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Burke; Nicolas Langlade; Stphane MuosInboxProject / Consortium meeting at PAG'ߎG{__substg1.0_0E02001F*__substg1.0_0E03001F*8:__substg1.0_0E04001F*Z`__substg1.0_0E05001F*9=\ __substg1.0_0E1D001F*]F__substg1.0_0E460102*<>___substg1.0_0E480102*`__substg1.0_0E4C0102*;Ca'ߎG{'ߎG{ *>h^«F *>h^«F *>h^«FBtD'A9 j 645FA64427214D498120FA2CD6839E51@ead.ubc.ca__substg1.0_0E530102*b__substg1.0_0E550102*@Bc__substg1.0_0E590102*d__substg1.0_0F030102*AEe__substg1.0_1000001F*J/__substg1.0_10090102*DFbt__substg1.0_1015001F*fV__substg1.0_1035001F*?Whdwith revised proposal attached Hi all, I made additional revisions to the Consortium proposal based on John s comments below. Please take a look and try to get back to me on Monday so we can get this to the companies. Note that I made some executive decisions to move things forward. Thus, take a close look and make changes as you feel necessary. Speed is important now. Nicolas - can you send me email contacts for the additional companies that might be interested? thanks, loren > On Oct 30, 2015, at 4:45 AM, John M. Burke <jmburke@uga.edu> wrote: > > Hi Loren, I'm very sorry but I haven't had time to look at this until just now. I am doubled up on teaching right now and also helping Ed to get his dissertation ready for his committee and those two things I've taken essentially all of my time. > > Anyway, it looks okay to me. A couple of comments: > > I think it would be fine to include he wild species data since we are producing it anyway. That could make it more attractive, though I'm honestly not sure how interested they would be. > > For stress responsive genes, I wonder if we shouldn't offer more since we will be doing more as part of the NSF and Genome Canada projects. All that is discussed at present is drought on Heliaphen but we will be doing RNAseq for all focal stresses in the NSF and Genome Canada projects so perhaps we could make this a bugger aspect of the proposal? > > Thanks, John > > On Wednesday, October 28, 2015, Rieseberg, Loren <loren.rieseberg@botany.ubc.ca> wrote: > Hi John, > > Have you had a chance to look at this? I wonder if we should include the sequences for the wild species we will be generating as part of the NSF / Genome Canada grant? I need to send something to the consortium tomorrow or Thursday announcing the Jan. 13 meeting. > > Thanks, loren > >> Begin forwarded message: >> >> From: Loren Rieseberg <lriesebe@mail.ubc.ca> >> Date: October 19, 2015 at 2:43:18 PM PDT >> To: Nicolas Langlade <Nicolas.Langlade@toulouse.inra.fr> >> Cc: "John M. Burke" <jmburke@uga.edu>, Stphane Muos <stephane.munos@toulouse.inra.fr>, Jerome Gouzy <Jerome.Gouzy@toulouse.inra.fr>, Hlne Bergs <Helene.Berges@toulouse.inra.fr> >> Subject: Re: Project / Consortium meeting at PAG >> >> Hi Nicolas, >> >> Thanks for putting this together. It is indeed very ambitious! I worry a bit about the costs of the PacBio sequencing proposed and think at least for the wilds it might make more sense to use the long read Illumina approach (more within my budget!). Also, I focused section 2 on further characterizing existing resources rather than additional data gathering. John and I have put a huge amount of effort into the SAM population and pre-bred lines, so I think our goal should be make these resources more useful rather than proposing additional sequencing. However, it is very exciting! >> >> Thanks, loren >> >> >> >>> >>> On Oct 18, 2015, at 10:07 AM, Nicolas Langlade <nicolas.langlade@toulouse.inra.fr> wrote: >>> >>> Hi Loren ad John, >>> Here is a draft project for the next 4-5 years of the consortium. >>> This is ambitious, but we think we should discuss on an ambitious ground with the private partners. >>> Please comment, complete, modify as you feel this document. >>> Looking forward to reading your comments, >>> Nicolas >>> >>> Le 15/10/2015 05:53, Rieseberg, Loren a crit : >>>> Hi John and Nicolas, >>>> >>>> We probably should send a note to the consortium and NSF/Genome Canada project about the PAG meeting soon so people can register and make travel arrangements. For consortium members, we probably should include a note about the Consortium plans over the next five years. >>>> >>>> Nicolas - have you had a chance to draft something? It looks like the short note I sent to Advanta with your initial thoughts was enough to change their minds. I had a meeting with Pioneer today where I mentioned that you had purchased a PacBio machine and sequenced and assembled two genomes. They were very impressed and will be sending someone to the consortium meeting to learn more. Also, you asked what we can contribute over the next several years. I think we can contribute an association mapping pipeline linked to the reference genome, WGS data for a wide array of wild diversity, genotypic data for the pre-bred lines at the USDA, as well as for the new