两个不同基因型小麦光抑制特性的比较

比较了两个不同基因型小麦(Triticum aestivum L.)"京411"和"小偃54"的原初光能转化效率、荧光猝灭参数和光合色素对强光胁迫的响应.在正常生长条件下"京411"的光合色素含量高于"小偃54";但在高光强下"京411"出现明显的光抑制,而"小偃54"对高光强的适应上优于"京411"."小偃54"适应高光强的原因是它在高光强下能大幅度地提高叶黄素循环的调控因子抗坏血酸的浓度及紫黄素脱环氧化酶(vDE)的活性,从而加速叶黄素循环对过多光能的耗散过程.     

不同品种小麦PSⅠ颗粒光抑制过程的光谱学比较研究

用不同品种小麦（Ttiticum aestivum L.）“京411”和“小偃54”中分离纯化的PSⅠ光破坏过程的光谱特性,并比较了两的异。结果表明,强光导致PSⅠ中色素的破坏,特别是683nm状态的Chl a分子对强光敏感。光照过程中,荧光光谱的变化表明,光破坏还导致了PSⅠ中能量传递过程的破坏。“小偃54”PSⅠ颗粒在光照初期,长波长状态的Chl a分子的吸收度值略有下降后,可保持较长时间的稳定水平,在40min后才开始明显下降,同时,在强光照射的初期荧光发射增强;而“京411”PSⅠ颗粒在光抑制过程中没有这些变化。推测“小偃54”PSⅠ可能通过将能量较多地分配给长波长状态的叶绿素分子和保持相对较少的天线色素分子,以避免过多的能量向P700反应中心传递,而起到保护作用。因而具有较强的抗光氧化能力。     

强光诱导小麦叶片花青素积累的研究

以'小偃54'、'京411'等67份小麦品种(系)为试材,对强光诱导小麦叶片花青素积累的现象及品种间变异进行分析,以探讨花青素在小麦响应强光过程中的生理与分子机制.结果显示,'小偃54'和'京411'分别经25℃强光处理24 h和48 h叶片花青素含量大幅增加,而15℃和30℃强光处理增幅较小;强光处理48 h后,'京411'的花青素含量高达7.2 μg·g-1FW,约为'小偃54'的10倍.对'京411'而言,花青素含量随叶位自下而上递减,并且按叶鞘、叶基部、叶中部和叶尖的部位依次递减.强光处理24 h和48 h后能诱导'小偃54'和'京411'的PAL活性增强.强光处理48 h后,紫色胚芽鞘类品种的花青素含量高于绿色胚芽鞘类品种,不同品种强光下的花青素吸收峰均在520 nm处.研究表明,强光下叶片花青素含量升高可能是紫色胚芽鞘类小麦品种抗强光的一种机制.     

烟草叶绿体NAD(P)H脱氢酶在抵御高温胁迫中的作用

经42℃高温处理48 h以上, 烟草(Nicotiana tabacum L.)ndhC-ndhK-ndhJ基因缺失突变体(ΔndhCKJ)植株较其野生型(WT)先出现茎部褐变、叶片萎蔫等氧化伤害症状. 作用光关闭后的叶绿素荧光动力学表明, WT植株中NAD(P)H脱氢酶(NDH)介导的PSI循环电子传递和叶绿体呼吸在高温胁迫时被促进了. 用甲基紫精(MV)处理叶圆片的结果显示, ΔndhCKJ光合机构更易受到光氧化伤害, 甚至首先发生叶绿素漂白. P700氧化还原分析表明, NDH介导的循环电子传递途径可能通过与MV竞争电子而减少活性氧(ROS)的积累. 将叶圆片于42℃处理6 h后, ΔndhCKJ光化学反应活性的下降比WT更显著, 与此一致, 可溶性Rubisco活化酶含量显著低于WT, 且电子传递链还原程度和非光化学能量耗散水平均显著高于WT. 叶绿素毫秒延迟发光慢相的测定结果显示NDH介导的循环电子传递有助于跨膜质子梯度(ΔpH)的形成, 但其耗用在DndhCKJ中受到严重抑制. 根据以上结果推测, NDH介导的循环电子传递在高温胁迫下运转加快, 并将过剩的电子分流至叶绿体呼吸途径, 此外, NDH途径提供的DpH可能在一定程度上有利于维持CO₂同化的进行, 从而能够减轻光氧化胁迫的伤害.     

