摘要：近些年来，灵芝产业的发展较为迅速，尤其是灵芝的药用市场前景广阔。灵芝酸作为灵芝中一种重要的次级代谢产物，具有很高的药用价值和经济效益。本课题比较了灵芝bHLH型转录因子沉默前后，灵芝酸的合成量的改变，从而预测bHLH型转录因子在灵芝酸合成调控中的作用。进而使用MeJA分别处理野生型和bHLH型转录因子30684沉默的灵芝菌丝，检测灵芝酸的含量变化以及灵芝菌丝活性氧（ROS）的含量改变，从而揭示bHLH型转录因子对灵芝酸的生物合成的调控机理。然后通过加入抗氧化剂清除沉默转化子中的ROS，同时测定灵芝酸合成含量的变化，初步猜测bHLH型转录因子通过调节ROS进而调控灵芝酸的合成。

关键词：灵芝；灵芝酸；bHLH型转录因子；ROS；清除剂

Study on the Regulation of Ganoderma lucidum bHLH Transcription Factor GL30684 on the Biosynthesis of Ganoderic acid

Abstract：In recent years, the industry of Ganoderma lucidum (G. lucidum) has a relativly rapid development, especially in the field of G. lucidum medicinal market. As an important secondary metabolite of G. lucidum, Ganoderic acid has a high medicinal value and economic benefits. In this study, we compared the changes of the Ganoderic acid content when bHLH transcription factor were silenced, so as to predict the role of bHLH transcription factor in the regulation of Ganoderic acid. And then we treated wild-type G. lucidum and GL30684 silence of transformants with MeJA. The content of Ganoderic acid and mycelium reactive oxygen species (ROS) changes in hyphae. It reveals the regulatory mechanism that bHLH-type transcription factor on the biosynthesis of Ganoderic acid. Then the ROS of the silenced transformants was removed by adding the antioxidant, and the changes of the synthesis of Ganoderic acid were detected. The bHLH transcription factor regulates the synthesis of Ganoderic acid by adjusting the ROS.

Key words：Ganoderma lucidum; Ganoderic acid; bHLH-type transcription factor; ROS; scavenger

目录

摘要 4

关键词 4

Abstract. 4

Key words 4

引言 5

1实验材料与方法 5

1.1实验材料 5

1.1.1菌种 5

1.1.2培养基 5

1.1.3试剂与溶液 6

1.1.4主要仪器设备 6

1.2实验方法 6

1.2.1菌种的活化和接种 6

1.2.2测定野生型和转化子中ROS的含量 6

1.2.3测定MeJA处理前后灵芝酸含量 6

1.2.4测定MeJA处理前后ROS含量 7

1.2.5清除沉默转化子中的ROS 7

1.2.6清除沉默转化子中ROS后，灵芝酸含量测定 7

2结果与分析 7

2.1 GL30684沉默前后ROS含量变化 7

2.2 MeJA处理后灵芝酸含量的变化 8

2.3 MeJA处理后ROS含量的变化 9