pre-bred lines we have generated, and probably one PacBio assembly (not sure what genotype). >>>> >>>> Best Wishes, loren >>>> >>>> >>>> >>>> >>>> >>>> Loren Rieseberg >>>> Botany Department >>>> University of British Columbia >>>> 3529-6270 University Blvd >>>> Vancouver, B.C. V6T 1Z4 >>>> Phone: 604-827-4540 >>>> Fax: 604-822-6089 >>>> >>> >>> -- >>> ================================================================== >>> >>> Nicolas Langlade >>> >>> Laboratoire Interactions Plantes Microorganismes >>> >>> INRA-CNRS >>> 24 Chemin de Borde Rouge - Auzeville >>> CS 52627 >>> 31326 Castanet Tolosan FRANCE >>> >>> Tel : +33 (0)5 61 28 57 78 (nouveau numro) >>> Fax : +33 (0)5 61 28 50 61 >>> Lab website : http://www2.toulouse.inra.fr/centre/lipm/ >>> Sunrise project website: http://www.sunrise-project.fr/ >>> Sunflower genomics webportal : http://www.heliagene.org/ >>> >>> ================================================================== >>> >>> <Ideas continuation international consortium 20151014.docx> >> >> Loren Rieseberg >> Botany Department >> University of British Columbia >> 3529-6270 University Blvd >> Vancouver, B.C. V6T 1Z4 >> Phone: 604-827-4540 >> Fax: 604-822-6089 >> > > Loren Rieseberg > Botany Department > University of British Columbia > 3529-6270 University Blvd > Vancouver, B.C. V6T 1Z4 > Phone: 604-827-4540 > Fax: 604-822-6089 > > > > -- > Sent from a mobile device. Loren Rieseberg Botany Department University of British Columbia 3529-6270 University Blvd Vancouver, B.C. V6T 1Z4 Phone: 604-827-4540 Fax: 604-822-6089 pvLZFuS rcpg1252Chtml11bid0 'ch set0 PVUUQW2U3FYkcm ;O05R `cgP d36 4r *\9 <>cg3 ead} "g4m/n?LpM- N__xOy_zo{~dw/hOcHiG0ll,LM/N?nLχ/?O"0I G0dPti GF3 top$QiumFbaF! Joh '962 mb be)w.+8&nbk`'a@qǞ v0$pakR okG0nFtryrg!cPM da EЏc"FaakqsO_lNot@ő יvupvϐ`ng 2gl>OO 303 15 Q4:t45M .| Bp׿< qjܲ@uga. uϖw`:oՏ֟ׯߖͅLʂ I'!߰bn'@qr$uvu0jdqad``FupGq!F #c@Ӛ$lpEQ!rF!$򛢓bEhbtw#җb1lЅᒀcmq/?O_o Anyy E@܀AQ0kf /?O_o PulFqfrauE qk`њpG`g!`!ʐF`@܀Q q3#GAbڐ!PFPkgܰ! 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Please take a look and try to get back to me on Monday so we can get this to the companies. Note that I made some exeRieseberg, Loren+nTRieseberg, LorenSMTPloren.rieseberg@botany.ubc.caJohn M. Burkeܧ@B+//O=EXCHANGELABS/OU=EXCHANGE ADMINISTRATIVE GROUP (FYDIBOHF23SPDLT)/CN=RECIPIENTS/CN=JMBURKESMTPloren.rieseberg@botany.ubc.ca__substg1.0_3FF90102*__substg1.0_3FFA001F*PR__substg1.0_3FFB0102*x__substg1.0_4022001F*QU__substg1.0_4023001F*:__substg1.0_4024001F*TV__substg1.0_4025001F*__substg1.0_4030001F*O_ EX/O=EXCHANGELABS/OU=EXCHANGE ADMINISTRATIVE GROUP (FYDIBOHF23SPDLT)/CN=RECIPIENTS/CN=JMBURKERieseberg, LorenRieseberg, LorenJMBURKEJMBURKE__substg1.0_4031001F* __substg1.0_4034001F*XZ__substg1.0_4035001F*__substg1.0_4038001F*Y] Rieseberg, LorenJohn M. BurkeJohn M. BurkeJohn M. Burkeloren.rieseberg@botany.ubc.caloren.rieseberg@botany.ubc.ca{QG?'q en-US, en-CA__substg1.0_4039001F*__substg1.0_405E001F*\^__substg1.0_4060001F*__substg1.0_5D01001F*[g:__substg1.0_5D02001F*:__substg1.0_64F00102*`bL__substg1.0_65E10102*__substg1.0_8003001F*aeReceived: from BLUPR02MB1650.namprd02.prod.outlook.com (10.162.213.16) by BY2PR02MB1655.namprd02.prod.outlook.com (10.163.27.17) with Microsoft SMTP Server (TLS) id 15.1.312.18 via Mailbox Transport; Fri, 30 Oct 2015 22:36:32 +0000 Received: from BY1PR0201CA0019.namprd02.prod.outlook.com (10.160.191.157) by BLUPR02MB1650.namprd02.prod.outlook.com (10.162.213.16) with Microsoft SMTP Server (TLS) id 15.1.312.18; Fri, 30 Oct 2015 22:36:30 +0000 Received: from BN1AFFO11FD048.protection.gbl (2a01:111:f400:7c10::109) by BY1PR0201CA0019.outlook.office365.com (2a01:111:e400:4814::29) with Microsoft SMTP Server (TLS) id 15.1.312.18 via Frontend Transport; Fri, 30 Oct 2015 22:36:30 +0000 Authentication-Results: spf=none (sender IP is 142.103.117.138) smtp.mailfrom=botany.ubc.ca; uga.edu; dkim=none (message not signed) header.d=none;uga.edu; dmarc=none action=none header.from=botany.ubc.ca; Received-SPF: None (protection.outlook.com: botany.ubc.ca does not designate permitted sender hosts) Received: from vmaprod4.mail-relay.ubc.ca (142.103.117.138) by BN1AFFO11FD048.mail.protection.outlook.com (10.58.53.63) with Microsoft SMTP Server (TLS) id 15.1.318.9 via Frontend Transport; Fri, 30 Oct 2015 22:36:29 +0000 Received: from vmaprod4.mail-relay.ubc.ca (localhost.localdomain [127.0.0.1]) by localhost (Email Security Appliance) with SMTP id BF06B26BA18_633F0ECB; Fri, 30 Oct 2015 22:36:28 +0000 (GMT) Received: from mx2.mail-relay.ubc.ca (lb_mx-out.mail-relay.ubc.ca [10.92.8.24]) (using TLSv1.2 with cipher ECDHE-RSA-AES256-GCM-SHA384 (256/256 bits)) (Client CN "mx2.mail-relay.ubc.ca", Issuer "mx2.mail-relay.ubc.ca" (not verified)) by vmaprod4.mail-relay.ubc.ca (Sophos Email Appliance) with ESMTPS id 24A5726B83D_633F0ECF; Fri, 30 Oct 2015 22:36:28 +0000 (GMT) Received: from smtp.mail.ubc.ca (s-itsv-hub04p.ead.ubc.ca [137.82.151.86]) (using TLSv1 with cipher AES256-SHA (256/256 bits)) (Client CN "rpc.mail.ubc.ca", Issuer "Entrust Certification Authority - L1K" (not verified)) by mx2.mail-relay.ubc.ca (Postfix) with ESMTPS id DD2B215F9A7; Fri, 30 Oct 2015 15:36:27 -0700 (PDT) Received: from EXCH-MBX05P.ead.ubc.ca ([169.254.5.167]) by s-itsv-hub04p.ead.ubc.ca ([137.82.151.86]) with mapi id 14.03.0248.002; Fri, 30 Oct 2015 15:36:27 -0700 From: "Rieseberg, Loren" To: "John M. 