强光下硫化氢通过促进光系统II的活性来缓解水稻的光抑制

以水稻品种‘II优084’为材料,测定了强光胁迫下,水稻光合速率、叶绿素荧光快速诱导曲线（OJIP）以及O2ˉ·和H2O2在水稻叶片中积累的影响。结果表明强光胁迫下,水稻的净光合速率及气孔导度下降;光系统II（PSII）反应中心关闭的比例以及电子传递链中光系统II受体侧原初醌受体（QA）的还原程度增加;PSII反应中心电子传递的量子产额、能量以及传递到下游电子链的比率下降;光抑制下PSII的过剩能量向PSI的状态装换减少;自由基的产生增加。而施加作为硫化氢（H2S）供体的外源硫氢化钠（NaHS）后,上述影响PSII活性的指标的负变化被缓解,捕光天线复合体LHC通过在两个光系统之间的移动,来调节两个光系统的能量分配。强光下H2S处理能促进LHC离开PSII,与PSI结合,从而减少PSII分配的激发能,增加PSI分配的激发能,缓解了PSII的过度还原。以上结果表明外源H2S通过促进PSII的光合活性来缓解水稻光抑制伤害。     

强光下硫化氢通过促进光系统Ⅱ的活性来缓解水稻的光抑制

以水稻品种‘II优084’为材料,测定了强光胁迫下,水稻光合速率、叶绿素荧光快速诱导曲线(OJIP)以及O2ˉ·和H2O2在水稻叶片中积累的影响。结果表明强光胁迫下,水稻的净光合速率及气孔导度下降;光系统II(PSII)反应中心关闭的比例以及电子传递链中光系统II受体侧原初醌受体(QA)的还原程度增加;PSII反应中心电子传递的量子产额、能量以及传递到下游电子链的比率下降;光抑制下PSII的过剩能量向PSI的状态装换减少;自由基的产生增加。而施加作为硫化氢(H2S)供体的外源硫氢化钠(NaHS)后,上述影响PSII活性的指标的负变化被缓解,捕光天线复合体LHC通过在两个光系统之间的移动,来调节两个光系统的能量分配。强光下H2S处理能促进LHC离开PSII,与PSI结合,从而减少PSII分配的激发能,增加PSI分配的激发能,缓解了PSII的过度还原。以上结果表明外源H2S通过促进PSII的光合活性来缓解水稻光抑制伤害。     

小麦叶片的状态转换涉及PSⅡ向PSⅠ激发能满溢的变化

叶片照远红光后,其叶绿素荧光参数Ｆｍ／Ｆｏ和两个光系统低温荧光产量Ｆ６８５／Ｆ７３５升高,照红光后,其Ｆｍ／Ｆｏ和Ｆ６８５＝Ｆ７３５降低;在照远荭光或红光过程中,与Ｆ６８５／Ｆ７３５的变化相比,Ｆｍ／Ｆｏ的变化幅度在较甜美的时间内最大;ＮａＦ预处理叶片经工光照射时,其Ｆｍ／Ｆｏ和Ｆ６８５／Ｆ７３５不增加ＤＣＭＵ预处理的叶片经红光照射时,这些结果表明,小麦叶片状态转换过程中两个光系统间能量分配贩变化     

两个不同基因型小麦光抑制特性的比较

比较了两个不同基因型小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）“京４１１”和“小偃５４”的原初光能转化效率、荧光猝灭参数和光合色素对强光胁迫的响应。在正常生长条件下“京４１１”的光合色素含量高于“小偃５４”;但在高光强下“京４１１”出现明显的光抑制,而“小偃５４”对高光强的适应上优于“京４１１”。“小偃５４”适应高光强的原因是它在高光强下能大幅度地提高叶黄素循环的调控因子抗坏血酸的浓度及紫黄     

