In vivo quality control of photosystem II in cyanobacteria Synechocystis sp. PCC 6803: D1 protein degradation and repair under the influence of light, heat and darkness

The cells of Synechocystis sp. PCC 6803 were subjected under photoinhibitory irradiation (600 micromolm(-2)s(-1)) at various temperatures (20-40 degrees C) to study in vivo quality control of photosystem II (PSII). The protease biogenesis and its consequences on photosynthetic efficiency (chlorophyll fluorescence ratio Fv/Fm) of the PSII, D1 degradation and repair were monitored during illumination and darkness. The loss in Fv/Fm value and degradation of D1 protein occurred not only under high light exposure, but also continued when the cells were subjected under dark restoration process after high light exposure. No loss in Fv/Fm value or D1 degradation occurred during recovery under growth/low light (30 micromol m(-2) s(-1)). Further, it helped the resynthesis of new D1 protein, essential to sustain quality control of PSII. In vivo triggering of D1 protein required high light exposure to switch-on the protease biosynthesis to maintain protease pool which induced temperature-dependent enzymatic proteolysis of photodamaged D1 protein during photoinhition and dark incubation. Our findings suggested the involvement and overexpression of a membrane-bound FtsH protease during high light exposure which caused degradation of D1 protein, strictly regulated by high temperature (30-40 degrees C). However, lower temperature (20 degrees C) prevented further loss of photoinhibited PSII efficiency in vivo and also retarded temperature-dependent proteolytic process of D1 degradation.     

Photoprotective Effects of D1 Protein Turnover and the Lutein Cycle on Three Ephemeral Plants under Heat Stress

To clarify the characteristics of photoinhibition and the primary defense mechanisms of ephemeral plant leaves against photodestruction under high temperature stress, inhibitors and the technology to determine chlorophyll fluorescence were used to explore the protective effects of D1 protein turnover and the lutein cycle in the high temperature stress of the leaves of three ephemeral plants. The results showed that the maximum light conversion efficiency (Fv/Fm) of the ephemeral plant leaves decreased, and the initial fluorescence (Fo) increased under 35°C ± 1°C heat stress for 1–4 h or on sunny days in the summer. Both Fv/Fm and Fo could be recovered after 8 h of darkness or afternoon weakening of the external temperature. Streptomycin sulfate (SM) or dithiothreitol (DTT) accelerated the decrease of Fv/Fm and the photochemical quenching coefficient (qP) in the leaves of three ephemeral plants at high temperature, and the decrease was greater in the SM than in the DTT treatment. When the high temperature stress was prolonged, the Y(II) values of light energy distribution parameters of PSII decreased, and the Y(NPQ) and Y(NO) values increased gradually in all the treatment groups of the three ephemeral plants. The results showed that the leaves of the three ephemeral plants had their own highly advanced mechanisms to protect against photodamage, which inhibited the turnover of D1 protein and xanthophyll cycle. This can damage the PSII reaction center in the leaves of the three ephemeral plants under high temperature. The protective effect of D1 protein turnover on heat stress in Erodium oxyrrhynchum and Senecio subdentatus was greater than that of the lutein cycle, while the protective effect of lutein cycle was greater than that of D1 protein turnover in Heliotropium acutiflorum subjected to heat damage.     

Photosynthetic Properties of Photosystem II in Arabidopsis thaliana lpa1 Mutant

In a previous study, we characterized a high chlorophyll fluorescence lpa1 mutant of Arabidopsis thaliana, in which approximately 20% photosystem (PS) II protein is accumulated. In the present study, analysis of fluorescence decay kinetics and thermoluminescence profiles demonstrated that the electron transfer reaction on either the donor or acceptor side of PSII remained largely unaffected in the lpa1 mutant. In the mutant, maximal photochemical efficiency (Fv/Fm, where Fm is the maximum fluorescence yield and Fv is variable fluorescence) decreased with increasing light intensity and remained almost unchanged in wild-type plants under different light conditions. The Fv/Fm values also increased when mutant plants were transferred from standard growth light to low light conditions. Analysis of PSII protein accumulation further confirmed that the amount of PSII reaction center protein is correlated with changes in Fv/Fm in lpa1 plants. Thus, the assembled PSII in the mutant was functional and also showed increased photosensitivity compared with wild-type plants.(Author for correspondence. Tel: +86 (0)10 6283 6256; Fax: +86 (0)10 8259 9384; E-mail: zhanglixin@ibcas.ac.cn)     

