比较形态学、聚丙酰胺凝胶电泳与二代测序在AM真菌群落研究中的应用

Application of comparative morphology, DGGE and NGS to the study of AM fungal community

采集内蒙古荒漠草原沙化梁地羊柴Hedysarum laeve根围土样,采用形态学方法、聚丙酰胺凝胶电泳(denatured gradient gel electrophoresis,DGGE)技术和二代测序(next generation sequencing,NGS)技术分析羊柴根围AM真菌群落组成和多样性。结果表明,应用NGS共鉴定AM真菌3纲4目10科18属,DGGE鉴定2纲2目2科5属,形态学鉴定1纲2目2科3属。AM真菌分类等级越低,NGS、DGGE和形态学鉴定结果差异越大,尤其在属水平上差异显著。与NGS技术相比,形态学和DGGE方法所反映AM真菌种类主要为优势菌种,显著低估了AM真菌物种组成并高估其丰度,分子测序是对形态学分类的补充和完善。

AM（arbuscular mycorrhiza） fungal community structure and diversity in the rhizosphere of Hedysarum laeve（Leguminosae） in desert steppe of Inner Mongolia were analysed by means of morphological identification, denaturing gradient gel electrophoresis（DGGE） fingerprinting technology and next generation sequencing（NGS） technology. Three classes, 4 orders, 10 families and 18 genera of AM fungi were revealed by NGS technique, and 2 classes, 2 orders, 2 families and 5 genera by DGGE, yet only 1 classes, 2 orders, 2 families and 3 genera were classified by morphological identification. There were significant difference between NGS, DGGE and morphological identification results, particularly in the genus level. Morphological method and DGGE fingerprint technology greatly underestimated AM fungal community composition and overestimated the relative abundance of dominant AM fungi, while molecular sequencing is a supplement and improvement over morphological classification.