多能硫杆菌1,5-二磷酸核酮糖羧化酶/加氧酶的酶学特征及基因检测

从固体平板的生长情况及RubisCO酶活性测定结果，表明多能硫杆菌是通过卡尔文循环固定的CO2，并具有该循环的关键酶──1，5－二磷酸核酮糖羧化酶／加氧酶．进一步将多能硫杆菌染色体DNA用各种限制性内切酸酶切，Southern转移到硝酸纤维素滤膜上，与光合细菌Rhodobactersphaeroides的RubisCO基因（rbcL－rbcS）探针杂交，显示出杂交带型，说明多能硫杆菌具有R．sphaeroides同源的羧化酶基因，并初步将该基因定位在3．0kb的PstⅠ片段内．     

光合细菌生物制氢反应器研究进展

光合细菌生物制氢技术,是将太阳能利用和环境治理结合起来的可再生能源生产技术,具有良好的环境效益、社会效益和经济效益.光合细菌制氢反应器研究是将该技术引向产业化的关键步骤.本文列举了目前光合细菌制氢研究中典型的反应器型式,分析了各自的优缺点;总结了反应器高效产氢所必需保证的运行操作条件,如光照强度、温度、pH值等,指出了目前研究中存在的不足;给出了规模化光合细菌生物制氢时需要用到的反应器性能评价指标.并针对目前反应器的研究现状,提出了后续研究应当遵循的方向.     

光合细菌对铜绿微囊藻生长的抑制效应

为了探讨光合细菌对水华藻类的控制作用,在实验室条件下,通过菌藻共同培养,研究了沼泽红假单孢菌(Rhodopseudomonas palustras)、球形红细菌(Rhodobacter sphaeroides)及其混合培养物对铜绿微囊藻(Microcystis aeruginosa)生长的抑制效应.实验结果表明,光合细菌混合培养物对铜绿微囊藻抑藻作用最强,培养5 d时,铜绿微囊藻生物量降低率达58.9%,培养时间、培养温度、菌体投加量及培养基pH值等对光合细菌混合培养物的抑藻作用均有不同程度的影响.光合细菌混合培养物发挥抑藻作用的适宜条件为培养温度25℃,培养时间≤5 d,光合细菌投加量(V_(光合细菌菌悬液):V_(藻培养液)=1:80),藻培养基pH为8.0左右.     

几株紫色非硫细菌的分离鉴定与产氢特性分析

从重庆地区不同环境淤泥、泥水样品中,经富集培养、分离纯化,获得5株紫色非硫细菌.根据菌体的菌落形态、染色特性、生理生化特征及活细胞光吸收峰对菌株进行常规鉴定,结合菌株16S rDNA扩增测序进行分子生物学分析验证,构建了菌株与数据库中近缘菌株的系统发育树.以优化的培养条件(营养、pH、接种量等参数)对供试菌株的生理生化特性和产氢能力做了比较分析.结果显示,5个菌株均为光合产氢细菌,菌株ANI、D1为沼泽红假单胞菌(Rhodopseudomonas palustris),AN2、AS1、BS1等3株为类球红细菌(Rhodobacter sphaeroides);其中类球红细菌菌株AN2在给定的培养条件下光合产氢能力最高,可达9.55μg/mL d-1,是一株有应用前景的光合产氢细菌.图4表2参15     

不同酸碱条件下光合细菌的生长产氢特性

为了解光合细菌的产氢机理,对沼泽红假单胞光合产氢菌CQK-01在光生物平板反应器中进行序批次培养,以470 nm LED灯提供连续光照,葡萄糖为碳源底物,研究不同初始酸碱条件下产氢光合细菌的生长特性、产氢特性以及能量转化效率.结果表明,在弱碱性条件下最适宜产氢光合细菌的生长;在反应液为酸性条件下,光合细菌具有较高的产氢量、产氢速率和能量转化效率,然而产氢纯度随着初始pH值的增大而升高;在温度30℃、光照强度1 000lx、底物浓度75 mmol/L的实验条件下,光合细菌的最佳产氢pH值为7.0,实验中最大累积产氢量为8.0 mmol,最大产氢速率为3.39 mmol g-1(cell dry weight)h-1,产氢纯度高达70%,底物能量转化效率最大为1.98%,光能转化效率最大为7.7%.     