Light inactivation of functional photosystem II in leaves of peas grown in moderate light depends on photon exposure

To determine the dependence of in vivo photosystem (PS) II function on photon exposure and to assign the relative importance of some photoprotective strategies of PSII against excess light, the maximal photochemical efficiency of PSII (Fv/Fm) and the content of functional PSII complexes (measured by repetitive flash yield of oxygen evolution) were determined in leaves of pea (Pisum satlvum L.) grown in moderate light. The modulation of PSII functionality in vivo was induced by varying either the duration (from 0 to 3 h) of light treatment (fixed at 1200 or 1800 mol photons · m^-2 · s^-1) or irradiance (from 0 to 3000 mol photons · m^-2 · s^-1) at a fixed duration (1 h) after infiltration of leaves with water (control), lincomycin (an inhibitor of chloroplast-encoded protein synthesis), nigericin (an uncoupler), or dithiothreitol (an inhibitor of the xanthophyll cycle) through the cut petioles of leaves of 22 to 24-day-old plants. We observed a reciprocity of irradiance and duration of illumination for PSII function, demonstrating that inactivation of functional PSII depends on the total number of photons absorbed, not on the rate of photon absorption. The Fv/Fm ratios from photoinhibitory light-treated leaves, with or without inhibitors, declined pseudo-linearly with photon exposure. The number of functional PSII complexes declined multiphasically with increasing photon exposure, in the following decreasing order of inhibitor effect: lincomycin > nigericin > DTT, indicating the central role of D1 protein turnover. While functional PSII and Fv/Fm ratio showed a linear relationship under high photon exposure conditions, in inhibitor-treated leaves the Fv/Fm ratio failed to reveal the loss of up to 25% of the total functional PSII under low photon exposure. The loss of this 25% of less-stable functional PSII was accompanied by a decrease of excitation-energy trapping capacity at the reaction centre of PSII (revealed by the fluorescence parameter, 1/Fo-1/Fm, where Fo and Fm stand for chlorophyll fluorescence when PSII reaction centres are open and closed, respectively), but not by a loss of excitation energy at the antenna (revealed by the fluorescence parameter, 1/Fm). We conclude that (i) PSII is an intrinsic photon counter under photoinhibitory conditions, (ii) PSII functionality is mainly regulated by D1 protein turnover, and to a lesser extent, by events mediated via the transthylakoid pH gradient, and (iii) peas exhibit PSII heterogeneity in terms of functional stability during photon exposure.Abbreviations D1 protein psbA gene product - DTT dithiothreitol - Fo chlorophyll fluorescence corresponding to open PSII reaction centres - Fv, Fm variable and maximum fluorescence after dark incubation, respectively - Fs, Fm steady-state and maximum fluorescence during illumination, respectively - P680 reactioncentre chlorophyll and primary electron donor of PSII - PS photosystem Financial support of this work by Department of Employment, Education and Training/Australian Research Council International Research Fellowships Program (Korea) is gratefully acknowledged.     

