Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana

Cotton blue disease is the most important viral disease of cotton in the southern part of South America. Its etiological agent, cotton leafroll dwarf virus (CLRDV), is specifically transmitted to host plants by the aphid vector (Aphis gossypii) and any attempt to perform mechanical inoculations of t...

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Autores principales:	Delfosse, V.C., Casse, M.F., Agrofoglio, Y.C., Kresic, I.B., Hopp, H.E., Ziegler-Graff, V., Distéfano, A.J.
Formato:	JOUR
Materias:	Arabidopsis thaliana CLRDV Gossypium hirsutum Infectious clone Nicotiana benthamiana Polerovirus capsid protein complementary DNA virus RNA agricultural inoculation article chlorosis cotton aphid cotton leafroll dwarf virus disease carrier disease transmission gene expression immunoblotting in vivo culture inoculation molecular cloning nonhuman priority journal replicon RNA virus viral plant disease virion virus detection virus genome Agrobacterium tumefaciens Animals Aphids Arabidopsis Cloning, Molecular Gene Expression Genome, Viral Gossypium Luteoviridae Plant Diseases South America Tobacco Transformation, Genetic Agrobacterium Aphididae Aphis gossypii Cauliflower mosaic virus Cotton leafroll dwarf virus Miridae RNA viruses
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_01681702_v175_n1_p64_Delfosse
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_01681702_v175_n1_p64_Delfosse
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spelling	todo:paper_01681702_v175_n1_p64_Delfosse2023-10-03T15:06:14Z Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana Delfosse, V.C. Casse, M.F. Agrofoglio, Y.C. Kresic, I.B. Hopp, H.E. Ziegler-Graff, V. Distéfano, A.J. Arabidopsis thaliana CLRDV Gossypium hirsutum Infectious clone Nicotiana benthamiana Polerovirus capsid protein complementary DNA virus RNA agricultural inoculation article chlorosis cotton aphid cotton leafroll dwarf virus disease carrier disease transmission gene expression immunoblotting in vivo culture inoculation molecular cloning Nicotiana benthamiana nonhuman priority journal replicon RNA virus viral plant disease virion virus detection virus genome Agrobacterium tumefaciens Animals Aphids Arabidopsis Cloning, Molecular Gene Expression Genome, Viral Gossypium Luteoviridae Plant Diseases South America Tobacco Transformation, Genetic Agrobacterium Agrobacterium tumefaciens Aphididae Aphis gossypii Arabidopsis thaliana Cauliflower mosaic virus Cotton leafroll dwarf virus Gossypium hirsutum Miridae Nicotiana benthamiana Polerovirus RNA viruses Cotton blue disease is the most important viral disease of cotton in the southern part of South America. Its etiological agent, cotton leafroll dwarf virus (CLRDV), is specifically transmitted to host plants by the aphid vector (Aphis gossypii) and any attempt to perform mechanical inoculations of this virus into its host has failed. This limitation has held back the study of this virus and the disease it causes. In this study, a full-length cDNA of CLRDV was constructed and expressed in vivo under the control of cauliflower mosaic virus 35S promoter. An agrobacterium-mediated inoculation system for the cloned cDNA construct of CLRDV was developed. Northern and immunoblot analyses showed that after several weeks the replicon of CLRDV delivered by Agrobacterium tumefaciens in Gossypium hirsutum plants gave rise to a systemic infection and typical blue disease symptoms correlated to the presence of viral RNA and P3 capsid protein. We also demonstrated that the virus that accumulated in the agroinfected plants was transmissible by the vector A. gossypii. This result confirms the production of biologically active transmissible virions. In addition, the clone was infectious in Nicotiana benthamiana plants which developed interveinal chlorosis three weeks postinoculation and CLRDV was detected both in the inoculated and systemic leaves. Attempts to agroinfect Arabidopsis thaliana plants were irregularly successful. Although no symptoms were observed, the P3 capsid protein as well as the genomic and subgenomic RNAs were irregularly detected in systemic leaves of some agroinfiltrated plants. The inefficient infection rate infers that A. thaliana is a poor host for CLRDV. This is the first report on the construction of a biologically-active infectious full-length clone of a cotton RNA virus showing successful agroinfection of host and non-host plants. The system herein developed will be useful to study CLRDV viral functions and plant-virus interactions using a reverse genetic approach. © 2013 Elsevier B.V. Fil:Hopp, H.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Distéfano, A.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_01681702_v175_n1_p64_Delfosse
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Arabidopsis thaliana CLRDV Gossypium hirsutum Infectious clone Nicotiana benthamiana Polerovirus capsid protein complementary DNA virus RNA agricultural inoculation article chlorosis cotton aphid cotton leafroll dwarf virus disease carrier disease transmission gene expression immunoblotting in vivo culture inoculation molecular cloning Nicotiana benthamiana nonhuman priority journal replicon RNA virus viral plant disease virion virus detection virus genome Agrobacterium tumefaciens Animals Aphids Arabidopsis Cloning, Molecular Gene Expression Genome, Viral Gossypium Luteoviridae Plant Diseases South America Tobacco Transformation, Genetic Agrobacterium Agrobacterium tumefaciens Aphididae Aphis gossypii Arabidopsis thaliana Cauliflower mosaic virus Cotton leafroll dwarf virus Gossypium hirsutum Miridae Nicotiana benthamiana Polerovirus RNA viruses
