| 摘要: |
| 【目的】分离筛选杜仲(Eucommia ulmoides Oliv.)树皮中产松脂醇二葡萄糖苷（Pinoresinol diglucoside，PDG）的内生真菌，为PDG的生产提供参考。【方法】采用表面灭菌法分离杜仲内生真菌，摇瓶培养分析各菌株产PDG的能力，结合形态学特征和rDNA内转录间隔区（Internal transcribed spacer, ITS）序列分析对高产菌株进行鉴定，研究培养温度、pH对其生长的影响及其对不同碳源和氮源的利用情况。【结果】共分离得到18株内生真菌，有3株具有产PDG能力，其中以菌株XP-8的PDG产量最高（11.65 mg/L），分类鉴定确定该菌株为拟茎点霉属(Phomopsis sp.)。菌株XP-8在20～35 ℃、pH 4～8的条件下均能较好生长，在28 ℃、pH=5时生长最快；菌株XP-8对麦芽糖和葡萄糖的利用能力较强，不能利用苹果酸；对硝酸钠的利用能力较强，但不能利用天门冬氨酸。【结论】获得了高产PDG的菌株XP-8，确定了其分类学地位，并研究了其生长特点。 |
| 关键词: 杜仲 松脂醇二葡萄糖苷 拟茎点霉属 分类鉴定 生长条件 |
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| 基金项目:陕西省科技攻关项目(2008K02-09)；全国大学生科技创新项目（G07036） |
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| Isolation,identification and growth conditions of endophytic fungi of Eucommia ulmoides Oliv.for production of PDG |
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| Abstract: |
| 【Objective】The study aimed at isolating pinoresinol diglucoside (PDG) producing endophytic fungi from Eucommia ulmoides Oliv. to identify the strain with the highest yield of PDG and to investigate the basic conditions for its growth.【Method】Endophytic fungi were isolated from the surface sterilized bark of Eucommia ulmoides Oliv.and then,cultivated in liquid broth at flask level to test the PDG production.The strain with the highest PDG production was selected and identified based on the morphological characteristics and the sequence of rDNA in the ITS district.Basic growth conditions of the strain were also studied in aspects of temperature,pH,carbon source and nitrogen source by the single factor experimental design.【Result】Eighteen strains of endophytic fungi were isolated from the bark of Eucommia ulmoides Oliv.Only three of them had the ability to produce of PDG.The strain XP-8,with the highest PDG production of 11.65 mg/L,was identified as Phomopsis sp.It could grow in a wide range of conditions from 20 ℃ to 35 ℃ and pH 4 to 8 with the best growth at 28 ℃ and pH 5.The strain XP-8 could use glucose and maltose,but not malic acid,among the tested carbon sources.Sodium nitrate showed to be the most favorable one among the tested nitrogen sources,whereas aspartic acid could not be used at all.【Conclusion】The PDG producing endophytic fungi were isolated from Eucommia ulmoides Oliv..The best strain having the highest PDG production was identified,and the basic conditions for its growth were also studied. |
| Key words: Eucommia ulmoides Oliv. pinoresinol diglucoside phomopsis sp. identification condition for colony growth |
