本文選題：硅藻 + 葉綠素��； 參考：《中國(guó)地質(zhì)大學(xué)》2016年博士論文

【摘要】：本研究以海洋中普遍存在的硅藻為主要研究對(duì)象,通過(guò)實(shí)驗(yàn)室藻類(lèi)培養(yǎng)實(shí)驗(yàn)的方式營(yíng)造出多種因素影響下的實(shí)驗(yàn)環(huán)境,將傳統(tǒng)的萃取法測(cè)定數(shù)據(jù)與采用活體熒光原理的傳感器測(cè)定結(jié)果進(jìn)行直接對(duì)比,獲取了以往依靠有限的現(xiàn)場(chǎng)比測(cè)數(shù)據(jù)無(wú)法充分展現(xiàn)的內(nèi)在規(guī)律；進(jìn)行了藻種、光照、溫度、鹽度、濁度等多種因素對(duì)葉綠素傳感器的影響研究,分析了不同藻種以及在不同的環(huán)境因素下的活體熒光變化規(guī)律,并根據(jù)影響機(jī)理將其進(jìn)行歸類(lèi)；在分析各種影響因素的基礎(chǔ)上,在葉綠素傳感器數(shù)據(jù)質(zhì)量保障方法上提出了新的見(jiàn)解,建立了一種基于硅藻液的葉綠素傳感器實(shí)驗(yàn)室標(biāo)定方法,建立了濁度影響修正方法和種間差異修正方法,并提出了一種簡(jiǎn)單可行的實(shí)際海水綜合因素影響整體修正方法。本論文主要研究?jī)?nèi)容和研究結(jié)果如下：1.光照對(duì)硅藻葉綠素活體熒光測(cè)量的影響光照是硅藻進(jìn)行光合作用的必要條件,光照強(qiáng)度較充分的條件下硅藻的葉綠素活體熒光也較強(qiáng),但過(guò)強(qiáng)的光照條件會(huì)引起硅藻自身的保護(hù)機(jī)制,從而引起短時(shí)間內(nèi)硅藻葉綠素活體熒光強(qiáng)度降低。經(jīng)過(guò)長(zhǎng)期培養(yǎng)后,最適光照條件下(60001x、12h亮/12h暗)的硅藻葉綠素活體熒光和萃取法葉綠素都比其他光照條件更高。短期光照強(qiáng)度連續(xù)變化時(shí),硅藻葉綠素活體熒光與光照強(qiáng)度之間存在較好的線性關(guān)系；在光照強(qiáng)度突變瞬間,硅藻葉綠素活體熒光會(huì)隨之出現(xiàn)一個(gè)明顯變化,但短期光照突變不影響硅藻葉綠素活體熒光的總體水平。短期不同光暗比對(duì)于硅藻的葉綠素活體熒光有著一些影響,光照時(shí)間占比越長(zhǎng),硅藻葉綠素活體熒光總體水平越高；經(jīng)過(guò)不同光暗比短期持續(xù)培養(yǎng)后,硅藻葉綠素活體熒光與萃取法葉綠素測(cè)定值之間的關(guān)系存在較大差異。2.主要環(huán)境參數(shù)對(duì)硅藻葉綠素活體熒光測(cè)量的影響溫度快速變化對(duì)硅藻葉綠素活體熒光的影響跟光照有關(guān)。在無(wú)光照條件下,溫度降低過(guò)程會(huì)略微提高硅藻的葉綠素活體熒光強(qiáng)度；在有光照條件下,溫度降低過(guò)程會(huì)顯著降低硅藻的葉綠素活體熒光強(qiáng)度；不管有無(wú)光照,溫度變化對(duì)萃取法葉綠素測(cè)定值的影響都很小。鹽度快速變化過(guò)程中萃取法葉綠素含量和葉綠素活體熒光變化規(guī)律與鹽度不變時(shí)都基本一致,表明短期鹽度變化對(duì)硅藻葉綠素活體熒光和萃取法葉綠素濃度的影響都很小。濁度對(duì)葉綠素?zé)晒獾挠绊懓ㄋ袘腋∥锏臒晒庑?yīng)和遮蔽效應(yīng)兩個(gè)方面。懸浮物的熒光效應(yīng)較為恒定,可以采用公式△Flu測(cè)=0.5109×Tu(R2=0.98)進(jìn)行修正。