木霉T-soybean ACCD酶功能分析
时间:2019-06-11 20:27 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：本实验以木霉T-soybean为研究对象，对木霉T-soybean 1-氨基环丙烷-1-羧酸脱氨酶 (ACCD) 功能进行了分析。2.0mM/L 1-氨基环丙烷-1-羧酸（ACC）能明显诱导ACCD酶活性。硫酸铵沉淀法对ACCD进行分离纯化，SDS-PAGE凝胶电泳结果表明，目的蛋白大小为37KDa，与预期结果一致。ACCD酶活性测定结果显示，ACC浓度为2mM/L时诱导木霉ACCD活性高达2.52U/mL；由ACC诱导的木霉ACCD粗酶液处理玉米后，玉米的发芽率 (95%)、鲜重 (0.902g)、株高 (2.74cm)、根长 (4.17cm)、根数 (7条) 较对照都有显著提高。对照玉米的发芽率为75%、平均鲜重为0.754g、平均株高为1.275cm、平均根长为2.71cm、平均根数为2.5条。结果表明ACCD酶对植物的发芽、生长具有明显的促进作用。36225 毕业论文关键词：木霉；1-氨基环丙烷-1-羧酸；功能分析 Functional Analysis of Trichoderma T-soybean ACCD Enzyme Abstract: The experiment with Trichoderma T-soybean as the research object, of Trichoderma T-soybean 1-aminoanthraquinone ring propane - 1-tightening carboxylic acid deaminase (ACCD) function were analyzed. 1- -1- 2.0mM/L carboxylic acid (ACC) can induce the activity of ACCD. The ammonium sulfate precipitation method was isolated and purified, SDS-PAGE gel electrophoresis showed that the size of the target protein was 37KDa, and the results were in agreement with the expected results of the ACCD gel electrophoresis.. ACCD enzymes activity was determined. The results showed, ACC concentration is 2mM/L induced by Trichoderma ACCD activity up to 2.52U/mL; by the ACC induced by Trichoderma ACCD crude enzyme liquid of corn and Corn Germination rate (95%), fresh weight 0.902g, height 2.74cm) and the root length (4.17), root number (7 A) have significantly improved than that of the control. The germination rate of maize was 75%, the average fresh weight was 0.754g, the average height was 1.275cm, the average root length was 2.71cm, the average number was 2.5,The results showed that ACCD had a significant effect on the germination and growth of plants. Key words: ACC; Trichoderma T-soybean; Functional analysis 目 录 摘 要 1 引言 2 1. 材料及方法 2 1.1 实验材料及仪器 2 1.1.1 实验材料及药品 2 1.1.2 实验仪器 2 1.2 实验方法 3 1.2.1药品的配制 3 1.2.2 木霉的活化 3 1.2.3 木霉ACCD的诱导 3 1.2.4 ACCD酶的分离纯化 3 1.2.5 ACCD酶活性测定 4 1.2.6 SDS-PAGE凝胶电泳 4 1.2.7 ACCD酶对玉米发芽和生长的影响 4 2. 结果与分析 4 2.1 ACCD酶活性测定 5 2.2 SDS-PAGE凝胶电泳结果 5 2.3 ACCD酶功能验证 6 2.3.1 分别用水和粗酶液处理对玉米种子发芽影响 6 2.3.2 分别用粗酶液和SM培养基处理对玉米发芽和生长影响 6 3. 结论与讨论 9 参考文献 10 致谢 12 木霉T-soybean ACCD酶功能分析 引言 在病害、盐碱、矿物质缺乏以及不利的pH等条件下植物会产生过量的乙烯。乙烯对植物的生长、发育、衰老、器官脱落等起着重要调节作用，过量的乙烯会抑制植物根系的生长，进而抑制植物的正常生长，使植物的抗病能力降低[1]。乙烯的生物合成途径：蛋氨酸→ S-腺苷蛋氨酸→ 1-氨基环丙烷-1-羧酸 (ACC) →乙烯[2-6]，催化S-腺苷蛋氨酸生成ACC的1-氨基环丙烷羧酸脱氨酶 (ACCD酶) 是植物乙烯合成的限速酶，决定着乙烯的产生，也影响着植物的生长[7]。而木霉所产生的ACCD酶能够切断乙烯的前体ACC产生酮戊二酸和氨，从而降低乙烯含量，促进植物的生长，提高植物的抗病性[8-11]。因此，木霉ACCD酶在生物防治方面具有良好及广泛的应用前景。 (责任编辑：qin) |