Please wait a minute...
Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front. Agric. China    2008, Vol. 2 Issue (2) : 125-130     DOI: 10.1007/s11703-008-0037-z
|
Inverted-repeat transgenic maize plants resistant to sugarcane mosaic virus
BAI Yunfeng1, YANG Hongchun2, QU Lin2, ZHENG Jun2, ZHANG Jinpeng2, WANG Maoyan2, XIE Wan2, ZHOU Xiaomei2, WANG Guoying3
1.Institute of Crop Genetics, Shanxi Academy of Agricultural Sciences;State Key Laboratory of Agrobiotechnology, China Agricultural University; 2.State Key Laboratory of Agrobiotechnology, China Agricultural University; 3.State Key Laboratory of Agrobiotechnology, China Agricultural University;Institute of Crop Sciences, Chinese Academy of Agricultural Sciences;
 Download: PDF(259 KB)   HTML
 Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract RNA silencing is a post-transcriptional gene-silencing phenomenon induced by double-stranded RNA (dsRNA). In an attempt to generate dsRNA-mediated transgenic maize plants resistant to sugarcane mosaic virus (SCMV), we cloned SCMV NIb gene-specific sequences and inserted it into the binary vector p3301 in the sense and antisense orientations (named SCMVirNIb), which could produce RNAs capable of duplex formation in plant cells. Maize immature embryos were co-cultured with Agrobacterium carrying two vectors, one marker-free vector harboring the SCMVirNIb and one vector harboring bar gene as the selective marker. Resistant calli were recovered by selection on medium containing Biolaphos. Among the regenerated plantlets from resistant calli, 14 plants have been certified to contain SCMVirNIb by PCR amplification and DNA dot blot. T1 plants derived from the 14 plants were challenged in a greenhouse with SCMV inoculums and the percentages of resistant plants in 11 T1 lines were higher than 60%. One plant in the T1 line was found to carry SCMVirNIb without bar gene by PCR assay.T2 plants derived from T1 SCMV resistant transgenic plants were challenged with SCMV inoculums in field. The percentages of resistant plants from 3 lines, including the line derived from the marker-free transgenic plant, were higher than 85%. The non-transgenic control plants were all susceptible. Further molecular analysis confirmed that the resistant plants from the marker-free transgenic line contained SCMVirNIb but not the bar gene.
Issue Date: 05 June 2008
 Cite this article:   
BAI Yunfeng,YANG Hongchun,QU Lin, et al. Inverted-repeat transgenic maize plants resistant to sugarcane mosaic virus[J]. Front. Agric. China, 2008, 2(2): 125-130.
 URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-008-0037-z
http://academic.hep.com.cn/fag/EN/Y2008/V2/I2/125
1 Anandalakshmi R Marathe R Ge X Herr J M Mau C Mallory A Pruss G Bowman L Vance V B 2000 A calmodulin-relatedprotein that suppresses posttranscriptional gene silencing in plantsScience290142144.
doi:10.1126/science.290.5489.142
2 Andrew P 2000 Suppression of posttranscriptional gene silencing by aplant viral protein localized in the nucleusEMBO J1916721680.
doi:10.1093/emboj/19.7.1672
3 Bai Y F Zhao J F Zheng J Zhang J P Wang M Y Gou M Y Dong Z G Yang H C Wang G Y 2006 SCMV-resistant transgenic maize mediated by antisense cp geneActa AgronicaSinica32(5)661665 (in Chinese)
4 Fire A Xu S Montgomery M K Kostas S A Driver S E Mellow C C 1998 Potent and specific genetic interference by double-strandedRNA in Caenorhabditis elegansNature391806811.
doi:10.1038/35888
5 Goldbach R Bucher E Prins R 2003 Resistance mechanisms to plant viruses:an overviewVirus Res92207212.
doi:10.1016/S0168‐1702(02)00353‐2
6 Guo H S Ding S W 2002 A viral proteininhibits the long range signaling activity of the gene silencing signalEMBO J21398407.
doi:10.1093/emboj/21.3.398
7 Ketting R F Haverkamp T H van Luenen H G Plasterk R H 1999 Mut-7 of C. elegans, requiredfor transposon silencing and RNA interference, is a homolog of Wernersyndrome helicase and RnaseD.Cell99133141.
doi:10.1016/S0092‐8674(00)81645‐1
8 Liu X H Zhang H W Liu X Liu X J Tan Z - B Rong T Z 2005 Isolation of the capsid protein gene of Maize Dwarf MosaicVirus and its transformation in maizeChinJ Biotechnol21(1)144148 (in Chinese)
9 Missiou A Kalantidis K Boutla1 A Tzortzakaki S Martin Tabler M Tsagris M 2004 Generation of transgenic potato plantshighly resistant to potato virus Y (PVY) through RNA silencingMol Breed14185197.
doi:10.1023/B:MOLB.0000038006.32812.52
10 Murry E Launie G 1993 Transgeniccorn plants expressing MDMV strain B coat protein are resistant tomixed infection of Maize Dwarf Mosaic Virus and Maize Chlorotic MottleVirusBio/technology1115591564.
doi:10.1038/nbt1293‐1559
11 Plasterk R H A 2002 RNA silencing: the genome's immune systemScience29612631265.
doi:10.1126/science.1072148
12 Silhavy D Molnar A Lucioli A Szittya G Hornyik C Tavazza M Burgyan J 2002 A viral proteinsuppresses RNA silencing and binds silencing-generated 21- to 25-nucleotidedouble-stranded RNAsEMBO J2130703080.
doi:10.1093/emboj/cdf312
13 Smith N A Singh S P Wang M B Stoutjesdijk P A Green A G Waterhou P A 2000 Total silencing by intron-spliced hairpinRNAsNature407319320.
doi:10.1038/35036500
14 Tabara H Sarkissian M Kelly W G Fleenor J Grishok A Timmons L Fire A Mello C C 1999 The rde-1 gene, RNA interference, and transposon silencing in C. elegans. Cell99123132.
doi:10.1016/S0092‐8674(00)81644‐X
15 Tenllado F Diaz-Ruiz J R 2001 Double-strandRNA-mediated interference with plant virus infectionJ Virol751228812297.
doi:10.1128/JVI.75.24.12288‐12297.2001
16 Wang M B Abbott D C Waterhouse P M 2000 A single copy of a virus-derived transgeneencoding hairpin RNA gives immunity to barley yellow dwarf virusMol Plant Pathol1347356.
doi:10.1046/j.1364‐3703.2000.00038.x
17 Waterhouse P M Graham M W Wang M B 1998 Virus resistance and gene silencing inplants can be induced by simultaneous expression of sense and antisenseRNAProc Natl Acad Sci USA951395913964.
doi:10.1073/pnas.95.23.13959
18 Zhang R Wang G Y Zhang X H Zhao H J 2001 Agrobacterium tumefaciens mediatedmaize transformationJ Agric Biotechno9(1)4548 (in Chinese)
19 Zhou X M 2001 Transfer of maize dwarf mosaic virus coat protein geneinto maize inbred line and the resistance of transgenic plant to MDMV.Dissertation for the Doctoral Degree.BeijingChina Agricultural University (in Chinese)
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed