title: Citrus leaf blotch virus invades meristematic regions in Nicotiana benthamiana and citrus.
abstract: To invade systemically host plants, viruses need to replicate in the infected cells, spread to neighbouring cells through plasmodesmata and move to distal parts of the plant via sieve tubes to start new infection foci. To monitor the infection of Nicotiana benthamiana plants by Citrus leaf blotch virus (CLBV), leaves were agroinoculated with an infectious cDNA clone of the CLBV genomic RNA expressing green fluorescent protein (GFP) under the transcriptional control of a duplicate promoter of the coat protein subgenomic RNA. Fluorescent spots first appeared in agroinfiltrated leaves 11-12 days after infiltration, indicating CLBV replication. Then, after entering the phloem vascular system, CLBV was unloaded in the upper parts of the plant and invaded all tissues, including flower organs and meristems. GFP fluorescence was not visible in citrus plants infected with CLBV-GFP. Therefore, to detect CLBV in meristematic regions, Mexican lime (Citrus aurantifolia) plants were graft inoculated with CLBV, with Citrus tristeza virus (CTV), a virus readily eliminated by shoot-tip grafting in vitro, or with both simultaneously. Although CLBV was detected by hybridization and real-time reverse transcription-polymerase chain reaction (RT-PCR) in 0.2-mm shoot tips in all CLBV-inoculated plants, CTV was not detected. These results explain the difficulty in eliminating CLBV by shoot-tip grafting in vitro.
authors.affiliation: Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias-IVIA, Moncada, Valencia 46113, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Agüero
authors.forename: María Carmen
authors.initials: MC
authors.lastname: Vives
authors.forename: Karelia
authors.initials: K
authors.lastname: Velázquez
authors.forename: Susana
authors.initials: S
authors.lastname: Ruiz-Ruiz
authors.forename: Jose
authors.initials: J
authors.lastname: Juárez
authors.forename: Luis
authors.initials: L
authors.lastname: Navarro
authors.forename: Pedro
authors.initials: P
authors.lastname: Moreno
authors.forename: José
authors.initials: J
authors.lastname: Guerri
mesh_terms: D002957:Citrus / Q000821:virology*; D035264:Flowers / Q000821:virology; D018519:Meristem / Q000821:virology*; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000472:pathogenicity*; D014026:Nicotiana / Q000821:virology*
references.citation: J Virol. 2008 Apr;82(8):4064-71
references.pmid: 18272576
references.citation: Plant Physiol. 1998 Oct;118(2):373-85
references.pmid: 9765523
references.citation: Virology. 2002 Apr 10;295(2):328-36
references.pmid: 12033792
references.citation: J Virol. 2005 Dec;79(24):15209-17
references.pmid: 16306592
references.citation: Virology. 2010 Oct 25;406(2):360-9
references.pmid: 20708769
references.citation: J Gen Virol. 2002 Jun;83(Pt 6):1493-1504
references.pmid: 12029165
references.citation: J Gen Virol. 2004 Mar;85(Pt 3):749-759
references.pmid: 14993661
references.citation: Plant J. 2000 Aug;23(3):349-62
references.pmid: 10929128
references.citation: Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6286-90
references.pmid: 8692807
references.citation: Methods Mol Biol. 2008;451:33-54
references.pmid: 18370246
references.citation: Virology. 2006 Jan 5;344(1):169-84
references.pmid: 16364748
references.citation: Plant Dis. 2004 Aug;88(8):906
references.pmid: 30812527
references.citation: Annu Rev Phytopathol. 2007;45:73-100
references.pmid: 17362202
references.citation: J Virol. 2008 Jul;82(13):6546-56
references.pmid: 18434397
references.citation: Plant J. 1992 Jul;2(4):549-57
references.pmid: 1344890
references.citation: Cell. 1999 Jun 11;97(6):743-54
references.pmid: 10380926
references.citation: Plant Cell. 1999 Mar;11(3):309-22
references.pmid: 10072393
references.citation: Protoplasma. 2001;218(1-2):57-66
references.pmid: 11732321
references.citation: J Gen Virol. 1972 Mar;14(3):273-6
references.pmid: 4112038
references.citation: Arch Virol. 2001;146(2):357-68
references.pmid: 11315643
references.citation: Virology. 2001 Aug 15;287(1):225-33
references.pmid: 11504557
references.citation: Virology. 1995 Jan 10;206(1):276-86
references.pmid: 7831782
references.citation: Mol Plant Microbe Interact. 2012 Oct;25(10):1326-37
references.pmid: 22670755
references.citation: J Gen Virol. 2009 Jul;90(Pt 7):1767-1774
references.pmid: 19282434
references.citation: Plant Cell. 1992 Nov;4(11):1405-1412
references.pmid: 12297638
references.citation: Virus Genes. 2012 Feb;44(1):131-40
references.pmid: 21948005
references.citation: J Virol Methods. 2009 Sep;160(1-2):57-62
references.pmid: 19406167
references.citation: Plant Cell. 2002 Jul;14(7):1497-508
references.pmid: 12119370
references.citation: Plant Physiol. 2001 Jan;125(1):209-18
references.pmid: 11154330
references.citation: Mol Plant Pathol. 2008 Nov;9(6):787-97
references.pmid: 19019007
references.citation: J Virol Methods. 2007 Nov;145(2):96-105
references.pmid: 17573130
references.citation: J Virol. 2010 Mar;84(5):2477-89
references.pmid: 20015979
references.citation: Virology. 2011 Mar 15;411(2):237-43
references.pmid: 21239029
references.citation: Trends Plant Sci. 2005 Aug;10(8):376-82
references.pmid: 16023398
references.citation: Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):3323-8
references.pmid: 9501261
references.citation: Plant J. 1995 Jun;7(6):1045-53
references.pmid: 7599646
references.citation: Plant Cell. 1997 Aug;9(8):1381-1396
references.pmid: 12237387
references.citation: Plant Physiol. 2005 Aug;138(4):1842-52
references.pmid: 16040651
references.citation: Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10208-12
references.pmid: 1438210