the xanthophyll-cycle pigments occurred over the course of 1 d.Abbreviations A antheraxanthin - -Car, -Car - and -carotene - EPS epoxidation state - PFD photon flux density, between 400 and 700 nm - V violaxanthin - Z zeaxanthin This work was supported by the U.S. Department of Agriculture, Competitive Research Grants Office, award No. 90-37130-5422, and a Faculty Development Award from the University of Colorado to W.W. Adams III.     

The interrelationship between the lower oxygen limit,chlorophyll fluorescence and the xanthophyll cycle in plants

The lower oxygen limit (LOL) in plants may be identified through the measure of respiratory gases [i.e. the anaerobic compensation point (ACP) or the respiratory quotient breakpoint (RQB)], but recent work shows it may also be identified by a sudden rise in dark minimum fluorescence (F _o). The interrelationship between aerobic respiration and fermentative metabolism, which occur in the mitochondria and cytosol, respectively, and fluorescence, which emanates from the chloroplasts, is not well documented in the literature. Using spinach (Spinacia oleracea), this study showed that F_o and photochemical quenching (q _P) remained relatively unchanged until O₂ levels dropped below the LOL. An over-reduction of the plastoquinone (PQ) pool is believed to increase F _o under dark + anoxic conditions. It is proposed that excess cytosolic reductant due to inhibition of the mitochondria’s cytochrome oxidase under low-O₂, may be the primary reductant source. The maximum fluorescence (F _m) is largely unaffected by low-O₂ in the dark, but was severely quenched, mirroring changes to the xanthophyll de-epoxidation state (DEPS), under even low-intensity light (≈4 μmol m⁻² s⁻¹). In low light, the low-O₂-induced increase in F _o was also quenched, likely by non-photochemical and photochemical means. The degree of quenching in the light was negatively correlated with the level of ethanol fermentation in the dark. A discussion detailing the possible roles of cyclic electron flow, the xanthophyll cycle, chlororespiration and a pathway we termed ‘chlorofermentation’ were used to interpret fluorescence phenomena of both spinach and apple (Malus domestica) over a range of atmospheric conditions under both dark and low-light.     

植物叶黄素循环的组成、功能和调节(综述)

对近几年来植物体内叶黄素循环的组成、功能、堇菜黄素脱环氧化酶和玉米黄素环氧化酶的结构、生化性质和调节,以及叶黄素的可转变性、定位等方面的研究进展作了综述。     

The xanthophyll cycle activity in kidney bean and cabbage leaves under salinity stress

Seven-day-old kidney bean and cabbage seedlings were treated with 0.1–0.3 M NaCl solutions for 3 days. Chlorophyll content decreased in NaCl-treated Phaseolus seedlings, but did not significantly decrease in Brassica seedlings. Photochemical efficiency of photosystem II at dark-adapted state was similar in both Phaseolus and Brassica. The de-epoxidation state of violaxanthin increased more than sixfold in Phaseolus but showed no significant change in Brassica seedlings during NaCl treatment under low light. Maximum de-epoxidation state of violaxanthin in vivo tested in high light (2000 μmol quanta/(m² s) increased in salt-stressed Phaseolus but decreased in Brassica seedlings. The nonphotochemical quenching (NPQ) also increased in Phaseolus but decreased in Brassica. This suggests that xanthophyll cycle pigments influence the NPQ in both Phaseolus and Brassica, but in an opposite way. The increase in the de-epoxidation state of violaxanthin in salt-stressed Phaseolus even under low light may be considered an early light signal to protect the pigment-protein complexes from salt-stress induced photodamage. It is proposed that in salt-stressed Brassica, the de-epoxidation is retarded and/or the epoxidation is accelerated leading to the accumulation of violaxanthin and a lower de-epoxidation state. Thus, light-induced violoxanthin cycle operation largely controls the photoprotection of photosynthetic apparatus in kidney bean leaves. Published in Russian in Fiziologiya Rastenii, 2006, Vol. 53, No. 1, pp. 113–121. The text was submitted by the authors in English.     

