Unité Mixte
de Recherche

Biologie et Génétique
des Interactions Plante-Parasite
 

CIRAD
UMR-BGPI TA A-54/K
Campus International
de Baillarguet
34398 Montpellier Cedex 5
FRANCE


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Copyright © CIRAD 2009
Group 2 : Interactions Virus Insect Plant (VIP) Research interest
 
The strange biology of multipartite viruses
Leader of the project: Stéphane Blanc
 

The way of life of multipartite viruses is poorly understood. Using Nanoviruses as biological models, we address the basic biology of multipartite viruses and experimentally investigate the possible benefits and costs in such biological systems.

Staff involved in the project

S. Blanc
Researcher
Leader of the project
M. Yvon
Technician
M.S. Vernerey
Engineer
G. Gallet
Post-doc
E. Pirolles
Assistant engineer
   
Master students
Geoffrey Carpentier (spring 2014, M2 univ. Montpellier, France)
Alexander Dale (winter 2015, Master 1 Erasmus, Oxford, United Kingdom)
Elise Découvreur (spring 2015, Master 1 Erasmus, Univ. Louvain, Belgium)
Major collaboration
Yannis Michalakis (Research director, CNRS, Montpellier)

 

Research project

Background and state of the art

Three main categories of virus genome structure and organization, independently of whether the nucleic acid is single- or double-stranded RNA or DNA, can be distinguished. The “monopartite” viruses have a single chromosome bearing all genetic information and packaged in a single virus particle. The “segmented” viruses have two or more chromosomes, which are all encapsidated together in a single virion. Finally, the “multipartite” viruses also have more than one chromosome, from 2 to 10 depending on the viral species, but each is encapsidated individually in distinct virus particles. Multipartite viruses are extremely frequent in plants, where they induce numerous diseases in major crops around the world. They also infect fungi and have recently been reported in insects.

Because of this puzzling compartmentalization of the genetic information of multipartite viruses into several virus particles, the functioning of their genome is intricately linked to that of their populations. Theoretical studies have proposed putative advantages of such viral systems, like enhanced stability, replication and/or recombination/shuffling of smaller nucleic acid segments; the counterpart costs being the reduced chances to infect new cells and new hosts with at least one copy of each, thus with no information loss. The current view of the way multipartite viral systems can be functional largely suffers from the lack of experimental data that would support any of the proposed benefits or even the actual existence of the above-mentioned cost at new cell and host infection.


The biological model and current results

We have recently demonstrated that the multipartite Faba bean necrotic stunt virus (FBNSV, Family Nanoviridae, Figure 1) reproducibly accumulates its eight genes (or genome segments) within host plants with different and specific relative frequencies, some genes being frequent and others rare (Figure 2 top panel). We here refer to the specific pattern of the frequencies of the eight segments as the “genome formula” (Figure 2 bottom panel). This discovery suggests an unforeseen putative benefit in multipartite viral systems, that is the capability to differentially control the copy number of each gene/segment, which in turn drives the virus population in the situation of maximum costs, due to the increased risk of losing rare segments.

Main questions of the project
We here develop a research program that experimentally addresses in parallel the relationship between the various genome segments and the population dynamics/genetics of multipartite viruses, using FBNSV as a model system. We investigate:

i) the link between the gene/segment copy number (genome formula), the regulation of gene expression, and the expression of viral phenotypes (accumulation, aggressiveness, transmission by aphid vectors);

ii) the actual distribution of distinct genomic segments in cells of the host and of the vectors, in order to reveal whether they are all always together in individual cells at all steps of the life cycle, or whether the genetic information can be temporarily separated in distinct locations of the host or vector;

iii) the way ssDNA segments, mRNA and proteins actually traffic within hosts and vectors to investigate whether the various segments can complement each other only within cells or also across cells;

