本文選題：氯磺隆 + Cd��； 參考：《山西大學》2016年博士論文

【摘要】：磺酰脲類除草劑是目前田間使用量較大的一類除草劑,常被用于田間闊葉雜草的防除。氯磺隆作為磺酰脲類除草劑的代表品種,在土壤中殘效期較長,容易對后茬敏感作物產生藥害。土壤長期污水和工業(yè)廢水混合灌溉、降塵污染、以及農藥和化肥的長期使用等,都會導致土壤鎘的積累。在農田土壤體系中殘留除草劑氯磺隆和重金屬同時存在,對作物的影響及其作用機制研究引起研究者廣泛關注,因此研究兩種污染物同時存在,對玉米幼苗和菠菜生長過程、抗氧化作用機制、光合作用機制、以及植株代謝產物和代謝途徑的影響和變化等,對食品安全評價、污染物治理以及環(huán)境污染與健康的關系至關重要。根據ISO11269-2(2013)的方法,將土壤分別用不同含量的氯磺隆單一處理、固定鎘含量(5.0mg/kg)單一處理、以及固定鎘含量(5.0mg/kg)改變氯磺隆含量的復合處理后,采用盆栽試驗的方法,對玉米幼苗和菠菜分別進行發(fā)芽和早期生長試驗。通過觀察生長過程、對植株相關抗氧化物酶的活性測定,評價氯磺隆和鎘對兩種作物生長及抗氧化作用機制的影響；采用葉綠素熒光技術評價氯磺隆和鎘對兩種作物光合作用的影響；從代謝組學的角度出發(fā),采用核磁技術、氣質聯(lián)用和液質聯(lián)用技術分析玉米幼苗和菠菜受到氯磺隆和鎘單一及復合脅迫后,代謝物及代謝途徑的變化。多種代謝組學技術、葉綠素熒光、以及對植株生理生化指標的研究結果顯示,氯磺隆、鎘及其復合污染對玉米幼苗和菠菜多種代謝途徑產生了影響。1、氯磺隆單一污染結果表明,土壤殘留氯磺隆,會誘導玉米幼苗和菠菜活性氧產生、使植株自身抗氧化系統(tǒng)和膜質過氧化自我修復機制被激活；光合作用系統(tǒng)能量的利用和傳遞受限,阻礙植株三羧酸循環(huán)的正常進行、導致部分氨基酸代謝異常、能量代謝和細胞壁的合成受到影響。首先,土壤殘留氯磺隆,誘導玉米幼苗和菠菜活性氧的產生,使其自我保護機制開啟,以阻礙氧化應激反應對植株的傷害,包括抗氧化系統(tǒng)的激活(SOD、CAT和GST酶活性顯著升高,P0.05)和膜質過氧化的修復(一定濃度的氯磺隆誘導丙二醛含量顯著升高,P0.05)。其次,氯磺隆會影響菠菜葉片的熒光產量及PSⅡ中QA、QB和質體醌之間的電子傳遞(OJIP曲線熒光強度下降,P0.05),而對玉米幼苗則沒有影響。第三,氯磺隆影響玉米幼苗和菠菜的光合反應中心,使得能量的利用和傳遞受到阻礙(ABS/RC顯著升高,P0.05),植株的防御系統(tǒng)也被激發(fā),將過多的能量以熱耗散的形式釋放(DI0/RC也顯著升高,P0.05),但對還原QA和電子傳遞沒有影響(TRo/RC和ET0/RC沒有顯著變化,P0.05)。第四,隨著氯磺隆含量的不斷增大,指示反應PSⅡ生理活性狀態(tài)的PIABS呈先上升后下降的趨勢,表明低含量氯磺隆誘導植株防御系統(tǒng)啟動,而高含量則導致作物光合作用系統(tǒng)受到不可逆轉的損害。第五,植株生理生化指標的改變,在代謝上表現(xiàn)為：阻礙了三羧酸循環(huán)的正常進行(檸檬酸和蘋果酸含量顯著升高,P0.05),造成了能量代謝從有氧呼吸到無氧呼吸的轉移(丙酮酸和乳酸含量的顯著升高,P0.05),植物體內糖類物質(蔗糖、葡萄糖、果糖等)的大量積累也反應植物能量代謝及細胞壁(半乳糖醛酸、鼠李糖)合成受到影響。第六,土壤中殘留氯磺隆還導致植株部分氨基酸代謝異常(支鏈氨基酸(Branch chain amino acids, BCAAs)、芳香族氨基酸(Aromatic amino acid, AAAs)、絲氨酸(Ser)、谷氨酸(Glu)等,與抗逆相關物質(谷氨酰胺、脯氨酸、腐胺、肌醇和GABA)的顯著升高。2、鎘單一污染結果指出,土壤鎘含量為5.0 mg/kg時,對玉米幼苗和菠菜的生長、光合作用過程沒有顯著影響；但造成了玉米幼苗和菠菜代謝途徑三羧酸循環(huán)的異常、其能量代謝、氨基酸和糖代謝在玉米幼苗和菠菜中的表現(xiàn)各異。首先,與空白對照組比,土壤鎘含量(5.0 mg/kg)沒有造成植株體內活性氧的積累,植株抗氧化的自我保護系統(tǒng)沒有被激活(SOD、POD、CAT和GST酶活性沒有顯著變化),丙二醛含量也沒有顯著積累；其次,鎘對玉米幼苗和菠菜的光合反應中心、能量反應及耗散的相關參數變化也沒有顯著影響。但是,對代謝產物和代謝途徑卻存在明顯的影響。第一,鎘造成了玉米幼苗和菠菜三羧酸循環(huán)的異常(玉米幼苗蘋果酸和檸檬酸含量顯著降低；菠菜蘋果酸和檸檬酸含量顯著升高,P0.05)；第二,能量代謝在玉米幼苗和菠菜中的表現(xiàn)各異,鎘誘導菠菜中能量代謝從有氧到無氧呼吸的轉移(乳酸含量顯著升高,P0.05),而玉米幼苗能量代謝卻沒有發(fā)生轉移(乳酸含量顯著(P0.05)下降,丙酮酸含量沒有顯著變化,P0.05)；與抗逆相關物質的含量變化也不同,鎘誘導玉米幼苗中腐胺和肌醇含量積累,脯氨酸沒有顯著變化,而菠菜則是脯氨酸和肌醇含量積累,腐胺含量下降。第三,鎘還誘導玉米幼苗和菠菜幾種氨基酸代謝異常(天冬氨酸、谷氨酸、絲氨酸、丙氨酸等)和糖類物質的積累。3、與氯磺隆和鎘的單一污染相比,復合污染結果顯示,植株葉片葉綠素含量顯著升高；高含量氯磺隆復合處理,菠菜葉片的熒光產量及PSⅡ中QA、QB和質體醌之間的電子傳遞受抑、對菠菜光合作用的影響二者存在協(xié)同作用,而對玉米幼苗沒有協(xié)同作用。對某些特定代謝物的聯(lián)合作用機制因植株種類而不同。首先,與單一污染相比,復合污染均導致玉米幼苗和菠菜植株葉片葉綠素含量顯著升高(P0.05),有利于光合作用的正常進行。同時,還能刺激植物體內自我保護機制,即抗氧化物酶活性更高(POD和CAT活性顯著升高,GST酶活性顯著降低,P0.05),有利于植物體內活性氧的清除。其次,不同含量的氯磺隆和鎘復合污染對玉米幼苗和菠菜熒光產量及PSⅡ中QA、QB和質體醌之間電子傳遞影響表現(xiàn)不同。具體表現(xiàn)為,低含量氯磺隆(0.001mg/kg)和Cd復合對玉米幼苗和菠菜植株光合作用均沒有影響(與各自對照相比,ABS/RC、DI0/RC和PIABS沒有顯著變化,P0.05)；而高含量氯磺隆和鎘復合污染,對玉米幼苗沒有協(xié)同作用(與單一污染相比較,ABS/RC、DI0/RC和PIABS沒有顯著變化,P0.05)；但菠菜葉片的熒光產量及PSⅡ中QA、QB和質體醌之間的電子傳遞則受到抑制,且隨著氯磺隆濃度越高,抑制作用越強；特別是對ABS/RC和DI0/RC值的影響較氯磺隆單一污染更為明顯(P0.05),表明鎘加重了氯磺隆對菠菜光合系統(tǒng)反應中心能量的影響,二者存在協(xié)同作用。同時,復合處理組PIABS顯著下降也表明二者對菠菜光合作用具有協(xié)同作用。第三,從代謝物含量的變化可以看出,氯磺隆和鎘復合污染對玉米幼苗某些特定代謝物如異亮氨酸、纈氨酸、絲氨酸、乳酸等含量具有拮抗作用,其余代謝物如丙酮酸、蘋果酸、檸檬酸和蛋氨酸在低含量復合處理組表現(xiàn)為拮抗作用,而高含量則為協(xié)同作用；氯磺隆和鎘復合污染對菠菜中異亮氨酸、谷氨酸、乳酸和檸檬酸等具有協(xié)同作用,其余代謝物纈氨酸、蘋果酸、γ-氨基丁酸在低含量復合處理組表現(xiàn)為協(xié)同作用,高含量則為拮抗作用。綜上所述,統(tǒng)計檢驗結合定量分析結果表明,土壤殘留氯磺隆會誘導玉米幼苗和菠菜活性氧產生,植物自身防御系統(tǒng)開啟(抗氧化系統(tǒng)的激活和膜質過氧的修復)；光合作用系統(tǒng)能量的利用和傳遞受限,植物代謝途徑三羧酸循環(huán)、部分氨基酸代謝、能量代謝及細胞壁合成受阻。土壤鎘(5mg/kg)雖然對玉米幼苗和菠菜的生長和光合作用過程沒有明顯影響,但對玉米幼苗和菠菜代謝物及代謝途徑產生了明顯影響。如玉米幼苗和菠菜三羧酸循環(huán)、氨基酸、糖和能量代謝發(fā)生異常。該研究結果揭示,氯磺隆與鎘復合污染存在交互作用,氯磺隆和鎘脅迫下,玉米幼苗和菠菜代謝中最大的不同表現(xiàn)在莽草酸途徑,推斷該途徑中多種主要代謝物變化可作為單子葉作物受脅迫的標記。