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--_000_BFBA7CE738D24ADF984C5D860D05FF28mailubcca_-- --_004_BFBA7CE738D24ADF984C5D860D05FF28mailubcca_ Content-Type: application/msword; name="Sunflower Genomic Resources Consortium Continuation_LR.doc" Content-Description: Sunflower Genomic Resources Consortium Continuation_LR.doc Content-Disposition: attachment; filename="Sunflower Genomic Resources Consortium Continuation_LR.doc"; size=107008; creation-date="Fri, 30 Oct 2015 22:36:26 GMT"; modification-date="Fri, 30 Oct 2015 22:36:26 GMT" Content-ID: Content-Transfer-Encoding: base64 --_004_BFBA7CE738D24ADF984C5D860D05FF28mailubcca_-- __substg1.0_8004001F*__substg1.0_8005001F*df__substg1.0_8006001F*:__substg1.0_8008001F*ckBT=1;II=[CID=4db8d86d-1824-4274-bd53-08d87e051d4a;IDXHEAD=01D1130868;IDXCOUNT=2];SBMID=20;S1=<022A2152-ADD0-4FF3-9081-CA521BBFAF9A@mail.ubc.ca>;FIXUP=22.6001;Version=Version 15.1 (Build 312.0), Stage=H10, TC, REFenBN1AFFO11FD048.protection.gblAnonymousNone (protection.outlook.com: botany.ubc.ca does not designate permitted sender hosts)<?xml version="1.0" encoding="utf-16"?> <EmailSet> <Version>15.0.0.0</Version> <Emails> __substg1.0_800A001F*__substg1.0_800C001F*hjj__substg1.0_800D001F*@__substg1.0_800E0048*io <Email StartIndex="536"> <EmailString>jmburke@uga.edu</EmailString> </Email> <Email StartIndex="1492" Position="Signature"> <EmailString>loren.rieseberg@botany.ubc.ca</EmailString> </Email> <Email StartIndex="1896" Position="Other"> <EmailString>lriesebe@mail.ubc.ca</EmailString> </Email> <Email StartIndex="1989" Position="Other"> <EmailString>Nicolas</EmailString> </Email> </Emails> </EmailSet><?xml version="1.0" encoding="utf-16"?> <ContactSet> <Version>15.0.0.0</Version> <Contacts> <Contact StartIndex="521"> <Person StartIndex="521"> <PersonString>John M. Burke</PersonString> </Person> <Emails> <Email StartIndex="536"> <EmailString>jmburke@uga.edu</EmailString> </Email> </Emails> <ContactString>John M. Burke &lt;jmburke@uga.edu</ContactString> </Contact> <Contact StartIndex="1485" Position="Signature"> <Person StartIndex="1485" Position="Signature"> <PersonString>Loren</PersonString> </Person> <Emails> <Email StartIndex="1492" Position="Signature"> <EmailString>loren.rieseberg@botany.ubc.ca</EmailString> </Email> </Emails> <ContactString>Loren &lt;loren.rieseberg@botany.ubc.ca</ContactString> </Contact> </Contacts> </ContactSet>shuJQU1ʒBzz{+Bias?SenderIsYou2DisplayName?$ConversationStarterIsYou2DisplayName?OnlyRecipie__substg1.0_800F0048*__substg1.0_80100102*ln__substg1.0_8014001F*__substg1.0_8016001F*mqntDisplayName?ConversationContributions?FlaggedBySender?MarkedImportantBySender?MarkedUnimportantBySender?SenderIsManagerDisplayName?SenderIsDirectReportDisplayName?SenderIsAutomated? 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NgramsInBody2I?DecisionTree1_22_55_12_Nov_14?DecisionTree2_22_55_12_Nov_148?DecisionTree3_22_55_12_Nov_14{?DecisionTree4_22_55_12_Nov_14 ?SenderCcDisplayNameSenderDListDisplayName ConversationStarterCcDisplayName#ConversationStarterDListDisplayNameRecipientsOnCcLineDisplayName2DistributionGroupsDisplayName["TeeEngineVersion","26.0.0","BuildNumber","Autopilot Exchange SDF_20151019.1","ChangesetNumber","C6151","Locale","en-US","MessageId","\u003cBFBA7CE7-38D2-4ADF-984C-5D860D05FF28@mail.ubc.ca\u003e"]<?xml version="1.0" encoding="utf-16"?> <TaskSet> <Version>15.0.0.0</Version> <Tasks>       !"#$%&'()*+,-./01234567;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[`bfginpqrstwxz{ <Task StartIndex="130"> <TaskString>Please take a look and try to get back to me on Monday so we can get this to the companies.</TaskString> <Assignees> <EmailUser Id="jmburke@uga.edu">John M. Burke</EmailUser> <EmailUser Id="Nicolas.Langlade@toulouse.inra.fr">Nicolas Langlade</EmailUser> <EmailUser Id="stephane.munos@toulouse.inra.fr">Stphane Muos</EmailUser> </Assignees> </Task> <Task StartIndex="378"> <TaskString>Nicolas - can you send me email contacts for the additional companies that might be interested?</TaskString> <Assignees> <EmailUser Id="jmburke@uga.edu">John M. Burke</EmailUser> <EmailUser Id="Nicolas.Langlade@toulouse.inra.fr">Nicolas Langlade</EmailUser> <EmailUser Id="stephane.munos@toulouse.inra.fr">Stphane Muos</EmailUser> </Assignees> </Task> <Task StartIndex="1546"> <TaskString>&gt; Have you had a chance to look at this?