本文選題：銅綠微囊藻 + 小檗堿��； 參考：《天津農(nóng)學(xué)院》2015年碩士論文

【摘要】：銅綠微囊藻(Microcystic aeruginosa)是藍(lán)藻水華的主要種類。小檗堿能夠有效抑制銅綠微囊藻的生長(zhǎng),在養(yǎng)殖池塘藍(lán)藻水華防控中具有廣闊的應(yīng)用前景。小檗堿能夠通過影響銅綠微囊藻的光合作用來(lái)抑制微囊藻的生長(zhǎng)。光照作為影響藻類光合作用最主要的環(huán)境因子,其條件變化勢(shì)必會(huì)影響小檗堿的抑藻效應(yīng)。為了探明小檗堿的抑藻作用機(jī)理,能夠科學(xué)有效地在實(shí)際生產(chǎn)中應(yīng)用小檗堿防治微囊藻水華,本研究以銅綠微囊藻生長(zhǎng)和葉綠素?zé)晒鈪?shù)(Fv/Fm、ΦPSⅡ、ETRmax、Yield′)作為檢測(cè)指標(biāo),探討了不同光照條件對(duì)小檗堿抑藻效應(yīng)的影響,并對(duì)其抑藻機(jī)理進(jìn)行初探,主要研究結(jié)果如下:1小檗堿對(duì)有毒銅綠微囊藻905、無(wú)毒銅綠微囊藻469抑藻效應(yīng)的比較在光照條件(70μmol·m-2·s-1)下,小檗堿能有效抑制有毒銅綠微囊藻905和無(wú)毒銅綠微囊藻469的生長(zhǎng),與無(wú)毒銅綠微囊藻469相比,有毒銅綠微囊藻905對(duì)小檗堿的抑殺效應(yīng)表現(xiàn)更為敏感;未受到小檗堿脅迫的對(duì)照組,有毒905和無(wú)毒銅綠微囊藻469的熒光參數(shù)基本保持恒定,并且相較于有毒銅綠微囊藻905,無(wú)毒銅綠微囊藻469的葉綠素?zé)晒鈪?shù)數(shù)值較高;添加小檗堿后,有毒銅綠微囊藻905和無(wú)毒銅綠微囊藻469的葉綠素?zé)晒鈪?shù)均顯著下降(P0.05),且有毒銅綠微囊藻905葉綠素?zé)晒鈪?shù)下降幅度更大。2光照強(qiáng)度對(duì)小檗堿抑藻效應(yīng)的影響在光照強(qiáng)度為50、100、150μmol·m-2·s-1的光照條件下,銅綠微囊藻的生長(zhǎng)均能受到小檗堿的有效抑制,4 mg/L小檗堿添加組銅綠微囊藻細(xì)胞密度隨著光照強(qiáng)度的升高逐漸下降,但各光照處理組間藻細(xì)胞密度差異不顯著(P0.05);添加4 mg/L小檗堿后,各光照強(qiáng)度處理組銅綠微囊藻的葉綠素?zé)晒鈪?shù)變化趨勢(shì)相同,均表現(xiàn)為在第1 d時(shí)降至0,后隨著培養(yǎng)時(shí)間的延長(zhǎng)保持為0。在持續(xù)黑暗(光照強(qiáng)度0μmol·m-2·s-1)條件下,添加不同濃度小檗堿后,銅綠微囊藻細(xì)胞密度與葉綠素?zé)晒鈪?shù)變化均與對(duì)照組相同,小檗堿無(wú)法發(fā)揮抑藻效應(yīng)。3黑暗脅迫對(duì)光恢復(fù)期小檗堿抑藻效應(yīng)的影響與光照條件下相比,經(jīng)歷黑暗脅迫恢復(fù)光照后小檗堿的抑藻能力減弱,具體表現(xiàn)在:黑暗脅迫2 d后恢復(fù)光照(70μmol·m-2·s-1)期間,2 mg/L小檗堿添加組銅綠微囊藻細(xì)胞密度隨培養(yǎng)時(shí)間延長(zhǎng)逐漸升高,葉綠素?zé)晒鈪?shù)均在恢復(fù)光照后第1 d下降至最低值后隨培養(yǎng)時(shí)間增加逐漸升高至穩(wěn)定值;5 mg/L和8 mg/L小檗堿添加組藻細(xì)胞密度逐漸下降,并分別在恢復(fù)光照后第8 d和第6 d下降至0,葉綠素?zé)晒鈪?shù)均在恢復(fù)光照后第1 d下降至0并保持為0。