NE1033: Biological Improvement of Chestnut through Technologies that Address Management of the Species, its Pathogens and Pests

(Multistate Research Project)

Status: Inactive/Terminating

SAES-422 Reports

Annual/Termination Reports:

[01/26/2010] [12/16/2010]

Date of Annual Report: 01/26/2010

Report Information

Annual Meeting Dates: 09/25/2009 - 09/26/2009
Period the Report Covers: 10/01/2008 - 09/01/2009

Participants

Anagnostakis, Sandra (Sandra.Anagnostakis@ct.gov),Connecticut Agricultural Experiment Station; Romero-Severson, Jeanne (Jeanne.Romero-Severson.1@nd.edu), University of Notre Dame; Rieske-Kinney, Lynne (lrieske@uky.edu), University of Kentucky; Nuss, Donald (nuss@umbi.umd.edu), University of Maryland Biotechnology Institute, Shady Grove; Shi, Diane (shid@umbi.umd.edu), University of Maryland Biotechnology Institute, Shady Grove; Jensen Ken (Jensen@umbi.umd.edu), University of Maryland Biotechnology Institute, Shady Grove; Fulbright, Dennis (fulbrig1@msu.edu), Michigan State University; Kubisiak, Tom (tkubisiak@fs.fed.us), USDA-Forest Service, Saucier, MS; Nelson, Dana (dananelson@fs.fed.us) USDA-Forest Service, Saucier, MS; Hillman, Bradley (Hillman@aesop.rutgers.edu), Rutgers University; Georgi, Laura (georgi@aesop.rutgers.edu), Rutgers University; Kaunzinger, Christina (cmkk@rci.rutgers.edu), Rutgers University; Sullivan, Raymond (raysully@gmail.com), Rutgers University; Handel, Steven (handel@aesop.rutgers.edu), Rutgers University; Dawe, Angus (dawe@nmsu.edu), New Mexico State University; Powell, William (wapowell@esf.edu), SUNY-ESF; Baier, Kathleen (kbaier@syr.edu), SUNY-ESF; Northern, Lilibeth (lcnorthern@esf.edu), SUNY-ESF; Zhang, Amelio Bo ((bozhang@syr.edu), SUNY-ESF; Newhouse, Andrew (andynewhouse@yahoo.com), SUNY-ESF; Sisco, Paul (paul@acf.org), TACF, Asheville; Carlson, John (jec16@psu.edu) Penn State University; Micsky, Gary (gwm6@psu.edu) , Penn State University; Barakat, Abdelali (aub14@psu.edu), Penn State University, Fitzsimmons, Sara (sara@acf.org), Penn State University; Craddock, Hill (hill-craddock@utc.edu),UT Chattanooga; Gurney, Kendra (Kendra@acf.org),TACF, Burlington; Hebard, Fred (fred@acf.org), TACF, Meadowview; MacDonald, William (macd@wvu.edu), West Virginia University; Paris, Robert (bob@acf.org), TACF, Beckley; Dale, Adam (adale@uoguelph.ca), University of Guelph

Brief Summary of Minutes

The meeting was called to order by Chairman Hillman at 9:00 am on September 25, 2009 at the Manchester Inn, Ocean Grove, NJ. As Senior Associate Director for the New Jersey Agriculture Experiment Station and Director of Cooperative Research, Hillman is now the Administrative Advisor for NE-1033.

2010 Business Meeting

Sandra Anagnostakis was elected Chair for 2011. Paul Sisco, chair-elect, stated that the 2010 meeting will be held September 16-19, 2010 at Cataloochee Ranch in Maggie Valley, NC. It's about 45 minutes west of Asheville in a gorgeous location at 5,000 feet adjoining the Great Smoky Mountains National Park. The website is: http://www.cataloocheeranch.com/

Note: Detailed accomplishment report is in the attached Minutes.

