Ascencio-Ibañez Jose Trinidad, Settlage Sharon B
North Carolina State University, Department of Molecular and Structural Biochemistry, Viral Genomics Group, CB 7651 Partners III, Room B04, Raleigh, NC 27695, USA.
J Virol Methods. 2007 Jun;142(1-2):198-203. doi: 10.1016/j.jviromet.2007.01.031. Epub 2007 Mar 6.
Geminiviruses belong to a rapidly growing group of plant pathogens that contribute to crop losses in tropical and subtropical areas of the world. Geminivirus infection is a model for plant DNA replication and virus/host interactions. Geminiviruses are also used as vectors to induce silencing of endogenous genes in several plant species. A method was analyzed for inoculating geminiviruses using plasmid DNA rubbed onto leaves in the presence of an abrasive (DNA abrasion). Although the use of DNA abrasion to inoculate geminiviruses has been described previously, the technique has fallen out of favor and has not been systematically optimized. However, consistent efficiencies of 100% infection rates can be achieved by DNA abrasion. The symptoms of Tomato Golden Mosaic Virus or Cabbage Leaf Curl Virus infection on Nicotiana benthamiana were similar in timing and appearance to the symptoms observed in plants inoculated using Agrobacterium as the delivery method. More importantly, silencing of an endogenous gene was highly efficient when a geminivirus silencing vector was inoculated by the DNA abrasion method. Other plant species successfully inoculated with geminiviruses by DNA abrasion were Nicotiana tabacum, Capsicum annuum and Nicandra physalodes. Unfortunately, Arabidopsis thaliana could not be infected with Cabbage Leaf Curl Virus using leaf abrasion, demonstrating limitation of the method. However, leaf abrasion to inoculate geminiviruses is an easy and inexpensive method that should be considered as an accessible technique to the growing number of researchers using geminiviruses.
双生病毒属于一类迅速增多的植物病原体，在世界热带和亚热带地区造成作物损失。双生病毒感染是植物DNA复制以及病毒与宿主相互作用的一个模型。双生病毒还被用作载体，在几种植物物种中诱导内源基因沉默。分析了一种在有研磨剂存在的情况下将质粒DNA涂抹在叶片上接种双生病毒的方法（DNA研磨接种法）。虽然此前已有关于使用DNA研磨接种法接种双生病毒的描述，但该技术已不再受欢迎且未得到系统优化。然而，通过DNA研磨接种法可实现100%的一致感染率。番茄金色花叶病毒或卷心菜曲叶病毒感染本氏烟草后的症状，在时间和外观上与使用农杆菌作为递送方法接种的植物中观察到的症状相似。更重要的是，当通过DNA研磨接种法接种双生病毒沉默载体时，内源基因的沉默效率很高。通过DNA研磨接种法成功接种双生病毒的其他植物物种有烟草、辣椒和假酸浆。遗憾的是，使用叶片研磨法无法用卷心菜曲叶病毒感染拟南芥，这证明了该方法的局限性。然而，叶片研磨接种双生病毒是一种简单且廉价的方法，对于越来越多使用双生病毒的研究人员来说，应将其视为一种可采用的技术。
