不同磷营养水平对烟草叶片光合作用和光呼吸的影响

随着磷营养水平的提高,烟草叶片的CO_2补偿点下降、光合速率上升。光呼吸在缺磷时最高。用光呼吸抑制剂处理烟草叶片后,光合的最适磷浓度提高。当CO_2浓度为560μl/L时,缺磷的烟草叶片在从21％O_2转入2％O_2时出现光合振荡,表明光呼吸与磷营养有密切关系。光呼吸在形成乙醇酸时所释放的磷,有回补叶绿体进行光合作用所需的磷的作用。     

不同氮素营养对黄麻叶片的光合作用、光呼吸的影响及光呼吸与硝酸还原的关系

不同硝态氮水平对黄麻叶片的光合强度、光呼吸强度、乙醇酸氧化酶及硝酸还原酶均产生显著的影响:除了光合强度以外,其余三者均随氮水平的增高而显著地增高;在低至正常氮水平范围内(1/4～1N),光合强度随氮水平的增高而增高,但在高硝态氮水平范围内(2～4N),光合强度有随氮水平的增高而略为下降的趋势。氨态氮水平的增高对光合强度、光呼吸强度、乙醇酸氧化酶活性也有影响:三者均随氮水平的增高而有所提高,但不如硝态氮处理的那样明显。在用氨态氮处理的植株中测不出硝酸还原酶的活性。用光呼吸抑制剂亚硫酸氢钠,α-羟基-2-吡啶甲烷磺酸(HPMS)及异烟肼(INH)真空渗入黄麻叶圆片以抑制光呼吸时,硝酸还原亦同时部分受抑。因此认为在乙醇酸途经中的甘氨酸氧化脱羧反应可能为硝酸还原提供还原剂NADH。     

光呼吸对光合过程中磷代谢的影响

与光呼吸受抑制的 2％O_2浓度下相比,在 21％O_2浓度下.离体甘薯叶细胞光合作用最适介质无机磷浓度较低.另外,在21％O_2浓度下,降低甘薯叶细胞介质 NaHCO_3浓度,叶细胞光下吸收介质~(32)Pi的量减少;降低完整菠菜叶绿体介质 NaHCO_3浓度,乙醇酸形成相对加强,而介质~(32)Pi掺入到有机磷化合物的量则相对减少.这些结果表明,有利于光呼吸的条件,可降低光合对外界Pi的需求量.     

光呼吸对光合过程中磷代谢的影响

与光呼吸受抑制的 2％O_2浓度下相比,在 21％O_2浓度下.离体甘薯叶细胞光合作用最适介质无机磷浓度较低.另外,在21％O_2浓度下,降低甘薯叶细胞介质 NaHCO_3浓度,叶细胞光下吸收介质~(32)Pi的量减少;降低完整菠菜叶绿体介质 NaHCO_3浓度,乙醇酸形成相对加强,而介质~(32)Pi掺入到有机磷化合物的量则相对减少.这些结果表明,有利于光呼吸的条件,可降低光合对外界Pi的需求量.     

关于光呼吸与光合作用关系的研究——Ⅲ.小麦叶片CO_2猝发及其与光合作用的关系

小麦叶片断光后CO_2猝发强度随断光前光强增加而增强。在空气中,小麦叶片CO_2猝发时间在1.5分钟左右,断光后0.5分钟内猝发最强。在无CO_2气流中,小麦CO_2猝发量显著增强,约为空气中猝发量的5～6倍。体外供给乙醇酸,能增加猝发量和延长猝发时间。随氧浓度提高,光合受抑,而猝发量增强,旺盛期猝发时间缩短。氧浓度对猝发的影响只有在断光前才起作用。氧浓度在2％以下,观察不到猝发。CO_2浓度虽可增加光合,增强猝发之势,但也有抑制光呼吸,降低猝发的效应,而后者一般是主要的。光呼吸,暗呼吸释放的CO_2都能在光下被光合作用再固定,再固定数量随外界CO_2浓度提高而增高,随氧浓度提高而下降。     

