宁夏苦豆子中产苦参碱内生真菌的分离与鉴定

doi:10.3864/j.issn.0578-1752.2013.13.003

Abstract

Abstract: 【Objective】An experiment was carried out to isolate and confirm whether the presence of fungal endophyte that could produce quinolizidine alkaloids including matrine, oxymatrine, oxysophocarpine or sophoridine from wild Sophora alopecuroides L in Ningxia. 【Method】The isolates were observed under the light microscope and scanning electron microscopy, respectively. The classification of each isolate was performed preliminarily according to the morphological characteristics. The chemical compositions of isolated fungi were detected by thin-layer chromatography, gas chromatograph-mass spectrometer and acid dye colorimetry to screen the quinolizidine alkaloids-producing fungal endophytes. The ITS and SSU rDNA sequences of the fungi were amplified and sequenced. Homology comparison was conducted and phylogenetic tree was established based on the sequences of the fungi. The alkaloids-producing fungal endophytes were classified according to the fungal morphology and phylogenetic analysis.【Result】Twenty-seven fungi were isolated from S. alopecuroides. Twenty-three of the 27 isolates were classified as Penicillium, Aspergillus, Alternaria and Fusarium based on morphology of conidia. The other four isolates could not produce conidia and were classified as mycelia sterile. However, only three fungal isolates, E1, E3 and E5 that belonged to Simplicillium, were conformed contain substantial amounts of matrine which is a kind of quinolizidine alkaloids. The contents of matrine were 20.4 μg•g-1, 17.4 μg•g-1 and 37.6 μg•g-1, respectively, in the mycelia of E1, E3 and E5. 【Conclusion】Homology and phylogenetic analysis results showed that the endophytes are more closely related to Simplicillium lanosoniveum. The morphologies of conidiophores and conidia in the three fungal endophytes were also highly similar to S. lanosoniveum. The results indicated that E1, E3 and E5 were S. lanosoniveum.

Key words: Sophora alopecuroides , matrine , fungal endophyte , Simplicillium lanosoniveum

YU Yong-Tao, HE Sheng-Hu, ZHAO Qing-Mei. Isolation and Identification of Matrine-Producing Fungal Endophytes from Sophora alopecuroides in Ningxia[J].Scientia Agricultura Sinica, 2013, 46(13): 2643-2654.

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Isolation and Identification of Matrine-Producing Fungal Endophytes from Sophora alopecuroides in Ningxia

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