iv) the plastictity of the genome formula in response to environmental changes and its evolution during adaptation to new host species.
Figure 1: Genome organization of nanoviruses on the example of Faba bean necrotic stunt virus (FBNSV)
Eight segments are circular ssDNAs each around 1000 bases : R encodes : Master-Rep assisting replication of all segments, S : coat protein CP separately encapsidating all segments, C : Clink resetting cell cycle, M : movement protein MP mediating in-plant virus migration, N : nuclear shuttle protein NSP regulating aphid transmission (B. Gronenborn, pers. com.), U1-U2-U4 have unknown functions. CR-SL: common region stem loop.
Figure 2 : Gene Copy Number regulation in Nanoviruses
Left Panel
The relative frequency of each FBNSV segment in Vicia faba plants systemically infected after inoculation by aphid vectors is plotted in dark blue and compared to that in agro-inoculated plants (light blue on the left). In Vicia faba, the frequency of the segments was not affected by the mode of inoculation (aphid- versus agro-inoculation), as indicated by ANOVA tests for each of the eight segments. The relative frequency of FBNSV segments in Medicago truncatula plants systemically infected after inoculation by aphid vectors is plotted in red. Vf-At1, Vf-At2, Vf-At3, Mt-At1 and Mt-At2 results from independent aphid-inoculation experiments and the number of infected plants in each set (n) is indicated on the right. A significant effect of the host species could be detected through ANOVA tests for 7 out of 8 segments. (From Sicard et al 2013)
Right Panel
The genome formulae of FBNSV in Vicia faba (GF Vf) and in Medicago truncatula (GFMt) were respectively calculated by pooling all Vf-At and Mt-At data sets, and are indicated below the graph.
The relative frequency of the segments (segment name below panels) was estimated by qPCR. The rarest is arbitrarily considered as 1 copy, and the copy number of the others is represented relative to it. Genome formula in Vicia faba (V.f. green left), Medicago truncatula (M.t. blue center) and Acyrtosiphon pisum (A.p. grey right). Adapted from Sicard et al 2013 and Sicard et al 2015.

This project provides key information on the basic “way of life” of multipartite viruses. In particular, the proposed experiments inform (or will inform) on both the putative benefits and costs in these enigmatic biological systems, for which decades of theoretical studies led to the recent conclusion that the current conceptual framework of virology cannot explain their existence.


Funding

The project is currently funded by INRA SPE division and by French ANR (ANR-nano project)

We are trying to attract additional external funds to further develop our project by investing more into:

-Structural approaches informing on the macromolecular complexes trafficking inside plants and aphid vectors
-Molecular and cellular interactions between virus and aphid vector
-Elucidate the specifics of the ecology of multipartite viruses


Forth coming events where results on the “nanovirus” program will be communicated (either virus-plant or virus-vector interactions)
Blanc S. Transmitted plant viruses can affect performances of starving aphid vectors. 13th International Plant Virus Epidemiology Symposium. 6-10 June 2016, Avignon, France.
Invited key note lecture.

Blanc S. The strange biology of plant-infecting multipartite viruses. New Frontiers in Plant Biology. 15-17 June 2016, Madrid, Spain.
Invited lecture

Blanc S. Vector transmission: commonalities and specificities in plant and animal viruses. 6th European Congress of Virology. 19-22 October 2016, Hamburg, Germany.
Invited Key note lecture

Blanc S. Nanoviruses have a non cell-autonomous replication cycle. 8th International Geminivirus Symposium and 6th International ssDNA comparative Virology Workshop. 7-10 November 2016, New Delhi, India.
Invited Key note lecture. Member of the International Advisory Committee.

Blanc S. Manipulation of insect vectors by nanoviruses. 11th European Congress of Entomology. 2-6 June 2018, Napoli, Itlay.
Invited Key note lecture. Co-convenor and co-chair of the session of “Symbiosis and Insect vector Biology”.


Publication list of the PI: S. Blanc (* corresponding author)

2016 (current work, article submitted and in preparartion)
Blanc S*. Manipulation of insect vectors by transmitted plant viruses. Current Opinion in Insect Science 2016. Invited review, in preparation (due February 2016).

