摘要灵芝作为大型药用真菌，富含较高的营养成分和极高的保健作用，在我国有着悠久的应用历史。灵芝三萜作为灵芝次级代谢产物之一，具有抗氧化、抗病毒等活性。转录因子SKN7在灵芝的生物学作用尚未进行解析，在本文中发现，SKN7沉默后能够显著提高灵芝三萜含量达25%，SKN7沉默后能够显著提高胞内ROS的含量至原来的1。5-2。0倍，并发现转化子灵芝三萜的上调能够被ROS清除剂恢复。水杨酸（salicylic acid，SA）能够诱导灵芝次级代谢产物的产生，然而转录因子SKN7沉默转化子在SA的诱导下灵芝三萜的生物合成进一步上调。初步揭示了SKN7确实参与SA对灵芝酸生物合成的调控，并作出大胆猜想：SKN7通过对SA处理引起的ROS爆发进行清除，而削弱加入SA对灵芝酸合成的影响。86091

毕业论文关键词：SKN7  活性氧  灵芝三萜  水杨酸

Research of the preliminary regulation mechanism ganoderma lucidum acid salicylic acid the transcription factors SKN7 participate in

Abstract As a large medicinal fungus, Ganoderma lucidum has high nutritional value and health care value。 It has a long history in China。 Ganoderic Acid as one of the secondary metabolites of G。lucidum, with antioxidant, antiviral and so on。 The biological role of transcription factor SKN7 in Ganoderma lucidum has not yet been resolved，in this paper, SKN7 silencing can significantly improve the GA content about 25%, SKN7 silencing could significantly improve the content of ROS to 1。5 to 2。0 times, and found raised GA content can be recovered by ROS scavenger。 Salicylic acid (SA) can induce secondary metabolites in Ganoderma lucidum, however, the transcription factor SKN7 silencing transformants are further upregulated in SA induced biosynthesis of GA content。 Preliminary reveals the SKN7 really participate in SA regulation of ganoderma lucidum acid biosynthesis and make a bold guess that SKN7 weaken the influence of ganoderma lucidum acid synthesis after SA was added through removing the outbreak of ROS。

Keywords:  SKN7   reactive oxygen species   ganoderic acid   salicylic acid

目  录

摘要 1

Abstract 1

前言 1

1  材料与方法 2

1。1 材料 2

1。1。1 菌株及试剂 2

1。1。2  菌株培养 2

1。2 方法 2

1。2。1 总灵芝酸含量测定 2

1。2。2 ROS检测测定 2

1。2。3 胞内H2O2含量测定 3

1。2。4 RNA提取和cDNA 制备 3

1。2。5 基因表达的qRT-PCR分析 4

1。2。6 结果分析 4

2  结果与分析 4

2。1  SKN7沉默后显著引起灵芝三萜的积累 4

2。2  SKN7沉默后显著引起灵芝胞内ROS的积累 5

2。3  还原剂处理SKN7转化子对灵芝三萜含量的影响 5

2。4  还原剂处理SKN7转化子对灵芝胞内ROS含量的影响 6

2。5  SKN7沉默转化子在SA处理条件下灵芝三萜的变化 7

2。6  SKN7沉默转化子在SA处理条件下胞内ROS含量的变化