DNA abrasion onto plants is an effective method for geminivirus infection and virus-induced gene silencing.
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.
Ascencio-Ibañez
Jose Trinidad
JT
North Carolina State University, Department of Molecular and Structural Biochemistry, Viral Genomics Group, CB 7651 Partners III, Room B04, Raleigh, NC 27695, USA. trino_ascencio@ncsu.edu
Settlage
Sharon B
SB
D017360:Arabidopsis / Q000821:virology*; D017554:Carbon Compounds, Inorganic / Q000008:administration & dosage*; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D017791:Geminiviridae / Q000145:classification / Q000235:genetics / Q000472:pathogenicity*; D020868:Gene Silencing; D008190:Lyases / Q000235:genetics / Q000378:metabolism*; D018515:Plant Leaves / Q000737:chemistry* / Q000821:virology; D012825:Silicon / Q000008:administration & dosage*; D014026:Nicotiana / Q000821:virology*; D014773:Virology / Q000379:methods
D017554:Carbon Compounds, Inorganic; D004279:DNA, Viral; C517455:carborundum; D008190:Lyases; C086788:magnesium chelatase; D012825:Silicon