Sicard A, Michalakis Y, Gutierrez S and S Blanc*. The strange biology of multipartite viruses. Submitted

Sicard A, Pirolles E, Gutierrez S, Yvon M, Michalakis Y and S Blanc*. A pluricellular way of life for a multipartite virus. In preparation

2015
Roumagnac P, Granier M, Bernardo P, Deshoux M, Ferdinand R, Galzi S, Fernandez E, Julian C, Abt I, Filloux D, Varsani A, Blanc S, Martin DP and M. Peterschmitt: Alfalfa leaf curl virus: An pahid transmitted geminivirus. J Virol. 2015 Sep 15;89(18):9683-8. doi: 10.1128/JVI.00453-15. Epub 2015 Jun 24.

Sicard A, Zeddam JL, Yvon M, Michalakis Y, Gutierrez S, Blanc* S: Circulative non propagative aphid-transmission of nanoviruses: an oversimplified view. J Virol. 2015 July 15 ;89(19) 9719-26. doi: 10.1128/JVI.00780-15

Gutiérrez S, Pirolles E, Yvon M, Baecker V, Michalakis Y and Blanc* S: The multiplicity of cellular infection changes depending on the route of cell infection in a plant virus. J Virol. 2015 Sep 15;89(18):9665-75. doi: 10.1128/JVI.00537-15.

Blanc* S and S Gutiérrez: The specifics of vector transmission of arboviruses of vertebrates and plants. Curr Opin Virol. 2015 Jul 30;15:27-33. doi: 10.1016/j.coviro.2015.07.003.


2014
Blanc* S, Drucker M, Uzest M: Localizing viruses in their insect vectors. Annu Rev Phytopathol 2014, 52:403-425.

2013
Bak A, Gargani D, Macia JL, Malouvet E, Vernerey MS, Blanc S, Drucker M: Virus factories of cauliflower mosaic virus are virion reservoirs that engage actively in vector transmission. J Virol 2013, 87:12207-12215.

Gutiérrez S, Michalakis Y, Van Munster M, Blanc* S: Plant feeding by insect vectors can affect life cycle, population genetics and evolution of plant viruses. Functional Ecology 2013, 27:610-622.

Martiniere A, Bak A, Macia JL, Lautredou N, Gargani D, Doumayrou J, Garzo E, Moreno A, Fereres A, Blanc* S and M. Drucker.: A virus responds instantly to the presence of the vector on the host and forms transmission morphs. Elife 2013, 2:e00183.

Sicard A, Yvon M, Timchenko T, Gronenborn B, Michalakis Y, Gutierrez S, Blanc* S: Gene copy number is differentially regulated in a multipartite virus. Nature Commununications 2013, 4:2248.

2012
Gutierrez S, Michalakis Y, Blanc* S: Virus population bottlenecks during within-host progression and host-to-host transmission. Curr Opin Virol 2012, 2:546-555.

Gutierrez S, Yvon M, Pirolles E, Garzo E, Fereres A, Michalakis Y, Blanc* S: Circulating virus load determines the size of bottlenecks in viral populations progressing within a host. PLoS Pathog 2012, 8:e1003009.

2011
Bak A, Irons SL, Martiniere A, Blanc S, Drucker M: Host cell processes to accomplish mechanical and non-circulative virus transmission. Protoplasma 2011.

Blanc* S, Drucker M: Functions of virus and host factors during vector-mediated transmission. In Recent Advances in Plant Virology. Edited by Caranta C, Aranda MA, Tepfer M, Lopez-Moya JJ: Caister Academic Press; 2011:103-120.

Martiniere A, Macia JL, Bagnolini G, Jridi C, Bak A, Blanc S, Drucker M: VAPA, an Innovative "Virus-Acquisition Phenotyping Assay" Opens New Horizons in Research into the Vector-Transmission of Plant Viruses. PLoS One 2011, 6:e23241.

