The Xanthophyll Cycle Modulates the Kinetics of Nonphotochemical Energy Dissipation in Isolated Light-Harvesting Complexes, Intact Chloroplasts, and Leaves of Spinach1

We analyzed the kinetics of nonphotochemical quenching of chlorophyll fluorescence (qN) in spinach (Spinacia oleracea) leaves, chloroplasts, and purified light-harvesting complexes. The characteristic biphasic pattern of fluorescence quenching in dark-adapted leaves, which was removed by preillumination, was evidence of light activation of qN, a process correlated with the de-epoxidation state of the xanthophyll cycle carotenoids. Chloroplasts isolated from dark-adapted and light-activated leaves confirmed the nature of light activation: faster and greater quenching at a subsaturating transthylakoid pH gradient. The light-harvesting chlorophyll a/b-binding complexes of photosystem II were isolated from dark-adapted and light-activated leaves. When isolated from light-activated leaves, these complexes showed an increase in the rate of quenching in vitro compared with samples prepared from dark-adapted leaves. In all cases, the quenching kinetics were fitted to a single component hyperbolic function. For leaves, chloroplasts, and light-harvesting complexes, the presence of zeaxanthin was associated with an increased rate constant for the induction of quenching. We discuss the significance of these observations in terms of the mechanism and control of qN.

Xanthophyll, carotene, and o-tocopherol stability in alfalfa as affected by pilot- and industrial-scale dehydration

Bibliography: p. 13-14.

Calculations on the end-group conformational barrier in carotenoids and a study of the bioavailability of xanthophyll esters in humans

The present study measures the increase in serum carotenoid concentration in 30 healthy individuals after supplementation with a low dose xanthophyll ester (3 and 6 mg of lutein equivalent/per day) when compared to a placebo. Serum levels of carotenoids were measured using HPLC and showed an increase in the concentration of lutein, zeaxanthin and four lutein metabolites proportional to dose. In order to further assess the importance of the end-group structure in carotenoids we have investigated the influence of the end-group type and functionality on the conformational energy barrier. We used the density functional method implemented on GAUSSIAN 98 to calculate the conformational energy curves for rotation of the P-ring or the E-ring relative to short polyene chains around the C6-C7 single bond. A large barrier is observed for the interconversion of conformers in the E-rings (8 kcal/mol) when compared to beta rings (2.3-3 kcal/mol).

Effect of lutein on the transport of Ca2+ across phospholipid bilayer and mitochondrial membrane

Lutein (3,3'-dihydroxy alpha-carotene), a xanthophyll present in plant chloroplasts, increases the permeability of phospholipid vesicles to Ca2+, even though the pigment does not bind the metal ion. Energy-dependent uptake of Ca2+ by mitochondria is inhibited by lutein, which permits a rapid efflux of the ion from Ca2+-loaded mitochondria. These results are consistent with the view that the deleterious action of lutein on mitochondrial oxidative phosphorylation results from its destabilizing action on membrane structure.

Carotenoids In 3 Stages Of Ripening Of Mango

The distribution of carotenoids, both qualitative and quantitative, during 3 stages of ripening of mango has been studied using chromatographic, spectroscopic and chemical methods. There was an increase in content as well as in number of carotenoids during ripening. The present study showed there were 15, 14 and 17 different carotenoids in the unripe, partially ripe and fully ripe mangoes, respectively. Even though phytofluene (39.26%) was the major carotenoid in the partially ripe mango, β-carotene constituted the major carotenoid in the unripe (37.47%) and fully ripe mango (50.64%). cis-β-Carotene was present only in the fully ripe mango. Only the unripe mango contained ζ-carotene, whereas γ-carotene was present in all the 3 stages of ripening. The major xanthophyll present in the unripe mango was mutatoxanthin (9.44%), whereas auroxanthin constituted the major hydroxylated carotenoid of the partially ripe (5.07%) and fully ripe (10.40%) mangoes. The percent of cryptoxanthin dropped to lower levels during ripening. As ripening proceeded, lutein completely is appeared. There were significant quantities of eaxanthin in the partially ripe and fully ripe mango. Epoxy carotenoids such as 5,6-monoepoxy-β-carotene, mutatochrome, cis-violaxanthin, luteoxanthin, mutatoxanthin and auroxanthin were observed in all 3 stages of ripening.