Exogenous progesterone alleviates heat and high light stress-induced inactivation of photosystem II in wheat by enhancing antioxidant defense and D1 protein stability

This experiment was conducted to test the effects of foliar application of progesterone on the photochemical efficiency of photosystem II (PSII) and photosynthetic rate in wheat flag leaves subjected to cross-stress of heat and high light during grain-filling stage. The results showed that progesterone pretreatment increased the activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase, and the contents of ascorbic acid and glutathione under the cross-stress. Meanwhile, the rate of O₂ ^? production, hydrogen peroxide (H₂O₂) and malondialdehyde contents in progesterone pretreated leaves were significantly lower under heat and high light stress. In parallel with the alleviation of oxidative stress, higher content of D1 protein in PSII reactive center was observed in progesterone pretreated leaves, resulting in a significant increase in the potential (Fv/Fm) and actual (ΦPS II) photochemical efficiency of PSII, and the net photosynthetic rate. In summary, this study suggested that foliar application of progesterone might protect the PSII complex from heat and high light stress-induced damage through enhancing antioxidant defense system and further facilitating D1 protein stability in the wheat leaves.     

不同光强下高锰对黄瓜光合作用特性的影响

采用营养液培养的方法,研究了不同光强下高锰对黄瓜植株生长、叶绿素含量、叶绿素荧光参数和光合作用的影响.结果表明,高锰处理抑制了黄瓜植株的生长,与弱光处理相比强光下抑制幅度更加显著.强光下,高锰处理显著降低叶绿素含量,但降低光强却增加其含量.强光下,高锰处理显著降低原初光能转换效率(F_v/F_m)、光合电子传递量子效率(ΦPSII)和光化学猝灭系数(qP);弱光下,高锰处理对F_v/F_m和qP无显著影响.高锰处理使净光合速率(P_n)和气孔导度(G_s)下降.尤其是在强光下下降幅度更大.高锰处理使细胞间CO₂浓度(C_i)在强光下升高,而在弱光下则下降.与C_i相反,高锰处理使气孔限制值(L_s)在强光下下降,而在弱光下上升.因此,强光下高锰胁迫使净光合速率下降可能是由非气孔限制引起的,而弱光下高锰处理使净光合速率下降可能是由气孔因子限制引起的.     

早熟桃夏季叶色转红过程中的光化学响应特征

比较研究了‘早美’和‘春蕾’2个早熟桃品种夏季叶色转红对太阳光能的利用和光系统Ⅱ的叶绿素荧光特征的影响。结果表明：早熟桃叶片色素组成的变化会显著影响其光合和叶绿素荧光特性。叶色转红后,早熟桃净光合速率（Pn）日均值、PSII最大光化学效率（Fv/Fm）、PSII实际光化学效率（ФPSII）均上升,无显著光抑制,而绿叶对照‘红花碧桃’的电子传递速率（ETR）、Fv／Fm和ФPSII值均显著下降,7月光合明显受抑制。叶色转红程度较深的‘早美’在夏季高温强光下表现优于‘春蕾’和对照。淬灭分析表明：叶片花色素苷的积累能在短时间内增加PSII天线色素吸收的光能用于光化学反应的份额（P）与用于反应中心热耗散的相对份额（D）。转红后的叶片光化学淬灭系数（qp）显著高于绿叶,PSII光化学效率较高,但耗散过剩激发能的能力显著低于绿叶对照。     

Xanthophyll Cycle and Inactivation of Photosystem Ⅱ Reaction Centers Alleviating Reducing Pressure to Photosystem Ⅰ in Morning Glory Leaves under Short-term High Irradiance

