内蒙古不同小花棘豆种群苦马豆素及其内生真菌关系的分析

doi:10.11843/j.issn.0366-6964.2022.01.030

摘要/Abstract

摘要： 小花棘豆是广泛分布于内蒙古草原和荒漠区的一种有毒植物，其主要毒性成分为有毒生物碱——苦马豆素（swainsonine，SW），牲畜采食后可引起中毒，甚至死亡，给当地畜牧业造成重大经济损失。本研究旨在探究内蒙古不同小花棘豆种群植株苦马豆素及其与内生真菌关系。采集内蒙古8个样地120株小花棘豆，利用萃取、离心、离子交换层析法提取和纯化植物单株和菌丝体的SW，用气相色谱-质谱联用法（GC-MS）检测SW水平，取茎和叶外植体分离培养内生真菌，提取植物和真菌的总DNA，扩增真菌特异序列，利用内生真菌微生物学特征和DNA序列比对进行鉴定。结果显示：8个种群共111株小花棘豆检测出SW，最高水平为369.05 μg·g^-1，平均水平32.78 μg·g^-1，从38株小花棘豆分离培养出内生真菌，纯培养下菌丝体呈松散白色绒毛状，菌落圆形、隆起、边缘整齐、辐射状生长，颜色逐渐呈现灰色、深灰色或褐色至深褐色，内生真菌均测出SW，其水平为0.83~2 573.24 μg·g^-1，经微生物学研究及5.8S rDNA/ITS序列比对分析，在属水平上鉴定该内生真菌为Alternaria。有Alternaria内生真菌的小花棘豆植株含SW，无该内生真菌的植株不含SW，培养的Alternaria内生真菌合成了SW。

关键词: 小花棘豆, 种群, 苦马豆素, 内生真菌

Abstract: Oxytropis glabra is a poisonous plant widely distributed in grassland and desert area of Inner Mongolia, its main toxic compound is an alkaloid (swainsonine, SW). Livestock is poisoned or even dead after feeding, resulting in a major loss of grassland animal husbandry in serious cases. This research explored the relationship between SW from Oxytropis glabra in diffe-rent populations and endophytic fungi in Inner Mongolia. One hundred and twenty plants of Oxytropis glabra from 8 populations in Inner Mongolia were collected, and the SW from indivi-dual plant and mycelia were isolated and purified by extraction, centrifugation and ion exchange chromatography. The SW levels were determined by GC-MS. The endophytic fungi were isolated from stems and leaves, and the total DNA of plants and fungi was extracted. The fungal specific sequences were amplified, and the fungus was identified both by microbiological characteristics and DNA sequence comparison. Results showed that SW was detected in 111 plants from 8 popula-tions of Oxytropis glabra with the highest level 369.05 μg·g^-1 and the average level 32.78 μg·g^-1. Endophytic fungi were isolated from 38 planets. Under pure culture condition, mycelia showed loose white villous, and the colonies were round, upheaval with neat edge and radial growth. The color of colonies gradually showed gray, dark gray or brown to dark brown. SW was detected in all endophytic fungi samples with the level range of 0.83-2 573.24 μg·g^-1. The endophytic fungus was identified as Alternaria by microbiological study and 5.8S rDNA/ITS sequence comparison. The plants with Alternaria endophytic fungi contained SW, while the plants without Alternaria endophytic fungi did not contain SW. The cultured Alternaria endophytic fungi synthesized SW.

Key words: Oxytropis glabra, populations, swainsonine, endophytic fungi

中图分类号:

S859.87
S452

王维夫, 钱亚光, 卢萍, 何珊, 杜玲, 李玉玲, 高峰. 内蒙古不同小花棘豆种群苦马豆素及其内生真菌关系的分析[J]. 畜牧兽医学报, 2022, 53(1): 304-314.

WANG Weifu, QIAN Yaguang, LU Ping, HE Shan, DU Ling, LI Yuling, GAO Feng. The Relationship between Swainsonine and Endophytic Fungi in Different Populations of Oxytropis glabra from Inner Mongolia[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 304-314.

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