苋菜AmMYB2基因密码子偏好性与进化分析

为了解苋菜AmMYB2基因的密码子使用偏好性、与不同物种中调控色素相关的R2R3-MYB基因的同源关系及异源表达受体,利用Codon W与EMBOSS程序及Excel、SPSS、Origin等软件,分析苋菜与其他物种中调控色素代谢的R2R3-MYB基因的密码子使用偏好性,比较苋菜AmMYB2基因与模式生物的密码子使用频率.结果显示:苋菜AmMYB2基因中,所有密码子的GC、GC1、GC2、GC3含量均低于50%,偏好性较强的密码子有17个(RSCU 1),偏好性最强的有7个(RSCU 2),其中以A/T结尾的密码子有19个,表明苋菜AmMYB2基因的密码子偏好以A/T结尾,一些氨基酸对密码子的使用具有较强的偏好性;苋菜AmMYB2基因密码子的有效密码子数(Effective number of codons,ENC)值为49.63,苋菜AmMYB2基因的密码子在ENC绘图中的基因位点更接近期望曲线,说明在进化过程中苋菜AmMYB2基因的密码子偏好性受突变压力的影响较大;苋菜基因与甜菜色素代谢相关的R2R3-MYB基因的密码子使用偏好性参数相近,同源关系较近物种密码子使用偏好性具有相似性;从密码子使用频率上分析得出,酵母菌表达系统更适合苋菜AmMYB2基因的异源表达,模式植物拟南芥、烟草、番茄均可作为苋菜AmMYB2基因的遗传转化受体,同苋菜代谢途径相似的甜菜也可以作为苋菜AmMYB2基因的瞬时表达受体.本研究可为苋菜AmMYB2基因功能的进一步研究提供重要参考.(图5表2参31)     

异养硝化-好氧反硝化菌Psracoccus pantotrophus ATCC 35512的研究进展

近年来发现了一类具有偶联异养硝化-好氧反硝化功能的细菌.Paracoccus pantotrophus ATCC 35512是其中最早发现的细菌.通过对该菌株16S rRNA序列和细胞色素c的氨基酸序列分析,确立其为一个新种Paracoccuspantotrophus,ATCC 35512被确定为模式株.该菌株的异养硝化-好氧反硝化偶联途径假说被提出,其反应途径中涉及的一系列酶和电子传递体也陆续被纯化研究.本文综述了对该菌株系统分类、形态和生理生化特征、硝化-反硝化偶联假说及氮循环酶系等的研究进展,并提出了进一步揭示异养硝化-好氧反硝化现象的研究方向.     

龙眼漆酶家族成员全基因组结构与功能分析

为了解龙眼漆酶(DlLAC)家族的生物学功能,采用生物信息学分析方法进行龙眼基因组LAC基因成员鉴定,蛋白结构域及特性、启动子顺式作用元件、分子进化树分析、体胚发生过程和组织器官特异表达的FPKM分析以及可能互作的mi RNA预测.结果显示:DlLAC家族基因包含42个基因成员,分为7大类;基因家族成员内含子数量1-8个不等,以5个为主,其中DlLAC15-1中存在600 bp短串联重复序列;DlLAC蛋白结构域共存在5种类型,均属于铜蓝蛋白,以3个铜蓝氧化酶结构域为主;DlLAC蛋白均为分泌型蛋白,亚细胞定位预测在胞外,包含2个以上糖基化位点,具有多个保守的motif;5端调控序列预测分析,DlLAC可能具有多个转录起始位点;DlLACp启动子序列均包含大量非生物胁迫应激响应元件、胚乳特异表达元件及MYB结合位点,推测MYB可能通过DlLAC进而调控龙眼胚胎发育.DlLAC9-1p、DlLAC12p、DlLAC17-2p含黄酮类化合物合成调控的MYB结合位点,可能参与龙眼生长发育过程中种子成熟和果皮成色的色素合成途径;DlLAC家族成员可能参与不同体胚发育过程和不同组织器官形态建成.42个龙眼漆酶成员共有28个受miRNA调控,13个成员受miR397调控,可能与木质素合成相关,8个成员受miR1535调控.非miR397调控的成员可能发挥其他重要的生物学功能.本研究表明,DlLAC家族成员在龙眼生长发育过程中除了参与木质素合成以外,还可能参与胚胎发育、胚乳发育、色素合成、组织器官特异表达等重要生物学功能,表现其功能上的多样性.     