Accomplishments

<b>Completion of proposed milestones:</b><br /> <br><b>2005:</b> <br /> <ul><li>Characterization of the role of hypovirus p29 in virus RNA accumulation in C. parasitica, and virus transmission through conidia of the fungus:<br /> <br>Completed early:<br /> <br>Suzuki, N., Maruyama, K., Moriyama, M. and Nuss, D. L. Hypovirus papain-like protease p29 functions in trans to enhance viral double-stranded RNA accumulation and vertical transmission. J. Virol. 77:11697-11707, 2003. <br /> <li>Generation of polyclonal antibodies against 5 overlapping regions of hypovirus ORF B, and construction of a C. parasitica EST database.<br /> <br>Both completed in 2004.<br /> </ul><br /> <p><b>2006:</b><br /> <ul><li>Publication of a C. parasitica EST database containing approximately 2500 ESTs. <br /> <br>Completed early.<br /> <br>Dawe, A.L., McMains, V.C., Panglao, M., Kasahara, S., Chen, B. and Nuss, D.L. An ordered collection of expressed sequences from Cryphonectria parasitica and evidence of genomic microsynteny with Neurospora crassa and Magnaporte grisea. Microbiology 149:2373-2384, 2003. <br /> </ul><br /> <p><b>2007:</b><br /> <ul><li>ORF B polyprotein processing pathway in C. parasitica confirmed, ORF B mature proteins responsible for altering fungal cell signaling pathways mapped and DNA microarray analysis of hypovirus-mediated alteration of fungal gene expression initiated. <br /> <br>ORF B polyprotein processing pathway not completed. Microarray analysis initiated giving publication in 2003. <br /> <br /> <p>Allen, T.D., Dawe, A.L. and Nuss, D.L. Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonectria parasitica to infection by virulence-attenuating hypovirus. Eukaryotic Cell 2:1253-1265, 2003. <br /> </ul><br /> <p><b>2008:</b><br /> <ul><li>Polyprotein processing maps completed for hypoviruses CHV1-EP713 and CHV1-Euro 7, and a detailed view compiled of the changes in cellular transcriptional profiles caused by infection of C. parasitica with mild and severe hypoviruses.<br /> <br>Polyprotein processing map has not been competed. Transcriptional profiles caused by mild and severe hypoviruse have been generated. The C. parasitica EST microarrays have also been used to examine the effect of hypovirus infection on G-protein signaling and to expose a linkage between mitochondrial and viral hypovirulence. <br /> <br /> <p>Allen, T.D. and Nuss, D.L. Specific and common alterations in host gene transcript accumulation following infection of the chestnut blight fungus by mild and severe hypoviruses. J. Virol. 78:4145-4155, 2004. <br /> <br /> <p>Dawe, A.L., Segers, G.C., Allen, T.D., McMains, V.C. and Nuss, D.L. Microarray analysis of Cryphonectria parasitica Ga- and Gbg- signaling pathways reveals extensive modulation by hypovirus infection. Microbiology 150:4033-4043, 2004.<br /> <br /> <p>Allen, T.D. and Nuss, D.L. Linkage between mitochondrial hypovirulence and viral hypovirulence in the chestnut blight fungus revealed by cDNA microarray analysis. Eukaryotic Cell 3:1227-1232, 2004. <br /> </ul><br /> <br /> <p><b>Completion of milestones not proposed:</b><br /> <ul><li>A proposal to sequence the C. parasitica genome was approved by the Department of Energy Community Sequencing Program in June of 2006. <br /> <br>The assembled 8.5 X C. parasitica genome sequence was released to the public on September 30, 2008. This is a tremendous resource for future studies on the chestnut blight fungus and its interaction with the chestnut tree. <br /> <li>Demonstration that hypovirus p29 suppresses RNA silencing in C. parasitica and in heterologous plant system. This is the first report of a mycovirus-encoded suppressor of RNA silencing.<br /> <br>Segers, G.C., van Wezel, R., Zhang, X., Hong, Y. and Nuss, D.L. Hypovirus Papain-like protease p29 suppresses RNA silencing in the natural fungal host and in a heterologous plant system. Eukaryotic Cell 5:896-904, 2006. <br /> <li>Demonstrated that RNA silencing serves as an antiviral defense mechanism in C. parasitica (first example for any fungus) against hypoviruses and mycoreoviruses.