Activation of violaxanthin cycle in darkness is a common response to different abiotic stresses : a case study in Pelvetia canaliculata

Background: In the violaxanthin (V) cycle, V is de-epoxidized to zeaxanthin (Z) when strong light or light combined with other stressors lead to an overexcitation of photosystems. However, plants can also suffer stress in darkness and recent reports have shown that dehydration triggers V-de-epoxidation in the absence of light. In this study, we used the highly stress-tolerant brown alga Pelvetia canaliculata as a model organism, due to its lack of lutein and its non-photochemical quenching independent of the transthylakoidal-ΔpH, to study the triggering of the V-cycle in darkness induced by abiotic stressors. Results: We have shown that besides desiccation, other factors such as immersion, anoxia and high temperature also induced V-de-epoxidation in darkness. This process was reversible once the treatments had ceased (with the exception of heat, which caused lethal damage). Irrespective of the stressor applied, the resulting de-epoxidised xanthophylls correlated with a decrease in Fv/Fm, suggesting a common function in the down-regulation of photosynthetical efficiency. The implication of the redox-state of the plastoquinone-pool and of the differential activity of V-cycle enzymes on V-de-epoxidation in darkness was also examined. Current results suggest that both violaxanthin de-epoxidase (VDE) and zeaxanthin-epoxidase (ZE) have a basal constitutive activity even in darkness, being ZE inhibited under stress. This inhibition leads to Z accumulation. Conclusion: This study demonstrates that V-cycle activity is triggered by several abiotic stressors even when they occur in an absolute absence of light, leading to a decrease in Fv/Fm. This finding provides new insights into an understanding of the regulation mechanism of the V-cycle and of its ecophysiological roles.

Photosynthetic responses of trees in high-elevation forests: comparing evergreen species along an elevation gradient in the Central Andes

Plant growth at extremely high elevations is constrained by high daily thermal amplitude, strong solar radiation and water scarcity. These conditions are particularly harsh in the tropics, where the highest elevation treelines occur. In this environment, the maintenance of a positive carbon balance involves protecting the photosynthetic apparatus and taking advantage of any climatically favourable periods. To characterize photoprotective mechanisms at such high elevations, and particularly to address the question of whether these mechanisms are the same as those previously described in woody plants along extratropical treelines, we have studied photosynthetic responses in Polylepis tarapacana Philippi in the central Andes (18 degrees S) along an elevational gradient from 4300 to 4900 m. For comparative purposes, this gradient has been complemented with a lower elevation site (3700 m) where another Polylepis species (P. rugulosa Bitter) occurs. During the daily cycle, two periods of photosynthetic activity were observed: one during the morning when, despite low temperatures, assimilation was high; and the second starting at noon when the stomata closed because of a rise in the vapour pressure deficit and thermal dissipation is prevalent over photosynthesis. From dawn to noon there was a decrease in the content of antenna pigments (chlorophyll b and neoxanthin), together with an increase in the content of xanthophyll cycle carotenoids. These results could be caused by a reduction in the antenna size along with an increase in photoprotection. Additionally, photoprotection was enhanced by a partial overnight retention of de-epoxized xanthophylls. The unique combination of all of these mechanisms made possible the efficient use of the favourable conditions during the morning while still providing enough protection for the rest of the day. This strategy differs completely from that of extratropical mountain trees, which uncouple light-harvesting and energy-use during long periods of unfavourable, winter conditions.