丹参提取液对球形红细菌菌体蛋白及几种酶的影响

为探索球形红细菌对丹参的生物转化机理,采用非变性聚丙烯酰胺凝胶电泳(N-PAGE)分别对丹参水提液、醇提液、醇水提液培养后球形红细菌菌体及纯球形红细菌(PRS)菌体蛋白质(Pro)、酯酶(EST)、细胞色素氧化酶(COD)、超氧化物歧化酶(SOD)和过氧化物酶(POD)进行分析,比较用丹参提取液培养前后球形红细菌菌体蛋白及4种酶的同工酶变化.结果表明:丹参提取液培养前后球形红细菌菌体蛋白及4种酶的同工酶谱带差别较大,同工酶的电泳迁移率、活性、所表达同工酶的数目及分布均有差异,培养d 2～6蛋白及酶表达量变化最大,d 14～20基本稳定.研究表明丹参能诱导球形红细菌生成新的蛋白质及酶,亦可抑制某些蛋白质和酶的合成;这些蛋白和酶可能参与了丹参化学成分的生物转化.图4参22     

光合细菌PCR检测技术的建立与应用

针对光合细菌传统的菌种鉴定方法和MPN定量方法存在费时、费力和准确性筹的问题,本研究建立了光合细菌特异PCR鉴定技术和实时荧光PCR定量的检测技术.以微生物肥料产品常用的光合细菌沼泽红假单胞菌和类球红细菌作为研究对象,分别依据16S rDNA序列和gyrB序列设计出具有种水平特异性的引物,优化并确定了PCR反应条件,其灵敏度达到100 pg/μL,对6个光合细菌产品鉴定的符合率为100%,结果表明建立的PCR鉴定技术具有特异性、灵敏性和实用性.再根据16S rDNA的保守序列设计了常用光合细菌通用引物,用其对系列稀释的已知菌含量样品的DNA模板进行荧光定量PCR,制作标准曲线.对10个光合细菌样品进行荧光定量PCR法测定,根据与标准曲线比较得出样品中的光合细菌含量,其结果与MPN法的相关系数为0.98,两者具有良好的相天性,结果表明建立的荧光定量PCR法可用于光合细菌的定量检测,并具有高特异性和快速等优点.     

Phycoerythrin and photosynthesis of the pelagic blue-green alga Trichodesmium thiebautii in the waters of Kuroshio,Japan

The photosynthetic function of phycoerythrin was investigated in the alga Trichodesmium thiebautii from the waters of Kuroshio, Japan. The spectroscopic characteristics of the in vivo and isolated T. thiebautii phycoerythrin pigments are identical, and have 3 absorption bands at 495, 547 and 562 nm. Light at the wavelengths corresponding to each absorption band of phycoerythrin is equally efficient in T. thiebautii photosynthesis, indicating that phycoerythrin is active in trapping light energy for photosynthesis. In the natural habitat, phycoerythrin is considered to be the main photosynthetic pigment in T. thiebautii photosynthesis.     