<br /> <br>Segers, G.C., Zhang, X., Deng, F., Sun, Q. and Nuss, D.L. Evidence that RNA silencing functions as an antiviral defense mechanism in fungi. PNAS USA 104:12902-12906, 2007. <br /> <li>First report of the cloning and sequence analysis of mycovirus-derived small RNAs (vsRNAs) generated by RNA silencing. The vsRNAs were shown to be produced, in a dicer dcl-2-dependent manner, from both positive and negative hypovirus RNA strands at a ratio of 3:2 and to be non-randomly distributed along the viral genome. C. parastiica was shown to respond to mycovirus infection with a 10-15 fold increase in dcl-2 transcript acuumulation while the expression of dcl-1 was modestly increased. The expression of dcl-2 was further increased (~35 fold) following infection by a CHV1-EP713 mutant that lacks the p29 suppressor of RNA silencing. A similar response in dicer gene expression following virus infection of plants or animals has not yet been reported. In this regard, it is anticipated that the evolutionary position of fungi relative to animals and plants will provide insights into additional novel mechanisms for the induction and suppression of RNA silencing pathways yet to be revealed in the other organisms. <br /> <br>Zhang, X., Segers, G.C., Sun, Q., Deng, F. and Nuss, D.L. Characterization of hypovirus-derived small RNAs generated in the chestnut blight fungus by an inducible DCL-2-dependnet pathway. Journal of Virology 82:2613-2619, 2008.<br /> <li>Virus RNA recombination is an important component of virus evolution that contributes to the emergence of new viruses and the generation of internally deleted mutant RNAs, termed defective interfering (DI) RNAs, that are derived from, and dependent on, the parental viral genomic RNA. We provided the first experimental evidence that a host RNA silencing pathway is required for DI RNA production and virus vector RNA instability for a single-strand, positive sense RNA virus.<br /> <br>Zhang, X. and Nuss, D.L. A host dicer is required for defective viral RNA production and recombinant virus vector instability for a positive sense RNA virus. Proc. Natl. Acad. Sci. USA. Early Edition, October 13, 2008.<br /> </ul><br /> <p><b>2009</b><br /> <ul><li>Determine specific genetic fingerprints of tested cultivars through the use of microsatellite markers and find loci useful for parentage analysis (Fulbright)<br /> <li>Refine the genetic linkage and genome sequence maps for map-based cloning of fungal vic and pathogenicity genes<br /> <li>Tom Kubisiak used the C. parasitica Genome sequence generated by the JGI Community sequencing Program to identify 689 simple sequence repeats (SSRs) and designed 141 primer pairs. One hundred and thirty Four of the 141 primer pairs amplified discrete products, with 96 of the 134 showing polymorphism for the JA17 and X17.8 parents of the mapping cross. Allele data were generated for 96 progeny from the mapping cross for 32 polymorphic SSRs and a total of 30 of these markers were placed within the context of the published C. parasitica linkage map (Kubisiak and Milgroom, 2006, FG&B 43:453-463). The new linkage data were used by the JGI finishing group to connect a number of scaffolds in Version 1 of the genome assembly to generate Version 2 that was released July 10, 2009.<br /> <li>Complete assembly and community manual annotation of the C. parasitica genome sequence<br /> <br>Nine hundred and ninety five of 11,251 transcripts have been manually curated by the C. parasitica annotation team to date. Version 2 of the genome assembly has been released by the JGI consisting of 26 scaffolds. Five of the new scaffolds contain two teleomers and are of a length consistent with a complete chromosome. Six additional scaffolds contain one teleomer and are in excess of 1 MB in length. <br /> <li>Use the C. parasitica genome sequence to develop new microarray chip and proteomics platforms for analysis of global gene expression in the blight fungus when challenged by viral pathogens<br /> <li>Orchard established in WV with advanced, back-cross chestnut trees from VA for assessment of host resistance with hypovirulence in the Cryphonectria parasitica populationreplanted in 2009.<br /> </ul><br /> <p><b>In progress.</b><br /> <p><b>2010</b><br /> <ul><li>Complete characterization of C. parasitica antiviral RNAsilencing pathways.<br /> Two Dicer genes were identified in the C. parasitica genome, cloned and disrupted. Dicer DCL2 was shown to be required for antiviral RNA silencing while Dicer DCL1 was not required (Segers et al., PNAS 2007, 104:12902-12906). Four Argonaute genes were identified in the C. parasitica genome, cloned and disrupted. Only Argonaute AGL2 was required for antiviral RNA silencing (Sun et al., PNAS, IN PRESS). Three RNA dependent RNA polymerases and an orthologue of the QIP exonuclease involved in transgene silencing in Neurospora crassa have been identified in the C. parasitica genome and are currently being cloned and disrupted. </ul>