</TaskString> <Assignees> <EmailUser Id="jmburke@uga.edu">John M. Burke</EmailUser> <EmailUser Id="Nicolas.Langlade@toulouse.inra.fr">Nicolas Langlade</EmailUser> <EmailUser Id="stephane.munos@toulouse.inra.fr">Stphane Muos</EmailUser> </Assignees> </Task> <Task StartIndex="1587"> <TaskString>I wonder if we should include the sequences for the wild species we will be generating as part of the NSF / Genome Canada grant?</TaskString> <Assignees> <EmailUser Id="jmburke@uga.edu">John M. 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Burkesip:jmburke@uga.eduܧ@B+//o=ExchangeLabs/ou=Exchange Administrative Group (FYDIBOHF23SPDLT)/cn=Recipients/cn=jmburke $ :<% :< __substg1.0_5FF70102*mx__properties_version1.00oX__recip_version1.0_#00000001:fjjfjj__substg1.0_0FF60102*u:<x:<x:<000 0_:<99@9  : @:_(_x:<_0t+nTNicolas LangladeSMTPNicolas.Langlade@toulouse.inra__substg1.0_0FF90102*v__substg1.0_0FFF0102*y__substg1.0_3001001F*| __substg1.0_3002001F*}.fr+nTNicolas LangladeSMTPNicolas.Langlade@toulouse.inra.frNicolas LangladeSMTPNicolas.Langlade@toulouse.inra.fr__substg1.0_3003001F*~B__substg1.0_300B0102*'__substg1.0_39FE001F*B__substg1.0_3A20001F* SMTP:NICOLAS.LANGLADE@TOULOUSE.INRA.FRNicolas.Langlade@toulouse.inra.frNicolas Langlade+nTNicolas LangladeSMTPNicolas.Langlade@toulouse.inra.fr  <__substg1.0_5FF70102*__properties_version1.00__recip_version1.0_#00000002:fjjEgjj__substg1.0_0FF60102*<<0"0 0D 0'<99D :" @:_<_0w+nTStphane MuosSMTPstephane.munos@toulouse.inra.fr+nTStphane MuosSMTP__substg1.0_0FF90102*__substg1.0_0FFF0102*__substg1.0_3001001F*__substg1.0_3002001F*stephane.munos@toulouse.inra.frStphane MuosSMTPstephane.munos@toulouse.inra.frSMTP:STEPHANE.MUNOS@TOULOUSE.INRA.FRstephane.munos@toulouse.inra.frStphane Muos+nTStphane MuosSMTP__substg1.0_3003001F*>__substg1.0_300B0102*%__substg1.0_39FE001F*>__substg1.0_3A20001F*__substg1.0_5FF70102*__properties_version1.00__attach_version1.0_#00000000<Egjjhjj__substg1.0_0FF90102*stephane.munos@toulouse.inra.fr  *<*<*<00 0@ 0%*<99@ : @:_*<_0Sunflower Genomic Resources Cons__substg1.0_3001001F*t__substg1.0_37010102*__substg1.0_3703001F*__substg1.0_3704001F*ortium Continuation_LR.doc.docSunflo~1.docSunflower Genomic Resources Consortium Continuation_LR.docapplication/mswordDE11F4F522DA7C499F27830B82567EB0@ead.ubc.ca      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~ࡱ>  |bjbj 4G3 &" .BBBBB0j(mW\0///////,25X/ mm /)0))) /)/))V0#@#AJbp# /000|#x6)6#)#$ //)06 :   Prospective project for the continuation of the international consortium for the development of genomics, genetics and bioinformatics resources for sunflower Partnership Public laboratories: University of British Columbia, University of Georgia, French National Institute of Agronomical Research (INRA) Private partners: up to now Biogemma (representing Soltis and RAGT2n), Syngenta, Pioneer, Dow, Advanta, KWS, Bayer. Potential new partners include MaisAdour, Caussade, NuSeeds, others?. Context The Sunflower Genomic Resources Consortium was created in 2012 in conjunction with a Genome Canada project led by L. Rieseberg lab in collaboration with UGA and INRA. The main goal was to pursue the production of the first high quality genomic sequence of sunflower genotype HA412-HO. In January 2015, the consortium partners decided to stop the assembly efforts and consider the bronze version of HA412-HO as the first reference sequence despite some assembly problems at the local scale. This reference sequence was made available to the consortium partners in January 2015 and 6 months later publicly at  HYPERLINK "https://www.heliagene.org/HA412.v1.1.bronze.20141015/" https://www.heliagene.org/HA412.v1.1.bronze.20141015/ . Concomitantly, the emergence of PacBio technology enabled INRA to initiate the sequencing of XRQ and PSC8 lines using this technology in the framework of the SUNRISE and HELIOR projects. In September 2015, initial results from PacBio sequencing of XRQ to 100X depth, indicate clearly that the sequence to be produced will be of much better quality. However, this genome will still be incomplete (about 20% of the sequence missing), and mis-assembly issues are likely due to scaffolding based on genetic markers only. Non-recombining regions and identical regions between parental lines of mapping populations will therefore likely contain local scaffolding errors. The production of pseudomolecules and annotation of the XRQ genome sequence is under way and should be made available to the consortium through the INRA website  HYPERLINK "http://www.heliagene.org" www.heliagene.org in early 2016. In addition, the sequencing of the PSC8 line is under way with a final coverage of around 50X. It has started to produce results that we have analyzed at the candidate locus level. Although gene order is fairly well conserved across both genotypes, intergenic regions, comprising very high levels of repeat elements, were very different and could hardly be aligned. The release date of the PSC8 sequence