investigated through chlorophyll fluorescence parameters in morning glory (Ipomoea setosa) leaves, which were dipped into water, dithiothreitol (DTT) and lincomycin (LM), respectively. During the stress, both the xanthophyll cycle and D1 protein turnover could protect PSI from photoinhibition. In DTT leaves, non-photochemical quenching (NPQ) was inhibited greatly and the oxidation level of P700 (P700+) was the lowest one. However, the maximal photochemical efficiency of PSII (Fv/Fm) in DTT leaves was higher than that of LM leaves and was lower than that of control leaves. These results suggested that PSI was more sensitive to the loss of the xanthophyll cycle than PSII under high irradiance. In LM leaves, NPQ was partly inhibited, Fv/Fm was the lowest one among three treatments under high irradiance and P700+ was at a similar level as that of control leaves. These results implied that inactivation of PSII reaction centers could protect PSI from further photoinhibition. Additionally, the lowest of the number of active reaction centers to one inactive reaction center for a PSII cross-section (RC/CSo), maximal trapping rate in a PSII cross-section (TRo/CSo), electron transport in a PSII cross-section (ETo/CSo) and the highest of 1-qP in LM leaves further indicated that severe photoinhibition of PSII in LM leaves was mainly induced by inactivation of PSII reaction centers, which limited electrons transporting to PSI. However, relative to the LM leaves the higher level of RC/CSo, TRo/CSo, Fv/Fm and the lower level of 1-qP in DTT leaves indicated that PSI photoinhibition was mainly induced by the electron accumulation at the PSI acceptor side, which induced the decrease of P700+ under high irradiance.     

Xanthophyll Cycle and Inactivation of Photosystem Ⅱ Reaction Centers Alleviating Reducing Pressure to Photosystem Ⅰ in Morning Glory Leaves under Short-term High Irradiance

Under 30-min high irradiance （1500μmol m^-2 s^-1）, the roles of the xanthophyll cycle and D1 protein turnover were investigated through chlorophyll fluorescence parameters in morning glory （Ipomoea setosa） leaves, which were dipped into water, dithiothreitol （DTT） and lincomycin （LM）, respectively. During the stress, both the xanthophyll cycle and D1 protein turnover could protect PSI from photoinhibition. In DTT leaves, non-photochemical quenching （NPQ） was inhibited greatly and the oxidation level of P700 （P700^＋） was the lowest one. However, the maximal photochemical efficiency of PSII （Fv/Fm） in DTT leaves was higher than that of LM leaves and was lower than that of control leaves. These results suggested that PSI was more sensitive to the loss of the xanthophyll cycle than PSII under high irradiance. In LM leaves, NPQ was partly inhibited, Fv/Fm was the lowest one among three treatments under high irradiance and P700^＋ was at a similar level as that of control leaves. These results implied that inactivation of PSII reaction centers could protect PSI from further photoinhibition. Additionally, the lowest of the number of active reaction centers to one inactive reaction center for a PSII cross-section （RC/CSo）, maximal trapping rate in a PSll cross-section （TRo/CSo）, electron transport in a PSll cross-section （ETo/CSo） and the highest of 1-qP in LM leaves further indicated that severe photoinhibition of PSII in LM leaves was mainly induced by inactivation of PSII reaction centers, which limited electrons transporting to PSh However, relative to the LM leaves the higher level of RC/CSo, TRo/CSo, Fv/Fm and the lower level of 1-qP in DTT leaves indicated that PSI photoinhibition was mainly induced by the electron accumulation at the PSI acceptor side, which induced the decrease of P700^＋ under high irradiance.     

The effects of salt stress on photosynthetic electron transport and thylakoid membrane proteins in the cyanobacterium Spirulina platensis

The response of Spirulina (Arthrospira) platensis to high salt stress was investigated by incubating the cells in light of moderate intensity in the presence of 0.8 M NaCl. NaCl caused a decrease in photosystem II (PSII) mediated oxygen evolution activity and increase in photosystem I (PSI) activity and the amount of P700. Similarly maximal efficiency of PSII (Fv/Fm) and variable fluorescence (Fv/Fo) were also declined in salt-stressed cells. Western blot analysis reveal that the inhibition in PSII activity is due to a 40 % loss of a thylakoid membrane protein, known as D1, which is located in PSII reaction center. NaCl treatment of cells also resulted in the alterations of other thylakoid membrane proteins: most prominently, a dramatic diminishment of the 47-kDa chlorophyll protein (CP) and 94-kDa protein, and accumulation of a 17-kDa protein band were observed in SDS-PAGE. The changes in 47-kDa and 94-kDa proteins lead to the decreased energy transfer from light harvesting antenna to PSII, which was accompanied by alterations in the chlorophyll fluorescence emission spectra of whole cells and isolated thylakoids. Therefore we conclude that salt stress has various effects on photosynthetic electron transport activities due to the marked alterations in the composition of thylakoid membrane proteins.     