Publications

2008-2009 Publications<br /> Aguilar, F.X., M.M. Cernusca, and M.A. Gold. 2009. A preliminary assessment of consumer preferences for chestnuts (Castanea spp.) using conjoint analysis. HortTechnology 19: 216-223. <br /> <br /> Andrade, G.M., C.J. Nairn, H.T. Le and S.A. Merkle. 2009. Sexually mature transgenic American chestnut trees via embryogenic suspension-based transformation. Plant Cell Reports 28:1385-97. <br /> <br /> Barakat, A., D.S. DiLoreto, Y. Zhang, C. Smith, K. Baier, W.A. Powell, N. Wheeler, R. Sederoff and J.E. Carlson. 2009. Comparison of transcriptome from cankers and healthy stems in American chestnut (Castanea dentata) and Chinese chestnut (Castanea mollissima). BMC Plant Biology, 9:51 11 pages.<br /> <br /> Cernusca, M.M., M.A. Gold and L.D. Godsey. 2008. Influencing consumer awareness through the Missouri chestnut roast. Journal of Extension 46(6). Published online. 8 p. www.joe.org/joe/2008december/rb7p.shtml <br /> <br /> Cooper, W.R. and L.K. Rieske. In review. Ecological interactions among Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) community associates. Ecological Entomology, submitted fall 2009.<br /> <br /> Cooper, W.R. and L.K. Rieske. In review. Chestnut species and jasmonic acid treatment influence development and community interactions of galls produced by Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae). Arthropod Plant Interactions, submitted summer 2009.<br /> <br /> Cooper, W.R. and L.K. Rieske. 2009. Woody stem galls interact with foliage to affect community associations. Environmental Entomology 38, 417-424.<br /> <br /> Cooper, W.R. and L.K. Rieske. 2008. Differential responses in American (Castanea dentata Marshall) and Chinese (C. mollissima Blume) chestnut (Fagales: Fagaceae) to foliar application of jasmonic acid. Chemoecology 18, 121-127.<br /> <br /> Cooper, W.R. and L.K. Rieske. 2009. Sustainable Management of the Invasive Asian Chestnut Gall Wasp? Interactions between native and introduced natural enemies. The Nutshell, June 2009.<br /> <br /> Dawe, A.L. W. A. Van Voorhies, T. A. Lau, A. V. Ulanov and Z. Li. 2009. Microbiology, in press (available online as doi: 10.1099/mic.0.029033-0).<br /> <br /> Gold, M.A., M.M. Cernusca and L.D. Godsey. 2008. A competitive market analysis of the U.S. shiitake mushroom marketplace. HortTechnology 18: 489-499. <br /> <br /> Gold, M.A., M.M. Cernusca and L.D. Godsey. 2009. Agroforestry Product Markets And Marketing. Chapter 11. In: Garrett, H.E. (ed). North American Agroforestry: An Integrated Science and Practice. 2nd Edition. Agronomy Society of America, Madison, WI.<br /> <br /> Gold, M.A. and H.E. Garrett. 2009. Agroforestry Nomenclature, Concepts and Practices. Chapter 3. In: Garrett, H.E. (ed). North American Agroforestry: An Integrated Science and Practice. 2nd Edition. Agronomy Society of America, Madison, WI. <br /> <br /> Gold, M.A. and M.H. Hall (eds.). 2009. Agroforestry Comes of Age: Putting Science into Practice. 11th North American Agroforestry Conference Proceedings. Columbia, MO. May 31-June 3, 2009. 517 p.<br /> <br /> Gonzalez, Donis I.R., M. Mandujano, C. Medina-Mora and D.W. Fulbright. 2009. Presence of mycotoxins after 90 days of storage in fresh chestnuts. 4th Annual Chestnut Symposium, Beijing, China, Oct. 2008. Acta Horticulturae (in press).<br /> <br /> Jacob-Wilk, D., M. Turina, P. Kazmierczak and N.K. Van Alfen. 2009. Silencing of Kex2 significantly diminishes the virulence of Cryphonectria parasitica. Molecular Plant-Microbe Interactions 22:211-221.<br /> <br /> Kim. M.-J., B.-R. Kwon, S.-M. Park, H.-J. Chung. M.-S. Yan, A.C. L. Churchill, N.K. Van Alfen and D.-H. Kim. 2008. Promoter analysis of the cell surface-abundant and hypoviral-regulated cryparin gene from Cryphonectria parasitica. Mol. Cells 26:496-502.<br /> <br /> Lan, Xiuwan; Ziting,Yao, Yan Zhou, Jinjie Shang, Haiyan Lin, D.L. Nuss, and B. Chen. 2008. Deletion of the cpku80 gene in the chestnut blight fungus, Cryphonectria parasitica, enhances gene disruption efficiency. Current Genetics 53:59-66. <br /> <br /> Milgroom M.G., K. Sotirovski, M. Risteski, and M.T. Brewer. 2009. Heterokaryons and recombinants of Cryphonectria parasitica in two clonal populations in southeastern Europe. Fungal Genet. Biol. 46: 849-854.<br /> <br /> Monteiro-Vitorello, C.B., G. Hausner, D.B. Searles, E.A. Gibb, D.W. Fulbright and H. Bertrand. 2009. The Cryphonectria parasitica mitochondrial ms gene: plasmid-like elements, introns and homing endonucleases. Fungal Genetics and Biology 46:837-848.<br /> <br /> Sun, Q., G.H. Choi and D.L. Nuss. 2009. Hypovirus-responsive transcription factor gene pro1 of the chestnut blight fungus Cryphonectria parasitica is required for female fertility, asexual spore development and stable maintenance of hypovirus infection. Eukaryotic Cell 8:262-270.<br /> <br /> Sun, Q., G.H. Choi and D.L. Nuss. 2009. A single Argonaute gene is required for induction of RNA silencing antiviral defense and promotes viral RNA recombination. Proc. Nat. Acad. Sci. (in press).<br /> <br /> Valdivia, C., L. Zabek, J. Arbuckle, M. Gold and C. Flora. 2009. Human and Institutional Dimensions of Agroforestry. Chapter 13. In: Garrett, H.E. (ed). North American Agroforestry: An Integrated Science and Practice. 2nd Edition. Agronomy Society of America, Madison, WI. <br /> <br /> Willyerd, K.L. A.M. Kemp and A.L. Dawe. 2009. Controlled gene expression in the plant pathogen Cryphonectria parasitica by use of a copper-responsive element. Appl. Environ. Microbiol. 75:5417-5420.<br /> <br /> Zhang, X. and D.L. Nuss. 2008. A host dicer is required for defective viral RNA production and recombinant virus vector RNA instability for a positive sense RNA virus. Proc. Nat. Acad. Sci. 105:16749-16754. <br />

Impact Statements

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Date of Annual Report: 12/16/2010

Report Information

Annual Meeting Dates: 09/17/2010 - 09/18/2010
Period the Report Covers: 10/01/2009 - 09/01/2010

