光照条件对小檗碱抑藻效应的影响及其机制初探

发布时间：2018-06-25 11:59

本文选题：铜绿微囊藻 + 小檗碱　；参考：《天津农学院》2015年硕士论文

【摘要】：铜绿微囊藻(Microcystic aeruginosa)是蓝藻水华的主要种类。小檗碱能够有效抑制铜绿微囊藻的生长,在养殖池塘蓝藻水华防控中具有广阔的应用前景。小檗碱能够通过影响铜绿微囊藻的光合作用来抑制微囊藻的生长。光照作为影响藻类光合作用最主要的环境因子,其条件变化势必会影响小檗碱的抑藻效应。为了探明小檗碱的抑藻作用机理,能够科学有效地在实际生产中应用小檗碱防治微囊藻水华,本研究以铜绿微囊藻生长和叶绿素荧光参数(Fv/Fm、ΦPSⅡ、ETRmax、Yield′)作为检测指标,探讨了不同光照条件对小檗碱抑藻效应的影响,并对其抑藻机理进行初探,主要研究结果如下:1小檗碱对有毒铜绿微囊藻905、无毒铜绿微囊藻469抑藻效应的比较在光照条件(70μmol·m-2·s-1)下,小檗碱能有效抑制有毒铜绿微囊藻905和无毒铜绿微囊藻469的生长,与无毒铜绿微囊藻469相比,有毒铜绿微囊藻905对小檗碱的抑杀效应表现更为敏感;未受到小檗碱胁迫的对照组,有毒905和无毒铜绿微囊藻469的荧光参数基本保持恒定,并且相较于有毒铜绿微囊藻905,无毒铜绿微囊藻469的叶绿素荧光参数数值较高;添加小檗碱后,有毒铜绿微囊藻905和无毒铜绿微囊藻469的叶绿素荧光参数均显著下降(P0.05),且有毒铜绿微囊藻905叶绿素荧光参数下降幅度更大。2光照强度对小檗碱抑藻效应的影响在光照强度为50、100、150μmol·m-2·s-1的光照条件下,铜绿微囊藻的生长均能受到小檗碱的有效抑制,4 mg/L小檗碱添加组铜绿微囊藻细胞密度随着光照强度的升高逐渐下降,但各光照处理组间藻细胞密度差异不显著(P0.05);添加4 mg/L小檗碱后,各光照强度处理组铜绿微囊藻的叶绿素荧光参数变化趋势相同,均表现为在第1 d时降至0,后随着培养时间的延长保持为0。在持续黑暗(光照强度0μmol·m-2·s-1)条件下,添加不同浓度小檗碱后,铜绿微囊藻细胞密度与叶绿素荧光参数变化均与对照组相同,小檗碱无法发挥抑藻效应。3黑暗胁迫对光恢复期小檗碱抑藻效应的影响与光照条件下相比,经历黑暗胁迫恢复光照后小檗碱的抑藻能力减弱,具体表现在:黑暗胁迫2 d后恢复光照(70μmol·m-2·s-1)期间,2 mg/L小檗碱添加组铜绿微囊藻细胞密度随培养时间延长逐渐升高,叶绿素荧光参数均在恢复光照后第1 d下降至最低值后随培养时间增加逐渐升高至稳定值;5 mg/L和8 mg/L小檗碱添加组藻细胞密度逐渐下降,并分别在恢复光照后第8 d和第6 d下降至0,叶绿素荧光参数均在恢复光照后第1 d下降至0并保持为0。而在未经黑暗胁迫的光照(70μmol·m-2·s-1)条件下,添加2 mg/L小檗碱后,铜绿微囊藻细胞密度随培养时间增加呈先下降后升高趋势,叶绿素荧光参数在第1 d下降至0后随培养时间增加逐渐升高至稳定值;添加5 mg/L和8mg/L小檗碱后,铜绿微囊藻细胞密度逐渐下降,并分别在第4 d和第3 d就下降至0并保持为0,叶绿素荧光参数在第1 d下降至0并保持为0。黑暗胁迫2 d后恢复光照(50、100、150μmol·m-2·s-1)期间,4 mg/L小檗碱添加组铜绿微囊藻细胞密度先下降后逐渐上升,并且细胞密度随着光照强度的升高逐渐下降,各光照组叶绿素荧光参数均在恢复光照后第1 d下降至最低值后随培养时间增加逐渐升高。而在未经过黑暗胁迫的光照(50、100、150μmol·m-2·s-1)条件下,添加4 mg/檗碱后,铜绿微囊藻细胞密度均随培养时间延长逐渐下降,在第5 d下降至0并保持为0,各光照组铜绿微囊藻叶绿素荧光参数均在第1 d下降至0并保持为0。黑暗胁迫不同时间(4 d、8 d和12d)恢复光照(45μmol·m-2·s-1)后,4 mg/L小檗碱添加组藻细胞密度变化趋势相同,均保持稳定一段时间后逐渐升高。其中黑暗胁迫12d相对于短时间(4 d、8 d)黑暗胁迫处理组藻细胞密度有所下降。不同黑暗胁迫时间处理组叶绿素荧光参数均表现为先下降后上升最终趋于平稳趋势。4施药前光照条件对小檗碱抑藻效应的影响将铜绿微囊藻在光照(40μmol·m-2·s-1)和黑暗条件下分别培养8 d后重新接种至相同初始密度,添加不同浓度小檗碱(2、4 mg/L)后置于光照(40μmol·m-2·s-1)条件下继续培养6 d,2 mg/L小檗碱不能有效抑杀铜绿微囊藻,藻细胞密度和叶绿素荧光参数均随培养时间延长先下降后升高,施药前的黑暗处理组藻细胞密度在6 d时显著高于施药前光照处理组的藻细胞密度(P0.05);4 mg/L小檗碱能够有效抑藻,6 d时藻细胞全部死亡,且施药前黑暗处理组与施药前光照处理组的藻细胞密度和叶绿素荧光参数具有相同的下降趋势。由以上研究结果可知,小檗碱抑藻效应存在光活化(依赖)现象,光照条件下小檗碱能够有效抑藻,但在黑暗培养条件下,小檗碱无法发挥抑藻效应,而黑暗胁迫恢复光照后小檗碱恢复抑藻能力,但抑藻效应下降,揭示小檗碱是光合作用抑制剂,通过参与铜绿微囊藻的光反应过程发挥抑藻效应。结合叶绿素荧光参数变化规律,推断小檗碱通过抑制PSII活性中心光合电子传递发挥作用。
[Abstract]:Microcystis aeruginosa (Microcystic aeruginosa) is the main species of cyanobacteria bloom. Berberine can effectively inhibit the growth of Microcystis aeruginosa, and has a broad application prospect in the prevention and control of cyanobacteria bloom in aquaculture ponds. Berberine can inhibit the growth of Microcystis by affecting the photosynthesis of Microcystis aeruginosa. Light is used as the influence of algae light. In order to explore the mechanism of berberine inhibiting algal inhibition, berberine can be used to prevent and control Microcystis. The growth of Microcystis aeruginosa and the fluorescence parameters (Fv/Fm, PS II, ETRmax, Yield ') of Microcystis aeruginosa are studied in this study. The effects of different illumination conditions on the inhibition of berberine on algae inhibition and its mechanism were discussed. The main research results were as follows: 1 berberine 905 Microcystis poisonous aeruginosa, non toxic Microcystis green Microcystis 469 inhibition effect in the light conditions (70 mu