The Chlorophyll <Emphasis Type="Italic">a</Emphasis> Fluorescence Characteristic in Different Types of Leaf Senescence in <Emphasis Type="Italic">Alhagi sparsifolia</Emphasis>

Senescence is both a highly controlled and a strictly regulated process that is gene dependent. To study the PSII reaction in different types of leaf senescence processes, stem girdling was performed on Alhagi sparsifolia to investigate the leaf status in the control, natural senescence, and girdling-induced senescence leaves. The results showed that during senescence, leaf soluble sugar content, starch content, and the energy absorbed by the unit reaction center (ABS/RC) increased; whereas leaf photosynthetic rate, photosynthetic pigment content, maximum photochemical efficiency (φ _Po), and energy used by the acceptor site in electron transfer (ET_o/RC) decreased. The result of the present research implied that stem girdling significantly accelerated leaf senescence, which was due to the accumulation of carbohydrate. Natural senescence is a highly controlled process, which is an ordered process played by genes, whereas girdling-induced senescence is a disordered one. In addition, natural senescence slightly inhibits the acceptor site of PSII but did not damage the donor site of PSII. Conversely, girdling-induced senescence not only damaged the donor site of PSII (for example, oxygen-evolving complex), but also significantly inhibited the acceptor site of PSII. Moreover, both types of senescence led to an increase in the energy absorbed by the unit reaction center (ABS/RC), which subsequently resulted in an increasing excitation pressure in the reaction center (DI_o/RC), as well as additional saved carotenoid for absorbing residual light energy and quenching reactive oxygen species during senescence.     

韧皮部环割诱导下的花花柴衰老机制

衰老是植物器官和组织发育的最后阶段, 是一个受到严格控制的高度协调过程, 其中碳水化合物浓度对衰老的影响十分显著。花花柴(Karelinia caspia)是塔克拉玛干沙漠南缘策勒绿洲的主要植物种, 为了研究花花柴在韧皮部环割后的碳水化合物变化和叶片衰老过程, 对其进行韧皮部环割, 测量叶片光合色素含量、光合速率、可溶性糖含量、淀粉含量、脱落酸(ABA)含量和叶水势。结果表明: (1)环割能够诱导花花柴叶片的衰老, 而诱导叶片衰老的主要因素有: 叶片碳水化合物的积累、叶片ABA含量的上升, 以及叶片水分状况的恶化。(2)相比于自然衰老, 环割诱导的衰老导致许多正常的生理过程受到破坏。(3)类胡萝卜素在衰老过程中主要起光保护的作用。(4)韧皮部半环割也导致花花柴各种生理指标显著下降, 表明植物无法通过增加剩余部分韧皮部筛管的运输通量而达到维持整个韧皮部运输系统顺畅的目的。     

Senescence mechanisms induced by phloem girdling in Karelinia caspia

AimsSenescence constitutes the final stage of a plant’s organ and tissue development, and is subject to gene control and strict regulation. Plant senescence is largely influenced by carbohydrate content and phloem girdling can induce leaf senescence. Our general objective is to study the effect of stem girdling on physiological conditions in Karelinia caspia. Specifically, we want to know the senescence processes after phloem girdling. In addition, we also want to know the possible mechanisms for the senescence processes. MethodsThree different types of girdling treatments, normal branch, semi-girdling, and full-girdling were performed on K. caspia. Twenty days after girdling, photosynthetic pigments content, photosynthetic rate, soluble sugar content, starch content, abscisic acid (ABA) content, and leaf water potential were measured.Important findings Phloem girdling can largely induce leaf senescence in K. caspia, and the reasons for leaf senescence may be as follows: girdling resulted in carbohydrate accumulation in leaf which subsequently led to “carbon feast” induced leaf senescence; girdling caused ABA accumulation in leaf and then resulted in senescence; girdling decreased water status, which may be another reason for leaf senescence. Compared with natural senescence, girdling induced senescence was a disorder and disorganized process, only a limited physiological process can be controlled by senescence related gene in the girdling induced senescence process. The most important role for carotenoids in the senescence process is to protect the photosynthetic apparatus from being damaged by excess light and reactive oxygen species. Many physiological indicators declined in the semi-girdled K. caspia leaves just like full-girdled leaves, indicating that portion (e.g. half) of the phloem cannot undertake the transport flux which was done by the whole phloem sieve.     

