中国农业科学 ›› 2009, Vol. 42 ›› Issue (10): 3662-3671 .doi: 10.3864/j.issn.0578-1752.2009.10.0035

• 畜牧·资源昆虫 • 上一篇    下一篇

西藏3种疯草中合成苦马豆素内生真菌的鉴定

余永涛,王建华,王 妍,宋毓民,耿果霞,李勤凡

  

  1. (西北农林科技大学动物医学院)
  • 收稿日期:2009-01-12 修回日期:2009-03-27 出版日期:2009-10-10 发布日期:2009-10-10
  • 通讯作者: 王建华

Identification of Swainsonine-Producing Fungal Endophytes from Three Species of Locoweeds in Tibet

YU Yong-tao, WANG Jian-hua, WANG Yan, SONG Yu-min, GENG Guo-xia, LI Qin-fan
  

  1. (西北农林科技大学动物医学院)
  • Received:2009-01-12 Revised:2009-03-27 Online:2009-10-10 Published:2009-10-10
  • Contact: WANG Jian-hua

摘要:

【目的】从西藏毛瓣棘豆、冰川棘豆、茎直黄芪中分离内生真菌。【方法】应用薄层色谱法、气相色谱、气相色谱-质谱联用法检测内生真菌中的苦马豆素(swainsonine,SW),筛选可生成SW的内生真菌;应用聚合酶链反应对真菌的ITS序列和IGS序列进行扩增,并根据ITS序列信息构建系统发育树,确定其种属分类;对IGS序列进行限制性酶切,并分析酶切产物。【结果】从3种疯草中分离到可以产生SW的4株内生真菌,各菌的形态、ITS序列的同源性与埃里格孢属真菌较为接近,系统发育分析表明4株真菌均为棘豆埃里格孢菌。ITS序列和IGS序列的限制性片段的分析表明,4株真菌遗传距离较近,但仍存在种间变异,可能为棘豆埃里格孢菌的不同亚种。【结论】本研究结果为探讨内生真菌合成SW机理研究奠定了基础,为动物疯草中毒病的防除和疯草资源的合理利用提供了新思路。

关键词: 疯草, 苦马豆素, 内生真菌, 棘豆埃里格孢

Abstract:

【Objective】 The fungal endophytes were isolated from three locoweeds Oxytropis sericopetala, Oxytropis glacialis and Astragalus strictus in Tibet. 【Method】 The swainsonine isolated from the endophytes was analyzed with thin-layer chromatography, gas chromatography and gas chromatography-mass spectrometry. The ITS and IGS regions of fungi were amplified by polymerase chain reactions. In addition to the sequences of ITS regions from endophytes, the phylogenetic tree based on the ITS sequences was constructed to determine the taxon of the endophytic fungi. The amplified products of IGS regions from endophytes were digested using restriction enzymes and the digested products were analyzed. 【Result】 Four swainsonine-producing fungal endophytes were isolated from three species of locoweeds. The morphology of fungi and homology of ITS sequences was closely related to that of Embellisia spp. Four isolates were named Embellisia oxytropis based on the phylogenetic tree. 【Conclusion】 Analysis of the ITS sequences and the digested IGS fragments indicated that four endophytes were closely related species. However, the intraspecific variation was present among the fungal endophytes from different host plants and the isolates may be different subspecies of Embellisia oxytropis. The research is important to explore the SW biosynthesis mechanism of endophytes and has provided a novel knowledge for the elimination of animal locoism and a reasonable utilization of locoweeds.

Key words: locoweed, swainsonine, fungal endophyte, Embellisia oxytropis