硅藻生长过程中活体荧光变化规律及其对叶绿素测量的影响

发布时间：2018-06-08 10:46

本文选题：硅藻 + 叶绿素　；参考：《中国地质大学》2016年博士论文

【摘要】：本研究以海洋中普遍存在的硅藻为主要研究对象,通过实验室藻类培养实验的方式营造出多种因素影响下的实验环境,将传统的萃取法测定数据与采用活体荧光原理的传感器测定结果进行直接对比,获取了以往依靠有限的现场比测数据无法充分展现的内在规律；进行了藻种、光照、温度、盐度、浊度等多种因素对叶绿素传感器的影响研究,分析了不同藻种以及在不同的环境因素下的活体荧光变化规律,并根据影响机理将其进行归类；在分析各种影响因素的基础上,在叶绿素传感器数据质量保障方法上提出了新的见解,建立了一种基于硅藻液的叶绿素传感器实验室标定方法,建立了浊度影响修正方法和种间差异修正方法,并提出了一种简单可行的实际海水综合因素影响整体修正方法。本论文主要研究内容和研究结果如下：1.光照对硅藻叶绿素活体荧光测量的影响光照是硅藻进行光合作用的必要条件,光照强度较充分的条件下硅藻的叶绿素活体荧光也较强,但过强的光照条件会引起硅藻自身的保护机制,从而引起短时间内硅藻叶绿素活体荧光强度降低。经过长期培养后,最适光照条件下(60001x、12h亮/12h暗)的硅藻叶绿素活体荧光和萃取法叶绿素都比其他光照条件更高。短期光照强度连续变化时,硅藻叶绿素活体荧光与光照强度之间存在较好的线性关系；在光照强度突变瞬间,硅藻叶绿素活体荧光会随之出现一个明显变化,但短期光照突变不影响硅藻叶绿素活体荧光的总体水平。短期不同光暗比对于硅藻的叶绿素活体荧光有着一些影响,光照时间占比越长,硅藻叶绿素活体荧光总体水平越高；经过不同光暗比短期持续培养后,硅藻叶绿素活体荧光与萃取法叶绿素测定值之间的关系存在较大差异。2.主要环境参数对硅藻叶绿素活体荧光测量的影响温度快速变化对硅藻叶绿素活体荧光的影响跟光照有关。在无光照条件下,温度降低过程会略微提高硅藻的叶绿素活体荧光强度；在有光照条件下,温度降低过程会显著降低硅藻的叶绿素活体荧光强度；不管有无光照,温度变化对萃取法叶绿素测定值的影响都很小。盐度快速变化过程中萃取法叶绿素含量和叶绿素活体荧光变化规律与盐度不变时都基本一致,表明短期盐度变化对硅藻叶绿素活体荧光和萃取法叶绿素浓度的影响都很小。浊度对叶绿素荧光的影响包括水中悬浮物的荧光效应和遮蔽效应两个方面。悬浮物的荧光效应较为恒定,可以采用公式△Flu测=0.5109×Tu(R2=0.98)进行修正。悬浮物遮蔽效应对荧光测定的影响与硅藻浓度有关,随着硅藻浓度的增加,屏蔽效应也逐渐增强。当浊度和藻类都达到一定浓度时,屏蔽效应会趋于一个相对稳定的状态。浊度遮蔽效应影响修正根据叶绿素浓度从低到高分别采用△F1u测=-0.08×Tu (R2=0.97)、ΔFlu测=0.0044×Tu2-1.5987×Tu (R2=0.99)和AFlu测=0.0186×Tu2-6.1739×Tu (R2=0.94)三个公式。3.叶绿素传感器数据质量保障方法可以基于硅藻液建立叶绿素传感器实验室标定方法。选取光照开始后4小时的硅藻液作为叶绿素传感器的标定溶液,将稀释后测得的8-10个浓度下标定溶液的叶绿素荧光测定值和叶绿素萃取法浓度值进行线性回归,获得叶绿素传感器的工作曲线。经过验证,标定后传感器法叶绿素a浓度和萃取法叶绿素a浓度偏差小于6%,表明标定结果具有较好的重现性。在分析藻种差异在海水藻类叶绿素活体荧光测量中的影响程度的基础上,可以建立种间差异影响修正方法。研究表明,不同纯种藻液活体荧光差异明显,藻液混合时的叶绿素活体荧光具有可叠加性。经过藻种间差异修正以后三种藻的叶绿素a浓度与萃取法叶绿素偏差小于±10%,表明该修正方法具有较好的效果。此外,还提出了一种简单可行的实际海水综合因素影响整体修正方法,以海水样品萃取法叶绿素a浓度为基准,对传感器法叶绿素a浓度进行曲线回归校正。4.叶绿素传感器研究方向展望叶绿素传感器数据溯源方面：应区分环境因素中对传感器自身荧光值测定的影响以及对藻类活体荧光的影响两个方面,并进行相应的专门研究；传感器数据影响研究应采用实验室和现场相结合的方式。叶绿素传感器数据指示意义方面：应充分研究叶绿素活体荧光本身具有的生态指示意义并进行应用,如将藻种的活体荧光差异应用到赤潮旺发过程优势藻种更替分析。叶绿素传感器研发方面：从传感器数据质量控制的角度推测,未来的海水叶绿素传感器可能是一个多通道光学测量仪,能够同时测定光照度、叶绿素活体荧光、浊度、CDOM以及藻种指示参数,同时需配合其他基础环境参数测量设备如CTD等一起使用。
[Abstract]:In this study, the main research object of this study is diatom, which is common in the ocean. Through the laboratory algae cultivation experiment, the experimental environment under the influence of various factors is created. The traditional extraction method is directly compared with the sensor measurement results using the principle of living fluorescence. The influence of various factors such as algae species, light, temperature, salinity and turbidity on the chlorophyll sensor was studied. The changes of different algae species and the change of living fluorescence under different environmental factors were analyzed, and the basis of the influence mechanism was classified, and the basis of various factors was analyzed. A new view is put forward on the method of data quality assurance for chlorophyll sensors. A calibration method for chlorophyll sensor based on diatom liquid is established, the correction method of turbidity influence and the method of interspecific correction are established, and a simple and feasible method for the overall correction of the effect of the actual sea water comprehensive factors is put forward. The main research contents and results are as follows: 1. the effect of light on the fluorescence measurement of diatom chlorophyll is the necessary condition for the photosynthesis of diatom, and the chlorophyll living fluorescence of diatom is stronger under the condition of full light intensity, but too strong light conditions will cause the protection mechanism of diatom itself, thus causing a short time. The fluorescence intensity of the chlorophyll living body of diatom diatom decreased. After a long period of culture, the chlorophyll in the chlorophyll living and extraction of diatom (60001x, 12h bright /12h dark) was higher than that of other light conditions. There was a good linear relationship between the fluorescence intensity and the light intensity of the chlorophyll living body of diatom. At the moment of light intensity mutation, the fluorescence of the chlorophyll living body of diatom has an obvious change, but the short term light mutation does not affect the overall level of the fluorescence of the chlorophyll in the diatom. The short term light and dark ratio has some influence on the chlorophyll living fluorescence of diatom, the longer the light time ratio is, the chlorophyll living body fluorescence is overall. The higher the level is, the relationship between the chlorophyll living fluorescence of diatom chlorophyll and the determination of chlorophyll after short-term continuous culture, the influence of.2. main environmental parameters on the fluorescence measurement of diatom chlorophyll living body is related to the illumination of the fluorescence of diatom chlorophyll live body. Under the conditions of temperature reduction, the fluorescence intensity of chlorophyll in diatom could be slightly increased. Under the conditions of light, the fluorescence intensity of the chlorophyll in the diatom was significantly reduced. The effect of temperature change on the determination of chlorophyll in the extraction method was very small. The changes of the content of the chlorophyll and the chlorophyll fluorescence were basically the same as the salinity, which showed that the effect of the short-term salinity changes on the chlorophyll concentration of the chlorophyll fluorescence and the extraction method was very small. The effect of turbidity on chlorophyll fluorescence included two aspects: fluorescence effect and masking effect of suspended matter in water. The fluorescence effect of suspended matter It should be relatively constant, and the formula Delta Flu =0.5109 x Tu (R2=0.98) can be used to modify it. The effect of the suspended substance shielding effect on the concentration of diatom is related to the concentration of diatom. With the increase of the concentration of diatom, the shielding effect is gradually enhanced. When the turbidity and algae reach a certain concentration, the shielding effect will tend to a relatively stable state. The correction of chlorophyll sensor data quality can be based on three formulas of =-0.08 * Tu (R2=0.97), Delta Flu, =0.0044, Tu2-1.5987 x Tu (R2=0.99) and AFlu =0.0186, AFlu =0.0186 * Tu2-6.1739 * *, based on the low to high chlorophyll concentration. The chlorophyll sensor data quality assurance method can be based on the diatom solution to establish the chlorophyll sensor laboratory standard. The diatom solution of 4 hours after light was selected as the calibration solution of the chlorophyll sensor. The chlorophyll fluorescence determination value of the diluted solution and the concentration value of the chlorophyll extraction method were linear regression to obtain the working curve of the chlorophyll sensor in the 8-10 concentration of the diluted solution after the dilution, and the chlorophyll a was verified by the calibrated sensor method. The concentration and extraction method of chlorophyll a concentration deviation is less than 6%, indicating that the calibration results have good reproducibility. On the basis of the analysis of the influence degree of algae species difference in the fluorescence measurement of algae chlorophyll in seawater, the interspecific effect correction method can be established. The chlorophyll living fluorescence of the three species was superimposed. After the interspecies difference, the chlorophyll a concentration and the chlorophyll deviation of the extraction method were less than 10%, indicating that the modified method had a good effect. In addition, a simple and feasible method for the overall modification of the actual marine synthetic factors was proposed, and the extraction of sea water samples was also proposed. Based on the concentration of chlorophyll a, curve regression correction of chlorophyll a concentration in sensor method, the research direction of.4. chlorophyll sensor is expected to look into the source of chlorophyll sensor data: the influence of environmental factors on the measurement of the fluorescence of the sensor and the effect on the fluorescence of algae in the environment should be distinguished from two aspects. The study of the influence of the sensor data should be combined with the laboratory and the field. The significance of the chlorophyll sensor data indicates that the ecological indication of the chlorophyll fluorescence itself should be fully studied and applied. For example, the difference of the living fluorescence of the algae should be used in the analysis of the dominance of the Dominant Algae in the process of red tide. Chlorophyll sensor research and development: from the point of view of sensor data quality control, the future sea water chlorophyll sensor may be a multi-channel optical measuring instrument, which can simultaneously determine light intensity, chlorophyll living fluorescence, turbidity, CDOM and algae indicator parameters, and other basic environmental parameters measurement equipment, such as CTD, and so on. Use it together.
【学位授予单位】：中国地质大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：Q945

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