外源Hg胁迫对3种烟草品种叶绿素含量和光合效应的影响

为研究不同浓度汞(Hg)对烟草光合作用的影响,选择福建省3个主栽烟草品种(翠碧1号、K326、云烟87)幼苗进行盆栽试验,设置0、1、2、4、8、16 mg·kg~(-1)共6个浓度组。结果表明:(1)3种烟草叶片叶绿素含量与对照比均无显著差异(P0.05),但F_v/F_m、F_v/F_o、PI_(ABS)随胁迫浓度增加均呈下降趋势,可见Hg胁迫下烟草叶绿素含量和光合强度没有必然的相关性。(2)在较高浓度(4 mg·kg~(-1))Hg胁迫下,OJIP曲线改变了形状,表明Hg胁迫对烟草叶片光合机构影响主要是PSⅡ及光系统反应中心受到损伤,光合电子传递过程受到抑制,这可能是烟草叶片光合效应显著下降的主要因素。(3)烟草叶片PSII单位反应中心比活性参数ABS/RC、TRo/RC、DIo/RC与胁迫浓度正相关,ETo/RC和RC/CSo与胁迫浓度呈负相关,在较高浓度Hg胁迫下烟草叶片通过耗散过剩的光能和提高多余能量消耗量来保护光合机构免受更大的伤害,表现出较强的自我调节能力。     

Time-course of photoadaptation in the photosynthesis-irradiance relationship of a dinoflagellate exhibiting photosynthetic periodicity

Cultures of the marine dinoflagellate Glenodinium sp. were light-shifted and rates of photoadaptation determined by monitoring changes in cell volume, growth rate, pigmentation, parameters of the photosynthesisirradiance (P-I) curves and respiration. To approximate physiological conditions of field populations, cells were cultured on an alternating light-dark cycle of 12hL:12hD, which introduced a daily periodicity of photosynthesis. One result of the present study was to demonstrate how specific parameters of the P-I relationship influenced by periodicity of the light: dark cycle are distinguished from photosynthetic parameters influenced by changes in light level. Under steady-state conditions, rates of both light-saturated (P_max) and light-limited photosynthesis changed in unison over the day; these changes were not related to pigmentation, and displayed their maxima midday. This close relationship between P_max and the slope (a) of the cellular P-I curves in steadystate conditions was quickly adjusted when growth illumination was altered. Rates of light-limited photosynthesis were increased under low light conditions and the periodicity of cellular photosynthesis was maintained. The short-term responses of the P-I relationship to changing light level was different, depending on (1) whether the light shift was from high to low light or vice versa, and (2) whether the high light levels were sufficient to promote maximal photosynthesis rates. Major increases in the photosynthetic carotenoid peridinin, associated with a single type of light-harvesting chromo protein in the chloroplast, was observed immediately upon shifting high light cultures to low light conditions. Following pigment synthesis, significant increases in rates of light-limited photosynthesis were observed in about one-tenth the generation time, while cellular photosynthetic potential was unaffected. it is suggested that general results were consistent with suggested that general results were consistent with earlier reports that the major photoadaptive strategy of Glenodinium sp. is to alter photosynthetic unit (PSU) size. Photoadaptive response times to high light were light-dependent, but appeared to be shower than photoadaptive responses to low light. If light intensities were bright enough to maximize growth rates, photosynthetic response times were on the order of a generation period and pigmentation fell quickly as cells divided at a faster rate. If light-intensities were not sufficient to maximize growth rates, then pigment content did not decline, while rates of light-limited photosynthesis declined quickly. In all cases, photoadaptation was followed best by monitoring fast changes in half saturation constants for photosynthesis, rather than fluctuating changes in pigmentation. Results compared well with time-course phenomena reported for other groups of phytoplankton. Overall, results suggest phytoplankton can bring about photo-induced changes in photosynthesis very quickly and thus accommodate widely fluctuating light regimes over short periods of time.     

