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Position: Home > Articles > Detection of Rubber Witches' Broom Disease with PCR and Nucleotide Hybridization Chinese Journal of Tropical Crops 2001,22 (2) 8-12

橡胶丛枝病分子检测研究

作  者:
邓晓东;费小雯;刘志昕;陈慕容;郑学勤
单  位:
中国热带农业科学院热带作物生物技术国家重点实验室;中国热带农业科学院植物保护研究所;中国热带农业科学院热带作
关键词:
聚合酶链式反应;核酸杂交;橡胶树丛枝病;植原体
摘  要:
感染橡胶丛枝病的长春花(菟丝子桥接),通过提取其总DNA,以植原体(phytoplas-ma)特异引物,进行 PCR扩增,结果得到860bp的 DNA片断,片断的大小与设计的相符。而健康长春花扩增得不到此片断。同样,将感染丛枝病的橡胶树总DNA,进行PCR扩增,结果与上述相同。该 860bp片断经地高辛标记,核酸杂交结果显示,感染丛核病的橡胶树阳性反应强烈,健康橡胶树仅有微弱反应。
译  名:
Detection of Rubber Witches' Broom Disease with PCR and Nucleotide Hybridization
作  者:
Deng Xiaodong, Fei Xiaowen, Liu Zhixin, Chen Murong, Zheng Xueqin (National Key Biotechnology Laboratory for Tropical Crops, CATAS, Haikou 571101; Plant Protection Research Institute, CATAS, Danzhou, Hainan 571737)
关键词:
PCR nucleotide hybridization rubber witches' broom disease phytoplasma
摘  要:
A novel method in which PCR and DNA hybridization techniques were utilized was established for detecting rubber witches' broom disease associated with phytoplasma. The total DNA extracted from infected periwinkle and diseased rubbers were amplified using special oligo nucleotide primers. The product of PCR was 860 hp fragment. The size of the fragment was consistent with the distance between the two primers in the sequence of the Oenothera phytoplasma (863 bp). The template DNA from healthy periwinkle and rubber couldn't amplified the 860 bp fragment. This 860 bp fragment was used as probe with Dig labeling and dot blot hybridization. The results showed that the DNA of diseased rubber could be strongly hybridized with this fragment.

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