而在未經(jīng)黑暗脅迫的光照(70μmol·m-2·s-1)條件下,添加2 mg/L小檗堿后,銅綠微囊藻細(xì)胞密度隨培養(yǎng)時(shí)間增加呈先下降后升高趨勢(shì),葉綠素?zé)晒鈪?shù)在第1 d下降至0后隨培養(yǎng)時(shí)間增加逐漸升高至穩(wěn)定值;添加5 mg/L和8mg/L小檗堿后,銅綠微囊藻細(xì)胞密度逐漸下降,并分別在第4 d和第3 d就下降至0并保持為0,葉綠素?zé)晒鈪?shù)在第1 d下降至0并保持為0。黑暗脅迫2 d后恢復(fù)光照(50、100、150μmol·m-2·s-1)期間,4 mg/L小檗堿添加組銅綠微囊藻細(xì)胞密度先下降后逐漸上升,并且細(xì)胞密度隨著光照強(qiáng)度的升高逐漸下降,各光照組葉綠素?zé)晒鈪?shù)均在恢復(fù)光照后第1 d下降至最低值后隨培養(yǎng)時(shí)間增加逐漸升高。而在未經(jīng)過黑暗脅迫的光照(50、100、150μmol·m-2·s-1)條件下,添加4 mg/檗堿后,銅綠微囊藻細(xì)胞密度均隨培養(yǎng)時(shí)間延長(zhǎng)逐漸下降,在第5 d下降至0并保持為0,各光照組銅綠微囊藻葉綠素?zé)晒鈪?shù)均在第1 d下降至0并保持為0。黑暗脅迫不同時(shí)間(4 d、8 d和12d)恢復(fù)光照(45μmol·m-2·s-1)后,4 mg/L小檗堿添加組藻細(xì)胞密度變化趨勢(shì)相同,均保持穩(wěn)定一段時(shí)間后逐漸升高。其中黑暗脅迫12d相對(duì)于短時(shí)間(4 d、8 d)黑暗脅迫處理組藻細(xì)胞密度有所下降。不同黑暗脅迫時(shí)間處理組葉綠素?zé)晒鈪?shù)均表現(xiàn)為先下降后上升最終趨于平穩(wěn)趨勢(shì)。4施藥前光照條件對(duì)小檗堿抑藻效應(yīng)的影響將銅綠微囊藻在光照(40μmol·m-2·s-1)和黑暗條件下分別培養(yǎng)8 d后重新接種至相同初始密度,添加不同濃度小檗堿(2、4 mg/L)后置于光照(40μmol·m-2·s-1)條件下繼續(xù)培養(yǎng)6 d,2 mg/L小檗堿不能有效抑殺銅綠微囊藻,藻細(xì)胞密度和葉綠素?zé)晒鈪?shù)均隨培養(yǎng)時(shí)間延長(zhǎng)先下降后升高,施藥前的黑暗處理組藻細(xì)胞密度在6 d時(shí)顯著高于施藥前光照處理組的藻細(xì)胞密度(P0.05);4 mg/L小檗堿能夠有效抑藻,6 d時(shí)藻細(xì)胞全部死亡,且施藥前黑暗處理組與施藥前光照處理組的藻細(xì)胞密度和葉綠素?zé)晒鈪?shù)具有相同的下降趨勢(shì)。由以上研究結(jié)果可知,小檗堿抑藻效應(yīng)存在光活化(依賴)現(xiàn)象,光照條件下小檗堿能夠有效抑藻,但在黑暗培養(yǎng)條件下,小檗堿無(wú)法發(fā)揮抑藻效應(yīng),而黑暗脅迫恢復(fù)光照后小檗堿恢復(fù)抑藻能力,但抑藻效應(yīng)下降,揭示小檗堿是光合作用抑制劑,通過參與銅綠微囊藻的光反應(yīng)過程發(fā)揮抑藻效應(yīng)。結(jié)合葉綠素?zé)晒鈪?shù)變化規(guī)律,推斷小檗堿通過抑制PSII活性中心光合電子傳遞發(fā)揮作用。
[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.
【學(xué)位授予單位】：天津農(nóng)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：X592;X173

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