条锈病对小麦(Triticum aestivum L.)叶片光合功能及光合功能蛋白D1表达的影响

测定了小麦(Triticum aestivum L.)感染小麦条锈病后的光合常数,以及叶绿素含量、类囊体膜光合电子传递速率和光合反应中心D1蛋白的变化.实验显示,条锈病侵染导致感病小麦叶片净光合速率与叶绿素含量降低;抗病小麦经侵染后净光合速率却有恢复过程,叶绿素含量先降后升.此外,感病小麦叶片被侵染后全链电子传递速率受到抑制,PSII电子传递速率的变化与全链电子传递速率的变化趋势相似,但PSI电子传递速率受到的影响较小;抗病小麦小麦叶片被侵染后电子传递速率所受影响较小.同时发现,病程中,感病和抗病小麦PSII的光合反应中心D1蛋白含量变化总是与PSII电子传递速率的变化类似,推测D1蛋白的表达量变化是引起PSII电子传递活性与全链电子传递速率变化的主要因素之一.     

条锈病对小麦（Triticum aestivum L.）叶片光合功能及光合功能蛋白D1表达的影响

测定了小麦（Triticum aestivum L.）感染小麦条锈病后的光合常数,以及叶绿素含量、类囊体膜光合电子传递速率和光合反应中心D1蛋白的变化。实验显示,条锈病侵染导致感病小麦叶片净光合速率与叶绿素含量降低;抗病小麦经侵染后净光合速率却有恢复过程,叶绿素含量先降后升。此外,感病小麦叶片被侵染后全链电子传递速率受到抑制,PSII电子传递速率的变化与全链电子传递速率的变化趋势相似,但PSI电子传递速率受到的影响较小;抗病小麦小麦叶片被侵染后电子传递速率所受影响较小。同时发现,病程中,感病和抗病小麦PSII的光合反应中心D1蛋白含量变化总是与PSII电子传递速率的变化类似,推测D1蛋白的表达量变化是引起PSII电子传递活性与全链电子传递速率变化的主要因素之一。     

Separation of light-induced linear, cyclic and stroma-sourced electron fluxes to P700+ in cucumber leaf discs after pre-illumination at a chilling temperature

Pre-illumination of cucumber leaf discs at 4 degrees C with low-irradiance white light (i) led to a marked decrease in the extent of photo-oxidation of P700 (the special chlorophyll pair in the PSI reaction center) in actinic light at room temperature and (ii) hastened the post-illumination re-reduction of P700+. Quantifying the linear, cyclic and stroma-sourced electron fluxes to P700+ in two actinic light regimes, we found that there was no increase in cyclic or linear electron fluxes to account for these changes. Rather, we observed a decrease in the maximum extent of P700 photo-oxidation assayed by a strong flash superimposed on continuous, background light of wavelength 723 nm, which we interpret to represent a loss of stable charge separation in PSI due to enhanced charge recombination as a result of the pre-illumination treatment. The funneling of electrons towards fewer non-damaged PSI complexes could explain the hastened post-illumination re-reduction of P700+, aided by a slight increase in a stroma-sourced electron flux after prolonged pre-illumination at 4 degrees C. Quantifying the separate fluxes to P700+ helps to elucidate the effects of chilling of cucumber leaf discs in the light and the reasons for the hastened post-illumination re-reduction of P700+.     

Quantification of cyclic electron flow around Photosystem I in spinach leaves during photosynthetic induction

The variation of the rate of cyclic electron transport around Photosystem I (PS I) during photosynthetic induction was investigated by illuminating dark-adapted spinach leaf discs with red + far-red actinic light for a varied duration, followed by abruptly turning off the light. The post-illumination re-reduction kinetics of P700⁺, the oxidized form of the photoactive chlorophyll of the reaction centre of PS I (normalized to the total P700 content), was well described by the sum of three negative exponential terms. The analysis gave a light-induced total electron flux from which the linear electron flux through PS II and PS I could be subtracted, yielding a cyclic electron flux. Our results show that the cyclic electron flux was small in the very early phase of photosynthetic induction, rose to a maximum at about 30 s of illumination, and declined subsequently to <10% of the total electron flux in the steady state. Further, this cyclic electron flow, largely responsible for the fast and intermediate exponential decays, was sensitive to 3-(3,4-dichlorophenyl)-1,1-dimethyl urea, suggesting an important role of redox poising of the cyclic components for optimal function. Significantly, our results demonstrate that analysis of the post-illumination re-reduction kinetics of P700⁺ allows the quantification of the cyclic electron flux in intact leaves by a relatively straightforward method.     

