己唑醇硝酸铜配合物对植物形态及ABA相关基因表达的影响

发布时间：2018-08-12 15:01

【摘要】：干旱胁迫对植物生长发育造成严重的损伤。小麦作为一种主要的粮食作物,如何有效缓解其在干旱胁迫下所受损伤,是解决人们生存大计的重要课题。三唑类化合物具有植物生长调节作用,同时可有效缓解非生物胁迫等对植物体的迫害。本文通过观察己唑醇硝酸铜配合物对植物气孔和根的形态影响,以及干旱胁迫下配合物对小麦幼苗ABA含量及 ABA 相关基因(ABA 合成关键基因 9-cis-epoxycarotenoid dioxigenase(jNCED)和ABA分解重要基因ABA8'-hydroxylase(CYP707A1))表达的影响,,探其是否可以缓解胁迫对植物造成的损害,提高植物体的耐旱性。(1)小麦和烟草叶片下表皮组织,进行不同浓度配合物溶液和不同时间处理。0-100 μM配合物处理1h,随着配合物浓度增加,烟草和小麦气孔开度逐渐减小,100μM时两者气孔开度达到最小值。设定的6个浓度(0μM、20μM、40μM、60μM、80μM、100 μM)下,烟草气孔开度平均值从2.37 μm减小到0.90 μm。而小麦气孔开度平均值大小为:1.06 μm、0.96 μm、0.75 μm、0.63 μm、0.59 μm、0.55 μm。0-9h 不同时间配合物处理下,烟草和小麦气孔孔径同样随着处理时间的延长,逐渐减小。烟草气孔孔径平均值在80 μM配合物处理后,从0 h的2.37 μm减小到9 h的0.94 μm。小麦经60 μM配合物处理,气孔孔径相比对照依次减小40.6%(1 h)、49.1%(3 h)、50.9%(6 h)、54.7%(9 h)。己唑醇硝酸铜配合物可降低植物气孔孔径。扫描电镜观察小麦根尖在60 μ己唑醇硝酸铜配合物处理下的变化。对照组小麦根尖根毛稀疏、短粗。处理组小麦根尖根毛浓密、纤长。两组根尖形态学上端无明显差别,而从距根尖形态学上端5mm处开始处理组根毛相比对照组浓密、纤长。(2)利用实时定量PCR(real-time quantitative PCR,RT-qPCR)方法探究己唑醇硝酸铜配合物在干旱胁迫下对小麦幼苗ABA含量和相关基因NCED和CYP7ED和表达量的影响。结果显示,干旱状态下,对照组和己唑醇硝酸铜配合物处理组NCED表达量均低于未处理0h时的表达量。但在失水过程中(24-144h)对照组表达量逐渐升高,而处理组NCED表达量先降低(24-48 h)后升高(96-144 h);复水后(144-194 h)两组表达量均下调表达。ABA分解关键基因CYP707A1表达量在整个处理过程中,相对0 h时都有所增高,对照组在144 h达到最高,处理组168 h表达量最高。同样,对照组和处理组CYP707A1表达趋势不同,对照组表达量在干旱阶段均高于处理组,复水后却低于处理组。实验发现,CYP707A1表达量的变化大于NCED。胁迫下,小麦幼苗叶片对照组和处理组ABA含量相对Oh都升高,96 h达到最大值,分别为Oh的4.38倍、4.64倍。整个过程中,处理组ABA高于对照组,所以,配合物具有提高小麦幼苗干旱耐受效能(3)通过对CYP707A1基因表达量的检测,验证了结合PCR扩增技术及金纳米星/石墨烯/沉积纳米金复合材料制备的电化学生物传感器的测定性能。结果表明,使用生物传感器检测的上节中不同处理小麦CYP707A1基因表达量与传统的RT-qPCR结果一致。并且,该生物传感器的检测范围宽至5个数量级,最低检测可达到13 pM,显示电化学生物传感器具有好的灵敏性、特异性。
[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.
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943.2

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