| VIGS实验技术体系在月季中的应用及优化 |
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| 引用本文： | 丁榕,梁晶,赵和文,张克中,崔金腾.VIGS实验技术体系在月季中的应用及优化[J].中国农学通报,2018,34(3):87-92. |
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| 作者姓名： | 丁榕 梁晶 赵和文 张克中 崔金腾 |
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| 作者单位： | 北京农学院园林学院,北京农学院园林学院,北京农学院园林学院,北京农学院园林学院,北京农学院园林学院 |
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| 基金项目： | 北京市教育委员会2015 年科技计划面上项目(KM201510020011)。 |
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| 摘 要： | 旨在月季中建立高效、快速的VIGS实验技术体系，以期能够改进VIGS技术体系在月季等木本植物表达效率低等缺陷。采用RT-PCR方法进行月季RhPDS 基因片段的克隆，在以pTRV2 为原载体，采用双酶切的方法构建了pTRV2-RhPDS 重组载体。经双酶切及测序验证后，通过电击法转化农杆菌GV3101，并分别采用高压喷枪法对月季植株进行侵染、采用真空渗透法对扦插苗以及组培苗进行侵染。采用qRT-PCR方法分析RhPDS 基因的表达。结果显示：构建的重组载体pTRV2-RhPDS，经双酶切验证，显示条带单一清晰，符合预期目标。在侵染方法和材料上，采用真空渗透法侵染组培苗是最为快速、高效的方式。该试验获得了较为高效、快速VIGS 技术体系，提高了VIGS 技术体系在月季上表达效率。
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| 关 键 词： | 贮存 贮存 沼液 污染 分析 评估 |
| 收稿时间： | 2017/9/27 0:00:00 |
| 修稿时间： | 2018/1/10 0:00:00 |
Application and Optimization of VIGS Experimental Technology System in Rosa Hybrida |
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| Abstract: | Objective] The aim of this study is to establish an efficient and fast VIGS experimental system in Rosa hybrida, so that can improve the efficiency of infection by VIGS which is relatively low for Rosa hybrida and other woody plants and provide an experimental basis for further functional study of gene.Method]The coding sequence of RhPDS was amplified from Rosa hybrida using RT-PCR method.Through double digested,pTRV2-RhPDS vectors were constructed.The test result was validated by gene sequencing and double digested,and then the VIGS expression vectors were transformed into Agrobacterium strain GV3101 by eletroporation.The whole plant was sprayed connected to a portable air compressor.The vacuum infiltration method was used for rose cuttings and tissue culture plant.The expression level of RhPDS gene was detected by RT-qPCR.Result]The results showed that the recombinant vector pTRV2-RhPDS, which was verified by double enzyme digestion, showed a single band and was in good agreement with the expected target. In the method and material of infection, it is the most rapid and efficient way to infect tissue culture seedlings by vacuum osmosis. Conclusion]In this research,we obtain an optimized technical system for VIGS which is improve the efficiency of infection by VIGS in Rosa hybrida and other woody plants and provide an solid experimental basis for further functional study of plant gene. |
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| Keywords: | rosa hybrida VIGS TRV RhPDS vacuum osmosis |
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