大气CO2浓度升高对植物光合作用的影响

大气CO2浓度不断升高以及由此带来的温室效应已成为全球变化研究的热点问题之一。CO2作为植物光合作用的底物,其浓度升高必然对植物的光合作用产生影响。大气CO2浓度升高对植物光合作用的影响主要体现在:对不同植物的光合色素含量均有影响,但结果有所差异;短期处理光合速率提高,而长期处理则可能出现光合适应,其适应机理目前尚存在分歧;不同光合类型植物的叶片形态结构有不同的响应结果,叶绿体超微结构也明显变化;生物量和产量提高。此外,CO2浓度升高与其它环境因子相互作用对植物的光合作用也具有重要影响。大气CO2浓度升高条件下对木本植物的研究、在分子水平上的深入研究以及在不同环境下的研究将成为未来研究的主要方向。     

Effect of light quality on photosynthesis of the reef coral Montipora verrucosa

Pieces of the reef coral Montipora verrucosa (Lam.), collected from Kaneohe Bay, Oahu, Hawaii in 1982, were grown in four low-light treatments (11% sunlight): blue, green, red and the full spectrum of photosynthetically active radiation (PAR); and at high-intensity full PAR (90% sunlight). These acclimated corals were then tested for photosynthetic ability in blue, green, red, and white light. The photosynthetic parameters that were measured were; ligh-saturated photosynthetic rate, the initial slope of the photosynthesis/irradiance curve, the light intensity where these two lines crossed, and dark respiration. While acclimation intensity had a pronounced effect, the results also showed that the color of the acclimation treatment influenced the photosynthetic responses of the corals. The color of the light used in the measurements of photosynthesis had much less effect on the photosynthetic responses of the corals.Contribution No. 729 of the Hawaii Institute of Marine Biology     

Humic substances. Part II†: Photophysical,photochemical and free radical characteristics

The effects of mercury (HgCl₂) on cell population, chlorophyll a concentration and rates of photosynthesis and excretion were investigated in the phytoplanktonic species Dunaliella minuta in laboratory cultures. Mercury, above 25ppb inhibited both cell population and chlorophyll a concentration approximately to the same extent, whereas the photosynthetic rate was inhibited to a significantly lesser degree. Although, the total photosynthetic rate of the tested organism was reduced, above a threshold concentration, the photosynthetic activity was not reduced under these conditions, but it was in fact significantly greater than that in the control culture. This may suggest that in D. minuta the inhibitory effect of mercury is primarily on cell division rather than cellular photosynthesis, which is enhanced by the fact mercury caused a significant increase of the mean cell volume. Mercury, also, decreased the growth rate and final cell yield. The excretory rate was markedly increased at concentrations ≥ 250 ppb of mercury, but at lower concentrations it tended to depend more on the physiological state of cells than on mercury concentration. In the different cultures, the photosynthetic activity showed variations which occurred without major changes in the chlorophyll a content per cell, which remained almost constant and independent of variations in cell size and growth conditions.     

Interactions between morphological and physiological plasticity optimize energy acquisition in corals

Morphological plasticity in response to environmental heterogeneity may be performance enhancing or may simply result from an intrinsic instability in morphology during development. Although patterns of morphological change are well documented for numerous taxa, it is often unclear whether this plasticity enhances the performance of organisms in the habitat to which they have acclimatized. Reef-building corals are an ideal model system in which to investigate this question. We here develop a three-dimensional geometric model and present a comprehensive photosynthesis data set with experimentally calibrated photosynthesis models that predicts energy acquisition by foliose corals as a function of colony shape. This allows us to assess the extent to which changes in colony morphology along an environmental gradient track the predicted optimal colony morphologies. Our results provide strong evidence that phenotypic plasticity in foliose corals optimizes photosynthetic energy acquisition and is not simply a mechanism to increase light capture. We show that the optimal morphology is constrained at the boundaries of the environmental gradient, with non-optimal morphologies in these habitats having greatly reduced energy acquisition. However, at the center of the environmental gradient, flexibility in photophysiology allows energy acquisition to be very similar for multiple morphologies. Our results highlight the importance of phenotypic plasticity at multiple scales. Variation in overall morphology is important at niche boundaries at which conditions are consistently more stressful, whereas physiological flexibility is important in intermediate and less predictable habitats in which a rapid and reversible response to environmental fluctuations is required.     