Participants

Anagnostakis, Sandra (Sandra.Anagnostakis@ct.gov),Connecticut Agricultural Experiment Station; Romero-Severson, Jeanne (Jeanne.Romero-Severson.1@nd.edu), University of Notre Dame; Rieske-Kinney, Lynne (lrieske@uky.edu), University of Kentucky; Nuss, Donald (nuss@umbi.umd.edu), University of Maryland Biotechnology Institute, Shady Grove; Fulbright, Dennis (fulbrig1@msu.edu), Michigan State University; Kubisiak, Tom (tkubisiak@fs.fed.us), USDA-Forest Service, Saucier, MS; Nelson, Dana (dananelson@fs.fed.us) USDA-Forest Service, Saucier, MS; Hillman, Bradley (Hillman@aesop.rutgers.edu), Rutgers University; Dawe, Angus (dawe@nmsu.edu), New Mexico State University; Powell, William (wapowell@esf.edu), SUNY-ESF; Baier, Kathleen (kbaier@syr.edu), SUNY-ESF; Newhouse, Andrew (andynewhouse@yahoo.com), SUNY-ESF; Sisco, Paul (phsisco@gmail.com), The American Chestnut Foundation, Asheville; Carlson, John (jec16@psu.edu) Penn State University; Micsky, Gary (gwm6@psu.edu), Penn State University; Barakat, Abdelali (abaraka@clemson.edu), Clemson University, Fitzsimmons, Sara (sara@acf.org), Penn State University; Craddock, Hill (hill-craddock@utc.edu),Unveristy of Tennessee Chattanooga; Gurney, Kendra (Kendra@acf.org),TACF, Burlington; Hebard, Fred (fred@acf.org), The American Chestnut Foundation, Meadowview; MacDonald, William (macd@wvu.edu), West Virginia University; Double, Mark (mdouble@wvu.edu), West Virginia University; Dane, Fenny (danefen@auburn.edu), Auburn University; Choi, Gil (gchoi12@umd.edu), University of Maryland, Shady Grove; Hughes, Keaton (keaton-hughes@utc.edu), Univeristy of Tennessee Chattanooga; Graziosi, Ignazio (i.grasiosi@uky.edu), University of Kentucky; Harris, Amelia (amelia-harris@utc.edu), University of Tennessee Chattanooga; Jarosz, Andrew (amjarosz@msu.edu), Michigan State University; Kazmierczak, Pam (pjkkaz@ucdavis.edu), University of Calfornia, Davis; Metaxas, Ana (ana-metaxas@utc.edu), University of Tennessee Chattanooga; Schlarbaum, Scott (tenntip@auk.edu), University of Tennessee Knoxville; Springer, Josh (srping47@msu.edu), Michigan State University; Wilk, Debora (dkwilk@ucdavis.edu), University of California Davis; Regnery, Russell (russellregnery@mac.com); Staton, Meg (mestato@yahoo.com) Clemson University; Abbott, Albert (aalbert@clemson.edu), Clemson University; Fang, Eric (gfang@clemson.edu), Clemson University; Bevins, David, The American Chestnut Foundation, Meadowview; Coughlin, Erin (erin.coughlin@vikings.berry.edu), Berry College; Pinchot, Leila (cpinchot@utk.edu) University of Tennessee Knoxville; D'Amico, Katherine (kmdamico@syr.edu), SUNY-ESF; Olukolu, Bode (bolukol@clemson.edu), Clemson University; Maddox, Jim (jjmaddox1@bellsouth.net), Tenness Valley Authority (retired); Jensen, Ken (jensenK@umd.edu), University of Maryland, Shady Grove