Inhibition of photosystems I and II and enhanced back flow of photosystem I electrons in cucumber leaf discs chilled in the light

Pre-illumination of cucumber leaf discs at a chilling temperature in low-irradiance white light resulted in accelerated re-reduction of P700(+) [the special Chl pair in the photosystem I (PSI) reaction centre] when the far-red measuring light was turned off. Measurements (in +/- methyl viologen or +/- DCMU conditions) of the re-reduction half time suggest that accelerated re-reduction of P700(+) appeared to be predominantly due to charge recombination and only partly due to reductants sustained by previous cyclic electron flow around PSI. Apparently, charge recombination in PSI was greatly enhanced by inhibition of forward, linear electron flow. Inhibition of PSII electron transport was observed to occur to a lesser extent than that of PSI, but only if the measurement of PSII functionality was free from complications due to downstream accumulation of electrons in pools. We suggest that promotion of controlled charge recombination and cyclic electron flow round PSI during chilling of leaves in the light may partly prevent further damage to both photosystems.     

Control of cytochrome b6f at low and high light intensity and cyclic electron transport in leaves

The light-dependent control of photosynthetic electron transport from plastoquinol (PQH(2)) through the cytochrome b(6)f complex (Cyt b(6)f) to plastocyanin (PC) and P700 (the donor pigment of Photosystem I, PSI) was investigated in laboratory-grown Helianthus annuus L., Nicotiana tabaccum L., and naturally-grown Solidago virgaurea L., Betula pendula Roth, and Tilia cordata P. Mill. leaves. Steady-state illumination was interrupted (light-dark transient) or a high-intensity 10 ms light pulse was applied to reduce PQ and oxidise PC and P700 (pulse-dark transient) and the following re-reduction of P700(+) and PC(+) was recorded as leaf transmission measured differentially at 810-950 nm. The signal was deconvoluted into PC(+) and P700(+) components by oxidative (far-red) titration (V. Oja et al., Photosynth. Res. 78 (2003) 1-15) and the PSI density was determined by reductive titration using single-turnover flashes (V. Oja et al., Biochim. Biophys. Acta 1658 (2004) 225-234). These innovations allowed the definition of the full light response curves of electron transport rate through Cyt b(6)f to the PSI donors. A significant down-regulation of Cyt b(6)f maximum turnover rate was discovered at low light intensities, which relaxed at medium light intensities, and strengthened again at saturating irradiances. We explain the low-light regulation of Cyt b(6)f in terms of inactivation of carbon reduction cycle enzymes which increases flux resistance. Cyclic electron transport around PSI was measured as the difference between PSI electron transport (determined from the light-dark transient) and PSII electron transport determined from chlorophyll fluorescence. Cyclic e(-) transport was not detected at limiting light intensities. At saturating light the cyclic electron transport was present in some, but not all, leaves. We explain variations in the magnitude of cyclic electron flow around PSI as resulting from the variable rate of non-photosynthetic ATP-consuming processes in the chloroplast, not as a principle process that corrects imbalances in ATP/NADPH stoichiometry during photosynthesis.     

Regulation of NAD(P)H dehydrogenase-dependent cyclic electron transport around PSI by NaHSO₃ at low concentrations in tobacco chloroplasts

Although bisulfite at low concentrations (L-NaHSO?) has been found to increase the cyclic electron transport around PSI (CET), its regulative mechanism remains unknown. In this work, the role of L-NaHSO? (0.1-500 μM) in NAD(P)H dehydrogenase-dependent CET (the NDH pathway) was investigated. After treatment of tobacco leaves with L-NaHSO?, the NDH pathway, as reflected by a transient post-illumination increase in Chl fluorescence, the dark reduction of P700+ after far-red light and the amount of NDH, was increased after the light-dark-light transition, but was slightly lowered under continuous light. Meanwhile, the linear electron transport (LET) was accelerated by L-NaHSO? under both the light regimes. Experiments in thylakoids further demonstrated that both LET, monitored by light-dependent oxygen uptake, and CET, as determined from the NADPH-dependent oxygen uptake and dark reduction of P700+, were enhanced by L-NaHSO? and the enhancements were abolished by superoxide dismutase. Furthermore, L-NaHSO?-induced CET was partially impaired in thylakoids of the ΔndhCKJ mutant, while L-NaHSO?-induced LET was not affected. Based on these results, we propose that the photooxidation of L-NaHSO? initiated by superoxide anions in PSI regulates NDH pathway to maintain efficient photosynthesis.     