Inhibition of root respiration induces leaf senescence in <Emphasis Type="Italic">Alhagi sparsifolia</Emphasis>

Leaf senescence can be induced by numerous factors. In order to explore the relationship between root respiration and leaf senescence, we utilized different types of phloem girdling to control the root respiration of Alhagi sparsifolia and its physiological response. Our results showed that both girdling and inhibition of root respiration led to a decline of stomatal conductance, photosynthesis, transpiration rate, chlorophyll (Chl) a, Chl b, carotenoid (Car) content, Chl a/b, Chl/Car, water potential, and Chl a fluorescence, as well as to an increase of abscisic acid (ABA), proline, and malondialdehyde content in leaves and to upregulation of senescence-associated gene expression. Our present work implied that both inhibition of root respiration and girdling can induce leaf senescence. In comparison with phloem girdling, the leaf senescence caused by inhibition of root respiration was less significant. The reason for girdling-induced senescence was ABA and carbohydrate accumulation. Senescence induced by inhibition of root respiration occurred due to leaf water stress resulting from inhibition of water absorption.     

Impact of phloem girdling on leaf gas exchange and hydraulic conductance in hybrid aspen

We investigated phloem-xylem interactions in relation to leaf hydraulic capacity in hybrid aspen (Populus tremula L. × P. tremuloides Michx.) by using phloem girdling method. Removal of bark tissues (phloem girdling) at the branch base resulted in a substantial decline in stomatal conductance (g_S), net photosynthetic rate (P_N), and leaf hydraulic efficiency, and in increase of leaf water potential (Ψ_L). Although gS declined more than P_N (83 versus 78 %), the ratio of intercellular to ambient CO₂ concentrations (c_i/c_a) increased from 0.67 to 0.87 in three days after girdling. Girdling induced a decrease in leaf hydraulic conductance (K_L) on average by 43 % (P = 0.006). The changes in gS and leaf conductance to water vapour were co-ordinated with K_L only in girdled branches whereas intrinsic water-use efficiency was invariant to K_L. The declines in K_L with girdling were not accompanied by changes in potassium ion concentration ([K⁺]), electrical conductivity, or pH of xylem sap. The results suggest that phloem girdling at the branch base does not influence the recirculation of ions between the phloem and xylem in hybrid aspen and the decrease of K_L in response to the manipulation is not related to changes in [K⁺] and total ionic content of xylem sap.     

Multiple sites of retardation of electron transfer in Photosystem II after hydrolysis of phosphatidylglycerol

Phosphatidylglycerol (PG), containing the unique fatty acid Δ3, trans-16:1-hexadecenoic acid, is a minor but ubiquitous lipid component of thylakoid membranes of chloroplasts and cyanobacteria. We investigated its role in electron transfers and structural organization of Photosystem II (PSII) by treating Arabidopsis thaliana thylakoids with phospholipase A₂ to decrease the PG content. Phospholipase A₂ treatment of thylakoids (a) inhibited electron transfer from the primary quinone acceptor Q_A to the secondary quinone acceptor Q_B, (b) retarded electron transfer from the manganese cluster to the redox-active tyrosine Z, (c) decreased the extent of flash-induced oxidation of tyrosine Z and dark-stable tyrosine D in parallel, and (d) inhibited PSII reaction centres such that electron flow to silicomolybdate in continuous light was inhibited. In addition, phospholipase A₂ treatment of thylakoids caused the partial dissociation of (a) PSII supercomplexes into PSII dimers that do not have the complete light-harvesting complex of PSII (LHCII); (b) PSII dimers into monomers; and (c) trimers of LHCII into monomers. Thus, removal of PG by phospholipase A₂ brings about profound structural changes in PSII, leading to inhibition/retardation of electron transfer on the donor side, in the reaction centre, and on the acceptor side. Our results broaden the simple view of the predominant effect being on the Q_B-binding site.     