Brief Summary of Minutes

Accomplishments

2009 Outcomes<br /> <br /> " Determine specific genetic fingerprints of tested cultivars through the use of microsatellite markers and find loci useful for parentage analysis (Fulbright)<br /> " Refine the genetic linkage and genome sequence maps for map-based cloning of fungal vic and pathogenicity genes<br /> " Tom Kubisiak used the C. parasitica Genome sequence generated by the JGI Community Sequencing Program to identify 689 simple sequence repeats (SSRs) and designed 141 primer pairs. One hundred and thirty Four of the 141 primer pairs amplified discrete products, with 96 of the 134 showing polymorphism for the JA17 and X17.8 parents of the mapping cross. Allele data were generated for 96 progeny from the mapping cross for 32 polymorphic SSRs and a total of 30 of these markers were placed within the context of the published C. parasitica linkage map (Kubisiak and Milgroom, 2006, FG&B 43:453-463). The new linkage data were used by the JGI finishing group to connect a number of scaffolds in Version 1 of the genome assembly to generate Version 2 that was released July 10, 2009.<br /> Ï Complete assembly and community manual annotation of the C. parasitica genome sequence<br /> Nine-hundred and ninety-five of 11,251 transcripts have been manually curated by the C. parasitica annotation team to date. Version 2 of the genome assembly has been released by the JGI consisting of 26 scaffolds. Five of the new scaffolds contain two teleomers and are of a length consistent with a complete chromosome. Six additional scaffolds contain one teleomer and are in excess of 1 MB in length. The assembly release Version 2 of whole genome shotgun reads was constructed with the Arachne assembler and improved with finishing reads. This release contains 26 main genome scaffolds totaling 43.9j Mb. Five scaffolds are considered complete teleomere on one end. The remaining 15 scaffolds are smaller and do not contain teleomers. Roughly half of the genome is contained in four scaffolds all at least 5.1 Mbp in length. Annotation of Version 2 assembly was produced by the JGI Annotation Pipeline using a variety of homology-based and ab initio predictors. The Version 1 Gene Catalog and its manual curations also were mapped to the Version 2 assembly and were included in the filtering procedure that determined the initial Version 2 Gene Catalog. After filtering for EST support, completeness and homology support, a total of 11, 609 genes were structurally and functionally annotated.<br /> Ï Use the C. parasitica genome sequence to develop new microarray chip and proteomics platforms for analysis of global gene expression in the blight fungus when challenged by viral pathogens<br /> " Orchard established in WV with advanced, back-cross chestnut trees from VA for assessment of host resistance with hypovirulence in the Cryphonectria parasitica populationreplanted in 2009.<br /> <br /> <br /> 2010 Outcomes<br /> Ï Complete characterization of C. parasitica antiviral RNA silencing pathways.<br /> Two Dicer genes were identified in the C. parasitica genome, cloned and disrupted. Dicer DCL2 was shown to be required for antiviral RNA silencing while Dicer DCL1 was not required (Segers et al., PNAS 2007, 104:12902-12906). Four Argonaute genes were identified in the C. parasitica genome, cloned and disrupted. Only Argonaute AGL2 was required for antiviral RNA silencing (Sun et al., PNAS, IN PRESS). Three RNA dependent RNA polymerases and an orthologue of the QIP exonuclease involved in transgene silencing in Neurospora crassa have been identified in the C. parasitica genome and are currently being cloned and disrupted. The C. parasitica gene oah, encoding the enzyme Oxaloacetate acetylhyrdolase (OAH), a member of the PEP mutase (PEPM)/isocitrate lysase (ICL) superfamily, that catalyzes the hydrolysis of oxaloacetate to oxalic acid and acetate, was cloned, characterized and disrupted. Knockout of the oah gene reduced the ability to form cankers on chestnut trees, which suggest that the enzyme plays a key role in virulence.<br /> <br /> Completion of milestones not proposed:<br /> " A proposal to sequence the C. parasitica genome was approved by the Department of Energy Community Sequencing Program in June of 2006. <br /> The assembled 8.5 X C. parasitica genome sequence was released to the public on September 30, 2008. This is a tremendous resource for future studies on the chestnut blight fungus and its interaction with the chestnut tree. <br /> " Demonstration that hypovirus p29 suppresses RNA silencing in C. parasitica and in heterologous plant system. This is the first report of a mycovirus-encoded suppressor of RNA silencing.<br /> Segers, G.C., van Wezel, R., Zhang, X., Hong, Y. and Nuss, D.L. Hypovirus Papain-like protease p29 suppresses RNA silencing in the natural fungal host and in a heterologous plant system. Eukaryotic Cell 5:896-904, 2006. <br /> " Demonstrated that RNA silencing serves as an antiviral defense mechanism in C. parasitica (first example for any fungus) against hypoviruses and mycoreoviruses.<br /> Segers, G.C., Zhang, X., Deng, F., Sun, Q. and Nuss, D.L. Evidence that RNA silencing functions as an antiviral defense mechanism in fungi. PNAS USA 104:12902-12906, 2007. <br /> " First report of the cloning and sequence analysis of mycovirus-derived small RNAs (vsRNAs) generated by RNA silencing. The vsRNAs were shown to be produced, in a dicer dcl-2-dependent manner, from both positive and negative hypovirus RNA strands at a ratio of 3:2 and to be non-randomly distributed along the viral genome. C. parastiica was shown to respond to mycovirus infection with a 10-15 fold increase in dcl-2 transcript acuumulation while the expression of dcl-1 was modestly increased. The expression of dcl-2 was further increased (~35 fold) following infection by a CHV1-EP713 mutant that lacks the p29 suppressor of RNA silencing. A similar response in dicer gene expression following virus infection of plants or animals has not yet been reported. In this regard, it is anticipated that the evolutionary position of fungi relative to animals and plants will provide insights into additional novel mechanisms for the induction and suppression of RNA silencing pathways yet to be revealed in the other organisms. <br /> Zhang, X., Segers, G.C., Sun, Q., Deng, F. and Nuss, D.L. Characterization of hypovirus-derived small RNAs generated in the chestnut blight fungus by an inducible DCL-2-dependnet pathway. Journal of Virology 82:2613-2619, 2008.<br /> " Virus RNA recombination is an important component of virus evolution that contributes to the emergence of new viruses and the generation of internally deleted mutant RNAs, termed defective interfering (DI) RNAs, that are derived from, and dependent on, the parental viral genomic RNA. We provided the first experimental evidence that a host RNA silencing pathway is required for DI RNA production and virus vector RNA instability for a single-strand, positive sense RNA virus.<br /> Zhang, X. and Nuss, D.L. A host dicer is required for defective viral RNA production and recombinant virus vector instability for a positive sense RNA virus. Proc. Natl. Acad. Sci. USA. Early Edition, October 13, 2008.<br /> <br />

Publications

2009-2010 Publications<br /> <br /> Aguilar, F.X., M.M. Cernusca, and M.A. Gold. 2009. A preliminary assessment of consumer preferences for chestnuts (Castanea spp.) using conjoint analysis. HortTechnology 19: 216-223.<p><br /> <br /> Anagnostakis, Sandra. 2010. Identification of chestnut trees. The Connecticut Agricultural Experiment Station. 29 Sept. 2010. <http://www.ct.gov/caes/lib/caes/documents/publications/fact_sheets/plant_pathology_and_ecology/identification_of_chestnut_trees.pdf>.<p><br /> <br /> Barakat, A., J. Carlson, M. Staton, T. Kubisiak, C. Smith, S. DiLoreto, K. Baier, M. Atkins, B. Blackmon, S. Ficklin, F. Hebard, P. Sisco, W. Powell, S. Anagnostakis, D. Nielsen, A. Abbott, N. Wheeler, and R. Sederoff. 2010. Analyses of the transcriptome of the Fagaceae species. Proceedings of Plant and Animal Genome XVIII Meeting, San Diego, California, January 9-13, 2010, W234.<p><br /> <br /> Barakat, A., D.S. DiLoreto, Y. Zhang, C. Smith, K. Baier, W.A. Powell, N. Wheeler, R. Sederoff, J. E. Carlson. 2009. Comparison of the transcriptomes of American chestnut (Castanea dentata) and Chinese chestnut (Castanea mollissima) in response to the chestnut blight infection. BMC Plant Biology. 9:51 (pp. 1-11).<p><br /> <br /> Carlson, J.E., A.G. Abbott, S. Anagnostakis, K. Baier, A. Barakat, J.B James, N. Islam-Faridi, S. Ficklin, F. Hebard, T. Kubisiak, C. Maynard, S.M. Merkle, W. Miller, C.J. Nairn, W. Powell, S.C. Schuster, L.P. Tomsho, T.K. Wagner and C.D. Nelson. 2010. Forest Health Initiative for American chestnut restoration. Proceedings of Plant and Animal Genome XVIII Meeting, San Diego, California, January 9-13, 2010, P523.<p><br /> <br /> Chen, Chen, Q. Sun, B. Narayanan, D.L. Nuss and O. Herzberg. 2010. Structure of oxaalacetate acetylhydrolase, a virulence factor of the chestnut blight fungus. J. Biol. Chem. 285:26685-26696.<p><br /> <br /> Costa, R., F. Tavares, H. Machado, S. Serrazina, L.T. Dinis, J. Gomez-Laranjo, M.S. Pais, and T. Kubisiak, T. 2009. Understanding disease resistance to Phytophthora cinnamomi in Castanea sp. Proceedings of the Genomics of Forest and Ecosystem Health in the Fagaceae (Beech Family), Nov 10-13, 2009, Research Triangle Park, NC, USA. p.32.<p><br /> <br /> Donis Gonzalez. I.R., M. Mandujano, C. Medina-Mora and D.W. Fulbright. 2009. Presence of mycotoxins after 90 days of storage in fresh chestnuts. 4th International Chestnut Symposium, Beijing, China. Acta Horticulturae 844: 69-74.<p><br /> <br /> Donis Gonzalez, I.R., E.T. Reyser, D. Guyer and D.W. Fulbright. 2010. Efficacy of postharvest treatments for reduction of molds and decay in fresh Michigan chestnuts. First European Chestnut Conference. Acta Horticulturae 866: 563-570.<p><br /> <br /> Donis Gonzalez, I.R., E.T. Reyser, D. Guyer and D.W. Fulbright. 2010. Shell mold and kernel decay of fresh chestnuts in Michigan. First European Chestnut Conference. Acta Horticulturae 866: 353-358.<p><br /> <br /> Fulbright, D.W. 2006. Chestnut culture in Italy. Annual Rept. Northern Nut Growers Association. 1-41 (published in 2010).<p><br /> <br /> Fulbright. D.W., M. Mandujan and S. Stadt. 2010. Chestnut production in Michigan. First European Chestnut Conference. Acta Horticulturae 866: 531-537.<br /> Gold, M.A., M.M. Cernusca and L.D. Godsey. 2009. Agroforestry Product Markets And Marketing. Chapter 11. In: Garrett, H.E. (ed). North American Agroforestry: An Integrated Science and Practice. 2nd Edition. Agronomy Society of America, Madison, WI.<p><br /> <br /> Gold, M.A. and H.E. Garrett. 2009. Agroforestry Nomenclature, Concepts and Practices. Chapter 3. In: Garrett, H.E. (ed). North American Agroforestry: An Integrated Science and Practice. 2nd Edition. Agronomy Society of America, Madison, WI.<p><br /> <br /> Gold, M.A. and M.H. Hall (eds.). 2009. Agroforestry Comes of Age: Putting Science into Practice. 11th North American Agroforestry Conference Proceedings. Columbia, MO. May 31-June 3, 2009. 517 p.<p><br /> <br /> Guyer, D. J. Xing, M. Mandjuano and D.W. Fulbright. 2010. Influence of selected factors on efficiency and effectiveness of a peeling machine for chestnut. First European Chestnut Conference. Acta Horticulturae 866: 595-603.<p><br /> <br /> Hunt, K.L., M.A. Gold and M.R. Warmund. 2008. Status of Chinese Chestnut (Castanea mollissima) Cultivar Performance in Missouri. 4th International Chestnut Congress Sept. 25-28, 2008, Beijing, China.<p><br /> <br /> Islam-Faridi, N., C.D. Nelson, P.H. Sisco, T.L. Kubisiak, F.V. Hebard, R.L. Paris and R.L. Phillips. 2009. Cytogenetic analysis of American chestnut (Castanea dentata) using fluorescent in situ hybridization. Acta Hort. (ISHS) 844:207-210.