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| 利用松茸ITS特异性引物对松茸分离物进行鉴定 | |
| 引用本文： | 沙涛,丁骅孙,张汉波,王敏艳,程立忠,赵之伟.利用松茸ITS特异性引物对松茸分离物进行鉴定[J].云南植物研究,2004,26(5):524-528. |
| 作者姓名： | 沙涛 丁骅孙 张汉波 王敏艳 程立忠 赵之伟 |
| 作者单位： | 1. 云南大学,生物资源保护与利用国家重点实验室培育基地,云南,昆明,650091 2. 云南大学生物学系,云南,昆明,650091 3. 云南大学,生物资源保护与利用国家重点实验室培育基地,云南,昆明,650091;云南大学生物学系,云南,昆明,650091 |
| 基金项目： | “十五”国家科技攻关计划“生物资源与生物安全技术研究开发”项目 (No . 2 0 0 1BA70 7B0 1)，中央级科研院所科技基础性工作专项———重要野生食用真菌种质资源收集与保藏研究 |
| 摘 要： | 利用一对ITS通用引物(YIS4-ITS5)和一对松茸物ITS特异性引物(TMF-TMR)对来源于云南省不同地区的6个松茸(Tricholoma matsutake)子实体及其6株分离物，3个假松茸(Tricholoma bakamatsutake)子实体及其3株分离物、侧耳(Pleurotus astreatus)、金针菇(Flammulina velutipes)和双孢蘑菇(Agaricus bisporus)的子实体进行了PCR扩增，琼脂糖凝胶电泳分析，结果表明ITS4-ITS5能将所有的样品扩增，并得到600bp左右的DNA扩增条带，TMFTMR扩增时，只有松茸子实体及其对应分离物有扩增条带，DNA片段大小在500bp左右。进一步对松茸子实体(TG25)及其分离物(TM25112．2)进行WS序列测定表明两者的DNA同源性为100％，从而证明所分离到的6个菌株确为松茸的纯培养物。 |
| 关 键 词： | 松茸 子实体 分离物 ITS DNA片段 云南 |
| 文章编号： | 0253-2700(2004)05-0524-05 |
| 修稿时间： | 2004年2月26日 |
Identification of Isolates from Fruit Bodies of Tricholoma matsutake by Specific ITS Primer |
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| SHA Tao,DING Hua-Sun,ZHANG Han-Bo.Identification of Isolates from Fruit Bodies of Tricholoma matsutake by Specific ITS Primer[J].Acta Botanica Yunnanica,2004,26(5):524-528. | |
| Authors: | SHA Tao DING Hua-Sun ZHANG Han-Bo |
| Institution: | SHA Tao~1,DING Hua-Sun~2,ZHANG Han-Bo~ |
| Abstract: | A pair of general primer (ITS4-ITS5) and a pair of specific primer (TMF-TMR) were used to test in the amplification of the internal transcribed spacer (ITS) region of the chromosomal DNA of fruit bodies of Tricholoma matsutake and its 6 isolates,Tricholoma bakamatsutake and its 3 isolates,fruit bodies of Flammulina velutipes,Pleurotus ostreatus and Agaricus bisporus.The primers of ITS4 and ITS5 amplified about 600?bp fragments for all samples.However,with TMF and TMR primers we can amplify about 500?bp fragments only from the fruit bodies of Tricholoma matsutake and their corresponding isolates.The ITS fragments of Tricholoma matsutake fruit body (TF25) and its corresponding isolate (TM25112.2) were sequenced and aligned.Their sequences showed a 100% homogeneity.Based on these facts,6 isolates designated as TM0611.2,TM15110.1,TM25112.2,TM28112.1,TM29112.1 and TM3012.2 were confirmed to be pure cultures of Tricholoma matsutake. |
| Keywords: | Tricholoma matsutake Internal transcribed spacer(ITS) Identification |
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