摘要齿兰环斑病毒是严重危害兰花植物的主要病毒之一，因其会造成观赏和经济价值的损失，并且目前也没有特效治疗措施，故抵御该兰花病毒势在必得。为了提高病毒RNA沉默载体的抗病毒有效性，提高后期转化效应和转基因植株的抗病效应，我们将齿兰环斑病毒ORSV沉默载体在模式寄主烟草中进行农杆菌浸润接种和瞬时表达，通过RT-PCR、sq RT-PCR等方法，对寄主感病症状、病毒体内富集以及病毒基因的表达进行分子鉴定和研究，以检测构建载体的抗病毒效应。本实验结果对兰花产业的抗病毒防治具有重要的理论指导意义和应用价值。本实验主要研究结果如下：ORSV沉默载体通过农杆菌侵润法，侵染本氏烟后，不影响本氏烟生长；ORSV沉默载体浸润本氏烟后，取叶片提取总RNA，经过RT-PCR检测后发现，本氏烟体内没有积累病毒基因组，即沉默载体的接种不影响植物生长；ORSV沉默载体与病汁液共接种本氏烟后，降低了本氏烟的发病率，推测构建的ORSV沉默载体具有沉默效应。Sq RT-PCR进一步检测后发现，ORSV沉默载体发挥了沉默功能，能够有效干涉病毒基因组的表达，抵御病毒的入侵。47214

Abstract：Odontoglossum ringspot virus is one of the major virus orchid serious harm plants，Because it will result in the loss of ornamental and economic value, and currently there is no specific treatment measures, so resist the orchid virus aspirations. In order to improve the effectiveness of antiviral RNA silencing vectors of the virus, increase the later transformation effect and   the resistance effect of transgenic plants, We will Odontoglossum ringspot virus ORSV silencing vectors for Agrobacterium infiltration inoculation and transient expression in tobacco in host mode, through the RT-PCR, sq RT-PCR and method, Molecular identification and study of the expression of host disease symptoms, the virus accumulation and virus gene, vector construction to detect the antiviral effect. These results have important theoretical significance and application value for the anti-virus control orchid industry. The main results of this experiment are as follows: ORSV silencing vector by Agrobacterium infiltration method, after infecting N. benthamiana, does not affect the growth of N. Benthamiana; ORSV silencing vector infiltration Nicotiana benthamiana, Total RNA was extracted from the leaves, after RT-PCR was found:Benthamiana is no accumulation of viral genome, namely the silencing vector inoculation did not affect plant growth; ORSV silencing vectors and disease juice co inoculation N.benthamiana, reduce the incidence rate of N.benthamiana. It is reasoned that the carrier has built ORSV silence silencing effect. Sq RT-PCR further showed that ORSV silencing plays silencing function, the expression of the viral genome can effectively interfere,  resist the invasion of virus.

毕业论文关键词：齿兰环斑病毒； RNA沉默； ORSV沉默载体；

Keyword: Odontoglossum ringspot virus；RNA silencing；ORSV silent carrier

 目    录

摘    要 II

目    录 III

引    言 1

1. 材料和方法 1

1.1. 实验材料 1

1.2. 载体菌复苏 1

1.3. ORSV沉默载体浸润寄主（ORSV的模式寄主—本氏烟） 1

1.4. ORSV沉默载体与病汁液共浸润本氏烟植株 2

1.5. RT-PCR检测ORSV沉默载体处理的本氏烟植株 2

2. 实验结果 3

2.1. ORSV沉默载体处理本氏烟后植株正常生长