Vuillaume F, Thebaud G, Urbino C, Forfert N, Granier M, Froissart R, Blanc S, Peterschmitt M: Distribution of the phenotypic effects of random homologous recombination between two virus species. PLoS Pathog 2011, 7:e1002028.

2010
Brault V, Uzest M, Monsion B, Jacquot E, Blanc S: Aphids as transport devices for plant viruses. C R Biol 2010, 333:524-538.

Gutierrez S, Yvon M, Thebaud G, Monsion B, Michalakis Y, Blanc* S: Dynamics of the multiplicity of cellular infection in a plant virus. PLoS Pathog 2010, 6.

Hoh F, Uzest M, Drucker M, Plisson-Chastang C, Bron P, Blanc S, Dumas C: Structural insights into the molecular mechanisms of cauliflower mosaic virus transmission by its insect vector. J Virol 2010, 84:4706-4713.

Khelifa M, Masse D, Blanc S, Drucker M: Evaluation of the minimal replication time of Cauliflower mosaic virus in different hosts. Virology 2010, 396:238-245.

Uzest M, Gargani D, Dombrovsky A, Cazevieille C, Cot D, Blanc* S: The "acrostyle": a newly described anatomical structure in aphid stylets. Arthropod Struct Dev 2010, 39:221-229.

2009
Martinière A, Gargani D, Uzest M, Lautredou N, Blanc S, Drucker M: A Role for Plant Microtubules in the Formation of Transmission-specific inclusion bodies of Cauliflower mosaic virus. Plant J 2009, 58:135-146.

Yvon M, Monsion B, Martin JP, Gutiérrez S, Blanc S: PCR-based amplification and analysis of specific viral sequences from individual plant cells. Journal of Virological Methods 2009, doi:10.1016/j.jviromet.2009.04.016

2008
Blanc* S: Vector transmission of plant viruses. In Encyclopedia of Virology, edn 3rd. Edited by Mahy BWJ, van regenmortel MHV: Elsevier Ltd.; 2008:274-282.

Monsion B, Duborjal H, Blanc S: Quantitative Single-letter Sequencing: a method for simultaneously monitoring numerous known allelic variants in single DNA samples. BMC Genomics 2008, 9:85.

Monsion B, Froissart R, Michalakis Y, Blanc S: Large bottleneck size in Cauliflower Mosaic Virus populations during host plant colonization. PLoS Pathog 2008, 4:e1000174.

Urbino C, Thebaud G, Granier M, Blanc S, Peterschmitt M: A novel cloning strategy for isolating, genotyping and phenotyping genetic variants of geminiviruses. Virol J 2008, 5:135.

2007
Blanc S: Virus transmission-getting out and in., vol 7. Edited by Heinlein EWaM. Berlin-Heidelberg: Springer-Verlag; 2007.

Brault V, Blanc S, Jacquot E: Comment les pucerons transmettent des maladies virales aux plantes. Biofutur 2007, 279:40-44.

Khelifa M, Journou S, Krishnan K, Gargani D, Esperandieu P, Blanc S, Drucker M: Electron-lucent inclusion bodies are structures specialized for aphid transmission of cauliflower mosaic virus. J Gen Virol 2007, 88:2872-2880.

Uzest M, Gargani D, Drucker M, Hebrard E, Garzo E, Candresse T, Fereres A, Blanc* S: A protein key to plant virus transmission at the tip of the insect vector stylet. Proc Natl Acad Sci U S A 2007, 104:17959-17964.

2006
Jridi C, Martin JF, Marie-Jeanne V, Labonne G, Blanc* S: Distinct viral populations differentiate and evolve independently in a single perennial host plant. J Virol 2006, 80:2349-2357.

2005
Ballut L, Drucker M, Pugniere M, Cambon F, Blanc S, Roquet F, Candresse T, Schmid HP, Nicolas P, Gall OL, et al.: HcPro, a multifunctional protein encoded by a plant RNA virus, targets the 20S proteasome and affects its enzymic activities. J Gen Virol 2005, 86:2595-2603.