mol. M-2. S-1), berberine can effectively inhibit the toxic aeruginosa microcapsules The growth of algae 905 and innoxious Microcystis of Microcystis aeruginosa was 469, compared with the innoxious Microcystis 469, toxic Microcystis 905 were more sensitive to berberine inhibition effect. No berberine stress control group, toxic 905 and non-toxic Microcystis 469 of the fluorescence parameters were kept constant, and compared to the toxic Microcystis 905, no The chlorophyll fluorescence parameters of Microcystis aeruginosa 469 were higher. After the addition of berberine, the chlorophyll fluorescence parameters of the toxic Microcystis aeruginosa 905 and the non toxic Microcystis aeruginosa were significantly decreased (P0.05), and the 905 chlorophyll fluorescence parameters of the toxic Microcystis aeruginosa were decreased more than that of the.2 light intensity on the effect of berberine on the algal inhibition effect. The growth of Microcystis aeruginosa could be effectively inhibited by berberine under the light conditions of 50100150 mol. M-2. S-1. The density of cell density of Microcystis in 4 mg/L berberine group decreased gradually with the increase of light intensity, but the difference of cell density between each light treatment group was not significant (P0.05); after adding 4 mg/L berberine, each light was light. The variation trend of chlorophyll fluorescence parameters of Microcystis aeruginosa in intensity treatment group was the same as that at first D, and then decreased to 0 when the incubation time was extended to 0. in the continuous dark (light intensity 0 mol. M-2. S-1). The changes of cell density and chlorophyll fluorescence parameters of Microcystis aeruginosa were all changed after adding different concentrations of berberine. The control group was the same, the effect of berberine on the effect of.3 dark stress on the effect of berberine on the inhibition of berberine in the light recovery period was compared with the light conditions. After the dark stress, the inhibition ability of berberine was weakened after the dark stress was restored. The effect of berberine after dark stress was 2 d after the dark stress was restored (70 mol. M-2. S-1), and 2 mg/L berberine was added to the group of aeruginosa. The cell density of Microcystis increased gradually with the incubation time, and the chlorophyll fluorescence parameters decreased to the lowest value after first D to the lowest value, and gradually increased to the stable value with the increase of culture time. 5 mg/L and 8 mg/L berberine added the algal cell density gradually, and decreased to 0 after the recovery of light and sixth D, respectively, and chlorophyll fluorescence. The parameters were decreased to 0 and maintained at 0. after the recovery of light, and the density of cell density of Microcystis aeruginosa increased first and then increased with the increase of 2 mg/L berberine without dark stress (70 mu mol. M-2. S-1). The chlorophyll fluorescence parameters decreased to 0 from first D to 0, and increased gradually with the increase of culture time. The cell density of Microcystis aeruginosa decreased gradually after adding 5 mg/L and 8mg/L berberine, and decreased to 0 and 0 respectively at fourth D and third D, and the