植物的乙醇酸氧化酶

乙醇酸氧化酶存在于植物细胞过氧物酶体中,是一种黄素蛋白,以FMN为辅基,含8个亚基。催化乙醇酸氧化为乙醛酸和乙醛酸氧化为草酸。受光活化。硫化钠和氰化物促进其活性:巯基抑制剂、α-羟基磺酸类、α-羟基丁炔酸抑制其活性。多种代谢物对其活性有调节作用。为光呼吸的关键酶之一,其活性随植物发育、矿质营养及感病而发生变化。     

菠菜叶片乙醇酸氧化酶的氧化甘油酸活性

首先从菠菜叶片中纯化了乙醇酸氧化酶（GO）。通过鉴定反应中氧的消耗以及反应产物H2O2的生成,证实菠菜GO具有氧化光呼吸途径中间代谢物甘油酸的活性。该氧化活性依赖于辅因子FMN和FAD,而不依赖核黄素和光黄素;其最适反应pH值为8．0,Km（甘油酸）值为7．14mmol／L,kcat值为1．04s^-1,活化能为17．29kJ／mol;草酸和丙酮酸对该氧化活性有明显的抑制作用,其中前者为典型的竞争性抑制。进一步通过两底物竞争作图表明：菠菜叶片GO氧化甘油酸反应和氧化乙醇酸反应为同一活性中心所催化。     

产乙醇酸氧化酶的重组毕赤酵母的构建及催化合成乙醛酸的研究

乙醇酸氧化酶（Go）是植物光呼吸途径中的一种关键酶,可以催化乙醇酸生产乙醛酸。从新鲜菠菜叶中提取总RNA,利用RT-PCR技术获得编码GO基因的cDNA片断。通过基因重组将GO基因克隆到载体pA0815中,构建了胞内表达载体pA0815／GO,重组质粒经电转整合至甲醇营养酵母GS115染色体。在混合碳源为10g/L山梨醇和0．5g/L甲醇的培养条件下,细胞的GO酶活达到474IU／g（DCW）。利用该重组毕赤酵母作为催化剂生产乙醛酸,结果表明：在乙醇酸浓度为0．25mol／L,重组酵母湿菌体为10dL,黄素单核苷酸（FMN）浓度为0．01mmol／L,pH8．0,20℃,反应18h后乙醛酸的产率达到51．8％。     

NaCl胁迫对西瓜幼苗生长和光合气体交换参数的影响

以早春红玉品种为材料,采用营养液水培法,研究了不同浓度NaCl胁迫对西瓜幼苗生长、叶片光合气体交换参数、质膜透性和脯氨酸含量的影响.结果表明:25 mmol·L-1NaCl处理9 d后对西瓜幼苗生长有促进作用,>75 mmol·L-1NaCl处理则显著抑制幼苗生长;NaCl处理显著提高了叶片光合色素含量,并在100 mmol·L-1NaCl处理下达到最大;叶片净光合速率、气孔导度和蒸腾速率均随NaCl浓度提高而显著降低;胞间CO2浓度随NaCl浓度提高呈先降低后升高的趋势,在75 mmol·L-1NaCl处理下降到最小;气孔限制值随NaCl浓度提高而增加,在75 mmol·L-1NaCl处理下达到最大值后趋于稳定;水分利用率随NaCl浓度提高呈先增加后降低的趋势,在75 mmol·L-1NaCl处理下达到最大;叶片质膜透性和脯氨酸含量均随NaCl浓度提高而显著增加.结果说明:NaCl胁迫显著抑制了西瓜叶片光合作用,且低浓度处理下光合速率降低的主要原因是气孔因素限制,高浓度胁迫下则转变为非气孔因素限制.     

