木霉T-soybean ACCD酶功能分析
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。
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