摘要
利用一对ITS通用引物 (ITS4 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 (Tricholomamatsutake)子实体及其 6株分离物 ,3个假松茸(Tricholomabakamatsutake)子实体及其 3株分离物、侧耳 (Pleurotusostreatus)、金针菇 (Flam mulinavelutipes)和双孢蘑菇 (Agaricusbisporus)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 6 0 0bp左右的DNA扩增条带 ,TMF TMR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 5 0 0bp左右。进一步对松茸子实体 (TG2 5 )及其分离物 (TM2 5 112 2 )进行ITS序列测定表明两者的DNA同源性为 10 0 % ,从而证明所分离到的 6个菌株确为松茸的纯培养物。
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.
出处
《云南植物研究》
CSCD
北大核心
2004年第5期524-528,共5页
Acta Botanica Yunnanica
基金
"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
关键词
松茸
子实体
分离物
ITS
DNA片段
云南
Tricholoma matsutake
Internal transcribed spacer(ITS)
Identification
