直立黄芪中产苦马豆素真菌的分离与鉴定

doi:10.11843/j.issn.0366-6964.2018.08.023

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (8): 1770-1780.doi: 10.11843/j.issn.0366-6964.2018.08.023

直立黄芪中产苦马豆素真菌的分离与鉴定

余永涛^1*, 李金荣¹, 赵清梅^2,3, 何生虎¹, 陈娟⁴, 杨奇⁴, 毛彦妮¹

1. 宁夏大学农学院, 银川 750021;
2. 北方民族大学生物科学与工程学院, 银川 750021;
3. 国家民委发酵酿造工程生物技术重点实验室, 银川 750021;
4. 宁夏回族自治区兽药饲料监察所, 银川 750011

收稿日期:2018-01-29 出版日期:2018-08-23 发布日期:2018-08-18
通讯作者: 余永涛,E-mail:yyt1211@163.com
作者简介:余永涛(1980-),男,宁夏平罗人,副教授,博士,硕士生导师,主要从事动物中毒病与营养代谢病研究
基金资助:
国家自然科学基金（31560713；31201962）

Isolation and Identification of Swainsonine-producing Fungi from Astragalus adsurgens Pall

YU Yong-tao^1*, LI Jin-rong¹, ZHAO Qing-mei^2,3, HE Sheng-hu¹, CHEN Juan⁴, YANG Qi⁴, MAO Yan-ni¹

1. School of Agriculture Ningxia University, Yinchuan 750021, China;
2. College of Biological Science and Engineering, North Minzu University, Yinchuan 750021, China;
3. Key Laboratory of Fermentation Brewing Engineering and Biotechnology State Nationalities Affairs Commission, Yinchuan 750021, China;
4. Ningxia Institute of Veterinary Drug and Feed Control, Yinchuan 750011, China

Received:2018-01-29 Online:2018-08-23 Published:2018-08-18

摘要/Abstract

摘要：

为了确定直立黄芪中是否存在能产生苦马豆素的真菌及该类真菌与链格孢属波状芽管孢组疯草内生真菌的遗传进化关系，对直立黄芪植物组织中的真菌进行了分离，应用α-甘露糖苷酶抑制法和超高效液相色谱-串联质谱法对分离菌株菌丝中的苦马豆素进行分析，筛选能够产生苦马豆素的菌株。PCR法扩增产苦马豆素真菌的内部转录间隔区（ITS）、甘油醛-3-磷酸脱氢酶（GPD）和β-酮脂酰合酶（β-ketoacyl synthase，KS）基因，测定其序列，进行序列一致性比对。基于ITS和GPD序列构建系统发育树，根据遗传进化分析结果和形态特征，对直立黄芪中的产苦马豆素真菌进行种属分类和命名。结果显示：从直立黄芪中共分离到43株真菌，其中有13株菌丝中含有苦马豆素，序列一致性和系统发育分析结果表明，该类真菌与已报道的直立黄芪病原真菌甘肃链格孢（Alternaria gansuense）和产苦马豆素疯草内生真菌（Alternaria Section Undifilum sp.）的亲缘关系最近。直立黄芪中存在能产生苦马豆素的真菌，根据形态特点和遗传进化分析结果，本研究将从直立黄芪中分离的产苦马豆素真菌划分到链格孢属波状芽管孢组中，并命名为甘肃波状芽管孢（Alternaria Section Undifilum gansuense）。

关键词: 直立黄芪, 甘肃波状芽管孢, 苦马豆素, 分离, 鉴定

Abstract:

The study was conducted to determine whether Astragalus adsurgens Pall is infected by the swainsonine-producing fungi, and the genetic evolution relationship between the Alternaria Section Undifilum species fungal endophytes from locoweeds and A. adsurgens Pall. Fungal isolates were isolated from the tissues of Astragalus adsurgens Pall. To screen the fungi that can produce swainsonine, the hyphal extract of each isolate was detected by means of the α-mannosidase Assay and ULPC-MS respectively. ITS, GPD, and KS genes from the swainsonine-producing fungi were amplified and sequenced for the analysis of homology and genetic evolution. The phylogenetic trees of fungi were built based on fungal ITS and GPD sequences. The taxon of the swainsonine-producing isolates from A. adsurgens Pall was determined according to morphology and the results of genetic evolution analysis. The present study showed that 43 isolates were cultured from the tissues of A. adsurgens Pall. The hyphae of thirteen isolates were confirmed containing swainonine. The results based on the analysis of homology and genetic evolution showed that swainonine-producing isolates were closely related to pathogenic fungus Alternaria gansuense from A. adsurgens Pall and the fungal endophytes Alternaria Section Undifilum sp. from locoweeds. This study determined the presence of swainonine-producing fungi inside A. adsurgens Pall. The isolates producing swainsonine are classified into Alternaria Section Undifilum sp and named Alternaria Section Undifilum gansuense based on the morphology and the results of genetic evolution analysis.

Key words: Astragalus adsurgens Pall, Alternaria Section Undifilum gansuense, swainsonine, isolation, identification

中图分类号:

S852.66

余永涛, 李金荣, 赵清梅, 何生虎, 陈娟, 杨奇, 毛彦妮. 直立黄芪中产苦马豆素真菌的分离与鉴定[J]. 畜牧兽医学报, 2018, 49(8): 1770-1780.

YU Yong-tao, LI Jin-rong, ZHAO Qing-mei, HE Sheng-hu, CHEN Juan, YANG Qi, MAO Yan-ni. Isolation and Identification of Swainsonine-producing Fungi from Astragalus adsurgens Pall[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1770-1780.

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直立黄芪中产苦马豆素真菌的分离与鉴定

Isolation and Identification of Swainsonine-producing Fungi from Astragalus adsurgens Pall

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