|
Special Issue:
植物病毒合辑Plant Virus
|
|
|
|
| Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants |
Pavel Cejnar1, Ludmila Ohnoutková2, Jan Ripl1, Tomáš Vl?ko2, Jiban Kumar Kundu1
|
1 Division of Crop Protection and Plant Health, Crop Research Institute, Prague 16106, Czech Republic
2 Department of Chemical Biology and Genetics, Centre of the Region Hana for Biotechnological and Agricultural Research Palacký University, Olomouc 78371, Czech Republic |
|
|
|
|
Abstract Wheat dwarf virus (WDV), an important cereal pathogen, is closely related to Maize streak virus (MSV), a model virus of the Mastrevirus genus. Based on its similarity to known MSV resistance strategies, a truncated part of the WDV replication-associated (RepA) gene (WDVRepA215) and the WDV RepA gene with a mutated retinoblastoma-related protein (RBR) interaction domain (WDVRepA215RBRmut) were cloned into the pIPKb002 expression vector and transformed into immature embryos of spring barley cv. Golden Promise plants through Agrobacterium-mediated transformation. A detailed study of T1-generation plants infected by leafhoppers (Psammotettix alienus) fed on infection sources of variable strength was performed over a 5-week period encompassing the initial stages of virus infection. A DNA WDV TaqMan qPCR assay normalized using the DNA puroindoline-b SYBR Green qPCR assay for samples on a per week basis revealed an approximately 2-week delay in WDVRepA215RBRmut plants to WDVRepA215 plants before significant increases in the WDV viral levels occurred. Both WDVRepA215 and WDVRepA215RBRmut plants showed similar levels of transgenic transcripts over the screened period; however, the transgenic plants also showed increased numbers of infected plants compared to the control plants.
|
|
Received: 10 October 2017
Accepted:
|
| Fund: This work was equally supported by the Czech Ministry of Education, Youth and Sports funding programme LH12161, and the Czech Ministry of Agriculture funding programme MZE RO0417. |
|
Corresponding Authors:
Correspondence Jiban Kumar Kundu, E-mail: jiban@vurv.cz
|
| About author: Pavel Cejnar, E-mail: cejnar@vurv.cz; |
Cite this article:
Pavel Cejnar, Ludmila Ohnoutková, Jan Ripl, Tomá? Vl?ko, Jiban Kumar Kundu.
2018.
Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants. Journal of Integrative Agriculture, 17(11): 2492-2500.
|
Boulton M I. 2002. Functions and interactions of mastrevirus gene products. Physiological and Molecular Plant Pathology, 60, 243–255.
Cejnar P, Ohnoutková L, Kostková M, Vl?ko T, Kumar J K. 2017. Transformation of barley with partial WDV replication protein. In: 1st COST iPLANTA Conference Creating a Plant RNAi Research Network. EU COST Action, Rome, Italy.
Dinant S, Ripoll C, Pieper M, David C. 2004. Phloem specific expression driven by wheat dwarf geminivirus V-sense promoter in transgenic dicotyledonous species. Physiologia Plantarum, 121, 108–116.
Edwards K, Johnstone C, Thompson C. 1991. A simple and rapid method for the preparation of plant genomic DNA for Pcr analysis. Nucleic Acids Research, 19, 1349–1349.
Galassi E, Gazzelloni G, Taddei F, Muccilli V, Gazza L, Pogna N. 2012. Kernel texture and hordoindoline patterns in barley (Hordeum vulgare). Molecular Breeding, 30, 1551–1562.
Gautier M F, Cosson P, Guirao A, Alary R, Joudrier P. 2000. Puroindoline genes are highly conserved in diploid ancestor wheats and related species but absent in tetraploid Triticum species. Plant Science, 153, 81–91.
Guiu-Aragones C, Diaz-Pendon J A, Martin-Hernandez A M. 2015. Four sequence positions of the movement protein of Cucumber mosaic virus determine the virulence against cmv1-mediated resistance in melon. Molecular Plant Pathology, 16, 675–684.
Gutierrez C. 1999. Geminivirus DNA replication. Cellular and Molecular Life Sciences, 56, 313–329.
Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer P, Kumlehn J. 2007. A set of modular binary vectors for transformation of cereals. Plant Physiology, 145, 1192–1200.
Horvath G V, Pettko-Szandtner A, Nikovics K, Bilgin M, Boulton M, Davies J W, Gutierrez C, Dudits D. 1998. Prediction of functional regions of the maize streak virus replication-associated proteins by protein-protein interaction analysis. Plant Molecular Biology, 38, 699–712.
Jarosova J, Kundu J K. 2010. Validation of reference genes as internal control for studying viral infections in cereals by quantitative real-time RT-PCR. BMC Plant Biology, 10, 146.
Kis A, Tholt G, Ivanics M, Varallyay E, Jenes B, Havelda Z. 2016. Polycistronic artificial miRNA-mediated resistance to Wheat dwarf virus in barley is highly efficient at low temperature. Molecular Plant Pathology, 17, 427–437.