Photochemical Efficiency of PSⅡand Membrane Lipid Peroxidation in Leaves of indica and japonica Rice (Oryza sativa) Under Chilling Temperature and Strong Light Stress Conditions

Relationships between fluorescence parameters and membrane lipid peroxidation in leaves of indica and japonica rice (Oryza sativa L.) during later growth stage were studied under chilling temperature and strong light stress conditions. Results showed that D1 protein contents of PSⅡ in photosynthetic app aratus dropped, the generation of antheraxanthin (A) and zeaxanthin (Z)of xanthophyll cycle were inhibited partly, PSⅡ photochemical efficiency （Fv/Fm）and non photochemical quenching (qN) were also decreased obviously. In addition, endogenous active oxygen scavenger—superoxide dismutase (SOD) reduced, superoxide anion radical (O[SX(B-*3)-[]·[SX]]2) and malondialdehyde (MDA) accumulated, as a result, photooxidation of leaves occurred under chilling temperature and strong light stress conditions. Obvious differences in the changes of the above mentioned physiological parameters between indica and japonica rice were observed. Experiments in leaves treated with inhibitors under chilling temperature and strong light conditions showed that indica rice was more sensitive to chilling temperature with strong light and subjected to photooxidation more than japonica rice. Notable positive correlation between D1 protein contents and Fv/Fm or (A+Z)/(A+Z+V), and a marked negative correlation between Fv/Fm and MDA contents were obtained by regression analysis in indica and japonica rice during chilling temperature and strong light conditions. According to the facts mentioned above, it was inferred that PSⅡ photochemical efficiency（Fv/Fm） was the key index to forecast for the prediction of photooxidation under stress circumstances and the physiological basis were the synthetic capacity of D1 protein and the protection of xanthophyll cycle.     

Recovery from Photoinhibition in Peas (Pisum sativum L.) Acclimated to Varying Growth Irradiances (Role of D1 Protein Turnover)

D1 protein turnover and restoration of the photochemical efficiency of photosystem II (PSII) after photoinhibition of pea leaves (Pisum sativum L. cv Greenfeast) acclimated to different light intensities were investigated. All peas acclimated to different light intensities were able to recover from photoinhibition, at least partially, at light intensities far above their growth light irradiance. However, the capacity of pea leaves to recover from photoinhibition under increasing high irradiances was strictly dependent on the light acclimation of the leaves; i.e. the higher the irradiance during growth, the better the capacity of pea leaves to recover from photoinhibition at moderate and high light. In our experimental conditions, mainly D1 protein turnover-dependent recovery was monitored, since in the presence of an inhibitor of chloroplast-encoded protein synthesis, lincomycin, only negligible recovery took place. In darkness, neither the restoration of PSII photochemical efficiency nor any notable degradation of damaged D1 protein took place. In low light, however, good recovery of PSII occurred in all peas acclimated to different light intensities and was accompanied by fast degradation of the D1 protein. The rate of degradation of the D1 protein was estimated to be 3 to 4 times faster in photoinhibited leaves than in nonphotoinhibited leaves under the recovery conditions of 50 [mu]mol of photons m-2 s-1. In moderate light of 400 [mu]mol of photons m-2 s-1, the photoinhibited low-light peas were not able to increase further the rate of D1 protein degradation above that observed in nonphotoinhibited leaves, nor was the restoration of PSII function possible. On the other hand, photoinhibited high-light leaves were able to increase the rate of D1 protein degradation above that of nonphotoinhibited leaves even in moderate and high light, ensuring at least partial restoration of PSII function. We conclude that the capacity of photoinhibited leaves to restore PSII function at different irradiances was directly related to the capacity of the leaves to degrade damaged D1 protein under the recovery conditions.     