高大气CO2浓度下氮素对小麦叶片光能利用的影响

关于氮素对高大气CO₂浓度下C₃植物光合作用适应现象的调节机理已有较为深入的研究, 但对其光合作用适应现象的光合能量转化和分配机制缺乏系统分析。该文以大气CO₂浓度和施氮量为处理手段, 通过测定小麦(Triticum aestivum)抽穗期叶片的光合作用-胞间CO₂浓度响应曲线以及荧光动力学参数来测算光合电子传递速率和分配去向, 研究了长期高大气CO₂浓度下小麦叶片光合电子传递和分配对施氮量的响应。结果表明, 与正常大气CO₂浓度处理相比, 高大气CO₂浓度下小麦叶片较多的激发能以热量的形式耗散, 增施氮素可使更多的激发能向光化学反应方向的分配, 降低光合能量的热耗散速率; 大气CO₂浓度升高后小麦叶片光化学淬灭系数无明显变化, 高氮叶片的非光化学猝灭降低而低氮叶片明显升高, 施氮促进PSII反应中心的开放比例, 降低光能的热耗散; 高大气CO₂浓度下高氮叶片通过PSII反应中心的光合电子传递速率(J_F)较高, 而且参与光呼吸的非环式电子流速率(J₀)显著降低, 较正常大气CO₂浓度处理的高氮叶片下降了88.40%, 光合速率增加46.47%; 高大气CO₂浓度下小麦叶片J_F-J₀升高而J₀/J_F显著下降, 光呼吸耗能被抑制, 更多的光合电子分配至光合还原过程。因此, 大气CO₂浓度增高条件下, 小麦叶片激发能的热耗散速率增加, 但增施氮素后小麦叶片PSII反应中心开放比例提高, 光化学速率增加, 进入PSII反应中心的电子流速率明显升高, 光呼吸作用被抑制, 光合电子较多地进入光化学过程, 这可能是高氮条件下光合作用适应性下调被缓解的一个原因。     

Regulation of photosynthesis in nitrogen deficient wheat seedlings

Nitrogen effects on the regulation of photosynthesis in wheat (Triticum aestivum L., cv Remia) seedlings were examined. Ribulose 1,5-bisphosphate carboxylase/oxygenase was rapidly extracted and tested for initial activity and for activity after incubation in presence of CO₂ and Mg²⁺. Freeze clamped leaf segments were extracted for determinations of foliar steady state levels of ribulose 1,5-bisphosphate, triose phosphate, 3-phosphoglycerate, ATP, and ADP. Nitrogen deficient leaves showed increased ATP/ADP and triose phosphate/3-phosphoglycerate ratios suggesting increased assimilatory power. Ribulose 1,5-bisphosphate levels were decreased due to reduced pentose phosphate reductive cycle activity. Nevertheless, photosynthesis appeared to be limited by ribulose 1,5-bisphosphate carboxylase/oxygenase, independent of nitrogen nutrition. Its degree of activation was increased in nitrogen deficient plants and provided for maximum photosynthesis at decreased enzyme protein levels. It is suggested that ribulose 1,5-bisphosphate carboxylase/oxygenase activity is regulated according to the amount of assimilatory power.     

植物中草酸积累与光呼吸乙醇酸代谢的关系

对几种C3 和C4 植物中草酸含量及相应的乙醇酸氧化酶活性测定结果表明 :叶片光呼吸强度及其关键酶活性大小与草酸积累量没有相关性 ;植物根中均能积累草酸 ,但未测出乙醇酸氧化酶活性。烟草根、叶中的草酸含量在不同生长时期差异明显 ,且二者呈极显著正相关 (y =2 .5 6 5lnx 2 .137,r =0 .749,P <0 .0 0 1) ,说明根中草酸可能来自叶片。氧化乙醇酸的酶的活性与氧化乙醛酸的酶的活性呈极显著线性正相关 (y =0 .2 41x 0 .0 0 6 ,r=0 .96 7,P <0 .0 0 0 1) ,进一步证实是乙醇酸氧化酶催化了两种底物的反应。烟草在不同生长期叶片中草酸总含量变化与相应的乙醇酸氧化酶活性变化亦没有相关性 ;低磷胁迫可显著诱导烟草根叶中的草酸形成和分泌 ,但并未影响乙醇酸氧化酶活性 ,进一步证明草酸积累与该酶活性大小无关     