chlorophyll fluorescence parameters decreased to 0 from first D and kept 0. dark stress 2 d after 2 D recovery (50100150 Mu mol. M-2. S-1), and 4 mg/L berberine was added to the Microcystis aeruginosa Microcystis. The cell density descended first and gradually increased, and the cell density gradually decreased with the increase of light intensity. The chlorophyll fluorescence parameters of each light group decreased to the lowest value after first D to the lowest value, and increased gradually with the culture time. The addition of 4 mg/ berberine under the condition of 50100150 Mu mol. M-2. The cell density of Microcystis aeruginosa decreased gradually with the incubation time, at fifth d to 0 and 0. The chlorophyll fluorescence parameters of Microcystis aeruginosa were decreased to 0 at first D in each light group and maintained at 0. dark times (4 D, 8 D and 12D), and 4 mg/L berberine was added to the density of algal cell density. The change trend was the same, and all remained stable for a period of time. The dark stress 12D relative to the short time (4 D, 8 d) decreased the density of algae cells in the treatment group. The chlorophyll fluorescence parameters in the treatment group of different dark stress treatment group were first descended and then the final trend was tending to a stable trend, and the light condition of the Berberis before.4 was applied to the Berberis The effect of alkaline algal inhibition on Microcystis aeruginosa in light (40 mol. M-2. S-1) and dark conditions was reinoculated to the same initial density after 8 D, respectively. After adding different concentrations of berberine (2,4 mg/L) to light (40 mol. M-2. S-1), 6 D was continued, and 2 mg/L berberine could not effectively inhibit the killing of Microcystis aeruginosa and the density of algae cells. The parameters of the degree and chlorophyll fluorescence decreased first and then increased with the incubation time. The density of the algal cells in the dark treatment group before 6 D was significantly higher than that of the algal cell density (P0.05) in the light treatment group. The 4 mg/L berberine could effectively inhibit the algae and all the algae cells died at 6 D, and the dark treatment group and the light treatment group before the application before the application of the drug The algal cell density and chlorophyll fluorescence parameters have the same downward trend. From the above results, it is known that berberine has light activation (dependence) phenomenon, berberine can effectively inhibit algae under light conditions, but berberine can not exert algae inhibition effect under dark culture conditions, and berberine recovery after the dark stress is restored to the berberine. The inhibition of algae inhibition, but the inhibition of algae inhibition, revealed that berberine was an inhibitor of photosynthesis. By participating in the light reaction process of Microcystis aeruginosa, the effect of algae inhibition was played. The effect of berberine was deduced by inhibiting the photoelectron transfer in the PSII active center by combining the variation of chlorophyll fluorescence parameters.
【学位授予单位】：天津农学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X592;X173

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