Leaf optical properties in higher plants: linking spectral characteristics to stress and chlorophyll concentration

A number of studies have linked responses in leaf spectral reflectance, transmittance, or absorptance to physiological stress. A variety of stressors including dehydration, flooding, freezing, ozone, herbicides, competition, disease, insects, and deficiencies in ectomycorrhizal development and N fertilization have been imposed on species ranging from grasses to conifers and deciduous trees. In all cases, the maximum difference in reflectance within the 400-850 nm wavelength range between control and stressed states occurred as a reflectance increase at wavelengths near 700 nm. In studies that included transmittance and absorptance as well as reflectance, maximum differences occurred as increases and decreases, respectively, near 700 nm. This common optical response to stress could be simulated closely by varying the chlorophyll concentration of model leaves (fiberglass filter pads) and by the natural variability in leaf chlorophyll concentrations in senescent leaves of five species. The optical response to stress near 700 nm, as well as corresponding changes in reflectance that occur in the green-yellow spectrum, can be explained by the general tendency of stress to reduce leaf chlorophyll concentration.     

Remote sensing of the xanthophyll cycle and chlorophyll fluorescence in sunflower leaves and canopies

Summary Sudden illumination of sunflower (Helianthus annuus L. cv. CGL 208) leaves and canopies led to excess absorbed PFD and induced apparent reflectance changes in the green, red and near-infrared detectable with a remote spectroradiometer. The green shift, centered near 531 nm, was caused by reflectance changes associated with the de-epoxidation of violaxanthin to zeaxanthin via antheraxanthin and with the chloroplast thylakoid pH gradient. The red (685 nm) and near-infrared (738 nm) signals were due to quenching of chlorophyll fluorescence. Remote sensing of shifts in these spectral regions provides non-destructive information on in situ photosynthetic performance and could lead to improved techniques for remote sensing of canopy photosynthesis.CIW Publication #1072     

Relationship between xanthophyll cycle and non-photochemical quenching in rice (Oryza sativa L.) plants in response to light stress

Thirty days old rice plants grown under low and moderate light conditions were transferred to full sunlight to observe the extent of photoinhibitory damage and protective mechanism, and the relationship between xanthophyll cycle and nonphotochemical quenching (qN) under changing light environment. Control plants (low, moderate and sun grown) exhibited similar Fv/Fm ratio, indicating similar photosynthetic efficiency prior to light stress. On exposure to the high light treatment, low light grown plants exhibited faster and higher degree of photoinhibition compared to moderate and high light grown plants. Moderate and high light grown plants showed relatively less photoinhibition and also showed higher qN, indicating better capacity of energy dissipation. Increase in qN in moderate light and sun grown plants was accompanied by conversion of violaxanthin (V) to antheraxanthin (A) and zeaxanthin (Z) indicating operation of Z-dependent thermal dissipation. Rice plants fed with ascorbate (AsA), a stimulator of the de-epoxidation state of V to Z, showed higher Fv/Fm ratio and qN than the plants fed with dithiothreitol (DTT) an inhibitor of xanthophyll cycle. This indicated that an increased amount of energy reached PS II reaction centre, due to absence of A and Z formation, thereby causing greater damage to photosynthesis in DTT fed rice plants. The present data confirmed the relationship between qN and Z in dissipating the excess light energy, thereby protecting plants against photodamage.     

The xanthophyll cycle and acclimation of Pinus ponderosa and Malva neglecta to winter stress

Seasonal differences in the efficiency of open PSII units (F _v/F _m), leaf pigment composition and xanthophyll cycle conversion (Z+A)/(V+A+Z), leaf adenylate status, and photosynthetic capacity were investigated in Pinus ponderosa (Ponderosa pine) and Malva neglecta. In P. ponderosa, acclimation to winter involved a lower photosynthetic capacity, higher carotenoid to chlorophyll ratio, persistent reductions in F _v/F _m corresponding to persistent retention of Z+A, and no change in foliar ATP/ADP ratios. In contrast, M. neglecta characterized in winter exhibited higher rates of photosynthesis than in summer with no change in carotenoid to chlorophyll ratio, while small nocturnally persistent reductions in F _v/F _m were observed exclusively on colder winter nights when nocturnal retention of Z+A, and high ATP/ADP ratios were also present. Upon removal of winter-stressed leaves or needles from the field to room temperature, a portion of F _v/F _m relaxed within 15 min of warming and recovery was completed within 5 h in M. neglecta but required 100 h in P. ponderosa. In M. neglecta, the entire recovery of F _v/F _m correlated with decreases in the foliar ATP/ADP ratio, while in P. ponderosa this ratio remained unchanged. Possible ATP-dependent forms of sustained (Z+A)-dependent energy dissipation are discussed including a nocturnally retained pH gradient on cold winter nights. The slow recovery in pine involved not only retention of Z+A, but apparently also a persistent engagement of Z+A for energy dissipation via an unidentified mechanism. Received: 15 May 1998 / Accepted: 9 November 1998     