紫黄质脱环氧化酶在烟草中的过量表达和反义抑制研究

叶黄素循环被发现具有热耗散的作用后，已引起人们广泛的关注。目前普遍认为叶黄素循环的色素定位于天线色素蛋白复合体上，在跨膜质子梯度（ΔpH）形成后，玉米黄质（zeaxanthin;Z）和环氧玉米黄质（antheraxanthin;A）能够从叶绿素中吸收过多的激发能，并以热能的形式耗散到体外，从而保护光合器官免受强光的破坏。紫黄质脱环氧化酶（Violaxanthin de-epoxidase;VDE）是叶黄素循环的关键酶，存在于植物类囊体内腔中，它催化紫黄质（violaxanthin）脱环氧化生成环氧玉米黄质(A)和玉米黄质(Z)。本文利用过量表达和反义抑制技术获得两种转基因烟草植株，并用于研究紫黄质脱环氧化酶在叶黄素循环中的作用。 首先我们从烟草中克隆了编码VDE酶的基因，分别以正向和反向插入到具有潮霉素抗性选择标记的双元载体pCAMBIA1301，构建了Tvde基因的过量表达载体pCBTO和反义抑制表达载体pCBTA。然后通过根癌农杆菌(Agrobacterium tumefaciens)介导法转化烟草(Nicotiana tabacum L.)，获得了过量表达和反义抑制两种转基因植株。PCR扩增潮霉素抗性基因hpt和Southern杂交检测结果表明，Tvde基因已整合到转基因烟草的基因组中，外源基因在转基因烟草基因组中以1个拷贝的形式存在。VDE酶活性测定表明，在反义抑制转化体中VDE酶活性被抑制60%，而在过量表达转化体中VDE酶活性提高了75%。通过色素的HPLC分析和荧光动力学测定结果表明，强光处理后，在反义抑制转化体和过量表达转化体中，Z的含量，DES，NPQ和Fv/Fm等数据说明转基因烟草中VDE含量与植物非光化学猝灭能力有直接关系，进而说明叶黄素循环具有热耗散的功能。

植物叶片生长过程中光合活性、光保护机制及光系统Ⅱ热稳定性的研究

叶片在成长进程中光饱和光合速率持续提高，尽管幼叶光呼吸的测定值较低，但幼叶光呼吸与总光合之比较高。叶片在成长初期就具有较高的最大光化学效率，但是仍略低于发育成熟的叶片。随着叶片的成长，光下叶片光系统II实际效率增加，而非光化学猝灭下降。幼叶叶黄素总量与叶绿素之比较高，随着叶而积的增加该比值下降;光下，幼叶脱环氧化程度较高。同时，我们也观察到叶片生长初期活性氧清除酶系的活性较高。叶片生长过程中提高的光破坏防御机制与叶片相对含水量呈现很好的负相关，而不是叶片水势。因此，推测叶片生长过程中光破坏防御机制的建立可能与叶片膨压有关。 自然状态下，不同展开程度的叶片均未发生明显的光抑制;但将所柏‘叶片平展并暴露在强光下时幼叶发生明显的光抑制，伴随叶丽积的增加光抑制程度减轻。自然条件下测量叶片角度，观察到在叶片展开过程中叶柄夹角逐渐增加：日动态过程中幼叶的悬挂角随光强增加而明显减小，而完全展丌叶的悬挂角变化幅度很小。叶片角度的变化使实际照射到幼叶叶表的光强减少。推测较强的光ll乎吸、依赖叶黄素循环的热耗散、活性氧清除酶系以及较大的叶角变化可能是自然状态下幼叶未发生严重光抑制和光破坏的原因。 与成熟叶片相比，高温严重地伤害新生叶片光系统IT的结构，并导致最大光化学效率和光系统II活性下降。高温对光系统II的伤害包括供体测和受体测;而进一步的研究和分析表明高温很可能影响放氧复合物活性，从而改变光系统II的结构并最终导致受体测电子传递受阻。叶片生长和光合机构的健全使得光系统II热稳定性逐步增强，因此推测叶片生长过程中光系统II热稳定性的增强可能主要与其放氧复合物结构和功能的完善有关。