Photosynthetic characteristics of giant kelp (Macrocystis pyrifera) determined in situ

Rates of net photosynthesis and nocturnal respiration by individual blades of the giant kelp Macrocystis pyrifera (L.) C. Agardh in southern California, were determined in situ by measuring oxygen production in polyethylene bags during spring/summer of 1983. Mature blades from different depths in the water column exhibited different photosynthetic characteristics. Blades from the surface canopy (0 to 1 m depth) exhibited higher photosynthetic capacity under saturating irradiance and higher photosynthetic efficiency at low irradiances than blades from 3 to 5 or 7 to 9 m depths. Saturating irradiance was lower for canopy blades than for deeper blades. Canopy blades showed no short-term photoinhibition, but photosynthetic rates of deeper blades were significantly reduced during 1 to 2 h incubations at high irradiances. Results of 1 to 2 wk acclimation experiments indicated that differences between photosynthetic characteristics of blades from different depths were primarily attributable to acclimation light conditions. Vertical displacement of blades within the kelp canopy occurred on a time-scale of 1 min to 1 h. Blades continually moved between the unshaded surface layer and deeper, shaded layers. Vertical movement did not maximize photosynthesis by individual blades; only a small proportion of blades making up a dense surface canopy maintained light-saturated photosynthetic rates during midday incubations. The relatively high photosynthetic rates exhibited by canopy blades over the entire range of light conditions probably resulted from acclimation to intermittent high and low irradiances, a consequence of vertical displacement. Vertical displacement also reduced the afternoon depression in photosynthesis of individual canopy blades. The overall effect of vertical displacement was optimization of total net photosynthesis by the kelp canopy and, therefore, optimization of whole-plant production.     

Photoadaptation of photosynthesis in Gonyaulax polyedra

Gonyaulax polyedra Stein exhibited a combination of photoadaptive strategies of photosynthesis when only a single environmental variable, the light intensity during growth, was altered. Which of several biochemical/physiological adjustments to the light environment were employed depended on the level of growth irradiance. The photoadaptive strategies employed over any small range of light levels appeared to be those best suited for optimizing photosynthetic performance and not photosynthetic capacity. (Photosynthetic performance, P _i, is defined as the rate of photosynthesis occurring at the level of growth irradiance.) Among all photosynthetic parameters examined, only photosynthetic performance showed a consistent correspondence to growth rates of G. polyedra. Above 3500 to 4000 W cm^-2, where photosynthetic performance was equal to photosynthetic capacity, cells were not considered light-limited in either photosynthesis or growth. At these higher light levels, photosynthetic perfomance, cell volume, growth rates and respiration rates remained maximal; photosynthetic pigment content varied only slightly, while the photosynthetic capacity of the cells declined. At intermediate light levels (3000 to 1500 W cm^-2), photosynthesis, not growth, was light-limited, and photoadaptive strategies were induced which enhance absorption capabilities and energy transfer efficiencies of chlorophyll a to the reaction centers of G. polyedra. Photosynthetic capacity remained constant at about 280 mol O₂ cm^-3 h^-1, while photosynthetic performance ranged from 100 to 130 mol O₂ cm^-3 h^-1. Major increases in photosynthetic pigments, especially peridinin-chlorophyll a-proteins and an unidentified chlorophyll c component, accompanied photoadaptation to low irradiances. Maximal growth rates of 0.3 divisions day^-1 were maintained, as were respiration rates of about-80 mol O₂ cm^-3 h^-1 and cell volumes of about 5.4×10^-8 cm^-3 cell^-1. Below about 1250 W cm^-2, photosynthesis in G. polyedra was so light-limited that photosynthetic performance was unable to support maximal growth rates. Under these conditions, G. polyedra displayed photostress responses rather than photoadaptive strategies. Photostress was manifested as reduced cell volumes, slower growth, and drastic reductions in pigmentation, photosynthetic capacity, and rates of dark respiration.