Phosphorylation of PSII proteins in maize thylakoids in the presence of Pb ions

Lead is potentially toxic to all organisms including plants. Many physiological studies suggest that plants have developed various mechanisms to contend with heavy metals, however the molecular mechanisms remain unclear. We studied maize plants in which lead was introduced into detached leaves through the transpiration stream. The photochemical efficiency of PSII, measured as an Fv/Fm ratio, in the maize leaves treated with Pb was only 10% lower than in control leaves. The PSII activity was not affected by Pb ions in mesophyll thylakoids, whereas in bundle sheath it was reduced. Protein phosphorylation in mesophyll and bundle sheath thylakoids was analyzed using mass spectrometry and protein blotting before and after lead treatment. Both methods clearly demonstrated increase in phosphorylation of the PSII proteins upon treatment with Pb²⁺, however, the extent of D1, D2 and CP43 phosphorylation in the mesophyll chloroplasts was clearly higher than in bundle sheath cells. We found that in the presence of Pb ions there was no detectable dephosphorylation of the strongly phosphorylated D1 and PsbH proteins of PSII complex in darkness or under far red light. These results suggest that Pb²⁺ stimulates phosphorylation of PSII core proteins, which can affect stability of the PSII complexes and the rate of D1 protein degradation. Increased phosphorylation of the PSII core proteins induced by Pb ions may be a crucial protection mechanism stabilizing optimal composition of the PSII complexes under metal stress conditions. Our results show that acclimation to Pb ions was achieved in both types of maize chloroplasts in the same way. However, these processes are obviously more complex because of different metabolic status in mesophyll and bundle sheath chloroplasts.     

草莓叶片光合作用对强光的响应及其机理研究

用便携式调制叶绿素荧光仪和光合仪研究了强光下草莓叶片荧光参数及表观量子效率的变化.结果表明,Fm、Fv/Fm、PSⅡ无活性反应中心数量和Q_A的还原速率在强光下降低,在暗恢复时升高;而PSⅡ反应中心非还原性Q_B的比例在强光下增加,在暗恢复时降低.上述荧光参数的变化幅度均以强光胁迫或暗恢复的前10 min最大.强光下Φ_PSII、ETR和qP先升高后降低,但qN先大幅度降低,然后小幅回升.强光处理4 h后,丰香和宝交早生的表观量子效率(AQY)分别降低了20.9%和37.5%;qE(能量依赖的非光化学猝灭)为NPQ(非光化学猝灭)的最主要成分.强光胁迫下丰香的Fo、Fm、Fv/Fm、Φ_PSII、ETR和AQY的变化幅度均明显比宝交早生小.DTT处理后,草莓叶片的Fm和Fv/Fm明显降低,Fo显著升高.可以认为,依赖叶黄素循环和类囊体膜质子梯度两种非辐射能量耗散在草莓叶片防御光损伤方面起着重要作用,丰香的光合机构比宝交早生更耐强光.     

水淹对水芹叶片结构和光系统II光抑制的影响

通过探讨在水淹条件下水芹(Oenanthe javanica)叶片结构的变化以及出水对其光系统II功能和光抑制的影响, 阐明水芹光合机构在水淹条件下及出水后死亡的可能原因。结果表明: 水淹条件下新生沉水功能叶光系统II(PSII)最大光化学效率(Fv/Fm) 、电子传递活性与对照叶片差异很小, 但水淹使气生功能叶的Fv/Fm显著降低; 植株总生物量呈负增长趋势; 活体弱光条件下, 沉水叶出水后2小时叶片相对含水量(RWC)和Fv/Fm无显著变化; 中等光强和强光条件下其RWC和Fv/Fm迅速降低; 离体条件下, 5小时的中等光强对沉水叶的Fv/Fm影响不显著, 在随后的弱光下能恢复到出水时的初始状态; 强光能使沉水叶的Fv/Fm大幅降低, 且弱光下不能恢复到出水时的初始水平; 在解剖结构上, 水芹沉水叶的叶片总厚度、上下表皮厚度和气孔大小都显著低于气生叶, 而且沉水叶没有明显的栅栏组织分化, 但是沉水叶上表皮的气孔密度显著高于气生叶。研究结果表明, 水淹使水芹原气生叶PSII功能迅速衰退, 但对新生沉水叶片影响很小。水芹植株出水后, 沉水叶片结构变化使其在光下保水能力下降, 而强光导致了光合机构的光抑制和反应中心失活。田间条件下两者共同作用则加剧了对叶片光合机构的破坏, 进而致使其死亡。     

