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【目的】揭示醉马草不同组织内生细菌和内生真菌种群组成和分布。【方法】采用液氮研磨法分别提取醉马草种子、叶、茎、根组织总DNA,采用通用引物扩增内生细菌16S rDNA和真菌ITS,通过限制性内切酶HhaⅠ和RsaⅠ对16S rDNA PCR产物酶切,HhaeⅢ和HinfⅠ对真菌rDNA ITS PCR产物酶切得到不同的TRFs片段。TRFs经T-RFLP分析程序结合基因文库比对后,分析醉马草不同组织中内生细菌和内生真菌的群落组成及内生菌群落相似性。【结果】研究表明醉马草根部内生细菌多样性最高,而种子内生真菌多样性最为丰富。醉马草各组织内生细菌优势菌属均为Bacillus(29%以上),种子、叶、茎、根内生真菌优势菌属分别为Mycosphaerella(6.5%),Teratosphaeria(4.5%),Fragum(1.1%),Sebacina(11.3%)。聚类分析表明茎和叶内生细菌群落结构相似,种子和其他组织内生细菌群落结构相似性较远,而茎和种子内生真菌群落结构相似,叶和其他组织内生真菌群落结构相似性较远。醉马草内生菌多样性丰富且存在尚未认识的新类群。
【Objective】 The objective of this study was to reveal the composition and distribution of endophytic fungi and endophytic fungi in different tissues. 【Method】 Total DNA was extracted from the seeds, leaves, stems and roots of Bicolor by liquid nitrogen grinding method. 16S rDNA PCR products digested, Hhae Ⅲ and Hinf Ⅰ fungi rDNA ITS PCR products digested different TRFs fragments. TRFs were analyzed by T-RFLP analysis program combined with gene library to analyze the community composition and endophytic community similarity of endophytic fungi and endophytic fungi in different tissues. 【Result】 The results showed that the endophytic bacteria diversity was the highest in drunken grass roots, while the endophytic fungi diversity was the most abundant. Bacillus (29% or more) were dominant endophytic bacteria in tissues of Mykonian grass and Mycosphaerella (6.5%), Teratosphaeria (4.5%) and Fragum (1.1) were the dominant species in the seeds, leaves, %), Sebacina (11.3%). Cluster analysis showed that the endophytic bacteria communities in stems and leaves were similar in structure, and the endophytic fungi community structure was similar between the seeds and other tissues, while the endophytic fungi community structure was similar between the stems and the seedlings. The endophytic fungi communities in leaves and other tissues were similar in structure Far. Drunken Endophytes rich in endophytic bacteria and there are new groups of unknown.