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首页> 外文期刊>FEMS Microbiology Ecology >Fungal endophytes in potato roots studied by traditional isolation and cultivation-independent DNA-based methods
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Fungal endophytes in potato roots studied by traditional isolation and cultivation-independent DNA-based methods

机译：马铃薯根系真菌内生菌的传统分离和不依赖培养的基于DNA的方法研究

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The composition and relative abundance of endophytic fungi in roots of field-grown transgenic T4-lysozyme producing potatoes and the parental line were assessed by classical isolation from root segments and cultivation-independent techniques to test the hypothesis that endophytic fungi are affected by T4-lysozyme. Fungi were isolated from the majority of root segments of both lines and at least 63 morphological groups were obtained with Verticillium dahliae, Cylindrocarpon destructans, Colletotrichum coccodes and Plectosporium tabacinum as the most frequently isolated species. Dominant bands in the fungal fingerprints obtained by denaturing gradient gel electrophoresis analysis of 18S rRNA gene fragments amplified from total community DNA corresponded to the electrophoretic mobility of the 18S rRNA gene fragments of the three most abundant fungal isolates, V. dahliae, C. destructans and Col. coccodes, but not to P. tabacinum. The assignment of the bands to these isolates was confirmed for V. dahliae and Col. coccodes by sequencing of clones. Verticillium dahliae was the most abundant endophytic fungus in the roots of healthy potato plants. Differences in the relative abundance of endophytic fungi colonizing the roots of T4-lysozyme producing potatoes and the parental line could be detected by both methods.
机译：田间转基因生产T4溶菌酶的马铃薯和亲本品系的根中内生真菌的组成和相对丰度通过根段经典分离和不依赖栽培的技术进行评估，以检验内生真菌受T4-溶菌酶影响的假说。从两个品系的大部分根段中分离出真菌，并获得了至少63个形态学组，其中最常见的是大黄萎病菌，Cylindrocarpon destructans，Colletotrichum coccodes和Plectosporium tabacinum。通过变性梯度凝胶电泳分析从总群落DNA扩增得到的18S rRNA基因片段获得的真菌指纹中的显性谱带对应于三种最丰富的真菌分离株V. dahliae，C。destructans和C.的18S rRNA基因片段的电泳迁移率。 Col.编码，但不包括烟草。通过克隆的测序证实了V. dahliae和Col. coccodes对这些分离物的条带分配。黄萎病菌是健康马铃薯植株根中最丰富的内生真菌。两种方法都可以检测到内生真菌相对丰度的差异，这些内生真菌定植在产生T4溶菌酶的马铃薯和亲本系的根部。

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