Heterogeneity of Photosystem I reaction centers in barley leaves as related to the donation from stromal reductants

The light-response curves of P700 oxidation and time-resolved kinetics of P700⁺ dark re-reduction were studied in barley leaves using absorbance changes at 820 nm. Leaves were exposed to 45 °C and treated with either diuron or diuron plus methyl viologen (MV) to prevent linear electron flow from PS II to PSI and ferredoxin-dependent cyclic electron flow around PSI. Under those conditions, P700⁺ could accept electrons solely from soluble stromal reductants. P700 was oxidized under weak far-red light in leaves treated with diuron plus MV, while identical illumination was nearly ineffective in diuron-treated leaves in the absence of MV. When heat-exposed leaves were briefly illuminated with strong far-red light, which completely oxidized P700, the kinetics of P700⁺ dark reduction was fitted by a single exponential term with half-time of about 40 ms. However, two first-order kinetic components of electron flow to P700⁺ (fast and slow) were found after prolonged leaf irradiation. The light-induced modulation of the kinetics of P700⁺ dark reduction was reversed following dark adaptation. The fast component (half time of 80–90 ms) was 1.5 larger than the slow one (half time of about 1 s). No kinetic competition occurred between two pathways of electron donation to P700⁺ from stromal reductants. This suggests the presence of two different populations of PSI. This revised version was published online in June 2006 with corrections to the Cover Date.     

Electron Fluxes through Photosystem I in Cucumber Leaf Discs Probed by far-red Light

Cucumber leaf discs were illuminated at room-temperature with far-red light to photo-oxidise P700, the chlorophyll dimer in Photosystem (PS) I. The post-illumination kinetics of P700(+) re-reduction were studied in the presence of inhibitors or cofactors of photosynthetic electron transport. The re-reduction kinetics of P700(+) were well fitted as the sum of three exponentials, each with its amplitude and rate coefficient, and an initial flux (at the instant of turning off far-red light) given as the product of the two. Each initial flux is assumed equal to a steady state flux during far-red illumination. The fast phase of re-reduction, with rate coefficient k (1) approximately 10 s(-1), was completely abolished by a saturating concentration of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU); it is attributed to electron flow to P700(+) from PS II, which was stimulated to some extent by far-red light. The intermediate phase, with rate coefficient k (1) approximately 1 s(-1), was only partly diminished by methyl viologen (MV) which diverts electron flow to oxygen. The intermediate phase is attributed to electron donation from reduced ferredoxin to the intersystem pool; reduced ferredoxin could be formed: (1) directly by electron donation on the acceptor of PS I; and/or (2) indirectly by stromal reductants, in line with only a partial inhibition of the intermediate phase by MV. Duroquinol enhanced the intermediate phase in the presence of DCMU, presumably through its interaction with thylakoid membrane components leading to the partial reduction of plastoquinone. The slow phase of P700(+) re-reduction, with rate coefficient k (1) approximately 0.1 s(-1), was unaffected by DCMU and only slightly affected by MV; it could be associated with electron donation to either: (1) the intersystem chain by stromal reductants catalysed by NAD(P)H dehydrogenase slowly; or (2) plastocyanin/P700(+) by ascorbate diffusing across the thylakoid membrane to the lumen. It is concluded that a post-illumination analysis of the fluxes to P700(+) can be used to probe the pathways of electron flow to PS I in steady state illumination.     

Control of cytochrome b6f at low and high light intensity and cyclic electron transport in leaves