In vivo chlorophyll forms in higher plants and algae sensitive to treatment with lutein

Lamella preparations of spinach, Chlorella, Phaeodactylum, Anabatnaand Porphyra were treated with a hydrophobic reagent, lutein,and the absorption and fluorescence spectra in the red regionbefore and after treatment were compared for changes causedby the treatment. Absorption spectra of all these preparationsunderwent the same spectral change, transformation of a bandat 684 nm into a band at 666 nm. The longer the maximum wavelengthof the red peak, the greater was the fractional absorbance decreaseat 684 nm. The content of C684 (the chlorophyll form responsiblefor the 684 nm band) in the lamellae was estimated from thefractional decreases as being progressively higher in the orderof Phaeodactylum, Porphyra, Anabatna, Chlorella and spinach.The fluorescence spectra at liquid nitrogen temperature beforetreatment showed two bands. The longer wavelength band was transformedby the treatment into a shorter wavelength band(s), as describedbelow, according to the maximum wavelengths: spinach, F735F695(or F686); Chlorella, F715F700 (or F686); Phaeodactylum, anunidentified componentF690; Anabaena, F732F685 (or F695); Porphyra,F726F683. These chlorophyll forms with fluorescence maxima between715 and 735 nm were, therefore, designated C684 based on absorptionspectrophotometry, and are considered to play a role in photosystemII. (Received August 15, 1972; )     

Analysis of xanthophyll cycle carotenoids and chlorophyll fluorescence in light intensity-dependent chlorophyll-deficient mutants of wheat and barley

Three light intensity-dependent Chl b-deficient mutants, two in wheat and one in barley, were analyzed for their xanthophyll cycle carotenoids and Chl fluorescence characteristics under two different growth PFDs (30 versus 600 mol photons·m^–2 s^–1 incident light). Mutants grown under low light possessed lower levels of total Chls and carotenoids per unit leaf area compared to wild type plants, but the relative proportions of the two did not vary markedly between strains. In contrast, mutants grown under high light had much lower levels of Chl, leading to markedly greater carotenoid to Chl ratios in the mutants when compared to wild type. Under low light conditions the carotenoids of the xanthophyll cycle comprised approximately 15% of the total carotenoids in all strains; under high light the xanthophyll cycle pool increased to over 30% of the total carotenoids in wild type plants and to over 50% of the total carotenoids in the three mutant strains. Whereas the xanthophyll cycle remained fairly epoxidized in all plants grown under low light, plants grown under high light exhibited a considerable degree of conversion of the xanthophyll cycle into antheraxanthin and zeaxanthin during the diurnal cycle, with almost complete conversion (over 90%) occurring only in the mutants. 50 to 95% of the xanthophyll cycle was retained as antheraxanthin and zeaxanthin overnight in these mutants which also exhibited sustained depressions in PS II photochemical efficiency (F_v/F_m), which may have resulted from a sustained high level of photoprotective energy dissipation activity. The relatively larger xanthophyll cycle pool in the Chl b-deficient mutant could result in part from the reported concentration of the xanthophyll cycle in the inner antenna complexes, given that the Chl b-deficient mutants are deficient in the peripheral LHC-II complexes.Abbreviations A antheraxanthin - Chl chlorophyll - F_o and F_m minimal yield (at open PS II reaction centers) and maximal yield (at closed centers) of chlorophyll fluorescence in darkness - F level of fluorescence during illumination with photosynthetically active radiation - F_m maximal yield (at closed centers) of chlorophyll fluorescence during illumination with photosynthetically active radiation - (F_m–F)/F_m actual efficiency of PS II during illumination with photosynthetically active radiation - F_v/F_m+(F_m–F_o)/F_m intrinsic efficiency of PS II in darkness - LHC_II light-harvesting chlorophyll-protein complex of Photosystem II - PFD photon flux density (between 400 and 700 nm) - PS I Photosystem I - PS II Photosystem II - V violaxanthin - Z zeaxanthin     

