title: Evaluation of DNA fragments covering the entire genome of a monopartite begomovirus for induction of viral resistance in transgenic plants via gene silencing.
abstract: Tomato-infecting begomoviruses, a member of whitefly-transmitted geminivirus, cause the most devastating virus disease complex of cultivated tomato crops in the tropical and subtropical regions. Numerous strategies have been used to engineer crops for their resistance to geminiviruses. However, nearly all have concentrated on engineering the replication-associated gene (Rep), but not on a comprehensive evaluation of the entire virus genome. In this study, Tomato leaf curl Taiwan virus (ToLCTWV), a predominant tomato-infecting begomovirus in Taiwan, was subjected to the investigation of the viral gene fragments conferring resistance to geminiviruses in transgenic plants. Ten transgenic constructs covering the entire ToLCTWV genome were fused to a silencer DNA, the middle half of N gene of Tomato spot wilt virus (TSWV), to induce gene silencing and these constructs were transformed into Nicotiana benthamiana plants. Two constructs derived from IRC1 (intergenic region flanked with 5' end Rep) and C2 (partial C2 ORF) were able to render resistance to ToLCTWV in transgenic N. benthamiana plants. Transgenic plants transformed with two other constructs, C2C3 (overlapping region of C2 and C3 ORFs) and Rep2 (3' end of the C1 ORF), significantly delayed the symptom development. Detection of siRNA confirmed that the mechanism of resistance was via gene silencing. This study demonstrated for the first time the screening of the entire genome of a monopartite begomovirus to discover viral DNA fragments that might be suitable for conferring virus resistance, and which could be potential candidates for developing transgenic plants with durable and broad-spectrum resistance to a DNA virus via a gene silencing approach.
authors: Department of Plant Pathology, National Chung Hsing University, Taichung, 402, Taiwan.
authors: Ching-Yi
authors: CY
authors: Lin
authors: Wen-Shi
authors: WS
authors: Tsai
authors: Hsin-Mei
authors: HM
authors: Ku
authors: Fuh-Jyh
authors: FJ
authors: Jan
references: Virology. 1994 Oct;204(1):82-90
references: 8091687
references: Virology. 1996 Oct 15;224(2):390-404
references: 8874500
references: Plant Cell. 1992 Oct;4(10):1321-31
references: 1446172
references: Biochim Biophys Acta. 2010 Mar-Apr;1799(3-4):337-51
references: 20079472
references: J Virol. 2004 Sep;78(17):9487-98
references: 15308741
references: Transgenic Res. 2007 Jun;16(3):385-98
references: 17103242
references: J Virol. 2004 Jul;78(14):7465-77
references: 15220420
references: Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8261-6
references: 9223349
references: Trends Plant Sci. 2005 Mar;10(3):144-51
references: 15749473
references: Phytopathology. 2004 May;94(5):490-6
references: 18943768
references: Transgenic Res. 2006 Jun;15(3):291-304
references: 16779645
references: Virus Res. 2000 Nov;71(1-2):123-34
references: 11137167
references: Transgenic Res. 2011 Apr;20(2):261-70
references: 20559871
references: Nature. 2000 Sep 21;407(6802):319-20
references: 11014180
references: Nature. 2004 Sep 16;431(7006):343-9
references: 15372041
references: Plant Biotechnol J. 2007 Mar;5(2):207-20
references: 17309676
references: Plant Biotechnol J. 2005 Jul;3(4):385-97
references: 17173627
references: Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5640-4
references: 7777563
references: Nature. 2000 Mar 16;404(6775):293-6
references: 10749213
references: Nature. 2001 Jan 18;409(6818):363-6
references: 11201747
references: Nucleic Acids Res. 2006 Jan 18;34(2):462-71
references: 16421273
references: Adv Virus Res. 2009;75:185-220
references: 20109667
references: J Gen Virol. 1993 Jan;74 ( Pt 1):147-51
references: 8423446
references: Annu Rev Phytopathol. 2010;48:269-91
references: 20687833
references: Mol Plant Microbe Interact. 2002 Mar;15(3):203-8
references: 11952122
references: Plant Mol Biol. 2004 Nov;56(4):601-11
references: 15630623
references: Nature. 2001 Jun 14;411(6839):834-42
references: 11459066
references: Phytopathology. 2003 Dec;93(12):1485-95
references: 18943612
references: Mol Plant Microbe Interact. 2007 Jun;20(6):717-26
references: 17555279
references: Virology. 2001 Dec 5;291(1):110-25
references: 11878881
references: Crit Rev Biochem Mol Biol. 2000;35(2):105-40
references: 10821479
references: J Gen Virol. 2000 Aug;81(Pt 8):2103-2109
references: 10900050
references: Plant Mol Biol. 1997 Jan;33(2):351-7
references: 9037152
references: Plant Mol Biol. 2009 Jun;70(3):265-72
references: 19300911
references: J Virol. 2005 Aug;79(15):9885-95
references: 16014949
references: Curr Biol. 1999 Aug 26;9(16):R599-601
references: 10469584
references: Trends Plant Sci. 2003 Mar;8(3):128-34
references: 12663223
references: J Gen Virol. 2004 Jun;85(Pt 6):1745-1749
references: 15166460
references: Arch Virol. 2006 Dec;151(12):2349-63
references: 16862387
references: Arch Virol. 2008;153(4):783-821
references: 18256781
references: Virus Res. 2004 Jun 1;102(1):85-96
references: 15068884
references: Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6721-5
references: 1862097
references: J Virol. 2005 Jun;79(12):7812-8
references: 15919934
references: Plant Mol Biol. 2007 Jul;64(5):549-57
references: 17492253
references: Oligonucleotides. 2007 Summer;17(2):251-7
references: 17638528
references: Acta Virol. 2008;52(3):143-9
references: 18999888
references: Plant Cell. 1990 Apr;2(4):279-289
references: 12354959
references: Science. 1999 Oct 29;286(5441):950-2
references: 10542148
references: Virology. 1994 Nov 1;204(2):847-50
references: 7941358
references: J Gen Virol. 2000 Jan;81(Pt 1):235-42
references: 10640563
references: Arch Virol. 2003 Dec;148(12):2341-52
references: 14648290
references: Trends Genet. 2001 Aug;17(8):449-59
references: 11485817
references: J Virol. 2003 Jun;77(12):6785-98
references: 12767999