<p><br /> <br /> Liang, H, P.A. Kumar, V. Nain, W.A. Powell, J.E. Carlson. Selection and Screening Strategies. Chapter 4. In: C. Kole, C.H. Michler, A.G. Abbott and T,C. Hall (eds) Transgenic Crop Plants: Volume 1: Principles and Development. Springer-Verlag  Berlin, Heidelberg, New York, Tokyo, 2010, pp 85-144.<p><br /> <br /> Lin, Y-C., J-M. Frigerio, C. Cabane, C. Bodenes, A. Daruvar, P. Rouze, M. Staton, A. Barakat, T. Kubisiak, A. Kremer, C. Plomion and Y. Van de Peer. 2009. In silico screening of conserved orthologous sequences (COS) from Fagaceae ESTs resources. Proceedings of the Genomics of Forest and Ecosystem Health in the Fagaceae (Beech Family), Nov 10-13, 2009, Research Triangle Park, NC, USA. Poster No.17, p.54.<p><br /> <br /> Medina-Mora, C.M. and D.W. Fulbright. 2010. Evaluation of simple sequence repeats (SSR) for genetic analysis of chestnut trees in Michigan orchards. First European Chestnut Conference. Acta Horticulturae 866: 127-133.<p><br /> <br /> Milgroom, M.G., K. Sotirovski, M. Risteski and M.T. Brewer. 2009. Heterokaryons and recombinants of Cryphonectria parasitica in two clonal populations in southeastern Europe. Fungal Genet. Biol.46: 849-854.<p><br /> <br /> Nelson, C.D., J.E. Carlson, S.A. Merkle, W. Powell, C.J. Nairn, C. Maynard, T.L. Kubisiak, F.V. Hebard, S. Anagnostakis and A. Abbott. 2009. Biotechnological tools for accelerating species restoration. Proceedings of the Genomics of Forest and Ecosystem Health in the Fagaceae (Beech Family), Nov 10-13, 2009, Research Triangle Park, NC, USA. p.39.<p><br /> <br /> Salamon, J.A., R. Acuña and A. L. Dawe. 2010. Phosphorylation of phosducin-like protein BDM-1 by protein kinase 2 (CK2) is required for virulence and Gb subunit stability in the fungal plant pathogen Cryphonectria parasitica. Mol. Microbiol. 76: 848-860.<p><br /> <br /> Sederoff, R., N. Wheeler, J. Carlson, A. Barakat, W. Powell, K. Baier, T. Kubisiak, P. Sisco, F. Hebard, S. Anagnostakis, J. Tomkins, A. Abbott, M. Staton, C. Smith, D. Nielsen and S. Ficklin. 2010. A genome platform for chestnut. Proceedings of Plant and Animal Genome XVIII Meeting, San Diego, California, January 9-13, 2010, W253.<p><br /> <br /> Sederoff, R., N. Wheeler, J. Carlson, A. Barakat, W. Powell, K. Baier, T. Kubisiak, P. Sisco, F. Hebard, S. Anagnostakis, J. Tomkins, A. Abbott, M. Staton, C. Smith, D. Nielson and S. Ficklin. 2009. The chestnut genome project: looking ahead, opportunities, and challenges. Proceedings of the Genomics of Forest and Ecosystem Health in the Fagaceae (Beech Family), Nov 10-13, 2009, Research Triangle Park, NC, USA. p.7.<p><br /> <br /> Sisco, P.H., R.R. Sederoff, J.P. Tomkins, J.E. Carlson, T.L. Kubisiak, M.E. Staton, F.V. Hebard, S.L. Anagnostakis, W.A. Powell and C.P. Smith. 2009. The United States National Science Foundation project on developing tools for the study of the Fagaceae: Castanea, Quercus, and Fagus. Acta Hort. (ISHS) 844:267-274.<p><br /> <br /> Spinger, J.C., A.L.D. Baines and A.M. Jarosz. 2009. Hypovirus mediated recovery of blight infected American chestnut trees in Michigan. Phytopathology 99: S123.<p><br /> <br /> Springer, J.C., M.T. Chansler, A.L.D. Baines and A.M. Jarosz. 2010. Vegetative incompatibility diversity in Michigan populations of the chestnut blight fungus, Cryphonectria parasitica. 95th Meeting of the Ecological Society of America, Pittsburgh, PA.<p><br /> <br /> Staton, M., D. Henry, M. Monsanto, S. Ficklin, C-H. Cheng, G., Fang, B. Blackmon, A.Barakat, J. Tomkins, F. Hebard, T. Kubisiak, J. Carlson, R. Sederoff, R. and A. Abbott. 2009. Comparative genome analysis of Chinese chestnut to model plant species and Fagaceae family species. Proceedings of the Genomics of Forest and Ecosystem Health in the Fagaceae (Beech Family), Nov 10-13, 2009, Research Triangle Park, NC, USA. p.20.<p><br /> <br /> Sun, Qihong, G.H. Choi and D.L. Nuss. 2009. A single argonaute gene is required for induction of RNA silencing antiviral defense and promotes viral RNA recombination. Proc. Nat. Acad. Sci. 106:17927-17932.<p><br /> <br /> Valdivia, C., L. Zabek, J. Arbuckle, M. Gold and C. Flora. 2009. Human and Institutional Dimensions of Agroforestry. Chapter 13. In: Garrett, H.E. (ed). North American Agroforestry: An Integrated Science and Practice. 2nd Edition. Agronomy Society of America, Madison, WI.<p><br /> <br /> Wheeler, N., D. Nielson, C. Smith, J. Tompkins, A. Abbott, M. Atkins, B. Blackmon, M. Staton, S. Ficklin, F. Hebard, P. Sisco, J. Carlson, A. Barakat, W. Powell, K. Baier, S. Anagnostakis, T. Kubsiak, C. Burdine and R. Sederoff. 2009. Annotated EST-based SSR and SNP genetic maps for chestnut. Proceedings of the Genomics of Forest and Ecosystem Health in the Fagaceae (Beech Family), Nov 10-13, 2009, Research Triangle Park, NC, USA. p.11.<br /> <br /> <br /> <br />

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