Production of plum pox virus HC-Pro functionally active for aphid transmission in a transient-expression system.
Potyviruses are non-persistently transmitted by aphid vectors with the assistance of a viral accessory factor known as helper component (HC-Pro), a multifunctional protein that is also involved in many other essential processes during the virus infection cycle. A transient Agrobacterium-mediated expression system was used to produce Plum pox virus (PPV) HC-Pro in Nicotiana benthamiana leaves from constructs that incorporated the 5' region of the genome, yielding high levels of HC-Pro in agroinfiltrated leaves. The expressed PPV HC-Pro was able to assist aphid transmission of purified virus particles in a sequential feeding assay, and to complement transmission-defective variants of the virus. Also, HC-Pro of a second potyvirus, Tobacco etch virus (TEV), was expressed and found to be functional for aphid transmission. These results show that this transient system can be useful for production of functionally active HC-Pro in potyviruses, and the possible uses of this approach to study the mechanism of transmission are discussed.
Goytia
Elisa
E
Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
Fernández-Calvino
Lourdes
L
Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
Martínez-García
Belén
B
Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
López-Abella
Dionisio
D
Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
López-Moya
Juan José
JJ
Consorcio CSIC-IRTA, Instituto de Biología Molecular de Barcelona (IBMB, CSIC), Jordi Girona 18-26, 08034 Barcelona, Spain.|Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.
D000818:Animals; D001042:Aphids / Q000821:virology*; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000502:physiology; D007303:Insect Vectors / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000378:metabolism; D017801:Plum Pox Virus / Q000737:chemistry / Q000472:pathogenicity*; D017800:Potyvirus / Q000737:chemistry / Q000472:pathogenicity; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012231:Rhizobium / Q000378:metabolism; D014026:Nicotiana / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis* / Q000502:physiology; D014774:Virulence
D011994:Recombinant Proteins; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C072775:HC-Pro protein, potyvirus