利用荧光淬灭作用研究川泽泻凝集素微环境与生色基团的空间分布

用不带电荷的极性淬灭剂丙烯酰胺和带电荷的I-离子、Cs 对川泽泻凝集素内源荧光进行淬灭研究,以揭示凝集素与小分子物质相互作用以及凝集素荧光生色基团的分布情况.天然川泽泻(Alisma plantago-aquatica Linn)凝集素在280nm和295nm波长下均显示λmax为328nm的荧光发射光谱,表明凝集素荧光生色基团位于相对疏水的环境中.中性淬灭剂丙烯酰胺对凝集素分子的荧光生色基团Tyr和Trp残基的可接近程度为96.81%,而对阴离子淬灭剂I-离子和阳离子淬灭剂Cs 离子的可接近程度分别为37.17%和21.96%.表明川泽泻凝集素荧光氨基酸残基主要位于分子内部的疏水核心.图3参22     

Effects of lindane on the photosynthetic apparatus of the cyanobacterium<Emphasis Type="Italic">anabaena</Emphasis>

INTENTION, GOAL, SCOPE, BACKGROUND: Cyanobacteria have the natural ability to degrade moderate amounts of organic pollutants. However, when pollutant concentration exceeds the level of tolerance, bleaching of the cells and death occur within 24 hours. Under stress conditions, cyanobacterial response includes the short-term adaptation of the photosynthetic apparatus to light quality, named state transitions. Moreover, prolonged stresses produce changes in the functional organization of phycobilisomes and in the core-complexes of both photosystems, which can result in large changes in the PS II fluorescence yield. The localization of ferredoxin-NADP+ reductase (FNR) at the ends of some peripheral rods of the cyanobacterial phycobilisomes, makes this protein a useful marker to check phycobilisome integrity. OBJECTIVE: The goal of this work is to improve the knowledge of the mechanism of action of a very potent pesticide, lindane (gamma-hexaclorociclohexane), in the cyanobacterium Anabaena sp., which can be considered a potential candidate for bioremediation of pesticides. We have studied the effect of lindane on the photosynthetic apparatus of Anabaena using fluorescence induction studies. As ferredoxin-NADP+ reductase plays a key role in the response to oxidative stress in several systems, changes in synthesis, degradation and activity of FNR were analyzed. Immunolocalization of this enzyme was used as a marker of phycobilisome integrity. The knowledge of the changes caused by lindane in the photosynthetic apparatus is essential for rational further design of genetically-modified cyanobacteria with improved biorremediation abilities. METHODS: Polyphasic chlorophyll a fluorescence rise measurements (OJIP) have been used to evaluate the vitality and stress adaptation of the nitrogen-fixing cyanobacterium Anabaena PCC 7119 in the presence of increasing concentrations of lindane. Effects of the pesticide on the ultrastructure have been investigated by electron microscopy, and FNR has been used as a marker of phycobilisome integrity. RESULTS AND DISCUSSION: Cultures of Anabaena sp. treated with moderate amounts of lindane showed a decrease in growth rate followed by a recovery after 72 hours of pesticide treatment. Concentrations of lindane below 5 ppm increased the photosynthetic performance and activity of the cells. Higher amounts of pesticide caused a decrease in these activities which seems to be due to a non-competitive inhibition of PS II. Active PS II units are converted into non-QA reducing, so called heat sink centers. Specific activity and amount of FNR in lindane-treated cells were similar to the values measured in control cultures. Release of FNR from the thylakoid after 48 hours of exposure to 5 ppm of lindane towards the cytoplasm was detected by immunogold labeling and electron microscopy. CONCLUSIONS: From these results, we conclude that the photosynthetic performance and activity of the cells are slightly increased in the presence of lindane up to 5 ppm. Moreover, in those conditions, lindane did not produce significant changes in the synthesis, degradation or activity of FNR. The high capability of Anabaena to tolerate lindane makes this cyanobacterium a good candidate for phytoremediation of polluted areas. RECOMMENDATION AND OUTLOOK: The results of this study show that cultures of Anabaena PCC 7119 tolerate lindane up to 5 ppm, without significant changes in the photosynthetic vitality index of the cells. However, a slight increase in phycobiliprotein synthesis is observed, which is related to total protein content. This change might be due to degradation of proteins less stable than phycobiliproteins. An identification of the proteins with altered expression pattern in the presence of the pesticide remains the subject of further work and will provide valuable information for the preparation of strains which are highly tolerant to lindane.     