[Abstract]:Sulfonylurea herbicides are one of the most widely used herbicides used in the field at present. They are often used for the prevention of broad leaved weeds in the field. Chlorsulfononide is a representative variety of sulfonylurea herbicides. It has a long residual effect in the soil and is easy to produce a drug damage to the later crop sensitive crops. The mixed irrigation of soil and industrial waste water, dust pollution, and agriculture are used in the field. The long-term use of drugs and chemical fertilizers will lead to the accumulation of cadmium in soil. The residual herbicide chlorsulfonide and heavy metals in the soil system in the farmland are simultaneous, the influence of the crop and the mechanism of its action have aroused widespread concern. Therefore, the study of two kinds of pollutants at the same time, the growth process of corn seedlings and spinach, antioxidant effect The mechanism, the photosynthesis mechanism, the effects and changes of the plant metabolites and metabolic pathways, and the relationship between the food safety assessment, the pollution control and the environmental pollution and health are essential. According to the method of ISO11269-2 (2013), the soil is treated with a single treatment with different content of chlorsulfonon, and the fixed cadmium content (5.0mg/kg) is treated by a single treatment, After a combined treatment of the content of chlorsulfonyl chloride with fixed cadmium content (5.0mg/kg), a pot experiment was used to test the germination and early growth of the maize seedlings and spinach. By observing the growth process, the activity of the related antioxidant enzymes was measured, and the growth of two crops and the antioxidants were evaluated by chlorononolonon and cadmium. The effects of the chlorophyll fluorescence technique were used to evaluate the effects of chlorsulfonyl and cadmium on the photosynthesis of two crops. From the perspective of metabolomics, the changes in metabolites and metabolic pathways of maize seedlings and spinach were analyzed by NMR technique, GC-MS and liquid chromatography-mass spectrometry. The results of metabonomics, chlorophyll fluorescence, and physiological and biochemical indexes of plant showed that chlorsulfonon, cadmium and its compound pollution had an effect on various metabolic pathways of maize seedlings and spinach. The single pollution result of chlorsulfonon showed that the residual chlorsulfonont in soil could induce the production of active oxygen of maize seedlings and spinach, and the plant itself could be induced by.1. The antioxidant system and the self repair mechanism of membranous