不同基因型小麦（Triticum aestivum）京411与小偃54抗光氧化特性及其机理的比较研究

对两种不同基因型小麦京411（北京地区高产小麦品种）和小偃54（在生产上已应用二十年的优良品种）幼苗及旗叶光抑制特性进行了比较研究，着重探讨了它们抗光氧化的差异及其机理，主要结果如下： 1. 强光条件下，同京411相比小偃54能保持较高的光合色素含量和放氧速率。其DCPIP光还原活性也较高。 2. 光谱特性分析表明，强光胁迫下小偃54在红区及蓝区的特征性吸收峰及F683荧光发射峰下降的幅度明显小于京411相应峰位的下降。 3. 色素蛋白复合物分析表明，强光条件下，京411色素蛋白复合物中LHCII聚合体大量的解聚，而小偃54在强光条件下仍能保持较高比例的LHCII聚合体。这可能在一定的程度上有利于小偃54在强光下维持较强的激发能耗散的能力。 4. 多肽SDS-PAGE分析表明，强光对不同基因型小麦中同PSII光抑制敏感性有关的两个外周蛋白23kD和17kD的影响不同。光抑制明显地降低了京411的23kD和17kD的含量，而对于小偃54中这两个外周蛋白23kD和17kD的影响不大。强光条件下小偃54旗叶PSII颗粒中捕光色素蛋白27kD含量提高，而京411捕光色素蛋白27kD含量明显的下降。 5. 上述结果表明，西北地区的优良小麦品种小偃54同北京地区的高产品种京411相比更耐强光的胁迫，其抗光氧化的能力较强。 6. 进一步分析表明，同京411相比，小偃54抗光氧化的主要原因是其不仅含有较大的叶黄素循环的色素库，而且在强光下能维持高的VDE酶活性及高水平的叶黄素循环的脱环化水平。 7. 叶黄素循环色素在两个小麦品种类囊体膜色素蛋白复合体中的分布存在差异，抗光氧化的小偃54大部分的VAZ分布在PSII上，其中绝大部分集中在LHCII聚合体上。LHCII上VAZ的集中分布可能有利于在强光下对过多光能的耗散，减少过多激发能对PSII的损伤。 8. 类囊体膜流动性分析表明，叶黄素循环参与了对类囊体膜流动性的调整，对维持强光下抗光氧化品种小偃54的类囊体膜相对稳定起重要作用。 9. 依赖于叶黄素循环的热耗散是抗光氧化品种小偃54的一个主要的光保护途径。