has not yet been determined. The consortium reached its goal to produce the first annotated genomic sequence of sunflower based on data produced by the Genome Canada project and INRA. It provided to the consortium members major resources such as BAC libraries ready to screen for fine-mapping, a physical map, reference transcriptomes with organ-specific expression patterns, a high quality and high density genetic map, and re-sequencing data for the 288 genotypes of the sunflower association mapping population. In addition, it secured the production of a second genomic sequence of higher quality in 2016. These resources will be a cornerstone for sunflower genetic and genomic research and breeding. Importantly, this consortium was a unique forum to bring together key international actors on one hand from the academic community and on the other hand from the biotech and seed companies. However, our current analysis of structural variation from high quality PacBio assemblies clearly shows that our knowledge of sunflower genomes is far from satisfactory, which will impede fast and efficient QTL fine-mapping and the introgression of advantageous alleles. We therefore believe that it is necessary for our community to continue these efforts and further develop resources in genomics, genetics and bioinformatics to take advantage of the post-genomic era in sunflower. This effort could be organized in four work packages: Produce genomic sequences of key sunflower genotypes Develop a novel genotyping tool that will include structural variation Develop and characterize genetic resources Produce an atlas of stress responsive genes for genome annotation and reverse genetics Work package 1. Produce genomic sequences of key sunflower genotypes Task 1.1 Sequence sunflower cultivated lines using PacBio technology Task leader: Nicolas Langlade, INRA Partners involved: INRA, UBC, UGA Aim: Produce the genomic sequences of sunflower lines chosen for their maximal diversity in the cultivated material. We aim to sequence 16 inbred lines representing a large genetic diversity in the cultivated pool originating both from USDA and INRA (Table 1). Strategy: We will use PacBio sequencing based on our successful experience to sequence the XRQ and PSC8 genomes. Our strategy will be to produce very-long insert size libraries to reach a contig N50 of circa 14kb with a total coverage around 60X. This should represent 200 SMRTCell with a throughput of 1Gb/SMRTCell. Bioinformatics analysis will consist in de novo assembly and annotation for each genome. Comparison with previously sequenced genomes will be performed to aid generation of pseudomolecules. Cost: Sequencing = 1700 k (500 / SMRTCell ( 100k per genotype) Bioinformatics = $165 k ($55 k / year x 3 years) Timeline: this will be spread across the first three years of the consortium. The sequencing and assembly will be performed at INRA Toulouse. Based on todays PacBio RSII sequencer outputs, we can sequence three to four sunflower genomes per platform per year. The new PacBio Sequel will allow us to sequence seven times more and therefore more easily reach our goal. Task 1.2 Sequence wild sunflower genomes using Illuminas Long-read Sequencing Technology Task leader: Loren Rieseberg, UBC Partners involved: INRA, UBC, UGA Aim: Produce the genomic sequences of wild sunflower relatives chosen for their diversity and source of resistance genes. We aim to sequence 3 H. annuus ecotypes carrying abiotic tolerance alleles and 6 Helianthus species (H. anomalus, H. argophyllus, H. exilis, H. paradoxus, H. petiolaris and H. debilis) carrying disease and orobanche resistance genes (Table 2). Strategy: Long-read sequencing technology generates large DNA fragments for sequencing, and assembles them into synthetic long reads (circa 10kb) that can be used for assembly. Our strategy will be to produce Tru-Seq synthetic long read libraries and sequence to circa 60X depth. This approach will produce less complete assemblies than PacBio, but also at much less expense. Cost: Sequencing = 90 k ($10 k / genome x 9 genomes); Bioinformatics = 165 k ($55 k / year x 3 years) Timeline: This will be spread across the first three years of the consortium. The sequencing would likely be performed at Genome Quebec in Montreal, whereas assembly would be performed at UBC. Work package 2. Develop and characterize genetic resources Task 2.1. Genotyping and phenotyping of the public cultivated sunflower association mapping (SAM) population, three wild sunflower association populations that are under development, as well as public pre-bred lines. Task leader: Loren Rieseberg, UBC and John Burke, UGA Partners involved: INRA, UBC, UGA Aim: Characterize genetic polymorphisms of wild cultivated and pre-bred material Strategy: During the first four years of the consortium, we re-sequenced the 288 cultivated genotypes making up the SAM