| 引用本文: | 龚攀,崔杰,李俊良,罗成飞,杨虎臣.VIGS技术在甜菜上的应用[J].生物信息学,2015,13(1):18-22. |
| GONG Pan,CUI Jie,LI Junliang,LUO Chengfei,YANG Huchen.Application of Virus-Induced Gene Silencing in Beta Vulgaris[J].Chinese Journal of Bioinformatics,2015,13(1):18-22. |
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| 摘要: |
| 病毒诱导基因沉默(VIGS)是一种应用于研究植物基因功能的反向遗传学手段。相对于基因敲除及转基因等方法,它具有时间短、成本低、高通量等优点。随着甜菜全基因组的测序完成,为尽早对大量序列信息进行注释和功能鉴定,急需建立甜菜VIGS体系。本文采用目前应用最广泛的烟草脆裂病毒载体在农杆菌的介导下侵染甜菜幼苗叶片,同时设立对照。提取甜菜幼苗叶片总RNA,并反转录成cDNA,设计特异引物进行RT-PCR检测。结果表明:在侵染植株上检测到了烟草脆裂病毒特异条带,证明该病毒可以对甜菜进行侵染,这一结果为下一步建立甜菜VIGS体系奠定了基础。 |
| 关键词: 甜菜 病毒诱导基因沉默 烟草脆裂病毒 RT-PCR |
| DOI：10.3969/j.issn.1672-5565.2015.01.04 |
| 分类号:TS242.2 |
| 基金项目:科技部技术开发研究专项(2013EG111232)；农业部甜菜产业技术体系项目(CARS-21010202)。 |
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| Application of Virus-Induced Gene Silencing in Beta Vulgaris |
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GONG Pan1,2, CUI Jie2,LI Junliang2,LUO Chengfei2,YANG Huchen2
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(1.School of Life Science and Technology, Harbin Institute of Technology, Harbin 150000,China; 2. School of Food Science and Engineering, Harbin Institute of Technology, Harbin 150090,China)
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| Abstract: |
| Virus-induced gene silencing is an effective method of reverse genetics research for gene function analysis in plants. Compared with gene knockout, transgene and some other methods, it has many advantages such as faster, low-cost and high-throughput. With the accomplishment of Beta vulgaris′ complete genome sequencing,the functional characterization of a large number of sequences need to be identified . It is important to develop a rapid method of VIGS functional analysis system of beet genes. In this paper, we mediated the TRV (Tobacco rattle virus) infection of sugar beet seedlings by Agrobacteriumwhile Tobacco rattle virusis currently the most widely used viral vectors. Total RNA was extracted from the beet leaf and reverse transcribed into cDNA, we designed specific primers used in RT-PCR to detect the virus on the infected beet. The results demonstrated that the virus could infect beet, which laid the foundation for the next VIGS system established in Beta vulgaris. |
| Key words: Beta vulgaris Virus-induced gene silencing Tobacco rattle virus RT-PCR |