外源ABA对玉米幼苗光抑制的调节

玉米幼苗经外源脱落酸(ABA)处理后，其生长与光合作用，如株高、干物质积累、净光合速率(Pn)、光合作用的量子效率(фC02)和羧化效率(CE)，以及光系统II (PSII)实际光化学效率(фPSII)等受到抑制，且该抑制程度与处理ABA的浓度呈相关性。PSII最大光化学活性(Fv/Fm)变化表明，以10和25μmol L-I ABA处理玉米幼苗7天，可明显提高其抗光抑制能力，而50μmol L-1ABA处理的玉米幼苗在相同条件下的抗光抑制能力下降。进一步以25μmol L-lABA处理玉米幼苗来研究，结果表明ABA处理可减缓强光下玉米叶片Pn、CE、фPS II和叶片吸收光能光化学猝灭(qP)的下降，同时增强叶片吸收光能的非光化学猝灭(NPQ)。另外，叶绿素荧光非光化学猝灭的中间组分(qm)增强，光抑制后Fv/Fm的恢复能力提高，这表明ABA处理高提高了强光下玉米幼苗的光系统状态转换能力和Psn循环修复作用。除此之外，ABA处理后玉米幼苗的叶黄素循环类色素，如紫黄质(V)、环氧玉米黄质(A)和玉米黄质(Z)的含量增加，叶黄素循环库(V+A+Z)增大，说明依赖于叶黄素循环的热耗散在ABA处理玉米幼苗中得到加强。另外，ABA处理幼苗在强光下保持较高фPsII／Pn活性，以及叶片抗氧化酶活性提高，如超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)、单脱氢抗坏血酸还原酶(DHAR)和谷胱甘肽还原酶(GR)，抗氧化物含量增加，如抗坏血酸(AsA)、脱氢抗坏血酸(DHAsA)、还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSH)，这说明ABA诱导Mehler-peroxidase反应的增强在提高玉米幼苗抗光抑制能力中也发挥重要作用。 玉米叶片光系统I和光系统II在相同强度(300μmolm-2 S-l)的红光（655nm）和远红光（700-770 nm）共同照射下，光系统I(PSI)和光系统II(PSII)吸收光能基本平衡，叶片光合作用处于状态1，此时Psn保持较高的光适应下最大荧光( Fml)。关闭远红光，使叶片只处在红光照射下，则会引起光下PSII最大荧光( Frri2)的降低。关闭远红光约20nun后，光下下降的Psn最大荧光基本达到稳定，叶片光合作用处于状态2。这种在状态l向状态2的转换过程中所发生的PSII最大荧光下降不受DTT（叶黄素循环抑制剂）的影响，且整个过程中PsII最大光化学效率( Fv/Fm)保持不变，而光下PSII初始荧光(F0')在前20min内迅速降低。另外，在PSII吸收的红光照射下，玉米叶片吸收光向PSII分配的量(B)不断减少，与此同时，吸收光能向PSI分配的量(a)不断增多。ABA预处理玉米幼苗7天，可进一步加强红光下PSII最大荧光(Fm2)的降低，使荧光参数Fm1/Fm2—1增大，而使β／α-1降低。另外，ABA处理较对照幼苗在红光下呈现更高的荧光非光化学猝灭中间组分(qm)。在引入叶绿体蛋白激酶抑制剂NEM的情况下，ABA处理与对照玉米叶片在红光下所表现的qm差异则消失。从状态1向状态2的转换过程中，ABA处理引起玉米叶片77K低温荧光F684/F732的下降幅度显著加大。以上结果说明ABA处理可提高玉米幼苗光合作用的状态转换能力。 用的25μmol L-l ABA对玉米幼苗进行长时间（根系浇灌7天，LT）和短时间（实验前一天晚上叶面喷施1次，ST）处理，研究叶片C02同化、PsII化学活性，以及叶黄素循环的变化。结果表明在非光抑制状态下，LT与ST对玉米叶片光化学活性( Fv/Fm)及叶片羧化效率(CE)没有明显影响，但二者都引起叶片净光合速率(Pn)与气孔导度(Gs)下降。LT处理增大玉米叶片叶黄素循环库，而ST处理对该库大小没有影响。1500μmol m-2 s-1强光可明显引起玉米幼苗叶片Fv/Fm降低，但与对照幼苗相比，LT处理能显著减缓Fv/Fm降低。经60min强光照射后，ST与对照在Fv/Fm、фPS II、Pn和CE等参数上没有明显差异，但这些参数在LT处理的玉米幼苗中仍保持较高水平。LT处理幼苗叶黄素循环类色素含量及非光化学荧光猝灭(NPQ)都显著高于对照，膜脂过氧化产物MDA含量比对照低。而ST处理与对照在叶黄素循环类色素含量、NPQ和MDA含量等方面没有明显差异。以上结果说明ST处理对玉米幼苗光抑制没有明显影响，而LT处理可增强玉米幼苗抗光抑制能力，这可能与ABA处理使玉米幼苗在强光下维持较高的C02同化作用，以及其诱导叶片叶黄素循环增大有关。