Responses of photosystem II of white elm to UV-B radiation monitored by OJIP fluorescence transients

Photosystem II (PSII) activities in both samara and leaf of white elm (Ulmus pumila L.) were significantly inhibited by enhanced UV-B radiation (UVBR). UVBR disturbed both the donor and acceptor sides of PSII. The plastoquinone (PQ) pool size on the acceptor side, the trapped excited energy for complete reduction of Q_A, and the proportion of closed PSII reaction centers (RCs) increased, with PSII RCs being transformed into dissipative sinks for excitation energy under UVBR. However, samara and leaf responded to UVBR in different ways. A decrease in the F ₀ for leaf induced by UV-B radiation suggests the formation of fluorescence-quenching centers. An increase in the V_I for leaf under UVBR might mean the accumulation of reduced Q_A and PQ. F ₀ and V_I for samara showed opposite change pattern. Leaf has the mechanism of regulation of the amount of light reaching the RC through decreasing the number of light-harvesting chlorophyll molecules under UVBR while samara may be unable to regulate the light-harvesting capacity. PSII in samara was more susceptible to UVBR than that in leaf, with PI_ABS for samara decreasing more rapidly by a factor of 6.4 than that for leaf. Samara can recover more easily from UVBR-induced damage to PSII than the leaf.     

Stem girdling influences concentrations of endogenous cytokinins and abscisic acid in relation to leaf senescence in cotton

Many studies have shown that root–shoot imbalance influences vegetative growth and development of cotton (Gossypium hirsutum L.), but few have examined changes in leaf senescence and endogenous hormones due to stem girdling. The objective of this study was to determine the correlation between some endogenous phytohormones, particularly cytokinins and abscisic acid (ABA), and leaf senescence following stem girdling. Field-grown cotton plants were girdled on the main stem 5 days after squaring (DAS), while the non-girdled plants served as control. Plant biomass, seed cotton yield, main-stem leaf photosynthetic (Pn) rate, chlorophyll (Chl) and malondialdehyde (MDA) concentrations, as well as levels of cytokinins and ABA in main-stem leaves and xylem sap were determined after girdling or at harvest. Main-stem girdling decreased the dry root weight and root/shoot ratio from 5 to 70 days after girdling (DAG) and reduced seed cotton yield at harvest. Main-stem leaf Pn and Chl concentration in girdled plants were significantly lower than in control plants. Much higher levels of MDA were observed in main-stem leaves from 5 to 70 DAG, suggesting that stem girdling accelerated leaf senescence. Girdled plants contained less trans-zeatin and its riboside (t-Z + t-ZR), dihydrozeatin and its riboside (DHZ + DHZR), and isopentenyladenine and its riboside (iP + iPA), but more ABA than control plants in both main-stem leaves and xylem sap. These results suggested that main-stem girdling accelerated leaf senescence due to reduced levels of cytokinin and/or increased ABA. Cytokinin and ABA are involved in leaf senescence following main-stem girdling.     

Photosynthesis and chlorophyllafluorescence during flag leaf senescence of field-grown wheat plants

The characteristics of photosynthetic gas exchange, chlorophyll a fluorescence, and xanthophyll cycle pigments during flag leaf senescence of field-grown wheat plants were investigated. With senescence progressing, the light-saturated net CO₂ assimilation rate expressed either on a basis of leaf area or chlorophyll decreased significantly. The apparent quantum yield of net photosynthesis decreased when expressed on a leaf area basis but increased when expressed on a chlorophyll basis. The maximal efficiency of PSII photochemistry decreased very little while actual PSII efficiency, photochemical quenching, and the efficiency of excitation capture by open PSII centers decreased considerably. At the same time, non-photochemical quenching increased significantly. A substantial decrease in the contents of violaxanthin and zeaxanthin, but a slight decrease in the content of antheraxanthin were observed. However, the de-epoxidation status of the xanthophyll cycle was positively correlated with progressive senescence. This increase was due mainly to a smaller decrease in zeaxanthin than in violaxanthin. Our results suggest that PSII apparatus remained functional, but a down-regulation of PSII occurred under the steady state of photosynthesis in senescent flag leaves. Such a down-regulation was associated with the closure of PSII centers and an enhanced xanthophyll cycle-related thermal dissipation in the PSII antennae.     

