Soybean mosaic virus Helper Component-Protease Alters Leaf Morphology and Reduces Seed Production in Transgenic Soybean Plants.

ABSTRACT Transgenic soybean (Glycine max) plants expressing Soybean mosaic virus (SMV) helper component-protease (HC-Pro) showed altered vegetative and reproductive phenotypes and responses to SMV infection. When inoculated with SMV, transgenic plants expressing the lowest level of HC-Pro mRNA and those transformed with the vector alone initially showed mild SMV symptoms. Plants that accumulated the highest level of SMV HC-Pro mRNA showed very severe SMV symptoms initially, but after 2 weeks symptoms disappeared, and SMV titers were greatly reduced. Analysis of SMV RNA abundance over time with region-specific probes showed that the HC-Pro region of the SMV genome was degraded before the coat protein region. Transgenic soybean plants that expressed SMV HC-Pro showed dose-dependent alterations in unifoliate leaf morphologies and seed production where plants expressing the highest levels of HC-Pro had the most deformed leaves and the lowest seed production. Accumulation of microRNAs (miRNAs) and mRNAs putatively targeted by miRNAs was analyzed in leaves and flowers of healthy, HC-Pro-transgenic, and SMV-infected plants. Neither expression of SMV HC-Pro nor SMV infection produced greater than twofold changes in accumulation of six miRNAs. In contrast, SMV infection was associated with twofold or greater increases in the accumulation of four of seven miRNA-targeted mRNAs tested.

[1]  Pengyin Chen,et al.  Soybean mosaic virus. , 2008 .

[2]  S. Ghabrial,et al.  Development of Bean pod mottle virus-based vectors for stable protein expression and sequence-specific virus-induced gene silencing in soybean. , 2006, Virology.

[3]  G. Rubin,et al.  Drosophila microRNAs exhibit diverse spatial expression patterns during embryonic development. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[4]  V. Ilardi,et al.  Hairpin RNA-Mediated Silencing of Plum pox virus P1 and HC-Pro Genes for Efficient and Predictable Resistance to the Virus , 2005, Transgenic Research.

[5]  Xuemei Chen,et al.  microRNA biogenesis and function in plants , 2005, FEBS letters.

[6]  Tobias Dezulian,et al.  Conservation and divergence of microRNA families in plants , 2005, Genome Biology.

[7]  G. Hartman,et al.  Soybean mosaic virus helper component-protease enhances somatic embryo production and stabilizes transgene expression in soybean. , 2005, Plant physiology and biochemistry : PPB.

[8]  David C. Baulcombe,et al.  RNA silencing. , 2005, Trends in biochemical sciences.

[9]  Adam M. Gustafson,et al.  microRNA-Directed Phasing during Trans-Acting siRNA Biogenesis in Plants , 2005, Cell.

[10]  John Quackenbush,et al.  The TIGR Gene Indices: clustering and assembling EST and known genes and integration with eukaryotic genomes , 2004, Nucleic Acids Res..

[11]  D. Headon,et al.  A Link Between mRNA Turnover and RNA Interference in Arabidopsis , 2004, Science.

[12]  Shou-Wei Ding,et al.  Three distinct suppressors of RNA silencing encoded by a 20-kb viral RNA genome. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Franck Vazquez,et al.  Endogenous trans-acting siRNAs regulate the accumulation of Arabidopsis mRNAs. , 2004, Molecular cell.

[14]  D. Baulcombe.  RNA silencing in plants , 2004, Nature.

[15]  Valerian V Dolja,et al.  Viral RNA silencing suppressors inhibit the microRNA pathway at an intermediate step. , 2004, Genes & development.

[16]  Olivier Voinnet,et al.  RETRACTED: Probing the MicroRNA and Small Interfering RNA Pathways with Virus-Encoded Suppressors of RNA Silencing[W] , 2004, Plant Cell.

[17]  S. Clough,et al.  Tissue-Specific Gene Silencing Mediated by a Naturally Occurring Chalcone Synthase Gene Cluster in Glycine max , 2004, The Plant Cell Online.

[18]  M. Senda,et al.  Patterning of Virus-Infected Glycine max Seed Coat Is Associated with Suppression of Endogenous Silencing of Chalcone Synthase Genes , 2004, The Plant Cell Online.

[19]  D. Bartel.  MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[20]  F. Nutter,et al.  Pathogen-derived transgenic resistance to soybean mosaic virus in soybean , 2001, Molecular Breeding.

[21]  S. Ghabrial,et al.  Production of transgenic soybean lines expressing the bean pod mottle virus coat protein precursor gene , 1996, Plant Cell Reports.

[22]  Neff Walker,et al.  A MicroRNA as a Translational Repressor of APETALA2 in Arabidopsis Flower Development , 2004 .

[23]  C. Llave,et al.  P 1 / HC-Pro , a Viral Suppressor of RNA Silencing , Interferes with Arabidopsis Development and miRNA , 2003 .

[24]  B. Reinhart,et al.  A viral suppressor of RNA silencing differentially regulates the accumulation of short interfering RNAs and micro-RNAs in tobacco , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  S. Pfeffer,et al.  P0 of Beet Western Yellows Virus Is a Suppressor of Posttranscriptional Gene Silencing , 2002, Journal of Virology.

[26]  L. Schwartz,et al.  Drosophila Morgue is an F box/ubiquitin conjugase domain protein important for grim-reaper mediated apoptosis , 2002, Nature Cell Biology.

[27]  H. Vaucheret,et al.  RNA Silencing and the Mobile Silencing Signal Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.001677. , 2002, The Plant Cell Online.

[28]  D. Baulcombe,et al.  RNA silencing , 2002, Current Biology.

[29]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[30]  J. Valkonen,et al.  Potyviral helper-component proteinase expressed in transgenic plants enhances titers of Potato leaf roll virus but does not alleviate its phloem limitation. , 2001, Virology.

[31]  A. Caudy,et al.  Role for a bidentate ribonuclease in the initiation step of RNA interference , 2001 .

[32]  C. Fagoaga,et al.  Transgenic citrus plants expressing the citrus tristeza virus p23 protein exhibit viral-like symptoms. , 2001, Molecular plant pathology.

[33]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[34]  A. Mallory,et al.  A viral suppressor of gene silencing in plants. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Carrington,et al.  Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. , 1997, The Plant cell.

[36]  R. Goldbach,et al.  Expression of the potato leafroll virus ORF0 induces viral-disease-like symptoms in transgenic potato plants. , 1997, Molecular plant-microbe interactions : MPMI.

[37]  H. Scholthof,et al.  Identification of tomato bushy stunt virus host-specific symptom determinants by expression of individual genes from a potato virus X vector. , 1995, The Plant cell.

[38]  H. A. Smith,et al.  Transgenic plant virus resistance mediated by untranslatable sense RNAs: expression, regulation, and fate of nonessential RNAs. , 1994, The Plant cell.

[39]  M. Westgate,et al.  A Flower and Pod Staging System for Soybean , 1992 .

[40]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[41]  M. E. Irwin,et al.  Soybean mosaic virus. , 1981 .