Site-specific mutations localized in the D-E loop of the D1 protein of photosystem II affect phototolerance in Synechocystis sp. PCC 6803 containing psbAII gene

Photosynthetic characteristics along with phototolerance and photoinhibition of photosystem II (PS II) were monitored in Synechocystis sp. PCC 6803 wild type (KC) and its psbAII mutants viz., I6 (N322I, I326F, and F328S), G6 (N267Y), and H7 (Y254C and I314V) that have up to three point mutations, localized in the D-E loop of the D1 polypeptide of PSII reaction centre. These strains exhibited entirely different growth trends upon shifting from 30 micormol m(-2)s(-1) to high irradiance (500 micromol m(-2)s(-1) , 30 degrees C). The I6 and H7 cells grew well, whereas KC and G6 cells showed inability for cell multiplication. The photosynthetic efficiency demonstrated about 50% loss in chlorophyll fluorescence of variable yield (Fv/Fm) within 20-30 min in all mutants, whereas the wild type (KC) cells could reach the same level of loss in 2 hr. I6 and H7 cells showed continuous cell growth and maintenance under long-term exposure of high light compared to G6 mutant and wild type cells. The wild type cells showed slow decrease in their photochemical activity and Fv/Fm values, compared to mutant cells. The recovery seemed to be almost identical, and also stimulated by growth light, inspite of differential photoinhibitory behaviours. Darkness and translational inhibitor lincomycin both were found to be unassociated with the restoration of photoinhibited process of PS II.     

氯化胆碱对低温弱光下黄瓜幼苗叶片细胞膜和光合机构的保护作用

1.07mmol／L氯化胆碱处理降低了低温弱光（6℃．PFD100μmol m^-2s^-1）下黄瓜幼苗叶片膜脂组分中主要是磷脂酰甘油（PG）的饱和脂肪酸含量,增加了膜脂不饱和度：减缓了膜透性的下降、MDA的产生速率、叶绿素的降解及PSII最大量子效率（Fv/Fm）、捕光效率（Fv＇／Fm＇）、光化学猝灭系数（qp）、实际光化学效率（ФPSII）和抗氧化酶POD、APX及CAT活性的下降;提高了非光化学猝灭系数（NPQ）和脯氨酸的含量。以上结果表明氯化胆碱处理保护了低温弱光对黄瓜叶片细胞膜和光合机构的伤害。     

Characterization of a phototolerant mutant of Synechocystis sp. PCC 6803 created by random mutagenesis of psbAII gene