spellingShingle	Arabidopsis thaliana CLRDV Gossypium hirsutum Infectious clone Nicotiana benthamiana Polerovirus capsid protein complementary DNA virus RNA agricultural inoculation article chlorosis cotton aphid cotton leafroll dwarf virus disease carrier disease transmission gene expression immunoblotting in vivo culture inoculation molecular cloning Nicotiana benthamiana nonhuman priority journal replicon RNA virus viral plant disease virion virus detection virus genome Agrobacterium tumefaciens Animals Aphids Arabidopsis Cloning, Molecular Gene Expression Genome, Viral Gossypium Luteoviridae Plant Diseases South America Tobacco Transformation, Genetic Agrobacterium Agrobacterium tumefaciens Aphididae Aphis gossypii Arabidopsis thaliana Cauliflower mosaic virus Cotton leafroll dwarf virus Gossypium hirsutum Miridae Nicotiana benthamiana Polerovirus RNA viruses Delfosse, V.C. Casse, M.F. Agrofoglio, Y.C. Kresic, I.B. Hopp, H.E. Ziegler-Graff, V. Distéfano, A.J. Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana
topic_facet	Arabidopsis thaliana CLRDV Gossypium hirsutum Infectious clone Nicotiana benthamiana Polerovirus capsid protein complementary DNA virus RNA agricultural inoculation article chlorosis cotton aphid cotton leafroll dwarf virus disease carrier disease transmission gene expression immunoblotting in vivo culture inoculation molecular cloning Nicotiana benthamiana nonhuman priority journal replicon RNA virus viral plant disease virion virus detection virus genome Agrobacterium tumefaciens Animals Aphids Arabidopsis Cloning, Molecular Gene Expression Genome, Viral Gossypium Luteoviridae Plant Diseases South America Tobacco Transformation, Genetic Agrobacterium Agrobacterium tumefaciens Aphididae Aphis gossypii Arabidopsis thaliana Cauliflower mosaic virus Cotton leafroll dwarf virus Gossypium hirsutum Miridae Nicotiana benthamiana Polerovirus RNA viruses
description	Cotton blue disease is the most important viral disease of cotton in the southern part of South America. Its etiological agent, cotton leafroll dwarf virus (CLRDV), is specifically transmitted to host plants by the aphid vector (Aphis gossypii) and any attempt to perform mechanical inoculations of this virus into its host has failed. This limitation has held back the study of this virus and the disease it causes. In this study, a full-length cDNA of CLRDV was constructed and expressed in vivo under the control of cauliflower mosaic virus 35S promoter. An agrobacterium-mediated inoculation system for the cloned cDNA construct of CLRDV was developed. Northern and immunoblot analyses showed that after several weeks the replicon of CLRDV delivered by Agrobacterium tumefaciens in Gossypium hirsutum plants gave rise to a systemic infection and typical blue disease symptoms correlated to the presence of viral RNA and P3 capsid protein. We also demonstrated that the virus that accumulated in the agroinfected plants was transmissible by the vector A. gossypii. This result confirms the production of biologically active transmissible virions. In addition, the clone was infectious in Nicotiana benthamiana plants which developed interveinal chlorosis three weeks postinoculation and CLRDV was detected both in the inoculated and systemic leaves. Attempts to agroinfect Arabidopsis thaliana plants were irregularly successful. Although no symptoms were observed, the P3 capsid protein as well as the genomic and subgenomic RNAs were irregularly detected in systemic leaves of some agroinfiltrated plants. The inefficient infection rate infers that A. thaliana is a poor host for CLRDV. This is the first report on the construction of a biologically-active infectious full-length clone of a cotton RNA virus showing successful agroinfection of host and non-host plants. The system herein developed will be useful to study CLRDV viral functions and plant-virus interactions using a reverse genetic approach. © 2013 Elsevier B.V.
format	JOUR
author	Delfosse, V.C. Casse, M.F. Agrofoglio, Y.C. Kresic, I.B. Hopp, H.E. Ziegler-Graff, V. Distéfano, A.J.
author_facet	Delfosse, V.C. Casse, M.F. Agrofoglio, Y.C. Kresic, I.B. Hopp, H.E. Ziegler-Graff, V. Distéfano, A.J.
author_sort	Delfosse, V.C.
title	Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana
title_short	Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana
title_full	Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana
title_fullStr	Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana
title_full_unstemmed	Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana
title_sort	agroinoculation of a full-length cdna clone of cotton leafroll dwarf virus (clrdv) results in systemic infection in cotton and the model plant nicotiana benthamiana
url	http://hdl.handle.net/20.500.12110/paper_01681702_v175_n1_p64_Delfosse
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Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana

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