peroxidation are activated; the utilization and transfer of energy in the photosynthesis system is limited, which hinders the normal progress of the three carboxylic acid cycle, resulting in abnormal metabolism of some amino acids, energy metabolism and the synthesis of cell walls. The production of oxygen, which opens the mechanism of self protection, prevents oxidative stress response to plants, including the activation of antioxidant systems (SOD, CAT and GST enzyme activity significantly increased, P0.05) and membranous peroxide repair (a certain concentration of chlorsulfonon induced malondialdehyde content increases significantly, P0.05). Secondly, chlorsulfonon will affect the fluorescence of spinach leaves Light yield and electron transfer between QA, QB and plastid quinone in PS II (OJIP curve fluorescence intensity decreased, P0.05), but no effect on maize seedlings. Third, chlorononolonon affects the photosynthetic reaction center of maize seedlings and spinach, which prevents the use and transmission of energy (ABS/RC significantly increased, P0.05), and the plant defense system is also stimulated. The excess energy was released in the form of heat dissipation (DI0/RC also increased significantly, P0.05), but had no effect on the reduction of QA and electron transfer (TRo/RC and ET0/RC did not change significantly, P0.05). Fourth, with the increase of the content of chlorsulfonyl methyl, the PIABS showing the physiological active state of PS II was rising and then decreasing, indicating the low content of chlorinated sulphur. The growth of plant defense system was induced, while high content resulted in irreversible damage to the photosynthesis system of crops. Fifth, the changes in physiological and biochemical indicators of plants were metabolized as hindering the normal cycle of the three carboxylic acid (citric acid and malic acid content increased significantly, P0.05), resulting in energy metabolism from aerobic respiration to A significant increase in the content of pyruvic acid and lactic acid, P0.05), the accumulation of carbohydrates (sucrose, glucose, fructose, etc.) in the plant also responds to the energy metabolism of plants and the synthesis of cell wall (galactoacid, Li Tang). Sixth, residual chlorosulonide in soil is also responsible for the abnormal metabolism of some amino acids. Chain amino acids (Branch chain amino acids, BCAAs), aromatic amino acids (Aromatic amino acid, AAAs), serine (Ser), glutamic acid (Glu), etc., with a significant increase in resistance related substances (glutamine, proline, proline, inositol and GABA). The cadmium single pollution results indicated that the soil cadmium content was 5, the growth of maize seedlings and spinach. Long, photosynthesis process was not significantly affected, but the metabolism of three carboxylic acid cycles in maize seedlings and spinach was abnormal, and its energy metabolism, amino acid and sugar metabolism in maize seedlings and spinach were different. First, the content of soil cadmium (5 mg/kg) did not cause the accumulation