Genetic engineering of polyamine and carbohydrate metabolism for osmotic stress tolerance in higher plants

Plant growth and productivity are greatly affected by various stress factors. The molecular mechanisms of stress tolerance in plant species have been well established. Metabolic pathways involving the synthesis of metabolites such as polyamines, carbohydrates, proline and glycine betaine have been shown to be associated with stress tolerance. Introduction of the stress-induced genes involved in these pathways from tolerant species to sensitive plants seems to be a promising approach to confer stress tolerance in plants. In cases where single gene is not enough to confer tolerance, metabolic engineering necessitates the introduction of multiple transgenes in plants.     

Fat metabolism in higher plants. Production of short- and medium-chain acyl-acyl carrier protein by spinach stroma preparations treated with cerulenin.

Incubation of stroma preparations from spinach chloroplasts with low concentrations of cerulenin (10 muM) resulted in severe inhibition of fatty acid synthesis but stimulated the release of medium-chain acids in very high proportions (60-70%). Preincubation of these preparations with cerulenin in the absence of substrate exerted no additional effect on subsequent fatty acid synthesis (as measured by incorporation of [14C]acetate into fatty acids) or the pattern of radioactive acids obtained. Acyl-protein, acyl-CoA, free fatty acids and lipids were resolved from each other and analysed for their distribution of 14C-labelled fatty acids. Acyl-protein derived from cerulenin-treated preparations was the only fraction which contained short- and medium-chain acids (C6--C12). The other fractions from both control and cerulenin-treated groups consisted exclusively of C16 and C18 acids. Acyl-protein was purified by gel filtration chromatography and was characterized as acyl-acyl carrier protein.     

Photoinhibition, xanthophyll cycle and in vivo chlorophyll fluorescence quenching of chilling-tolerant Oxyria digyna and chilling-sensitive Zea mays

The relationship between susceptibility to photoinhibition, zeaxanthin formation and chlorophyll fluorescence quenching at suboptimal temperatures was studied in chilling-sensitive maize and in non-acclimated and cold-acclimated Oxyria digyna , a chilling-tolerant plant of arctic and alpine habitats. In maize, zeaxanthin formation was strongly suppressed by chilling. Zeaxanthin formed during preillumination at 20°C did not protect maize leaves from photoinhibition during a subsequent high-light, low-temperature treatment, as judged from the ratios of variable to maximal fluorescence, F_v/F_m. However, such preillumination significantly increased non-photochemical quenching (q_N) at low temperatures, mainly due to an enhancement of the fast-relaxing q_N component (i.e., of energy-dependent quenching. q_E). In O. digyna , cold-acclimation resulted in an increased zeaxanthin formation in the temperature range of 2.5–20°C. Cold-acclimation substantially decreased the susceptibility towards photoinhibition at 4°C, but q_N remained nearly unchanged between 2 and 38°C, as compared to control plants. Effects of cold acclimation on photosynthesis, photochemical quenching and quantum efficiency of photosystem II were small and indicated a slight amelioration only of the function of the photosynthetic apparatus at suboptimal temperatures (2–20°Ct. I) is concluded, that the xanthophyll cycle is strongly influenced by cold acclimation, while effects on the photosynthetic carbon assimilation only play a minor role in O. digyna.     

The xanthophyll cycle is induced by light irrespective of water status in field-grown lavender (Lavandula stoechas) plants

The water relations, the photosynthetic capacity and the pigment content of leaves, i.e. chlorophylls, carotenes and xanthophylls, were analysed during the summer drought and recovery after autumn rainfalls in lavender ( Lavandula stoechas L.) plants grown in Mediterranean field conditions. Summer drought caused photoinhibition of photosynthesis and significant decreases in chlorophylls (by ca 75%), β -carotene (by ca 65%), and lutein and neoxanthin (by ca 50%), although their contents remained unaltered between predawn and midday, suggesting a progressive decrease in response to drought. In contrast, the levels of violaxanthin decreased from predawn to midday, giving rise to enhanced formation of zeaxanthin and antheraxanthin in high light. Zeaxanthin and antheraxanthin formation was not induced by water deficit. Although the levels of photosynthetic pigments were severely affected by water deficit, carotenoids decreased less than chlorophylls, which resulted in increased levels of carotenoids per unit of chlorophyll. We conclude that the enhanced formation of zeaxanthin in high light and the increased levels of carotenoids per unit of chlorophyll observed in water-stressed plants may help to avoid photoinhibitory damage to the photosynthetic apparatus.     

