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On-line Access: 2011-02-04

Received: 2010-04-28

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Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.2 P.83-92


Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite

Author(s):  Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang, Xue-ping Zhou

Affiliation(s):  State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China

Corresponding email(s):   zzhou@zju.edu.cn

Key Words:  Virus induced gene silencing, Geminivirus, Alphasatellite, Nicotiana tabacum

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Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang, Xue-ping Zhou. Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite[J]. Journal of Zhejiang University Science B, 2011, 12(2): 83-92.

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author="Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang, Xue-ping Zhou",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

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%T Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite
%A Chang-jun Huang
%A Tong Zhang
%A Fang-fang Li
%A Xin-yue Zhang
%A Xue-ping Zhou
%J Journal of Zhejiang University SCIENCE B
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%P 83-92
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%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000157

T1 - Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite
A1 - Chang-jun Huang
A1 - Tong Zhang
A1 - Fang-fang Li
A1 - Xin-yue Zhang
A1 - Xue-ping Zhou
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 2
SP - 83
EP - 92
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1000157

Virus-induced gene silencing (VIGS) is a recently developed technique for characterizing the function of plant genes by gene transcript suppression and is increasingly used to generate transient loss-of-function assays. Here we report that the 2mDNA1, a geminivirus satellite vector, can induce efficient gene silencing in Nicotiana tabacum with Tobacco curly shoot virus. We have successfully silenced the β-glucuronidase (GUS) gene in GUS transgenic N. tabacum plants and the sulphur desaturase (Su) gene in five different N. tabacum cultivars. These pronounced and severe silencing phenotypes are persistent and ubiquitous. Once initiated in seedlings, the silencing phenotype lasted for the entire life span of the plants and silencing could be induced in a variety of tissues and organs including leaf, shoot, stem, root, and flower, and achieved at any growth stage. This system works well between 18–32 °C. We also silenced the NtEDS1 gene and demonstrated that NtEDS1 is essential for N gene mediated resistance against Tobacco mosaic virus in N. tabacum. The above results indicate that this system has great potential as a versatile VIGS system for routine functional analysis of genes in N. tabacum.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Becker, A., Lange, M., 2010. VIGS—genomics goes functional. Trends Plant. Sci., 15(1):1-4.

[2]Briddon, R.W., Stanley, J., 2006. Subviral agents associated with plant single-stranded DNA viruses. Virology, 344(1):198-210.

[3]Burch-Smith, T.M., Anderson, J.C., Martin, G.B., Dinesh-Kumar, S.P., 2004. Applications and advantages of virus-induced gene silencing for gene function studies in plants. Plant J., 39(5):734-746.

[4]Bustin, S.A., Benes, V., Garson, J.A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M.W., Shipley, G.L., et al., 2009. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin. Chem., 55(4):611-622.

[5]Cai, X.Z., Wang, C.C., Xu, Y.P., Xu, Q.F., Zheng, Z., Zhou, X.P., 2007. Efficient gene silencing induction in tomato by a viral satellite DNA vector. Virus Res., 125(2): 169-175.

[6]Carrillo-Tripp, J., Shimada-Beltran, H., Rivera-Bustamante, R., 2006. Use of geminiviral vectors for functional genomics. Curr. Opin. Plant Biol., 9(2):209-215.

[7]Chellappan, P., Vanitharani, R., Ogbe, F., Fauquet, C.M., 2005. Effect of temperature on geminivirus-induced RNA silencing in plants. Plant Physiol., 138(4):1828-1841.

[8]Cogoni, C., Macino, G., 1997. Isolation of quelling-defective (qde) mutants impaired in posttranscriptional transgene-induced gene silencing in Neurospora crassa. PNAS, 94(19):10233-10238.

[9]Cogoni, C., Macino, G., 2000. Post-transcriptional gene silencing across kingdoms. Curr. Opin. Genet. Dev., 10(6):638-643.

[10]Cui, X.F., Tao, X.R., Xie, Y., Fauquet, C.M., Zhou, X.P., 2004. A DNAβ associated with Tomato yellow leaf curl China virus is required for symptom induction. J. Virol., 78(24):13966-13974.

[11]Ding, C.J., Qing, L., Li, Z.H., Liu, Y., Qian, Y.J., Zhou, X.P., 2009. Genetic determinants of symptoms on viral DNA satellites. Appl. Environ. Microbiol., 75(16):5380-5389.

[12]Ding, X.S., Schneider, W.L., Chaluvadi, S.R., Mian, M.A., Nelson, R.S., 2006. Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Mol. Plant Microbe Interact., 19(11):1229-1239.

[13]English, J.J., Mueller, E., Baulcombe, D.C., 1996. Suppression of virus accumulation in transgenic plants exhibiting silencing of nuclear genes. Plant Cell, 8(2):179-188.

[14]Falk, A., Feys, B.J., Frost, L.N., Jones, J.D., Daniels, M.J., Parker, J.E., 1999. EDS1, an essential component of R gene-mediated disease resistance in Arabidopsis has homology to eukaryotic lipases. PNAS, 96(6):3292-3297.

[15]Fauquet, C.M., Briddon, R.W., Brown, J.K., Moriones, E., Stanley, J., Zerbini, M., Zhou, X.P., 2008. Geminivirus strain demarcation and nomenclature. Arch. Virol., 153(4):783-821.

[16]Fire, A., Xu, S., Montgomery, M.K., Kostas, S.A., Driver, S.E., Mello, C.C., 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391(6669):806-811.