冬小麦抗寒性和光合作用对增强UV-B辐射与温度交互作用的响应

人类活动产生的氯氟烃化合物破坏了大气臭氧层，导致了到达地球表面的UV-B辐射大幅度增加。UV-B辐射增强可以影响到植物的生长、形态与发育等各个方面，因此有关增强UV-B辐射对植物的影响，及其与许多环境因子复合作用的研究都已经广泛开展。但是增强UV-B辐射与温度，特别是与低温的相互作用的研究报道很少。在北半球的晚秋至早春这段时期里，一些越冬生长的植物将面临着UV-B辐射增强和低温的双重胁迫，因此，迫切需要进行UV-B辐射和低温生长环境下植物的响应及其机制的研究。 以人工气候生长室中生长的冬小麦(Triticum aestivum)幼苗为试验材料，研究了低剂量（4.2 kJ m-2 d-1 UV-BBE，LUVB）和较高剂量（7.0 kJ m-2 d-1 UV-BBE，HUVB）UV-B辐射处理对20/16℃条件下幼苗抗寒力的交叉适应性及其抗氧化系统的反应;同时还研究了在两种生长温度（25/20℃和10/5℃）条件下，低剂量（4.2 kJ m-2 d-1 UV-BBE，LUVB）和超高剂量（10.3 kJ m-2 d-1 UV-BBE，SHUVB）UV-B辐射处理幼苗的生长速率、光合与荧光参数、叶黄素循环色素、抗氧化系统、以及抗寒性和酚类物质等生理反应，以期阐明不同温度条件下生长的冬小麦对UV-B辐射的生长、光合作用以及抗寒性响应与适应机制。主要结果如下： 1.在LUVB辐射处理下，在20/16℃和25/20℃条件下生长的冬小麦幼苗LT50值都显著降低，HUVB辐射处理对在20/16℃条件下生长的幼苗LT50值也可以显著降低，而SHUVB辐射对25/20℃条件下生长的幼苗LT50值没有显著影响。但是，LUVB和SHUVB辐射处理都导致了10/5℃条件下生长的幼苗LT50值的显著增加。表明适当的UV-B辐射能增强较高温度（20/16℃或25/20℃）条件下冬小麦幼苗的抗寒力，即表现出对冷冻低温的交叉适应性，但低温（10/5℃）生长条件却削弱了UV-B辐射下冬小麦的抗寒能力。 2.在20/16℃条件下接受UV-B辐射预处理的幼苗在-6℃条件下冷冻胁迫6 h再缓慢恢复6 h后，与未进行UV-B辐射处理的对照相比，其叶片过氧化氢酶（CAT）、愈创木酚过氧化物酶（GPX）、谷胱甘肽还原酶（GR）活性，谷胱甘肽氧化还原比例（GSH/GSSG）都显著提高，而由硫代巴比妥酸反应物质（TBARS）代表的膜质过氧化程度显著低于对照。此外，UV-B辐射期间处理幼苗的H2O2含量较对照显著增加，而冷冻恢复以后却明显低于对照。表明UV-B辐射诱导的抗寒力的提高应该与冷冻恢复后植株体内抗氧化系统的上调表达有关，H2O2可能参与了UV-B辐射对低温的交叉适应的信号传导。 3.除25/20℃生长条件下的LUVB处理的小麦幼苗外，UV-B辐射显著降低幼苗的相对生长速率（RGR）、净光合速率（Pn）、光系统II最大量子产量（Fv/Fm）、光系统II实际量子产量（(F΄m−Fs)/F΄m）以及光化学淬灭（qP），但是UV-B辐射并不影响叶片胞间CO2浓度（Ci），而且冬小麦幼苗生长和光合作用的抑制被增加的UV-B辐射剂量和降低的温度加强。UV-B辐射引起的光抑制由非气孔限制所导致，而且主要与PS II光化学效率降低有关。 4.UV-B辐射显著增加了两个温度条件（20/16℃或25/20℃）下生长的冬小麦幼苗叶黄素循环过程中紫黄素（V）的合成，但抑制了V向玉米黄质（Z）的转化，从而造成了对照与LUVB辐射处理幼苗之间的叶片中脱环氧化比例（DEPS）和NPQ无显著性差异，但SHUVB辐射处理幼苗叶片中DEPS和NPQ显著降低。因此，在本试验条件下，增强UV-B辐射处理的冬小麦可能并不通过热耗散形式形成光保护机制，光抑制形成的过剩激发能的耗散可能更多地通过代谢途径来实现。 5.UV-B辐射处理提高了在25/20℃条件下幼苗的超氧化物歧化酶（SOD）、抗坏血酸过氧化物酶（APX）和GR等活性，以及抗坏血酸氧化还原比例（AsA/DHA）和GSH/GSSG;但是在10/5℃下，UV-B辐射除了导致SOD和CAT活性升高之外，对APX活性和AsA/DHA并不产生明显影响，但GPX和GSH/GSSG则显著降低。说明UV-B辐射幼苗的抗氧化系统在较高生长温度下显著地增强，而在低温10/5℃下被严重地削弱或降低，即低温阻止了代谢途径的光保护机制的正常运转。 6.多酚物质在UV-B辐射或低温10/5℃条件下都能显著地累积，且在UV-B辐射和低温复合作用下增加尤其显著，表明多酚物质在两个温度生长条件下特别是低温条件下都参与了对UV-B辐射幼苗的保护。 7.在高温条件下仅仅SHUVB处理的幼苗TBARS含量显著增加，而低温10/5℃条件下两个UV-B辐射处理都非常显著地上升，说明与高温生长条件相比较，低温加重了UV-B辐射引起的氧化胁迫，低温10/5℃条件下幼苗多酚的增加以及抗氧化系统的部分增强都没有能阻止UV-B辐射对幼苗的伤害。