Trace metal levels in freshwater fish, sediment and water

The trace metal concentrations in water, sediment and aquatic organisms, such as fish, could indicate the level and tendency of the pollution. This is important not only for the protection of the environment, but for evaluation of the quality of fish meat either captured from natural waters or cultured in fishponds. The total trace metal concentrations in samples of fish from different regions of Hungary and from different species have been determined by using an X-ray fluorescence technique (EDXRF). Water, sediment and fish samples from fishpond systems with different feeding and stocking has also been analyzed. In the case of zinc contents, differences have been traced between the cultured and wild common carp. In the case of common carp reared under different feeding conditions, differences were also observed in the zinc concentration. The retention of the trace metals in the fish has been studied by measuring the levels in sediment, water and feed. The different retention can be explained by the different availability of zinc in the applied feeds, which can be related to the presence of different metal species in the feeds.     

Sensitivity analyses of woody species exposed to air pollution based on ecophysiological measurements

GOAL, SCOPE AND BACKGROUND: Air pollution has been of a major problem in the Pearl River Delta of south China, particularly during the last two decades. Emissions of air pollutants from industries have already led to damages in natural communities and environments in a wide range of the Delta area. Leaf parameters such as chlorophyll fluorescence, leaf area (LA), dry weight (DW) and leaf mass per area (LMA) had once been used as specific indexes of environmental stress. This study aims to determine in situ if the daily variation of chlorophyll fluorescence and other ecophysiological parameters in five seedlings of three woody species, Ilex rotunda, Ficus microcarpa and Machilus chinensis, could be used alone or in combination with other measurements for sensitivity indexes to make diagnoses under air pollution stress and, hence, to choose the correct tree species for urban afforestation in the Delta area. METHODS: Five seedlings of each species were transplanted in pot containers after their acclimation under shadowing conditions. Chlorophyll fluorescence measurements were made in situ by a portable fluorometer (OS-30, Opti-sciences, U.S.A). Ten random samples of leaves were picked from each species for LA measurements by area-meter (CI-203, CID, Inc., U.S.A). DW was determined after the leaf samples were dried to a constant weight at 65 degrees C. LMA was calculated as the ratio of DW/LA. Leaf N content was analyzed according to the Kjeldhal method, and the extraction of pigments was carried out according Lin et al. RESULTS AND DISCUSSION: The daily mean Fv/Fm (Fv is the variable fluorescence and Fm is the maximum fluorescence) analysis showed that Ilex rotunda and Ficus microcarpa were more highly resistant to pollution stress, followed by Machilus chinensis, implying that the efficiency of photosystem II in I. rotunda was less affected by air pollutants than the other two species. Little difference in daily change of Fv/Fm in I. rotunda between the polluted and the clean site was also observed. However, a relatively large variation of Fv/Fm appeared in the other two species, particularly in M. chinensis, suggesting that they were more sensitive to air pollutants than I. rotunda. The mean LA was reduced for all species growing at the polluted site. The mean LMA for all species exceeded the sclerophylly threshold given by Cowling and Campbell and increased for those under pollution stress, which could be explained as one of the acclimation strategies for plants to air pollution stress. Little difference in leaf chlorophyll content was observed in F. microcarpa and M. chinensis, while remarkable differences were found in I. rotunda growing at the polluted and the clean site. Content of leaf carotenoids was largely reduced in I. rotunda growing at the polluted site, but increased in F. microcarpa and M. chinensis, compared with plants growing at the clean site. Plants growing at the clean site had a lower leaf N content than those growing at the polluted site. In addition, species with a higher resistance to pollution stress showed less difference in leaf N content than those sensitive species. CONCLUSION: Based on Fv/Fm measurements of the three woody species, I. rotunda showed the highest resistance to air pollutants from ceramic industries, followed by F. microcarpa. M. chinensis was the most sensitive species to air pollution, had lowest capacities to cope with the air pollution stress, which was consistent with visual injury symptoms observed in the crown profiles of plants at the polluted site. Fv/Fm, LAM, LA, leaf pigments and N content could be used alone or in combination to diagnose the extent of the physiological injury. The ratio of Fv/Fm, however, was the best and most effective parameter. RECOMMENDATION AND OUTLOOK: Tree species which have higher air-pollutant resistance, as diagnosed by such ecophysiological parameters, should be considered first and planted widely for urban afforestation or forest regeneration in areas where the forest was seriously degraded or forest health was markedly effected by the same kind of air pollutants.     

