【摘要】：干旱脅迫對植物生長發(fā)育造成嚴重的損傷。小麥作為一種主要的糧食作物,如何有效緩解其在干旱脅迫下所受損傷,是解決人們生存大計的重要課題。三唑類化合物具有植物生長調(diào)節(jié)作用,同時可有效緩解非生物脅迫等對植物體的迫害。本文通過觀察己唑醇硝酸銅配合物對植物氣孔和根的形態(tài)影響,以及干旱脅迫下配合物對小麥幼苗ABA含量及 ABA 相關(guān)基因(ABA 合成關(guān)鍵基因 9-cis-epoxycarotenoid dioxigenase(jNCED)和ABA分解重要基因ABA8'-hydroxylase(CYP707A1))表達的影響,,探其是否可以緩解脅迫對植物造成的損害,提高植物體的耐旱性。(1)小麥和煙草葉片下表皮組織,進行不同濃度配合物溶液和不同時間處理。0-100 μM配合物處理1h,隨著配合物濃度增加,煙草和小麥氣孔開度逐漸減小,100μM時兩者氣孔開度達到最小值。設(shè)定的6個濃度(0μM、20μM、40μM、60μM、80μM、100 μM)下,煙草氣孔開度平均值從2.37 μm減小到0.90 μm。而小麥氣孔開度平均值大小為:1.06 μm、0.96 μm、0.75 μm、0.63 μm、0.59 μm、0.55 μm。0-9h 不同時間配合物處理下,煙草和小麥氣孔孔徑同樣隨著處理時間的延長,逐漸減小。煙草氣孔孔徑平均值在80 μM配合物處理后,從0 h的2.37 μm減小到9 h的0.94 μm。小麥經(jīng)60 μM配合物處理,氣孔孔徑相比對照依次減小40.6%(1 h)、49.1%(3 h)、50.9%(6 h)、54.7%(9 h)。己唑醇硝酸銅配合物可降低植物氣孔孔徑。掃描電鏡觀察小麥根尖在60 μ己唑醇硝酸銅配合物處理下的變化。對照組小麥根尖根毛稀疏、短粗。處理組小麥根尖根毛濃密、纖長。兩組根尖形態(tài)學上端無明顯差別,而從距根尖形態(tài)學上端5mm處開始處理組根毛相比對照組濃密、纖長。(2)利用實時定量PCR(real-time quantitative PCR,RT-qPCR)方法探究己唑醇硝酸銅配合物在干旱脅迫下對小麥幼苗ABA含量和相關(guān)基因NCED和CYP7ED和表達量的影響。結(jié)果顯示,干旱狀態(tài)下,對照組和己唑醇硝酸銅配合物處理組NCED表達量均低于未處理0h時的表達量。但在失水過程中(24-144h)對照組表達量逐漸升高,而處理組NCED表達量先降低(24-48 h)后升高(96-144 h);復水后(144-194 h)兩組表達量均下調(diào)表達。ABA分解關(guān)鍵基因CYP707A1表達量在整個處理過程中,相對0 h時都有所增高,對照組在144 h達到最高,處理組168 h表達量最高。同樣,對照組和處理組CYP707A1表達趨勢不同,對照組表達量在干旱階段均高于處理組,復水后卻低于處理組。實驗發(fā)現(xiàn),CYP707A1表達量的變化大于NCED。脅迫下,小麥幼苗葉片對照組和處理組ABA含量相對Oh都升高,96 h達到最大值,分別為Oh的4.38倍、4.64倍。整個過程中,處理組ABA高于對照組,所以,配合物具有提高小麥幼苗干旱耐受效能(3)通過對CYP707A1基因表達量的檢測,驗證了結(jié)合PCR擴增技術(shù)及金納米星/石墨烯/沉積納米金復合材料制備的電化學生物傳感器的測定性能。結(jié)果表明,使用生物傳感器檢測的上節(jié)中不同處理小麥CYP707A1基因表達量與傳統(tǒng)的RT-qPCR結(jié)果一致。并且,該生物傳感器的檢測范圍寬至5個數(shù)量級,最低檢測可達到13 pM,顯示電化學生物傳感器具有好的靈敏性、特異性。
[Abstract]:Drought stress causes serious damage to plant growth and development. As a major food crop, how to effectively alleviate the damage caused by drought stress in wheat is an important issue to solve people's livelihood. In this paper, the effects of copper hexazole nitrate complex on the stomatal and root morphology of plants, ABA content and ABA related genes (9-cis-epoxycarotenoid dioxigenase (jNCED) and ABA decomposition important genes (CYP707A1) expression in Wheat Seedlings under drought stress were studied. (1) The stomatal openings of tobacco and wheat decreased gradually with the increase of the concentration of the complexes, and reached to 100 mu M when the lower epidermal tissues of wheat and tobacco leaves were treated with different concentrations of complex solution and different time. The average stomatal aperture of tobacco and wheat decreased from 2.37 micron to 0.90 micron at 6 concentrations (0, 20, 40, 60, 80, 100 micron). The average stomatal aperture of wheat was 1.06 micron, 0.96 micron, 0.75 micron, 0.63 micron, 0.59 micron, 0.55 micron, 0.0-9 h, respectively. The average stomatal diameter of tobacco decreased from 2.37 micron in 0 h to 0.94 micron in 9 h after treatment with 80 micron complexes. The stomatal diameter of wheat treated with 60 micron complexes decreased by 40.6% (1 h), 49.1% (3 h), 50.9% (6 h), 54.7% (9 h) compared with the control in turn. The root hairs of the control group were thinner, shorter and thicker than those of the control group. The root hairs of the treatment group were dense and slender. There was no significant difference between the two groups in morphology, but the root hairs of the treatment group were denser and slender than those of the control group from 5 mm away from the top. Real-time quantitative PCR (RT-qPCR) was used to investigate the effects of copper hexazole nitrate complex on ABA content, NCED and CYP7ED expression in Wheat Seedlings under drought stress. However, during dehydration (24-144h), the expression of NCED in the control group increased gradually, and the expression of NCED in the treatment group decreased first (24-48h) and then increased (96-144h); after rehydration (144-194h), the expression of CYP707A1 in both groups was down-regulated. Similarly, the expression of CYP707A1 in control group and treatment group was higher than that in treatment group at drought stage, but lower than that in treatment group after rehydration. The change of expression of CYP707A1 was greater than that in NCED. Under stress, the ABA content in control group and treatment group of wheat seedling leaves increased relative to Oh. In the whole process, the ABA of treatment group was higher than that of control group, so the complex could improve drought tolerance of wheat seedlings (3) The electrochemical synthesis of Au nano-star/graphene/deposited Au nanocomposites was verified by detecting the expression of CYP707A1 gene. The results showed that the expression of CYP707A1 gene in wheat treated with different treatments in the previous section detected by biosensor was consistent with the results of traditional RT-q PCR. Specificity.
【學位授予單位】：西北大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q943.2

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