Photosensitivity and photosynthetic characteristics have been analyzed in wild type (KC) and its psbAII mutant (I6) of Synechocystis having three point amino acid substitutions, i.e., N322I, I326F and F328S, which are localized in the C-terminal extension of D1 protein of the photosystem II reaction center. Wild type and mutant cells show almost an identical growth pattern under normal/low light (30 mumol m-2s-1, 30 degrees C) liquid culture (BG-11) condition. However, upon shifting the cultures to high light (500 mumol m-2s-1, 30 degrees C), these two types of cells exhibit entirely different growth characteristics, i.e., the mutant cells continue to grow normally whereas, the control cells fail to adapt the light stress and eventually resulting in complete loss of the photosynthetic pigments. On the other hand, a quick loss in the Fv/Fm value with half--decay time of about 30 min is observed in the mutant, in contrast to 120-130 min in case of control, upon shifting to high light conditions. In spite of this, mutant cells are able to adapt and grow well under prolonged high light exposure even after losing a major part of the variable yield of chlorophyll fluorescence (Fv/Fm). The high light treatment also induced decrease in the level of D1 protein in the mutant. However, half-decay time for D1 is much longer (approximately 10 hr) than that of variable fluorescence. Thus, the mutant cells have shown an unique way for cell growth and maintenance under high light even after losing Fv/Fm and photosynthetic oxygen evolving capacity as well as D1 content to a great extent. Therefore, these results could extend an interesting insight to understand the coordination of physiological, biochemical and molecular mechanisms regulating phototolerance of the photosynthetic organisms.     

Increased photoinhibition in dehydrated leaves of hot pepper (Capsicum annuum L.) is not accompanied by an incremental loss of functional PSII

We examined the photosynthetic responses to photoinhibition in dehydrated leaves of hot pepper (Capsicum annuum L.). Stress was induced by immersing the roots of whole plants in Hoaglands solution containing polyethylene glycol (PEG) under high light (900 μmol photons m^-2 · s^-1). This PEG-treatment lowered the leaf water potential and the maximal rate of photosynthetic O₂ evolution (Pmax) linearly, in a time-dependent manner, to about 50% inhibition after 6 h. Pmax also decreased linearly as the period of high-light treatment lengthened. That inhibitory response was not as extreme, showing about 30% inhibition after 6 h. However, when the treatments of dehydration and high light were simultaneously administered, Pmax decreased more rapidly, in a synergistic fashion, showing about 90% inhibition within 2 h. Dehydration, in contrast to the light treatment, did not lower the maximal photochemical efficiency (Fv/Fm). Furthermore, this decline in Fv/Fm for light-treated, dehydrated leaves was almost identical to the response of photoinhibited leaves that were not dehydrated. Similar changes were observed in the number of functional PSII complexes. The decrease in Pmax and the amount of functional PSII was linearly correlated in photoinhibited leaves, but not in dehydrated leaves, regardless of light treatment. Therefore, we have demonstrated that exacerbated photoinhibition in dehydrated leaves occurs without an incremental loss of functional PSII.     

外源ATP对NaCl胁迫下菜豆叶片叶绿素荧光特性的调节

盐胁迫是影响植物生长的主要逆境因子之一,外源ATP被发现可作为信号分子参与植物对逆境胁迫生理反应的调节。为了探明外源ATP在植物盐胁迫响应中的作用,以增强植物对土壤盐渍化的耐性,更好地应用于土壤盐渍化修复。该研究以菜豆( Phaseolus vulgaris)为材料,通过叶绿素荧光技术探讨了外源ATP 对菜豆叶片在NaCl胁迫下叶绿素荧光特性的变化规律。结果表明：在NaCl胁迫下,叶片光系统Ⅱ( PSⅡ)潜在最大光化学量子效率( Fv/Fm)、光适应下最大光化学效率( Fv′/Fm′)、PSⅡ光适应下实际光化学效率[ Y (Ⅱ)]、光化学荧光猝灭( qP)、电子传递速率( ETR)与对照组相比均有显著性下降,而非光化学猝灭( NPQ)和( qN)较对照组有显著性增加,这表明NaCl胁迫导致菜豆叶片光系统Ⅱ光化学效率的下降和光能耗散的增加。而外源ATP(eATP)的处理能有效缓解NaCl胁迫所造成的Fv/Fm、Fv′/Fm′、Y(Ⅱ)、qP、ETR下降和NPQ、qN的上升。该研究结果表明在NaCl胁迫下外源ATP可以有效地提高菜豆幼苗光系统Ⅱ( PSⅡ)的光化学反应效率。