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采用组织PCR-末端限制性片段长度多态性分析(T-RFLP)免培养技术,从南药植物高良姜道地产地植株根、根茎、茎、叶4个不同组织总共检测到28个不同的末端限制性片段(T-RFs),说明高良姜内生真菌群体至少组成28个不同的真菌种;不同组织其T-RFLP图谱、T-RFs数目、优势T-RFs组成不同,内生真菌群体Shannon多样性指数和Shannon均匀度指数也随组织而异;采用水蒸气蒸馏法、甲醇抽提-高效液相色谱法测定高良姜主要活性成分挥发油、高良姜素含量,结果显示,该2类活性组分在其根、根茎、茎、叶4个不同组织均有积累,且均以根茎最高;相关性分析显示2类所测活性成分与该宿主内生真菌群体Shannon多样性指数及其均匀度呈极显著的负相关,与325 bp对应于拟盘多毛孢菌属Pestalotiopsis的优势T-RFs呈极显著的正相关。
Tissue PCR-based restriction fragment length polymorphism (T-RFLP) culture-free technique was used to detect a total of 28 different tissues from 4 different tissues of roots, rhizomes, stems and leaves of native plant Galangal. (T-RFs), indicating that at least 28 different fungal species were composed in endophytic fungi. The T-RFLP patterns, the number of T-RFs and the dominant T-RFs in different tissues were different. Shannon diversity index and Shannon evenness index also varied with the organization. The contents of volatile oil and galangin in the main active components of Alpinia officinarum were determined by steam distillation and methanol extraction - high performance liquid chromatography. The results showed that these two kinds of activities Rhizomes, stems and leaves, and all had the highest rhizomes. The correlation analysis showed that the Shannon’s diversity index and its evenness of the two tested active components and the host endophytic fungi Showed a very significant negative correlation with the 325 bp corresponding to the dominant plate Pestalotiopsis dominant T-RFs showed a significant positive correlation.