The light-dependent control of photosynthetic electron transport from plastoquinol (PQH₂) through the cytochrome b₆f complex (Cyt b₆f) to plastocyanin (PC) and P700 (the donor pigment of Photosystem I, PSI) was investigated in laboratory-grown Helianthus annuus L., Nicotiana tabaccum L., and naturally-grown Solidago virgaurea L., Betula pendula Roth, and Tilia cordata P. Mill. leaves. Steady-state illumination was interrupted (light-dark transient) or a high-intensity 10 ms light pulse was applied to reduce PQ and oxidise PC and P700 (pulse-dark transient) and the following re-reduction of P700⁺ and PC⁺ was recorded as leaf transmission measured differentially at 810-950 nm. The signal was deconvoluted into PC⁺ and P700⁺ components by oxidative (far-red) titration (V. Oja et al., Photosynth. Res. 78 (2003) 1-15) and the PSI density was determined by reductive titration using single-turnover flashes (V. Oja et al., Biochim. Biophys. Acta 1658 (2004) 225-234). These innovations allowed the definition of the full light response curves of electron transport rate through Cyt b₆f to the PSI donors. A significant down-regulation of Cyt b₆f maximum turnover rate was discovered at low light intensities, which relaxed at medium light intensities, and strengthened again at saturating irradiances. We explain the low-light regulation of Cyt b₆f in terms of inactivation of carbon reduction cycle enzymes which increases flux resistance. Cyclic electron transport around PSI was measured as the difference between PSI electron transport (determined from the light-dark transient) and PSII electron transport determined from chlorophyll fluorescence. Cyclic e⁻ transport was not detected at limiting light intensities. At saturating light the cyclic electron transport was present in some, but not all, leaves. We explain variations in the magnitude of cyclic electron flow around PSI as resulting from the variable rate of non-photosynthetic ATP-consuming processes in the chloroplast, not as a principle process that corrects imbalances in ATP/NADPH stoichiometry during photosynthesis.     

Evidence for active cyclic electron flow in twig chlorenchyma in the presence of an extremely deficient linear electron transport activity

Fast and slow chlorophyll fluorescence induction curves at high and low actinic visible light, post-illumination changes in fluorescence yield and reflectance changes at 820 nm induced by far-red light were used to characterize the state of PSII and PSI and their electron transport capabilities in chlorophyllous twig cortices of Eleagnus angustifolius L., while corresponding leaves served as controls. Twigs displayed low dark-adapted PSII photochemical efficiencies and particularly low linear electron transport rates when illuminated. In addition, their PSII population was characterized by a high proportion of inactive, non-Q_B-reducing centers and an incomplete quenching of fluorescence during the slow induction phase. It is suggested that PSII in twigs is an inefficient electron donor to PSI and/or the reductive pentose phosphate cycle. Yet, in spite of this apparent PSII deficiency, pools of intermediate electron carriers and potential PSI activity were more than sufficient to support the observed linear electron transport rates. Moreover, the rate of PSI reduction upon far-red/dark transitions and the magnitude of fluorescence yield increase upon white light/dark transitions were compatible with an efficient electron flow to PSI from stromal donors in the absence of PSII activity. We conclude that corticular chlorenchyma may be actively engaged in cyclic at the expense of a linear electron flow and discuss the possible physiological significance of this finding in conjunction with the particular microenvironmental conditions encountered within twigs.     

Cyclic electron flow around photosystem I in C(3) plants. In vivo control by the redox state of chloroplasts and involvement of the NADH-dehydrogenase complex

Cyclic electron flow around photosystem (PS) I has been widely described in vitro in chloroplasts or thylakoids isolated from C(3) plant leaves, but its occurrence in vivo is still a matter of debate. Photoacoustic spectroscopy and kinetic spectrophotometry were used to analyze cyclic PS I activity in tobacco (Nicotiana tabacum cv Petit Havana) leaf discs illuminated with far-red light. Only a very weak activity was measured in air with both techniques. When leaf discs were placed in anaerobiosis, a high and rapid cyclic PS I activity was measured. The maximal energy storage in far-red light increased to 30% to 50%, and the half-time of the P(700) re-reduction in the dark decreased to around 400 ms; these values are comparable with those measured in cyanobacteria and C(4) plant leaves in aerobiosis. The stimulatory effect of anaerobiosis was mimicked by infiltrating leaves with inhibitors of mitochondrial respiration or of the chlororespiratory oxidase, therefore, showing that changes in the redox state of intersystem electron carriers tightly control the rate of PS I-driven cyclic electron flow in vivo. Measurements of energy storage at different modulation frequencies of far-red light showed that anaerobiosis-induced cyclic PS I activity in leaves of a tobacco mutant deficient in the plastid Ndh complex was kinetically different from that of the wild type, the cycle being slower in the former leaves. We conclude that the Ndh complex is required for rapid electron cycling around PS I.