低浓度硫酸甲酯吩嗪（PMS）对叶片光合放氧的促进作用

外加低浓度循环光合磷酸化电子递体硫酸甲酯吩嗪(PMS)对菠菜、大豆、水稻和小麦叶片光合放氧有促进作用,与此同时叶片ATP含量也得到增加。PMS对经8 mmol L~(-1)NH_4Cl处理过的菠菜叶片的光合放氧也有促进,最适促进浓度比未经NH_4Gl处理的叶片高,促进的幅度也大。幼龄叶与成长叶相比,幼龄叶的光合磷酸化活性和P/O比值低于成长叶片,其光合放氧速率受PMS促进的幅度大于成长叶片。因此光合磷酸化也可以成为光合作用的一个重要限制因素。     

Evidence for the presence in tobacco leaves of multiple enzymes for the oxidation of glycolate and glyoxylate

The enzymic oxidation of glycolate to glyoxylate and glyoxylate to oxalate by preparations purified from tobacco (Nicotiana tabacum var Havana Seed) leaves was studied. The K_m values for glycolate and glyoxylate were 0.26 and 1.0 millimolar, respectively. The ratio of glycolate to glyoxylate oxidation was 3 to 4 in crude extracts but decreased to 1.2 to 1.5 on purification by (NH₄)₂SO₄ fractionation and chromatography on agarose A-15 and hydroxylapatite. This level of glyoxylate oxidation activity was higher than that previously found for glycolate oxidase (EC 1.1.3.1). The ratio of the two activities was changed by reaction with the substrate analog 2-hydroxy-3-butynoate (HBA) which at all concentrations inhibited glyoxylate oxidation to a greater extent than glycolate oxidation. The ratio of the two activities could also be altered by changing the O₂ concentration. Glycolate oxidation increased 3.6-fold when the O₂ atmosphere was increased from 21 to 100%, whereas glyoxylate oxidation increased only 1.6-fold under the same conditions. These changes in ratio during purification, on inhibition by HBA, and under varying O₂ concentrations imply that tobacco leaves contain at least two enzymes capable of oxidizing glycolate and glyoxylate.     

Short-term feedback inhibition of photosynthesis in wheat leaves supplied with sucrose and glycerol at two temperatures

The inhibition of photosynthesis by reduced sink demand or low rates of end product synthesis was investigated by supplying detached wheat (Triticum aestivum L. cv. Tauro) leaves with 50 mM sucrose, 50 mM glycerol or water through the transpiration stream for 2 h, either at 23 or 12 °C. Lowering the temperature and sucrose and glycerol feeding decreased photosynthetic oxygen evolution at high irradiance and saturating CO₂. The decrease in temperature reduced the pools of sucrose and starch, and the ratio glucose 6-phosphate (G6P)/fructose 6-phosphate (F6P), while it increased the concentrations of G6P and F6P (hexose phosphates). Sucrose feeding, in contrast to glycerol feeding, increased sucrose, glucose and fructose contents and the G6P/F6P ratio. Sucrose and glycerol incubations at 23 °C, as well as decreasing the temperature in leaves incubated in water, increased the concentration of triose-phosphates (glyceraldehyde 3-phosphate and dihydroxyacetone phosphate, TP) and decreased the glycerate 3-phosphate (PGA) content, thus increasing the TP/PGA ratio; they also tended to increase the ribulose 1,5-bisphosphate (RuBP) content and the RuBP/PGA ratio. Sucrose and glycerol feeding at 12 °C and the decrease in temperature of leaves incubated in these solutions decreased TP and RuBP contents and the TP/PGA and RuBP/PGA ratios. The results suggest that the phosphate limitation caused by accumulation of end products, restriction of their synthesis and sequestration of cytosolic phosphate can inhibit photosynthesis through decreased carboxylation of RuBP or, with increased phosphate limitation, through lowered supply of ATP. This revised version was published online in August 2006 with corrections to the Cover Date.     