of active oxygen in the plant compared with the blank control group. The antioxidant self-protection system was not activated (SOD, POD, CAT and GST activity did not change significantly), and MDA content was not significantly accumulated; secondly, there was no significant influence on the photosynthetic reaction center of maize seedlings and spinach, energy response and dissipation related parameters. However, the metabolic products and metabolic pathways were present. First, cadmium resulted in abnormal cycling of three carboxylic acids in maize seedlings and spinach (the content of malic acid and citric acid in maize seedlings decreased significantly; the content of malic acid and citric acid in spinach increased significantly, P0.05); second, energy metabolism in maize seedlings and spinach were different. Cadmium induced energy metabolism in spinach from aerobic to no The transfer of oxygen respiration (the content of lactic acid increased significantly, P0.05), but the energy metabolism of maize seedlings did not change (P0.05), the content of pyruvic acid was not significantly changed, P0.05), and the content of anti inverse related substances was also different. The content of putrescine and inositol in the seedlings of cadmium induced jade rice seedlings was not significantly changed. The contents of proline and inositol were accumulated and the content of putrescine decreased. Third, cadmium also induced the accumulation of several amino acids in maize seedlings and spinach (aspartic acid, glutamic acid, serine, alanine, etc.) and the accumulation of carbohydrates. Compared with the single pollution of chlorosinolide and cadmium, the compound pollution results showed that the chlorophyll content of plant leaves was contained in.3. The amount of the high content chlorsulfonon combined treatment, the fluorescence yield of spinach leaves and the electron transfer between QA, QB and plastid quinone in PS II were inhibited. The synergistic effects on the photosynthesis of spinach were synergistic, but there was no synergistic effect on the maize seedlings. The joint action mechanism of some specific metabolites was different from the plant species. First, compared with the single pollution, the compound pollution resulted in a significant increase in the chlorophyll content of the leaves of the maize seedlings and spinach plants (P0.05), which was beneficial to the normal photosynthesis. At the same time, it could also stimulate the mechanism of self protection in the plant, that is, the activity of antioxidant enzymes is higher (the activity of POD and CAT increased significantly, the activity of GST decreased significantly, P0.05). The effects of chlorsulfonylon and cadmium on the fluorescence yield of maize seedlings and spinach and the electron transfer between QA, QB and plastid quinones in PS II were different. The specific performance was that low content of chlorononolonon (0.001mg/kg) and Cd had no effect on the photosynthesis of maize seedlings and spinach plants. Compared with the control, ABS/RC, DI0/RC