Expression of telomerase activity is closely correlated with the capacity for cell division in tobacco plants

Telomerase is a specialized RNA-directed DNA polymerase that adds telomeric repeats to the ends of linear eukaryotic chromosomes. This activity is developmentally regulated in mammals. Here, we investigated the expression of telomerase activities in various cell types of tobacco plants using the telomere repeat amplification protocol (TRAP) assay. The greatest telomerase activity was detected in BY-2 suspension culture cells, while a relatively high level of activity was also found in roots. Because these two cell types contain a high proportion of actively dividing cells, our results indicate a close correlation between telomerase activity and the capacity for division in tobacco cells. Consistent with this observation was the very low level of telomerase activity in stems, leaves, and flowers, all tissues that had negligible activity of cell division. The specific expression of telomerase in actively dividing plant cells suggests that the pattern of telomerase regulation is largely conserved between higher plants and mammals.     

Enhanced glutathione metabolism is correlated with sulfur-induced resistance in Tobacco mosaic virus-infected genetically susceptible Nicotiana tabacum plants

Sulfur-induced resistance, also known as sulfur-enhanced defense (SIR/SED) was investigated in Nicotiana tabacum cv. Samsun nn during compatible interaction with Tobacco mosaic virus (TMV) in correlation with glutathione metabolism. To evaluate the influence of sulfur nutritional status on virus infection, tobacco plants were treated with nutrient solutions containing either sufficient sulfate (+S) or no sulfate (-S). Sufficient sulfate supply resulted in a suppressed and delayed symptom development and diminished virus accumulation over a period of 14 days after inoculation as compared with -S conditions. Expression of the defense marker gene PR-1a was markedly upregulated in sulfate-treated plants during the first day after TMV inoculation. The occurrence of SIR/SED correlated with a higher level of activity of sulfate assimilation, cysteine, and glutathione metabolism in plants treated with sulfate. Additionally, two key genes involved in cysteine and glutathione biosynthesis (encoding adenosine 5'-phosphosulfate reductase and γ-glutamylcysteine synthetase, respectively) were upregulated within the first day after TMV inoculation under +S conditions. Sulfate withdrawal from the soil was accelerated at the beginning of the infection, whereas it declined in the long term, leading to an accumulation of sulfur in the soil of plants grown with sulfate. This observation could be correlated with a decrease in sulfur contents in TMV-infected leaves in the long term. In summary, this is the first study that demonstrates a link between the activation of cysteine and glutathione metabolism and the induction of SIR/SED during a compatible plant-virus interaction in tobacco plants, indicating a general mechanism behind SIR/SED.     

Resistance to chytridiomycosis varies among amphibian species and is correlated with skin peptide defenses

Innate immune mechanisms of defense are especially important to ectothermic vertebrates in which adaptive immune responses may be slow to develop. One innate defense in amphibian skin is the release of abundant quantities of antimicrobial peptides. Chytridiomycosis is an emerging infectious disease of amphibians caused by the skin fungus, Batrachochytrium dendrobatidis . Susceptibility to chytridiomycosis varies among species, and mechanisms of disease resistance are not well understood. Previously, we have shown that Australian and Panamanian amphibian species that possess skin peptides that effectively inhibit the growth of B. dendrobatidis in vitro tend to survive better in the wild or are predicted to survive the first encounter with this lethal pathogen. For most species, it has been difficult to experimentally infect individuals with B. dendrobatidis and directly evaluate both survival and antimicrobial peptide defenses. Here, we demonstrate differences in susceptibility to chytridiomycosis among four Australian species ( Litoria caerulea, Litoria chloris, Mixophyes fasciolatus and Limnodynastes tasmaniensis ) after experimental infection with B. dendrobatidis , and show that the survival rate increases with the in vitro effectiveness of the skin peptides. We also observed that circulating granulocyte, but not lymphocyte, counts differed between infected and uninfected Lit. chloris . This suggests that innate granulocyte defenses may be activated by pathogen exposure. Taken together, our data suggest that multiple innate defense mechanisms are involved in resistance to chytridiomycosis, and the efficacy of these defenses varies by amphibian species.