Strigolactone signaling regulates rice leaf senescence in response to a phosphate deficiency

Strigolactones (SLs) act as plant hormones that inhibit shoot branching and stimulate secondary growth of the stem, primary root growth, and root hair elongation. In the moss Physcomitrella patens, SLs regulate branching of chloronemata and colony extension. In addition, SL-deficient and SL-insensitive mutants show delayed leaf senescence. To explore the effects of SLs on leaf senescence in rice (Oryza sativa L.), we treated leaf segments of rice dwarf mutants with a synthetic SL analogue, GR24, and evaluated their chlorophyll contents, ion leakage, and expression levels of senescence-associated genes. Exogenously applied GR24 restored normal leaf senescence in SL-deficient mutants, but not in SL-insensitive mutants. Most plants highly produce endogenous SLs in response to phosphate deficiency. Thus, we evaluated effects of GR24 under phosphate deficiency. Chlorophyll levels did not differ of in the wild-type between the sufficient and deficient phosphate conditions, but increased in the SL-deficient mutants under phosphate deficiency, leading in the strong promotion of leaf senescence by GR24 treatment. These results indicate that the mutants exhibited increased responsiveness to GR24 under phosphate deficiency. In addition, GR24 accelerated leaf senescence in the intact SL-deficient mutants under phosphate deficiency as well as dark-induced leaf senescence. The effects of GR24 were stronger in d10 compared to d17. Based on these results, we suggest that SLs regulate leaf senescence in response to phosphate deficiency.     

Chlorophyll a fluorescence kinetics of mung bean (Vigna radiata L.) grown under artificial continuous light

Continuous light can be used as a tool to understand the diurnal rhythm of plants and it can also be used to increase the plant production. In the present research, we aimed to investigate the photosynthetic performance of V. radiata under continuous light as compared with the plants grown under normal light duration. Chlorophyll a fluorescence transient (OJIP test) technique was used to understand the effect on various stages of photosynthesis and their consequences under continuous light condition. Various Chl a Fluorescence kinetic parameters such as Specific energy fluxes (per Q_A-reducing PSII reaction center (RC)) (ABS /RC; TR₀/RC; ET₀/RC; DI₀/RC), phenomenological fluxes, leaf model, (ABS/CSm; TR/CSm; ETo/CSm), Quantum yields and efficiencies (φPo; φEo; Ψo) and Performance index (PI_abs) was extracted and analysed in our investigation. Conclusively, our study has revealed that continuous light alters the photosynthetic performance of V. radiata at a different point but also improve plant productivity.     

Girdling interruption between source and sink in Quercus pubescens does not trigger leaf senescence

Metabolite changes and senescence behaviour after mechanical phloem girdling were studied in leaf tissue of Quercus pubescens. Sugar accumulation is not only considered to be an important part of several developmental signalling pathways, but is also seen as one of the basic triggers for senescence induction, or at least an obligatory accessory phenomenon. Our survey showed that an accumulation of the soluble sugars, glucose and fructose, was not on its own obligatorily connected with the induction of leaf senescence, since no indication or even an onset of senescence could be observed during the course of the experiment. Instead, we observed an inhibition of leaf development with a decrease of photosynthesis and a slow-down of development in nearly all chlorophyll a fluorescence analysis parameters using the JIP-test. We detected a change of metabolites linked to oxidative stress, possibly due to an overexcitation of the developmentally inhibited photosynthetic apparatus.     

Solar UV-B effects on PSII performance in Betula nana are influenced by PAR level and reduced by EDU: results of a 3-year experiment in the High Arctic

The long-term and diurnal responses of photosystem II (PSII) performance to near-ambient UV-B radiation were investigated in High Arctic Betula nana. We conducted an UV exclusion experiment with five replicated blocks consisting of open control (no filter), photosynthetic active radiation and UV-B transparent filter control (Teflon), UV-B-absorbing filter (Mylar) and UV-AB-absorbing filter (Lexan). Ethylenediurea (EDU), a chemical normally used to protect plants against ozone injury, was sprayed on the leaves both in the field and in an additional laboratory study to investigate if EDU mitigated the effects of UV-B. Chlorophyll-a fluorescence induction curves were used for analysis of OJIP test parameters. Near-ambient UV-B radiation reduced across season maximum quantum yield (TR(o) /ABS = F(v) /F(m)), approximated number of active PSII reaction center (RC/ABS) and the performance index (PI(ABS)), despite improved leaf screening against UV-B with higher content of UV-B-absorbing compounds and a lower specific leaf area. EDU application counteracted the negative impact of UV-B on TR(o) /ABS, RC/ABS and PI(ABS) . This indicates that the mechanisms behind UV-B and ozone damage share some common features. The midday depression was present in all treatments, but TR(o) /ABS and PI(ABS) were persistently lower in near-ambient UV-B compared to UV-B reduction. The recovery phase was particularly impaired in near-ambient UV-B and interactive effects between treatment × hour raised TR(o) /ABS, RC/ABS and PI(ABS) higher in reduced UV-B compared to near-ambient UV-B. This demonstrates current solar UV-B to reduce the PSII performance both on a daily as well as a seasonal basis in this High Arctic species.     