population to 5-10x depth. This is a very large data set (>10 TB), so variant calling was a challenge. However, the SNP calls are now complete, which we will release to the consortium later this year. The SNPs were called against the bronze genome, which is not as complete or accurate as the XRQ sequence. Also, structural variants have not been called. Therefore, over the next two years, we will re-align these sequences against the XRQ reference sequence and call both SNPs and structural variants. An association mapping pipeline will be developed and provided along with phenotypic data for many agriculturally relevant traits, including drought, low nutrient, salt, and flooding tolerance. The association population has very low LD throughout much of the genome, so it is possible in many cases to map QTLs down to individual genes. Therefore, we expect this resource to be of conservable value to the sunflower community. In addition to the SAM population resources, in the context of a new Genome Canada project, we are developing association populations for wild populations of three species: H. annuus, H. argophyllus, and H. petiolaris. Approximately 500 individuals of each species will be sequenced to 5-7x depth and their genomes will be scanned for associations with drought, low nutrient, salt, and flooding tolerance. Lastly, in the context of an ongoing project with the Global Crop Diversity Trust on adapting crops to climate changes, we will provide phenotypic and high-resolution genotypic data for more than 400 public pre-bred lines developed by the USDA and UBC (Table 3). This will facilitate the introgression of valuable alleles from wild sunflower accessions. Cost: Bioinformatics = $220 k ($55 k / year x 4 years); Sequencing, genotyping, and phenotyping costs covered by NSF, Genome Canada, and Crop Trust grants Timeline: The genotypic data for the pre-bred lines will be made available in year 1, whereas the new set of variant calls and association mapping pipeline for the SAM population with not be available until near the end of year 2. Association mapping in the wild species association populations will be carried out in years 3 and 4. Both field- and greenhouse-derived phenotypic data will be added to the association mapping website/pipeline over the next four years. Task 2.2. Produce a mutagenized population Task leader: Nicolas Langlade, INRA Partners involved: public partners + private Aim: produce a very large mutant population in an already sequenced reference maintainer line (e.g. XRQ) for forward and reverse genetics. Produce DNA pools for screening for reverse genetics. Strategy: Two strategies will be used: EMS and irradiation (gamma-ray) mutagenesis. The target for EMS would be 20,000 M2 families. EMS mutagenesis and production of M1 seeds can be difficult and the number of EMS families would depend on M1 seed production. Each partner would multiply 1000 families over 2 years and provide the DNA to INRA-CNRGV. Our target for irradiation mutagenesis would be 20,000 M2 families as well. Each partner would be in charge of producing M1 and M2 generations for 2000 families. M2 seeds for both strategies will be pooled per family (>8 plants) and sent to one or several partner(s) for DNA extraction. Cost: Irradiation cost to be determined (quote under way), M1 and M2 multiplications by private partners. EMS: either sub-contracted or mutagenesis shared between public labs. M2 production would be shared between private partners. 20k for DNA extraction and 10k for pooling Timeline: Irradiation strategy - mutagenesis in 2016, M1 in 2016 (if early agreement), otherwise in 2017. M2 in 2017 or 2018. EMS strategy: mutagenesis and M1 in 2017 and M2 in 2018. Note regarding legal issues: IP for the genetic material will be shared between all WP partners. All WP partners will have access to it. Use of this genetic material in collaboration with public labs may be permitted unless a non-member company is involved. Work package 3. Develop a novel genotyping tool that can assay both SNP and structural variants and genotype MAGIC population Task 3.1. Develop a 500k AXIOM array for assaying both SNP and structural variants. Task leader: Stphane Muos, INRA Partners involved: INRA, UBC, UGA Aim: Produce a high-throughput genotyping tool such as Affymetrix AXIOM chip to genotype SNP and structural variants across the sunflower genome. Strategy: identify SNPs based on resequencing data already produced by the consortium including the SAM population and genotyping by sequencing produced in WP2. The goal will be to cover every gene with at least 3 SNPs. Based on PacBio de novo sequencing from WP1, structural polymorphisms will be identified and markers developed to type them. Our goal is develop approximately 500k markers based on the AXIOM technology. Cost: 40 k to cover the bioinformatics design. We expect a genotyping price of