桃和苹果库源关系中源叶光合作用，光抑制及其光保护机制的研究

以‘早久保’（Prunus persica (L.) Batch.）为试材，在果实最后迅速生长期，通过去果处理降低库力，同时设留果对照，并通过环剥和保留相同数量叶片严格控制库源关系，进行了源叶净光合速率（Pn）、叶绿素荧光、叶黄素循环、抗氧化酶及抗氧化同化物日变化的研究。结果表明，和留果对照相比，去果处理显著降低了源叶Pn、气孔导度（gs）和蒸腾速率（E），但显著增加了胞间二氧化碳浓度（Ci）、叶面饱和蒸汽压亏缺（VPDl）和叶片温度（Tl）。光系统II光化学效率（ΦPSII）以及羧化速率（CE）与Pn平行降低。中午去果降低Pn主要归因于非气孔限制。在低库需条件下，开放的PSII反应中心捕获能量的降低以及关闭的PSII反应中心的增加导致了ΦPSII的降低。去果处理叶片中依赖于叶黄素循环的热耗散以及抗氧化系统的上调保护叶片免受光氧化破坏。和留果对照相比，去果处理的叶片有更大的叶黄素循环库，更高的脱环氧化状态以及更高的抗氧化酶活性，包括超氧化物歧化酶（SOD）、抗坏血酸过氧化物酶（APX）、单脱氢抗坏血酸还原酶（MDAR）和脱氢抗坏血酸还原酶（DHAR）的活性以及更高的还原型抗坏血酸（AsA）和还原型谷胱甘肽（GSH）的含量。但与此同时，去果显著增加了过氧化氢（H2O2）以及丙二醛（MDA）的含量，这意味着在去果处理的叶片中可能会发生光氧化破坏。 以一年生‘皇家嘎拉’苹果（Malus domestica Borkh.）组培苗为试材，通过环剥降低库力，进行了源叶Pn、叶绿素荧光、核酮糖-1,5-二磷酸羧化酶/氧化酶（Rubisco）以及光系统II（PSII）复合体关键蛋白PsbA和PsbO含量日变化的研究。和对照相比，环剥显著降低了源叶Pn、gs和E，但是却显著增加了Ci、Tl和淀粉的含量。在低库需下，开放的PSII反应中心捕获能量的降低以及关闭的PSII反应中心的增加导致了ΦPSII的降低。另一方面，环剥降低了光合作用关键酶Rubisco以及PSII复合体PsbA和放氧复合体PsbO的含量。以上结果表明，环剥降低Pn主要归因于非气孔限制。

Modelling a light-driven phytoplankton succession

We used a numerical model to investigate if and to what extent cellular photoprotective capacity accounts for succession and vertical distribution of marine phytoplankton species/groups. A model describing xanthophyll photoprotective activity in phytoplankton has been implemented in the European Regional Sea Ecosystem Model and applied at the station L4 in the Western English Channel. Primary producers were subdivided into three phytoplankton functional types defined in terms of their capacity to acclimate to different light-specific environments: low light (LL-type), high light (HL-type) and variable light (VL-type) adapted species. The LL-type is assumed to have low cellular level of xanthophyll-cycling pigments (PX) relative to the modelled photosynthetically active pigments (chlorophyll and fucoxanthin (FUCO) = PSP). The HL-type has high PX content relative to PSP while VL-type presents an intermediate PX to PSP ratio. Furthermore, the VL-type is capable of reversibly converting FUCO to PX and synthesizing new PX under high-light stress. In order to reproduce phytoplankton community succession with each of the three groups being dominant in different periods of the year, we had also to assume reduced grazing pressure on HL-adapted species. Model simulations realistically reproduce the observed seasonal patterns of pigments and nutrients highlighting the reasonability of the underpinning assumptions. Our model suggests that pigment-mediated photophysiology plays a primary role in determining the evolution of marine phytoplankton communities in the winter-spring period corresponding to the shoaling of the mixed layer and the increase of light intensity. Grazing selectivity however contributes to the phytoplankton community composition in summer.