Root hypoxia reduces leaf growth : role of factors in the transpiration stream

This study examined the potential role of restricted phloem export, or import of substances from the roots in the leaf growth response to root hypoxia. In addition, the effects of root hypoxia on abscisic acid (ABA) and zeatin riboside (ZR) levels were measured and their effects on in vitro growth determined. Imposition of root hypoxia in the dark when transpirational water flux was minimal delayed the reduction in leaf growth until the following light period. Restriction of phloem transport by stem girdling did not eliminate the hypoxia-induced reduction in leaf growth. In vitro growth of leaf discs was inhibited in the presence of xylem sap collected from hypoxic roots, and also by millimolar ABA. Disc growth was promoted by sap from aerated roots and by 0.1 micromolar ZR. The flux of both ABA and ZR was reduced in xylem sap from hypoxic roots. Leaf ABA transiently increased twofold after 24 hours of hypoxia exposure but there were no changes in leaf cytokinin levels.     

Differential changes in degradation of chlorophyll-protein complexes of photosystem I and photosystem II during flag leaf senescence of rice.

The stability of chlorophyll-protein complexes of photosystem I (PSI) and photosystem II (PSII) was investigated by chlorophyll (Chl) fluorescence spectroscopy, absorption spectra and native green gel separation system during flag leaf senescence of two rice varieties (IIyou 129 and Shanyou 63) grown under outdoor conditions. During leaf senescence, photosynthetic CO(2) assimilation rate, carboxylase activity of Rubisco, chlorophyll and carotenoids contents, and the chlorophyll a/b ratio decreased significantly. The 77 K Chl fluorescence emission spectra of thylakoid membranes from mature leaves had two peaks at around 685 and 735 nm emitting mainly from PSII and PSI, respectively. The total Chl fluorescence yields of PSI and PSII decreased significantly with senescence progressing. However, the decrease in the Chl fluorescence yield of PSI was greater than in the yield of PSII, suggesting that the rate of degradation in chlorophyll-protein complexes of PSI was greater than in chlorophyll-protein complexes of PSII. The fluorescence yields for all chlorophyll-protein complexes decreased significantly with leaf senescence in two rice varieties but the extents of their decrease were significantly different. The greatest decrease in the Chl fluorescence yield was in PSI core, followed by LHCI, CP47, CP43, and LHCII. These results indicate that the rate of degradation for each chlorophyll-protein complex was different and the order for the stability of chlorophyll-protein complexes during leaf senescence was: LHCII>CP43>CP47>LHCI>PSI core, which was partly supported by the green gel electrophoresis of the chlorophyll-protein complexes.     

Photosynthetic characterization of Jerusalem artichoke during leaf expansion

Gas exchange, chlorophyll a fluorescence and modulated 820 nm reflection were investigated to explore the development of photosynthesis in Jerusalem artichoke (Helianthus tuberosus L.) leaves from initiation to full expansion. During leaf expansion, photosynthetic rate (Pn) increased and reached the maximal level when leaves were fully expanded. The same change pattern was also found in the stomatal conductance and chlorophyll content. Lower Pn could not be ascribed to the higher stomatal resistance in developing leaves, as intercellular CO₂ concentration was not significantly lower in these leaves. Lower Pn partly resulted from the lower actual photochemical efficiency of PSII in developing leaves, as more excited energy was dissipated through non-photochemical quenching. The development of primary photochemical reaction and electron transport in the donor side of PSII was completed in the initiating leaves. However, the development of electron transport in the acceptor side of PSII was not accomplished until leaves were fully expanded, indicated by the change in probability that an electron moves further than primary quinone (ψo). PSI activity changed in parallel with ψo suggesting that PSI cooperated well with PSII during leaf expansion. It should be stressed that the development of carbon fixation process was later than primary photochemical reaction but earlier than photosynthetic electron transport during leaf expansion. The later development of photosynthetic electron transport may reduce the production of reactive oxygen species from Mehler reaction, particularly under low carbon fixation.     