应用蝶啶荧光分析法推断巨尾阿丽蝇成虫日龄

为探明蝶啶荧光分析法在尸食性蝇类成虫日龄推断中的应用潜力,采用荧光分光光度计对4种恒温和自然变温下巨尾阿丽蝇(Aldrichina grahami)雌雄成虫头壳蝶啶含量与日龄的关系进行了研究.结果表明,性别、温度和日龄对成虫蝶啶含量均有显著影响,其中日龄为最主要的影响因子.在16℃、20℃、24℃和28℃下,雌雄成虫头壳蝶啶含量与日龄均呈显著的线性关系(P<0.01,R2>0.90).蝶啶积累速率与温度也呈显著的线性关系.应用蝶啶荧光分析法对自然变温下巨尾阿丽蝇雌雄成虫日龄的估计表明,估计日龄与实际日龄呈显著的线性关系,其中雌成虫日龄估计误差平均为2.68d,雄成虫为±1.56d.以上研究结果表明,蝶啶荧光分析法在巨尾阿丽蝇成虫尤其是雄成虫日龄的推断中具有较大的应用价值.图4表2参19     

用荧光光谱研究腐殖酸与金属离子Al^3＋的配合作用

用同步荧光法对两种来源不同的腐殖酸与金属离子Ａｌ＾３＋的配合作用进行了研究,采用非线性回归分析,用荧光强度变化数据,拟合出金属配合物（１：１）的条件稳定常数Ｋ,在配合滴定进行的同时,探讨了溶液在５００ｎｍ处的瑞利散射现象。     

基于分子信标探针的荧光定量PCR方法检测转基因食品

选择了内源基因大豆植物凝集素(lectin)、玉米转化酶(invertase)和外源基因花椰菜花叶病毒35S(CaMV35S)启动子的分子信标探针,确定了探针浓度和镁离子浓度等反应条件,分别对转基因大豆和转基因玉米系列标准品进行内源基因和外源基因的荧光PCR扩增,在PCR反应过程中分别以两种荧光通道信号分别追踪同一样品DNA内源基因和外源基因的扩增动力学变化,并依此绘制了循环阈值与转基因食品百分比含量之间的标准曲线,建立了转基因大豆和转基因玉米的分子信标探针-荧光定量PCR检测方法,该方法具有特异性强、敏感性高的特点,实现了对转基因食品的定量分析.图7表1参11     

荧光猝灭法测定菲、苊、芘与腐殖酸的结合常数

用荧光猝灭法测定了胡敏酸（Fluka）与苊、菲和芘相互作用的结合常数（KDOC），其lgKDOC值分别为4．4，4．48和4．89．用同样方法测得长江水生FA以及黑龙江水生FA和HA的KDOC之间有显著差别．HA的KDODC一般高于FA．黑龙江水生FA的KDOC高于长江．对同一来源的腐殖酸，三种多环芳烃的KDOC与KOW呈线性相关．     

干热河谷主要造林树种光合作用光抑制的防御机制

比较研究了干热河谷8个造林树种气孔导度、光合色素、叶绿索荧光的日变化和季节变化，结果表明：①气孔导度雨季较高，多呈“午睡”现象的双峰曲线日进程，旱季气孔导度很低，全呈单峰曲线日进程；叶绿素荧光动力学参数Fv／Fm呈反正态分布曲线日进程，qP和φPsⅡ呈正态分布曲线日进程，白昼qNP随时间呈上升趋势，表明树种启动了过剩激发能耗散功能．②叶绿素总含量、叶绿素a／b值及类胡萝卜素／叶绿素值从雨季到旱季都呈下降趋势，减少了对光能的吸收．③旱季马占相思(Acacia mangium)、刺槐(Robinia pseudoacacia)的Fv／Fm值明显低于雨季值，差异显著，发生了严重的光抑制；qP和qNP值除赤桉(Eucalyptus dulebsis)变化不明显外，其余树种旱季qP值下降而qNP值上升．④对干旱、高光强、高温光保护的主动适应方式是保持较高的光化学猝灭、PSII线性电子传递效率和维持一定耗散过剩光能的能力，如降低光化学猝灭而增加非光化学猝灭来适应逆境，则是一种对生长不利的内损耗被动适应方式．图4表2参14     

Detennination of trace uranium in environment by laser induced fluorescence method

ＤｅｔｅｎｎｉｎａｔｉｏｎｏｆｔｒａｃｅｕｒａｎｉｕｍｉｎｅｎｖｉｒｏｎｍｅｎｔｂｙｌａｓｅｒｉｎｄｕｃｅｄｆｌｕｏｒｅｓｃｅｎｃｅｍｅｔｈｏｄＷｕＪｉｎｈａｉ，ＹｕａｎＺｈｅｎｇａｎ，ＬｉＪｉｎｑｕａｎ，ＺｈａｎｇＣｈｕａｎｚｕｏ，ＲｅｎＬｉｈｕａ...