光和底物对乙醇酸氧化酶的基因表达的诱导

从菠菜中提纯了乙醇酸氧化酶并制备其抗体,经免疫双扩散、Ｗｅｓｔｅｒｎｂｌｏｔ和Ｎｏｒｔｈｅｒｎｂｌｏｔ证实水稻和豌豆黄化苗中不存在乙醇酸氧化酶。在黑暗中,底物可促进该酶基因的表达,而在黄化苗光照初期,推测光可能是不经过底物促进该酶基因的表达。     

Expression of spinach glycolate oxidase in Saccharomyces cerevisiae: purification and characterization.

Glycolate oxidase from spinach has been expressed in Saccharomyces cerevisiae. The active enzyme was purified to near-homogeneity (purification factor approximately 1400-fold) by means of hydroxyapatite and anion-exchange chromatography. The purified glycolate oxidase is nonfluorescent and has absorbance peaks at 448 (epsilon = 9200 M-1 cm-1) and 346 nm in 0.1 M phosphate buffer, pH 8.3. The large bathochromic shift of the near-UV band indicates that the N(3) position is deprotonated at pH 8.3. A pH titration revealed that the pK of the N(3) is shifted from 10.3 in free flavin to 6.4 in glycolate oxidase. Glycolate oxidase is competitively inhibited by oxalate with a Kd of 0.24 mM at 4 degrees C in 0.1 M phosphate buffer, pH 8.3. Three pieces of evidence demonstrate that glycolate oxidase stabilizes a negative charge at the N(1)-C(2 = O) locus: the enzyme forms a tight sulfite complex with a Kd of 2.7 x 10(-7) M and stabilizes the anionic flavosemiquinone and the benzoquinoid form of 8-mercapto-FMN. Steady-state analysis at pH 8.3, 4 degrees C, yielded a Km = 1 x 10(-3) M for glycolate and Km = 2.1 x 10(-4) M for oxygen. The turnover number has been determined to be 20 s-1. Stopped-flow studies of the reductive (k = 25 s-1) and oxidative (k = 8.5 x 10(4) M-1 s-1) half-reactions have identified the reduction of glycolate oxidase to be the rate-limiting step.     

Early Inhibition of Photosynthesis during Development of Mn Toxicity in Tobacco

Early physiological effects of developing Mn toxicity in young leaves of burley tobacco (Nicotiana tabacum L. cv KY 14) were examined in glass-house/water cultured plants grown at high (summer) and low (winter) photon flux. Following transfer of plants to solutions containing 1 millimolar Mn²⁺, sequential samplings were made at various times for the following 9 days, during which Mn accumulation by leaves increased rapidly from ~70 on day 0 to ~1700 and ~5000 microgram per gram dry matter after 1 and 9 days, respectively. In plants grown at high photon flux, net photosynthesis declined by ~20 and ~60% after 1 and 9 days, respectively, and the onset of this decline preceded appearance (after 3 to 4 days) of visible foliar symptoms of Mn toxicity. Intercellular CO₂ concentrations and rates of transpiration were not significantly affected; moreover, the activity of the Hill and photosystem I and II partial reactions of chloroplasts remained constant despite ultimate development of severe necrosis. Though the activity of latent or activated polyphenol oxidase increased in parallel with Mn accumulation, neither leaf respiration nor the activity of catalase [EC 1.11.1.6] and peroxidase [EC 1.10.1.7] were greatly affected. These effects from Mn toxicity could not be explained by any changes in protein or chlorophyll abundance. Additionally, they were not a consequence of Mn induced Fe deficiency. Therefore, inhibition of net photosynthesis and enhancement of polyphenol oxidase activity are early indicators of excess Mn accumulation in tobacco leaves. These changes, as well as leaf visual symptoms of Mn toxicity, were less severe in plants cultured and treated at low photon flux even though the rates of leaf Mn accumulation at high and low photon flux were essentially equivalent.     