Froissart R, Roze D, Uzest M, Galibert L, Blanc S, Michalakis Y: Recombination every day: abundant recombination in a virus during a single multi-cellular host infection. PLoS Biol 2005, 3:e89.

Moreno A, Hebrard E, Uzest M, Blanc* S, Fereres A: A single amino acid position in the helper component of cauliflower mosaic virus can change the spectrum of transmitting vector species. J Virol 2005, 79:13587-13593.

Plisson C, Uzest M, Drucker M, Froissart R, Dumas C, Conway J, Thomas D, Blanc S, Bron P: Structure of the mature P3-virus particle complex of cauliflower mosaic virus revealed by cryo-electron microscopy. J Mol Biol 2005, 346:267-277.

2004
Blanc* S: Insect transmission of viruses. In Microbe-vector interactions in vector-borne diseases. Edited by Gillespie SH, Smith GL, Osbourn A: Cambridge University Press; 2004:42-61. [Symposium S (Series Editor), vol 63.]

Froissart R, Uzest M, Ruiz-Ferrer V, Drucker M, Hebrard E, Hohn T, Blanc* S: Splicing of Cauliflower mosaic virus 35S RNA serves to downregulate a toxic gene product. J Gen Virol 2004, 85:2719-2726.

2003
Plisson C, Drucker M, Blanc S, German-Retana S, Le Gall O, Thomas D, Bron P: Structural characterization of HC-Pro, a plant virus multifunctional protein. J Biol Chem 2003, 278:23753-23761.

2002
Drucker M, Froissart R, Hebrard E, Uzest M, Ravallec M, Esperandieu P, Mani JC, Pugniere M, Roquet F, Fereres A, and Blanc* S : Intracellular distribution of viral gene products regulates a complex mechanism of cauliflower mosaic virus acquisition by its aphid vector. Proc Natl Acad Sci U S A 2002, 99:2422-2427.

Drucker M, German-Retana S, Espérandieu P, Le Gall O, Blanc* S: Purification of a viral protein from infected plant tissues using the strep tag II affinity tag. BioTech Intl. 2002, 14:16-18.

Froissart R, Michalakis Y, Blanc* S: Helper component-transcomplementation in the vector transmission of plant viruse. Phytopathology 2002, 92:576-579.

Lett JM, Granier M, Hippolyte I, Grondin M, Royer M, Blanc S, Reynaud B, Peterschmitt M: Spatial and temporal distribution of geminiviruses in leafhoppers of the genus Cicadulina monitored by conventional and quantitative polymerase chain reaction. Phytopathology 2002, 92:65- 74.

Palacios I, Drucker M, Blanc S, Leite S, Moreno A, Fereres A: Cauliflower mosaic virus is preferentially acquired from the phloem by its aphid vectors. J Gen Virol 2002, 83:3163-3171.

2001
Blanc* S Hébrard E, Drucker M, Froissart R: Molecular basis of vector transmission : Caulimoviruses. In Virus-Insect-Plant interactions. Edited by Harris K, Smith OP, Duffus JE: Academic Press; 2001:143-166.

Hebrard E, Drucker M, Leclerc D, Hohn T, Uzest M, Froissart R, Strub JM, Sanglier S, van Dorsselaer A, Padilla A, C. Dumas and S Blanc* : Biochemical characterization of the helper component of Cauliflower mosaic virus. J Virol 2001, 75:8538-8546.

Leh V, Jacquot E, Geldreich A, Haas M, Blanc S, Keller M, Yot P: Interaction between the open reading frame III product and the coat protein is required for transmission of cauliflower mosaic virus by aphids. J Virol 2001, 75:100-106.

Raccah B, Huet H, Blanc S: Potyviruses. In Virus-Insect-Plant interactions. Edited by Harris K, Duffus JE, Smith OP: Academic Press; 2001:181-206.