Koonin E V, Ilyina T V. 1992. Geminivirus replication proteins are related to prokaryotic plasmid rolling circle DNA-replication initiator proteins. Journal of General Virology, 73, 2763–2766.
Kundu J K, Gadiou S, ?ervená G. 2009. Discrimination and genetic diversity of Wheat dwarf virus in the Czech Republic. Virus Genes, 38, 468–474.
Larkin M A, Blackshields G, Brown N P, Chenna R, McGettigan P A, McWilliam H, Valentin F, Wallace I M, Wilm A, Lopez R, Thompson J D, Gibson J, Higgins D G. 2007. Clustal W and Clustal X version 2.0. Bioinformatics, 23, 2947–2948.
Lindblad M, Sigvald R. 2004. Temporal spread of Wheat dwarf virus and mature plant resistance in winter wheat. Crop Protection, 23, 229–234.
Lindsten K, Vacke J. 1991. A possible barley adapted strain of Wheat dwarf virus (WDV). Acta Phytopathologica et Entomologica Hungarica, 26, 175–180.
Missich R, Ramirez-Parra E, Gutierrez C. 2000. Relationship of oligomerization to DNA binding of Wheat dwarf virus RepA and Rep proteins. Virology, 273, 178–188.
Nygren J, Shad N, Kvarnheden A,Westerbergh A. 2015. Variation in susceptibility to Wheat dwarf virus among wild and domesticated wheat. PLoS ONE, 10, e0121580.
Ohnoutkova L, Zitka O, Mrizova K, Vaskova J, Galuszka P, Cernei N, Smedley M A, Harwood W A, Adam V, Kizek R. 2012. Electrophoretic and chromatographic evaluation of transgenic barley expressing a bacterial dihydrodipicolinate synthase. Electrophoresis, 33, 2365–2373.
Pinel-Galzi A, Dubreuil-Tranchant C, Hebrard E, Mariac C, Ghesquiere A, Albar L. 2016. Mutations in Rice yellow mottle virus polyprotein P2a involved in RYMV2 gene resistance breakdown. Frontiers in Plant Science, 7, 1779.
Shepherd D N, Martin D P, McGivern D R, Boulton M I, Thomson J A, Rybicki E P. 2005. A three-nucleotide mutation altering the Maize streak virus Rep pRBR-interaction motif reduces symptom severity in maize and partially reverts at high frequency without restoring pRBR-Rep binding. Journal of General Virology, 86, 803–813.
Shepherd D N, Mangwende T, Martin D P, Bezuidenhout M, Kloppers F J, Carolissen C H, Monjane A L, Rybicki E P, Thomson J A. 2007a. Maize streak virus-resistant transgenic maize: A first for Africa. Plant Biotechnology Journal, 5, 759–767.
Shepherd D N, Mangwende T, Martin D P, Bezuidenhout M, Thomson J A, Rybicki E P. 2007b. Inhibition of Maize streak virus (MSV) replication by transient and transgenic expression of MSV replication-associated protein mutants. Journal of General Virology, 88, 325–336.
Shepherd D N, Martin D P, Thomson J A. 2009. Transgenic strategies for developing crops resistant to geminiviruses. Plant Science, 176, 1–11.
Shepherd D N, Dugdale B, Martin D P, Varsani A, Lakay F M, Bezuidenhout M E, Monjane A L, Thomson J A, Dale J, Rybicki E P. 2014. Inducible resistance to Maize streak virus. PLoS ONE, 9, e105932.
Steger G. 1994. Thermal denaturation of double-stranded nucleic acids: Prediction of temperatures critical for gradient gel electrophoresis and polymerase chain reaction. Nucleic Acids Research, 22, 2760–2768.
Taylor S, Wakem M, Dijkman G, Alsarraj M, Nguyen M. 2010. A practical approach to RT-qPCR-Publishing data that conform to the MIQE guidelines. Methods, 50, S1–S5.
Triant D A, Whitehead A. 2009. Simultaneous extraction of high-quality RNA and DNA from small tissue samples. Journal of Heredity, 1002, 246–250.
Vacke J. 1961. Wheat dwarf virus disease. Biologia Plantarum, 3, 228–233.
Virus Collection. 2016. The virus collection of the crop research institute, Prague 2016. http://www.vurv.cz/collections/vurv.exe/search?lang=cz&org=VI
Xie Q, Suarez-Lopez P, Gutierrez C. 1995. Identification and analysis of a retinoblastoma binding motif in the replication protein of a plant DNA virus: Requirement for efficient viral DNA replication. The EMBO Journal, 14, 4073–4082.
Zadoks J C, Chang T T, Konzak C F. 1974. Decimal code for growth stages of cereals. Weed Research, 14, 415–421.
Zaidi S S, Mansoor S, Ali Z, Tashkandi M, Mahfouz M M. 2016. Engineering plants for geminivirus resistance with CRISPR/Cas9 system. Trends in Plant Science, 21, 279–281. |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