Dynamic changes in photosynthesis and chlorophyll fluorescence in Nicotiana tabacum infested by Bemisia tabaci (Middle East–Asia Minor 1) nymphs

Bemisia tabaci Middle East–Asia Minor 1 (MEAM1) is a worldwide pest. To determine whether MEAM1 nymphs produce the same symptoms in different host plants, we measured the plant growth and chlorophyll content of tobacco and cotton plants that were infested by MEAM1 nymphs. Furthermore, to investigate the spatial and temporal changes in photosynthesis caused by MEAM1 nymphs, the net photosynthetic rate (Pn) and chlorophyll a fluorescence of local and systemic tobacco leaves were assayed at 8, 11, 14, and 20 days after MEAM1 adult removal, which represent the stages of 1st, 2nd, 3rd, and 4th instar nymphs, respectively. The results showed that MEAM1 nymph infestation reduced the plant height and internode length of tobacco at 14 and 20 days, as well as the dry weight of infested and systemic tobacco leaves. However, MEAM1 nymph infestation did not affect the plant height or internode length of cotton. Also, the dry weight and chlorophyll and carotenoid content of infested and systemic leaves of cotton plants were not influenced by MEAM1 nymph infestation. However, the contents of chlorophyll a and b and carotenoids in infested tobacco leaves decreased over time; the chlorophyll a content of systemic tobacco leaves decreased at 11, 14, and 20 days. The chlorophyll and carotenoid contents in infested and systemic leaves of cotton plants were not influenced by MEAM1 nymph infestation. In addition, the Pn of infested tobacco leaves decreased at 14 and 20 days, while the Pn in systemic tobacco leaves decreased after 11 days. The greatest decrease in performance index on absorption basis (PI _ABS ) of infested and systemic tobacco leaves occurred on day 14. The fluorescence intensity at 2 ms (J peak) and 300 μs (K peak) increased on day 14, which indicates that 3rd instar nymphs caused serious damage to the primary photochemical reactions and donor side of PSII. These results suggest that MEAM1 nymph infestation had different effects on tobacco and cotton plants. The infestation caused spatial and temporal changes in photosynthesis in tobacco plants. The lower chlorophyll a content may have been related to the lower net photosynthetic rate of systemic and infested tobacco leaves. The decreased stability of the oxygen-evolving complex and the reaction center of PSII and the decrease in electron transport were the main reasons for the decrease in the level of photosynthesis in tobacco leaves caused by MEAM1 nymphs during various stages of infestation.     

Effect of nitrogen nutrition on the carbohydrate repression of photosynthesis in leaves of Phaseolus vulgaris L.

When carbohydrates accumulate in leaves, photosynthesis is repressed. Limited nitrogen nutrition is thought to enhance this repressing effect. However, the interaction between carbohydrate and nitrogen limitation in leaf photosynthesis has not been examined intensively. In this study, we grew Phaseolus vulgaris L. plants at three different nitrogen levels, and examined the effects of sucrose feeding to the roots on the nitrogen content, carbohydrate content and photosynthetic properties of the primary leaves. Nitrogen content and photosynthetic rate were lower and the carbohydrate content was greater in plants grown with limited nitrogen than in well-fertilized plants. Sucrose feeding to the plants increased carbohydrate content and decreased photosynthetic rate and nitrogen content. The increase in carbohydrate content and the decreases in nitrogen content and photosynthetic rate occurred at the same time, and the negative relationship between the carbohydrate content and photosynthetic rate did not differ among nitrogen nutrition levels. These results show that carbohydrate accumulation in the leaves leads to a decrease in photosynthetic rate. At low nitrogen nutrition levels, carbohydrates accumulated markedly, which accelerated this effect. It appears that the nitrogen nutrition level influences leaf photosynthesis through changing the carbohydrate level rather than through modifying sensitivity of the leaf to the carbohydrate level.