[17]Gossele, V., Fache, I., Meulewaeter, F., Cornelissen, M., Metzlaff, M., 2002. SVISS—a novel transient gene silencing system for gene function discovery and validation in tobacco plants. Plant J., 32(5):859-866.

[18]Huang, C.J., Xie, Y., Zhou, X.P., 2009. Efficient virus-induced gene silencing in plants using a modified geminivirus DNA1 component. Plant Biotechnol. J., 7(3):254-265.

[19]Igarashi, A., Yamagata, K., Sugai, T., Takahashi, Y., Sugawara, E., Tamura, A., Yaegashi, H., Yamagishi, N., Takahashi, T., Isogai, M., et al., 2009. Apple latent spherical virus vectors for reliable and effective virus-induced gene silencing among a broad range of plants including tobacco, tomato, Arabidopsis thaliana, cucurbits, and legumes. Virology, 386(2):407-416.

[20]Jefferson, R.A., Kavanagh, T.A., Bevan, M.W., 1987. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J., 6(13):3901-3907.

[21]Koncz, C., Mayerhofer, R., Koncz-Kalman, Z., Nawrath, C., Reiss, B., Redei, G.P., Schell, J., 1990. Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana. EMBO J., 9(5):1337-1346.

[22]Kumagai, M.H., Donson, J., Dellacioppa, G., Harvey, D., Hanley, K., Grill, L.K., 1995. Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. PNAS, 92(5):1679-1683.

[23]Li, Z.H., Xie, Y., Zhou, X.P., 2005. Tobacco curly shoot virus DNAβ is not necessary for infection but intensifies symptoms in a host-dependent manner. Phytopathology, 95(8):902-908.

[24]Liu, Y.L., Schiff, M., Marathe, R., Dinesh-Kumar, S.P., 2002a. Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to Tobacco mosaic virus. Plant J., 30(4):415-429.

[25]Liu, Y.L., Schiff, M., Dinesh-Kumar, S.P., 2002b. Virus-induced gene silencing in tomato. Plant J., 31(6):777-786.

[26]Mansoor, S., Briddon, R.W., Zafar, Y., Stanley, J., 2003. Geminivirus disease complexes: an emerging threat. Trends Plant Sci., 8(3):128-134.

[27]Mansoor, S., Zafar, Y., Briddon, R.W., 2006. Geminivirus disease complexes: the threat is spreading. Trends Plant Sci., 11(5):209-212.

[28]Napoli, C., Lemieux, C., Jorgensen, R., 1990. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell., 2(4):279-289.

[29]Peart, J.R., Cook, G., Feys, B.J., Parker, J.E., Baulcombe, D.C., 2002. An EDS1 orthologue is required for N-mediated resistance against Tobacco mosaic virus. Plant J., 29(5):569-579.

[30]Purkayastha, A., Dasgupta, I., 2009. Virus-induced gene silencing: a versatile tool for discovery of gene functions in plants. Plant Physiol. Biochem., 47(11-12):967-976.

[31]Ramachandran, V., Chen, X., 2008. Small RNA metabolism in Arabidopsis. Trends Plant Sci., 13(7):368-374.

[32]Ryu, C.M., Anand, A., Kang, L., Mysore, K.S., 2004. Agrodrench: a novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse Solanaceous species. Plant J., 40(2):322-331.

[33]Szittya, G., Silhavy, D., Molnar, A., Havelda, Z., Lovas, A., Lakatos, L., Banfalvi, Z., Burgyan, J., 2003. Low temperature inhibits RNA silencing-mediated defence by the control of siRNA generation. EMBO J., 22(3):633-640.

[34]Tao, X.R., Zhou, X.P., 2004. A modified viral satellite DNA that suppresses gene expression in plants. Plant J., 38(5):850-860.

[35]Tuttle, J.R., Idris, A.M., Brown, J.K., Haigler, C.H., Robertson, D., 2008. Geminivirus-mediated gene silencing from Cotton leaf crumple virus is enhanced by low temperature in cotton. Plant Physiol., 148(1):41-50.

[36]Voinnet, O., 2001. RNA silencing as a plant immune system against viruses. Trends Genet., 17(8):449-459.

[37]Whitham, S., Dinesh-Kumar, S.P., Choi, D., Hehl, R., Corr, C., Baker, B., 1994. The product of the Tobacco mosaic virus resistance gene N: similarity to toll and the interleukin-1 receptor. Cell, 78(6):1101-1115.

[38]Wiermer, M., Feys, B.J., Parker, J.E., 2005. Plant immunity: the EDS1 regulatory node. Curr. Opin. Plant Biol., 8(4):383-389.

[39]Wu, P.J., Zhou, X.P., 2005. Interaction between a nanovirus-like component and the Tobacco curly shoot virus/ satellite complex. Acta Biochim. Biophys. Sin., 37(1):25-31.

[40]Xie, Z.X., Johansen, L.K., Gustafson, A.M., Kasschau, K.D., Lellis, A.D., Zilberman, D., Jacobsen, S.E., Carrington, J.C., 2004. Genetic and functional diversification of small RNA pathways in plants. PLoS Biol., 2(5):e104.

[41]Zhou, X.P., Liu, Y.L., Calvert, L., Munoz, C., Otim-Nape, G.W., Robinson, D.J., Harrison, B.D., 1997. Evidence that DNA-A of a geminivirus associated with severe cassava mosaic disease in Uganda has arisen by interspecific recombination. J. Gen. Virol., 78(8):2101-2111.

[42]Zhou, X.P., Xie, Y., Zhang, Z.K., 2001. Molecular characterization of a distinct begomovirus infecting tobacco in Yunnan, China. Arch. Virol., 146(8):1599-1606.

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