2000
Héricourt F, Blanc S, Redeker V, Jupin I: Evidence for phosphorylation and ubiquitinylation of turnip yellow mosaic virus RNA-dependent RNA polymerase domain expressed in a baculovirus-insect cell system. Biochem. J. 2000, 349:417-425.

1999
Blanc S, Dolja VV, Llave C, T.P. P: Histidine-tagging and purification of tobacco etch potyvirus helper component protein. Journal of Virological Methods 1999, 77:11-15.

Hébrard E, Froissart R, Louis C, Blanc* S: Les modes de transmission des virus phytopathogènes par vecteurs. Virologie 1999, 3:35-48.

Leh V, Jacquot E, Geldreich A, Hermann T, Leclerc D, Cerutti M, Yot P, Keller M, Blanc* S: Aphid transmission of cauliflower mosaic virus requires the viral PIII protein. Embo J 1999, 18:7077-7085.

1998
Blanc S, Ammar ED, Garcia-Lampasona S, Dolja VV, Llave C, Baker J, Pirone TP: Mutations in the potyvirus helper component protein: effects on interactions with virions and aphid stylets. J Gen Virol 1998, 79 ( Pt 12):3119-3122.

1997
Blanc S, Lopez-Moya JJ, Wang R, Garcia-Lampasona S, Thornbury DW, Pirone TP: A specific interaction between coat protein and helper component correlates with aphid transmission of a potyvirus. Virology 1997, 231:141-147.

1996
Blanc S, Schmidt I, Vantard M, Scholthof HB, Kuhl G, Esperandieu P, Cerutti M, Louis C: The aphid transmission factor of cauliflower mosaic virus forms a stable complex with microtubules in both insect and plant cells. Proc Natl Acad Sci U S A 1996, 93:15158-15163.

Pirone TP, Blanc S: Helper-dependent vector transmission of plant viruses. Annu Rev Phytopathol 1996, 34:227-247.

1995
Kiss-Laszlo Z, Blanc S, Hohn T: Splicing of cauliflower mosaic virus 35S RNA is essential for viral infectivity. Embo J 1995, 14:3552-3562.

1994
Schmidt I, Blanc S, Esperandieu P, Kuhl G, Devauchelle G, Louis C, Cerutti M: Interaction between the aphid transmission factor and virus particles is a part of the molecular mechanism of cauliflower mosaic virus aphid transmission. Proc Natl Acad Sci U S A 1994, 91:8885-8889.

1993
Blanc S, Cerutti M, Chaabihi H, Louis C, Devauchelle G, Hull R: Gene II product of an aphid-nontransmissible isolate of cauliflower mosaic virus expressed in a baculovirus system possesses aphid transmission factor activity. Virology 1993, 192:651-654.

Blanc S, Cerutti M, Usmany M, Vlak JM, Hull R: Biological activity of cauliflower mosaic virus aphid transmission factor expressed in a heterologous system. Virology 1993, 192:643-650.

Blanc S, Schmidt I, Kuhl G, Esperandieu P, Lebeurier G, Hull R, Cerutti M, Louis C: Paracrystalline structure of cauliflower mosaic virus aphid transmission factor produced both in plants and in a heterologous system and relationship with a solubilized active form. Virology 1993, 197:283-292.

 


 

   
 
 
   
   
   
 
 
 
asques et ascospores de Magnaporthe orizae - copyright : JL Notteghem spores Magnaporthe oryzae - copyright : JL Notteghem bactéries Xanthomonas pseudoalbilineans (gauche) et Xanthomonas albilineans (droite). Les deux produisent l'antibiotique albicidine (structure en haut de la photo - copyright : S. Cociancich/A. Mainz
  champignon Magnaporthe (vert) en train d'attaquer une feuille de riz - copyright : A. Delteil/JB Morel test d'anticorps sur puceron (Mysus persicae) - copyright : MS Vernerey/M. van Munster/M. Uzest