Chlorophyll a fluorescence analysis of high-yield rice (Oryza sativa L.) LYPJ during leaf senescence

Photosystem II (PSII) photochemistry was examined by chlorophyll (Chl) a fluorescence analysis in high-yield rice LYPJ flag leaves during senescence. Parameters deduced from the JIP-test showed that inhibition of the donor side of PSII was greater than that of the acceptor side in hybrid rice LYPJ. The natural senescence process was accompanied by the increased inactivation of oxygen-evolving complex (OEC) and a lower total number of active reaction centers per absorption. It indicated that the inhibition of electron transport caused by natural senescence might be caused partly by uncoupling of the OEC and/or inactivation of PSII reaction centers. Chl fluorescence parameters analyzed in this study suggested that energy dissipation was enhanced in order to protect senescent leaves from photodamage. Nevertheless, considerably reduced PSI electron transport activity was observed at the later senescence. Thus, natural senescence inhibited OEC-PSII electron transport, but also significantly limited the PSII-PSI electron flow.     

Soybean leaf responses to threecornered alfalfa hopper petiole girdling

Specific leaf weight, percent moisture, and free sugar, starch, and amino nitrogen content of soybean (Glycine max (L.) Merrill) leaves were measured at 1, 3, 7, and 10 day(s) after petiole girdling by threecornered alfalfa hopper,Spissistilus festinus (Say) (Homoptera: Membracidae) nymphs. Leaf starch was increased at 1, 3, and 7 day(s) after girdling and largely accounted for corresponding increases in specific leaf weight and decreases in percent moisture, free sugars, and amino nitrogen. Specific leaf weight was increased at 10 days after girdling despite no increase in starch. Amino nitrogen content was decreased 10 days after girdling. When leaf dry weights were corrected for starch, free sugar content was not affected by girdling, and amino nitrogen content was reduced only at 3 and 10 days. The amino nitrogen: free sugar ratio was reduced only at 10 days after girdling. Changes in leaf starch indicated a rapid but reversible effect of girdling on leaf carbohydrate metabolism.     

Photo- and antioxidant protection and salicylic acid accumulation during post-anthesis leaf senescence in Salvia lanigera grown under Mediterranean climate

Post-anthesis leaf senescence is a key developmental process in the life of plants as it is the time during which material built up by the plant during its growth phase is mobilized into reproductive tissues. Here we aimed to study the extent of photo- and antioxidant protection and salicylic acid (SA) accumulation during post-anthesis leaf senescence in a perennial plant, Salvia lanigera Poir. grown under Mediterranean field conditions. SA levels increased sharply (up to 2.7-fold) during early stages of leaf senescence until fruit and seed formation occurred (i.e. 4 weeks after anthesis). Later on, SA levels kept at constant high levels until leaf abscission occurred (i.e. 7 weeks after anthesis). Reductions in chlorophyll and carotenoid (lutein, violaxanthin and β-carotene) levels occurred progressively during leaf senescence. In contrast, xanthophyll cycle de-epoxidation increased during early stages of leaf senescence and remained constant later, similar to SA accumulation. Indeed, xanthophyll cycle de-epoxidation strongly positively correlated with SA levels (r² = 0.92). The maximum efficiency of PSII (F_v/F_m ratio) kept around 0.80 throughout the experiment, except during the latest stage of leaf senescence (i.e. after fruit and seed formation), when this ratio decreased to 0.72, thus indicating damage to PSII. It is concluded that endogenous SA levels increase sharply during early stages of post-anthesis leaf senescence and concomitantly with activation of photoprotection mechanisms, such as xanthophyll cycle-dependent excess energy dissipation, thus avoiding damage to PSII until fruit and seed formation have been accomplished.