利用两种方法检测苹果锈果类病毒

Detection of Apple scar skin viroid using two methods

以克隆ASSVd的部分序列,通过RT-PCR成功合成了地高辛标记的cDNA探针,提取苹果和梨树枝条的总RNA,用斑点杂交技术对其进行了检测试验,结果表明,探针具有很高的灵敏度和特异性。地高辛标记的cDNA探针不与阴性对照枝条RNA以及感染PBCVd、AFCVd、ADFVd枝条总RNA发生杂交,仅与感染ASSVd样品的总RNA杂交。

In order to clone partial sequence of Apple scar skin viroid （ASSVd）, total RNA was extracted from the shoots of apple and pear trees, and the cDNA probe labeled by digoxigenin was synthesized by reverse transcription polymerase chain reaction （RT-PCR）. Detection of ASSVd was conducted using blot hybridization. Results showed that the cDNA probe was sensitive and specific, hybridizing only with the total RNA from ASSVd infected shoots, but not with those from negative control shoots or the shoots infected by Pear blister canker viroid （PBCVd）, Apple fruit crinkle viroid （AFCVd） and Apple dimple fruit viroid （ADFVd）.