采用组织块法从马尾松(Pinus massoniana Lamb.)根、茎和叶片中分离获得内生真菌,初步筛选出产油脂菌株且对其油脂含量进行了分析;并采用形态学及分子生物学方法对产油脂菌株进行了分类鉴定。结果显示:从马尾松植株中共获得21 株内生真菌菌株,其中分离自根、茎和叶片的菌株分别为4 株、6 株和11 株。有14 株菌株的菌丝中有油滴;其中,分离自叶片的菌株ZP-1、分离自根的菌株ZP-2 和分离自茎的菌株ZP-3 的菌丝内油滴较多且油脂含量较高,平均油脂含量分别达到29. 12%、25. 03% 和30. 56%,差异极显著(P<0. 01)。不同菌株的菌落颜色、菌丝和分生孢子形态特征明显不同,菌株ZP-1、ZP-2 和ZP-3 的形态分别与拟青霉属(Paecilomyces Bainier)、生赤壳菌属(Bionectria Speg.)和镰刀菌属(Fusarium Link)菌种的形态相似。与GenBank 中相关真菌ITS 序列的比对以及NJ 系统树分析结果显示:菌株ZP-1 与拟青霉属、菌株ZP-2 与生赤壳菌属、菌株ZP-3 与镰刀菌属间的ITS片段序列相似性均达到99%,在各自的NJ 系统树上它们也分别聚在一起。初步确定菌株ZP-1、ZP-2 和ZP-3 分别属于拟青霉属、生赤壳菌属和镰刀菌属。

By means of tissue block method, endophytic fungi were obtained from root, stem and leaf of Pinus massoniana Lamb., and producing lipid strains were preliminarily selected out and their lipid contents were analyzed. And these lipid strains were also classified and identified by morphological and molecular biological methods. The results show that 21 strains of endophytic fungi have been obtained from P. massoniana, in which, the number of strains isolated from root, stem and leaf is 4, 6 and 11 strains, respectively. There are oil bodies in mycelia of 14 strains, in which, the number of oil body is more and lipid content is higher in mycelia of strain ZP-1 from leaf, strain ZP-2 from root and strain ZP- 3 from stem, and average lipid content reaches to 29. 12%, 25. 03% and 30. 56%, respectively, with the extremely significant difference (P<0. 01). Colony color, morphological characteristics of mycelium and conidium of different strains are obviously different, and morphology of strains ZP-1, ZP-2 and ZP-3 is similar to that of fungus species in genera of Paecilomyces Bainier, Bionectria Speg. and Fusarium Link, respectively. The results of alignment with related fungus ITS sequences in GenBank and analysis of NJ phylogenetic tree show that the sequence similarity of ITS fragment of strain ZP-1 with Paecilomyces, strain ZP-2 with Bionectria, strain ZP-3 with Fusarium all reaches to 99%, and they are gathered respectively in their own NJ phylogenetic tree. It is preliminarily determined that strains ZP-1, ZP-2 and ZP-3 belong to genera of Paecilomyces, Bionectria and Fusarium, respectively.