Localization of the mechanism of pH-dependent non- photochemical fluorescence quenching in thylakoid membranes

Higher plants have evolved a well-conserved set of photoprotective mechanisms, collectively designated Non-Photochemical Quenching of chlorophyll fluorescence (qN), to deal with the inhibitory absorption of excess light energy by the photosystems. Their main contribution originates from safe thermal deactivation of excited states promoted by a highly-energized thylakoid membrane, detected via lumen acidification. The precise origins of this energy- or LlpH-dependent quenching (qE), arising from either decreased energy transfer efficiency in PSII antennae (~ Young & Frank, 1996; Gilmore & Yamamoto, 1992; Ruban et aI., 1992), from alternative electron transfer pathways in PSII reaction centres (~ Schreiber & Neubauer, 1990; Thompson &Brudvig, 1988; Klimov et aI., 1977), or from both (Wagner et aI., 1996; Walters & Horton, 1993), are a source of considerable controversy. In this study, the origins of qE were investigated in spinach thylakoids using a combination of fluorescence spectroscopic techniques: Pulse Amplitude Modulated (PAM) fluorimetry, pump-probe fluorimetry for the measurement of PSII absorption crosssections, and picosecond fluorescence decay curves fit to a kinetic model for PSII. Quenching by qE (,..,600/0 of maximal fluorescence, Fm) was light-induced in circulating samples and the resulting pH gradient maintained during a dark delay by the lumenacidifying capabilities of thylakoid membrane H+ ATPases. Results for qE were compared to those for the addition of a known antenna quencher, 5-hydroxy-1,4naphthoquinone (5-0H-NQ), titrated to achieve the same degree of Fm quenching as for qE. Quenching of the minimal fluorescence yield, F0' was clear (8 to 130/0) during formation of qE, indicative of classical antenna quenching (Butler, 1984), although the degree was significantly less than that achieved by addition of 5-0H-NQ. Although qE induction resulted in an overall increase in absorption cross-section, unlike the decrease expected for antenna quenchers like the quinone, a larger increase in crosssection was observed when qE induction was attempted in thylakoids with collapsed pH gradients (uncoupled by nigericin), in the absence of xanthophyll cycle operation (inhibited by DTT), or in the absence of quenching (LlpH not maintained in the dark due to omission of ATP). Fluorescence decay curves exhibited a similar disparity between qE-quenched and 5-0H-NQ-quenched thylakoids, although both sets showed accelerated kinetics in the fastest decay components at both F0 and Fm. In addition, the kinetics of dark-adapted thylakoids were nearly identical to those in qEquenched samples at F0' both accelerated in comparison with thylakoids in which the redox poise of the Oxygen-Evolving Complex was randomized by exposure to low levels of background light (which allowed appropriate comparison with F0 yields from quenched samples). When modelled with the Reversible Radical Pair model for PSII (Schatz et aI., 1988), quinone quenching could be sufficiently described by increasing only the rate constant for decay in the antenna (as in Vasil'ev et aI., 1998), whereas modelling of data from qE-quenched thylakoids required changes in both the antenna rate constant and in rate constants for the reaction centre. The clear differences between qE and 5-0H-NQ quenching demonstrated that qE could not have its origins in the antenna alone, but is rather accompanied by reaction centre quenching. Defined mechanisms of reaction centre quenching are discussed, also in relation to the observed post-quenching depression in Fm associated with photoinhibition.