北京地区设施草莓8种病毒的酶联免疫吸附测定检测

Detection of eight viruses infecting greenhouse strawberry in Beijing by enzyme-linked immunosorbent assay

摘要:

【背景】病毒可以随同草莓无性繁殖材料传播扩散，导致产量和品质下降。选育无病毒种苗是草莓病毒病防治的主要措施，高效、灵敏的检测技术可为草莓病毒病防治提供技术保障。【目的】为明确8种能够侵染草莓的病毒在北京地区设施草莓上的发生情况，应用酶联免疫吸附测定(Enzyme-Linked Immunosorbent Assay，ELISA)进行检测。【方法】将待检病毒的阳性对照样品分别进行间接法和双抗夹心法(Double-Antibody Sandwich，DAS) ELISA检测，以空白对照和阴性对照作参照比较，确定应用DAS-ELISA检测体系结合田间症状对北京地区采集的草莓样品进行病毒检测，并利用逆转录-聚合酶链式反应和序列测定方法对阳性样品进一步验证，确定检测结果的可靠性。【结果】侵染北京地区设施草莓的病毒为草莓轻型黄边病毒(Strawberry Mild Yellow Edge Virus，SMYEV)。‘红颜’品种的病毒检出率为8.1%，‘天香’品种的检出率为2.7%，其他品种均未检出病毒。【结论】研究结果为北京地区设施草莓病毒的检测以及草莓病毒病的科学防治提供理论依据。

Abstract:

[Background] Viruses could be disseminated through asexual reproductive materials and severely affect the output and quality of strawberry. The production of virus-free seedling is one of the main methods to control strawberry viral diseases. Therefore, the efficient and sensitive detection of viruses is important for viral disease control. [Objective] To investigate the occurrence of eight important viruses commonly infect greenhouse strawberry in Beijing, the different strawberry plants collected from different districts were tested by enzyme-linked immunosorbent assay (ELISA). [Methods] We established a double-antibody sandwich ELISA (DAS-ELISA) method to detect eight viruses. Compared with the negative control, the positive control samples showed expected results in indirect ELISA and double-antibody sandwich ELISA (DAS-ELISA) analyses. Combined?with the symptom observation in the field, DAS-ELISA was used to detect viruses in the greenhouse strawberry samples. Virus infected samples were further verified by reverse transcription-polymerase chain reaction (RT-PCR) and sequence analysis. [Results] The results showed that only strawberry mild yellow edge virus (SMYEV) was detected in strawberry plants collected in Beijing. 8.1% of ‘Benihoppe’ and 2.7% of ‘Tianxiang’ cultivar plants tested could detect the infection by SMYEV, whereas there were no virus detected in other varieties. [Conclusion] This study presents important information and techniques for virus detection and strawberry viral disease management in Beijing area.