摘要三叶青是一种珍稀的药用植物。大量的药理实验证明三叶青具有抗肿瘤、保肝、抗病毒、抗炎、调节免疫作用等药理作用。由于近年来随着对三叶青的需求量不断增加，三叶青野生资源日益稀缺。植物内生真菌与植物是相互依存的共生关系，其对植物的生长发育和抗逆性有着重要影响。植物扩展蛋白存在于细胞壁中，是一类能使植物细胞壁舒张的细胞壁活性蛋白，其与植物的生长发育有着密切的关系。本研究利用荧光定量PCR等生物技术手段探究三叶青块根中内生真菌对三叶青扩展基因表达含量以及药用化学成分的影响。72398

实验结果表明：经9。7-2和11。23-1内生真菌代谢产物处理的三叶青植株生长较快，植株粗壮，同时扩展蛋白相对表达含量较对照组有增加，而经8。16-8内生真菌代谢产物处理的三叶青植株生长缓慢，植株矮小，同时扩展蛋白相对表达含量较对照组有降低，这一现象表明植株的生长状况与扩展蛋白的含量具有一定的相关性。使用紫外分光光度计测定叶片中药用成分总黄酮的含量，经过内生真菌代谢产物处理的三叶青植株中总黄酮含量普遍增加，其中Th-9。7-5三叶青植株中总黄酮含量高达39。6194 mg/g。DW，是对照组的1。93倍。

Abstract Tetrastigma hemsleyanum is a precious medicinal plant。 A large number of pharmacological experiments show that Tetrastigma hemsleyanum has pharmacological effect，include anti-tumor, liver protection, anti-viral, anti-inflammatory, and regulating immune function。 In recent years， wild Tetrastigma hemsleyanum resources are constantly reducing with increasing demand。  The endophytic fungi have built a interdependent symbiotic relationship with host plants, which have important influence on plant growth and stress resistance。 Expansin exists in the plant cell wall, which is a kind of  active protein prompting cell wall relaxation。 Expansin is closely related to plant growth。 In this study, the influence of endophytic fungi from Tetrastigma hemsleyanum on expansin gene expression and medicinal chemical composition were explored by applying biological techniques such as quantitative PCR。

The experimental results show that Tetrastigma hemsleyanum plants which were processed by 9。7-2 and 11。23-1 endophytic fungi metabolites were sturdy and grow fast, while the expression of expansin gene in those plants was upregulated。 Similarly, Tetrastigma hemsleyanum plants which were processed by 8。16-8 endophytic fungi metabolites were dwarf and grow slowly, while the expression of expansin gene in those plants was downregulated。 This phenomenon showed that the growth of plants was related to the expression of expansin gene。 The flavonoid contents were detected by the method of UV spectrophotometry at 500 nm。 The results showed that the flavonoid contents were increased in Tetrastigma hemsleyanum plants which were processed by endophytic fungi metabolites。 Among them, the flavonoid content in Th-9。7-5 was up to 39。6194 mg/g。DW, which was three times more than the control group’s。

毕业论文关键词：三叶青; 内生真菌; 扩展蛋白; 荧光定量PCR; 总黄酮含量

Key Word:  Tetrastigma hemsleyanumts;  endophytic fungus;  expansin;  fluorescence quantitative PCR;  total flavonoids

目   录 4

引言 5

1  材料与方法 6

1。1 实验材料与试剂 6

1。2 实验方法 6

1。2。1 三叶青内生真菌的分离及鉴定 6

1。2。2 内生真菌代谢产物的获得 6