Agroinfiltration Transient Assay to Evaluate the Potential of IHPRNA Constructs for Lily Mottle Virus Resistance. | [International Journal of Agriculture and Biology • 2019]

Author(s):

1. Pinsan Xu: Plants, Bacterial Strains, Plasmids and other Reagents, School of Life Science and Medicine, Dalian University of Technology, Panjin, China: School of life science and biotechnology, Dalian University of Technology, Dalian, China

2. Yue Qi: Plants, Bacterial Strains, Plasmids and other Reagents, School of Life Science and Medicine, Dalian University of Technology, Panjin, China

3. Xinlei Wang: Plants, Bacterial Strains, Plasmids and other Reagents, School of Life Science and Medicine, Dalian University of Technology, Panjin, China

4. Zhengyao Zhang: Plants, Bacterial Strains, Plasmids and other Reagents, School of Life Science and Medicine, Dalian University of Technology, Panjin, China

Abstract:

In order to obtain a resistant strain of Lily mottle virus (LMoV), three RNA interference (RNAi) vectors with intron hairpin RNA structure (ihpRNA) were constructed and transformed into Agrobacterium GV 3101 strain. The gene hotspots of LMoV coated protein (CP) and cylindrical inclusion (CI) were selected, and the positive and negative fragments of the target gene were inserted into both sides of the Pdk intron fragment of plant expression vector pRNAi-LIC by one-step, zero-background ligation-independent cloning (OZ-LIC) method. Antiviral analysis was performed using Agrobacterium-mediated transientexpression assay. RT-PCR and indirect ELISA results showed that the hairpin RNA from the reverse repeats of CI-517 bp and CI-295 bp could interfere the virus effectively, while the CP-400 bp hairpin's impact was not significant. The work illustrates that the genetically modified genome could LMoV.

Page(s): 67-70

DOI: 10.17957/IJAB/15.0863

Published: Journal: International Journal of Agriculture and Biology, Volume: 21, Issue: 1, Year: 2019

Keywords:

Keywords are not available for this article.

References:

[1] Asjes , C.J., 2000.Control of aphid-borne Lily symptomless virus and Lily mottle virus in Lilium in the Netherlands,Vir. Res. 71 23 -32

[2] Chen , Y.,W.T.Shen,P.Yan, 2011.Fast construction of dsRNA prokaryotic expression vector of Papaya ringspot virus CP gene,J. Trop. Org. 2 113 -116

[3] Ding , Y.J.,Z.S.Zhou,Y.F.Dong, 2011.Construction of the RNAi vector of GVA MP gene,Chin. Agric. Sci. Bull 27 256 -259

[4] Helliwell , C.,P.Waterhouse, 2003.Constructs and methods for highthroughput gene silencing in plants,Methods 30 289 -295

[5] Hirai , S. , S.,Oka , E.,Adachi and H.,Kodama, 2007.The effects of spacer sequences on silencing efficiency of plant RNAi vectors,Plant Cell Rep 26 651 -659

[6] Kusaba , M., 2004.RNA interference in crop plants,Curr. Opin. Biotechnol 15 139 -143

[7] Sherman , J.H.,S.Choudhuri,J.L.Vicini, 2015.RNAi technologies in agricultural biotechnology,Regul 73 671 -680

[8] Tenllado , F.,C.Llave,J.R.Díaz-Ruíz, 2004.RNA interference as a new biotechnological tool for the control of virus diseases in plants,Vir. Res. 102 85 -96

[9] Urcuqui-Inchima , S.,A.L.Haenni,F.Bernardi, 2001.Potyvirus proteins: a wealth of functions,Vir. Res. 74 157 -175

[10] Wesley , S.V.,C.A.Helliwell,N.A.Smith,M.Wang,D.T.Rouse,Q.Liu,P.S.Gooding,S.P.Singh,D.Abbott,P.A.Stoutjesdijk,S.P.Robinson,A.P.Gleave,A.G.Green and P.M. Waterhouse, 2001.Construct design for efficient, effective and high-throughput gene silencing in plants,Plant 27 581 -590

[11] Xu , G.Y.,N.Sui,Y.Tang, 2010.One-step, zero-background ligationindependent cloning intron-containing hairpin RNA constructs for RNAi in plants, 187 240 -250

[12] Xu , P.S.,C.W.Zhao,Z.Y.Zhang, 2017.Identification of an RNA silencing suppressor encoded by lily symptomless virus,Intl. J. Agric. Biol 19 1259 -1264

[13] Xu , P.S.,J.F.Bai,J.W.Liu, 2011.Construction of LSV and LMoV binary virus resistant RNAi vector using Gateway technology,Chin. Agric. Sci. Bull 27 144 -147

[14] Yan , P.,W.T.Shen,X.Y.Li, 2013.Progress in construction of hpRNA vector for plant RNAi, 9 7 -12

[15] Zhang , Z.Y.,X.Y.Xia,J.Song,P.S.Xu, 2017.Accumulation of Viral Coat Protein in Chloroplasts of Lily Leaves Infected with Lily Mottled Virus, Intl, 19 1265 -1269

Citations

Downloads

Views