circa 200 /per sample if we have enough pre-orders. Partners involved in chip development and pre-orders will have access to the chip. Anonymous results of genotyping will be shared between partners to evaluate the tool design and marker quality. Timeline: Chip development would take place in 2018 after most of the de novo sequencing was completed. We anticipate that it will take circa three months to finalize markers. Task 3.2. Genotype MAGIC populations Task leader: Stphane Muos, INRA Partners involved: INRA, UBC, UGA Aim: Genotype 1200 RILs from MAGIC populations currently under development Strategy: To facilitate the efficient genetic analysis of complex trait variation in Helianthus and to produce materials containing exotic alleles that can be readily deployed in breeding programs, we have partnered with industry to develop modified Multiparent Advanced Generation Inter-Cross (MAGIC) mapping populations that include both cultivated and wild sunflower donors. Such populations combine the high power to detect QTLs offered by biparental populations with the ability to assay a broader spectrum of diversity and improved resolution afforded by association mapping. Because sunflower is a hybrid crop, we are developing separate male and female populations. For each population, our crossing design involves four elite breeding lines and four wild donors, which will be intermated and selfed to develop 600 RILs. Our plan is to use the Axiom array to assay the 1200 RILS for SNP and structural variants. Cost: 240 k . Of this, approximately 120 is available for genotyping from recently funded NSF and Genome Canada projects. Timeline: We will genotype the female MAGIC population in year 3 and the male population in year WP 4. Produce an atlas of stress responsive genes for genome annotation and reverse genetics Task leaders: Nicolas Langlade, INRA and John Burke, UGA Partners involved: INRA, UBC, UGA Aim: aid genome annotation through the identification of expressed genes and the annotation of genes as involved in tolerance to abiotic stresses and pathogens. This will be key to identifying the most likely candidate genes underlying QTLs and for targeting them for reverse genetics validation using the mutant population and/or through transformation and RNAi. Strategy: Expression levels will be assessed by RNA-Seq. For each stress, 3-5 genotypes will be used, including reference one (e.g. XRQ), sensitive and tolerant/resistant lines. Stresses include: Drought stress at seedling stage: in roots and leaves at three timepoints (UGA) Drought stress at vegetative stress: in roots and leaves at one stage (managed on the Heliaphen platform at INRA) Drought stress at seed filing stage in roots, leaves and seeds at 3 stages (managed on the Heliaphen platform at INRA) Salt stress at seedling stage: in roots and leaves at three timepoints (UGA) Flooding stress at seedling stage: in roots and leaves at three timepoints (UBC) Low nutrient stress at seedling stage: in roots and leaves at three timepoints (UGA) Cold stress at seedling stage: in roots and leaves (INRA) Orobanche: roots infected by orobanche or not (INRA) Phoma: stems at the level and aound the infection by Phoma macdonaldii (INRA) Downy mildew: roots, hypocotyles and cotyledons after primary infection of Plasmopara halstedii (INRA) Cost: ~200k (corresponds to ~200 per sample) for data production. Bioinformatics = $220 k ($55 k / year x 4 years); Note that ~100k for data production is available from NSF and Genome Canada projects. 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Principal component analysis of the genetic diversity of the public lines in the INRA core-collection using 8844 SNPs. Already sequenced lines are represented by blue circles and lines to be sequenced by PacBio are represented in red. Table 1. Lines already sequenced (grey) and to be sequenced LineOther nameSourcePedigreeTraitSF009ADVINRAADVENT.CE.A.7.3.3.2.1.2.1.1. R100?SF092IRINRAIMPIRA SC4.194.8.6.C11.1.1.1.3.1SF1092603INRAM.H.2.1..SF193XRQINRAQPD(HA89 PROGR)B.1.2.7.C3.2.1.2.1SF279OQP8/OPB8INRAOQPCR OQP(PAA1 RHA345)A.1.9.18.1.2SF281RHA438USDAOQPCR OQPRHA 438 (RHA340xRHA344 OLEIQUE)SF31783HR4INRAPCR P(V6540 G509 RHA274) +RM+OL MTPSF326PSC8INRAPCR PISCMR5.313.6.7.7.5+RMSF342PW3RMINRAQCR PW3RM (Pl7 ex YEQ) S304SF230noneINRAUSSCL 23.4.1.2(1+2)(RF-015.21.2)Downy MildewRHA 358USDARHA 274*3/DDRShort statureRHA 408USDARomania R-line Sclerotinia pop-1Sclerotinia stalk rot resistantRHA 426USDARHA 409//RHA 376*2/IMI res. H. annuusIMISUN source, Sclerotinia stalk rot resistantRHA 455USDARHA 440/HO IS R-lineHO, Sclerotinia resistantHA 467USDAHA 411/ROMPH//HA425/87CAEB/3/HA 434/HA 412Sclerotinia and Phomopsis resistant, HO, IMISUN, moderate statureHA 821USDAHA 300 selectionHA 404USDAHA 821/RASSVETEarly maturityHA89USDALR1INRAOrobanche resistance Table 2. List of wild ecotypes to sequence using Illuminas