懸浮物遮蔽效應(yīng)對(duì)熒光測(cè)定的影響與硅藻濃度有關(guān),隨著硅藻濃度的增加,屏蔽效應(yīng)也逐漸增強(qiáng)。當(dāng)濁度和藻類(lèi)都達(dá)到一定濃度時(shí),屏蔽效應(yīng)會(huì)趨于一個(gè)相對(duì)穩(wěn)定的狀態(tài)。濁度遮蔽效應(yīng)影響修正根據(jù)葉綠素濃度從低到高分別采用△F1u測(cè)=-0.08×Tu (R2=0.97)、ΔFlu測(cè)=0.0044×Tu2-1.5987×Tu (R2=0.99)和AFlu測(cè)=0.0186×Tu2-6.1739×Tu (R2=0.94)三個(gè)公式。3.葉綠素傳感器數(shù)據(jù)質(zhì)量保障方法可以基于硅藻液建立葉綠素傳感器實(shí)驗(yàn)室標(biāo)定方法。選取光照開(kāi)始后4小時(shí)的硅藻液作為葉綠素傳感器的標(biāo)定溶液,將稀釋后測(cè)得的8-10個(gè)濃度下標(biāo)定溶液的葉綠素?zé)晒鉁y(cè)定值和葉綠素萃取法濃度值進(jìn)行線性回歸,獲得葉綠素傳感器的工作曲線。經(jīng)過(guò)驗(yàn)證,標(biāo)定后傳感器法葉綠素a濃度和萃取法葉綠素a濃度偏差小于6%,表明標(biāo)定結(jié)果具有較好的重現(xiàn)性。在分析藻種差異在海水藻類(lèi)葉綠素活體熒光測(cè)量中的影響程度的基礎(chǔ)上,可以建立種間差異影響修正方法。研究表明,不同純種藻液活體熒光差異明顯,藻液混合時(shí)的葉綠素活體熒光具有可疊加性。經(jīng)過(guò)藻種間差異修正以后三種藻的葉綠素a濃度與萃取法葉綠素偏差小于±10%,表明該修正方法具有較好的效果。此外,還提出了一種簡(jiǎn)單可行的實(shí)際海水綜合因素影響整體修正方法,以海水樣品萃取法葉綠素a濃度為基準(zhǔn),對(duì)傳感器法葉綠素a濃度進(jìn)行曲線回歸校正。4.葉綠素傳感器研究方向展望葉綠素傳感器數(shù)據(jù)溯源方面：應(yīng)區(qū)分環(huán)境因素中對(duì)傳感器自身熒光值測(cè)定的影響以及對(duì)藻類(lèi)活體熒光的影響兩個(gè)方面,并進(jìn)行相應(yīng)的專(zhuān)門(mén)研究；傳感器數(shù)據(jù)影響研究應(yīng)采用實(shí)驗(yàn)室和現(xiàn)場(chǎng)相結(jié)合的方式。葉綠素傳感器數(shù)據(jù)指示意義方面：應(yīng)充分研究葉綠素活體熒光本身具有的生態(tài)指示意義并進(jìn)行應(yīng)用,如將藻種的活體熒光差異應(yīng)用到赤潮旺發(fā)過(guò)程優(yōu)勢(shì)藻種更替分析。葉綠素傳感器研發(fā)方面：從傳感器數(shù)據(jù)質(zhì)量控制的角度推測(cè),未來(lái)的海水葉綠素傳感器可能是一個(gè)多通道光學(xué)測(cè)量?jī)x,能夠同時(shí)測(cè)定光照度、葉綠素活體熒光、濁度、CDOM以及藻種指示參數(shù),同時(shí)需配合其他基礎(chǔ)環(huán)境參數(shù)測(cè)量設(shè)備如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.
【學(xué)位授予單位】：中國(guó)地質(zhì)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：Q945

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