and PIABS did not significantly change, P0.05); while the high content of chlorononolonon and cadmium pollution had no synergistic effect on maize seedlings (compared with single pollution, ABS/RC, DI0/RC and PIABS had no significant changes, P0.05), but the fluorescence yield of spinach leaves and the electron transfer between QA, QB and plastid quinones in PS II. It was inhibited, and with the higher concentration of chlorsulfonon, the stronger the inhibitory effect; especially the effect on ABS/RC and DI0/RC value was more obvious than that of chlorsulfonon single pollution (P0.05), indicating that cadmium aggravated the effect of chlorsulfonon on the central energy of the photosynthetic system reaction of spinach, the two had synergistic effect. At the same time, the PIABS of the compound treatment group decreased significantly. The two groups have a synergistic effect on the photosynthesis of spinach. Third, from the change of metabolite content, it can be seen that the compound pollution of chlorsulfonyl and cadmium has antagonistic effects on some specific metabolites such as isoleucine, valine, serine and lactic acid in maize seedlings, while the remaining metabolites such as pyruvic acid, malic acid, citric acid and methionine are low The compound treatment group showed antagonism, while the high content was synergistic, and the compound pollution of chlorsulfonon and cadmium had synergistic effect on isoleucine, glutamic acid, lactic acid and citric acid in spinach. The other metabolites, valine, malic acid and gamma aminobutyric acid, were synergistic in the low content compound treatment group and the high content was antagonistic. To sum up, the statistical test combined with quantitative analysis showed that the residual chlorsulfonon in soil could induce the production of active oxygen of maize seedlings and spinach, the plant self defense system opened (the activation of antioxidant system and the repair of membrane oxygen); the utilization and transmission of the photosynthesis system energy was limited, the plant metabolism pathway three carboxylic acid cycle, part The metabolism of amino acids, energy metabolism and cell wall synthesis were blocked. Although soil cadmium (5mg/kg) had no obvious influence on the growth and photosynthesis process of maize seedlings and spinach, it had a significant influence on the metabolites and metabolic pathways of maize seedlings and spinach, such as the cycle of three carboxylic acids, amino acids, sugar and energy metabolism in maize seedlings and spinach. The results revealed that the compound pollution of chlorsulfonon and cadmium was interactive. Under the stress of chlorsulfonyl and cadmium, the biggest difference in the metabolism of maize seedlings and spinach was shown in the way of shikimic acid. It was concluded that many major metabolites in this pathway could be used as a marker for the stress of mono cotyledon crops.
【學位授予單位】：山西大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：X592

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