Long-read Sequencing Technology SpeciesPopulation localityTraits of interestHelianthus annuusWestern NebraskaDrought toleranceHelianthus annuusSouthern Manitoba, CanadaEarliness (day neutral), cold hardinessHelianthus annuusGreat Salt Lake, UtahSalt toleranceHelianthus anomalusLittle Sahara Sand Dunes, UtahLarge seeds, nitrogen use efficiencyHelianthus argophyllusTexas Coastal PlainDrought tolerance, disease resistanceHelianthus exilisNorth Coast Range, CaliforniaHeavy metal toleranceHelianthus paradoxusSanta Rosa, New MexicoSalt ToleranceHelianthus petiolarisGreat Sand Dunes National Park, ColoradoLarge Seeds, nitrogen use efficiencyHelianthus debilisSource of Orobanche resistance Ten lines in Figure 1, but 20 lines in Table 2. uuDujukuuuuuuuv vYvZvvvvvvwww7w8wCwvwww~wwwwwwwwwwwwwwwwx&xUxVx^x_xdxxxxϳϳϳڦϦڦϦڦϦڦϦڦhTh1PJnH tH  hTh1PJmH nH sH tH hTh1PJnH tH hTh1B*mH phsH hTh16B*phhTh1B*ph hThGhTh15B*phh1mH sH h15mH sH 5uDuIuTu[uduju$d$Ifa$gdTgd1jukuqu+$d$Ifa$gdTkd3$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTquuuzuuu$d$Ifa$gdTuuu+$d$Ifa$gdTkd4$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTuuuuu$d$Ifa$gdTuuu+$d$Ifa$gdTkd5$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTuuuuu$d$Ifa$gdTuuu+$d$Ifa$gdTkd6$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTuuuvv v&v0v5vXvYvFf8$d$Ifa$gdT YvZv`v+$d$Ifa$gdTkd:$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytT`vgvlvvv$d$Ifa$gdTvvv+$d$Ifa$gdTkd;$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTvvvvv$d$Ifa$gdTvvv+$d$Ifa$gdTkd<$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTvvvwww www6w7wFf>$d$Ifa$gdT 7w8w>w+$d$Ifa$gdTkd@$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytT>wCwHwiwvw$d$Ifa$gdTvwwww+$d$Ifa$gdTkdA$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTwwwww$d$Ifa$gdTwww+$d$Ifa$gdTkdB$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTwwwww$d$Ifa$gdTwww+$d$Ifa$gdTkdC$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTwwx&xUx$d$Ifa$gdTUxVx^x+$d$Ifa$gdTkdD$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytT^x_xdxyxx$d$Ifa$gdTxxx+$d$Ifa$gdTkdE$$Ifl,rn ^#ffj ~ t0H$644 lBap2ytTxxxxyyyyyy.y/y5y6y7y;y*mH sH hzMhzMmH sH &yyzzz*z;zMzRkdK$$IflF &#n $  t0j$6    44 lapytTd$IfgdTMzNz`zzzz`RRRd$IfgdTkd7L$$IflF &#n $  t0j$6    44 lapytTzzzzz`RRRd$IfgdTkdL$$IflF &#n $  t0j$6    44 lapytTzzz{3{`RRRd$IfgdTkdM$$IflF &#n $  t0j$6    44 lapytT3{4{K{_{{`R<R<d$If^`#[U^'EM4iU|neU]6/W@&rP2Ɓf `mb&We6Z$WW-U%ekoW@*;$\LM IYFfAU.uv2>%W?4WZ*`K57+ar.M٣#W+Q_w/uW}*r (N! wTkTU)6p\=RS/ڗsWrtDMu WA,eYX%WArխSrdf1A'Usr$N}} 5I(+\yR_A'*@'WATQ_AVr4L4B>r4I !WA{ ee|Z'WAK=P_A*h7+h\ [L} :5+O,Y" "B@rcCBrx4D WAj*P_AErԧ`8T!WAej*n'UP"Ǩrԡ J) p #QA(K#2P\ \$WTH}UpgN}GUp 5`' sfOVVg*(F 0GŒ\W9es+Up F+$J}E*8CM?\8CWtETpUSm(&W )U U94r R2UK)N"gp·yT׉ՠ(䡦 /(ܽ!+WLHByZN_d'U4$QnUP_F"w@2Vg\E *0`Vr9oG25* jLCM]rOsErRSj\Co\aq&UN(`dr2^øGNE)4WUBMa UY"( M}U\EIj*nSUz**pbMXarWIT4xgU\b+\3>x+PPUn"WqRUP״r`N6 "N5e6 Urq`\oj*L$Ul$TVr$*d W @2ϐd؟$Wg38?Lb^\lkl ~9D@}Z7M[V38UCD9P{(rUG@&B{9$Wlh'̜@V.䪴d"Q5^/ z}{sTw`O_I+EY mq`o\ 3"ɞ%0r`o\x@jطB/s썒lDٳ5s`o\0g3W>=:ʵ!i|]Krs`O@_g3PE*siU1E4ITUͰAuρR*#W5@nKp`J etT.=1*.g3^*}&;ykB䪸z9:B5~+\q>;Е$QuEjCY!U-Q_D-=j0X! =f| Gc Tb2Q_THJH}5SrUZ+HfyfrteIO}-ZmEVg(*lpr/oWLQnLqnOrUJDfzu2GoLYD%T8`LoM4n68ί~qxd??LoDϙ^$x92!rP_^](ANʛL.5R=%ˈ-RSq\Ul2cn}fʈq OlJym n2c.,~DurUIF,ou1ӏcI 1+?l<$T8FArVWTh~j*ʒ(@}\lB?*J}Ցoٝ~ΗJC2RWL}U}vLUa2aWS' nfu%}ގտeh}TL{>>*enmt#*xHベwӍk77 r<*ǰ-a|WDՐUMl hbPZlUj*+o >k% BP_5>i8lj*X%WA +ºOz~n 6UѴz ,TIT\qO8智Ooz5"qy3"~N=Ub$]#Ud,r HTp\}8 6ĵKm]p\ S_s*h*ot j?M"8OzNݻwPMuAMUP욫ieq sҭN -Ud+W}Y J~<*C,L?oVZ.:ue +^ߥў za|թT m -! c|a% \.`C*xܷx`|gUSA?* 1-vvUSAW*3i'sw^Mre9麍{4wt>쭝7BTp (;#bj+] 5 v\4)X#||xyξJga_$#WA5 >VWgJϦ: 9A&WAMψZN'z^I_w̦jhssLUPGBo|x޺w c_QUS, WAf!+(S/D5 ng~dvh;=sߤ-@dr$m=֭^%`!PI]wOʞ 4UY?e5fB4*Ȭk:1%l"qrTf `8BeN-  cTZ & 3Qqa3!2 *s\Fӄl3\dvKdUL< 61]-U}Cg .in ҡP5 ͝ @[*%$M5S/ܙE]rLYα)BVW @3Lw9>DL'm\uDyf&qz6E8^{r}>5 2z\PSqK@4r@Mw-U%}JPhJ3 *$Vӥ m450 0&=M9'pO-*DyϝʲPk;U)U|Am6 ~NF~>N4〷{Of#R±vW6h5?<*"}UL~_|*'νez  J{Λm*j3VE B!,>'?; > z[MME WA׺62T yW@4rt*B+YjZPmnb*Qʡqo^%֠Q*Kݚ*um'rz W#fSUUQ1Fx9@ qnJ*{V0UPӌD'T |?wR `*xlQj䴡{쿻i83uUPݡ2 묯 Wg꒫[h5U-+,ͦ\82ܹ|dH_55^ja򓫞`MӌTS_Ӣseu a@+\YF!Kk"']3~ TAUQlfa-"泯, &W߻^8^RThMyyo,Zy-NryzJf[l@)_/>qU$W7}q}U =388=%C bِo3Wx 6ܷ$Xu^{ϼu` *2@ݡIDATାAMե1M6'WE4k~Ocl!\IT=9i$WA8 ȣ8*$AP}zK[T}UE*-" P0*o{4gST!WAeAj=ꢲkIrTH(J=ڭO5NW~tic*3]Qwo?q*Tսt }ph-faPTPW/vp 3Q}xs1 %WA??OĞwϊۡ Ĕ+H}UP't5U I}Upg^o =TSBU+IœO2:$WA1'UH. |#WAb@*\pAઋB5 Fr\re~4DK0@BU) =R(LKO)B}E*8)xZ|s R_ (ڧZ\qC%7 ~0xw0>HJrlq?A2`shx*XZW4MtxyWh\鳦{?WEa8zuN?u/ Utρ˾}bE(aUtJM5{:'A葚!f= AUImsr9={*:"1TIWE N^ lex\E~joLnTq_'7z^BՆϴ˧O=.cWgDB*rR41mQ{ޱ^01>r [fG|ǶW 3>Hqr8iOr3>\U= eYU *T4K=W U\(]Bj*oW풫B*]u2.R_Hlمbv (UCf[ krpE Wq; パUM$Wq'RP_"Wq 5WU "U]r%P_U!WQ#0$*>Cm1>x+K}u3T9dyZ-r8HƁ T8%Wd WL$Ul$TVr$*d W @2ϐdM$8Hfz`wU ݽk Wf>wYrUT?;Aꗚ U=$T9/@2Uq6Lk_Cꂳd8V]f,#?UyL_$WeH"䪴T8Un\} ʭRq`o\d؛#Wel FUs6LlمO)>af~`h1GU r6@) U~6@YiUq6D\ @2N Q$\dr6{zbHCꁾ*ώg'H`z0W0wlOI Nl63p{Em)UܻWM؟ɓXFgrU'vB8#We`|:5\ G Ñ9rUB+&Wbz±f><ۨnʒ\Uտ|ʺ)Q\U^̽ Eܚ~٠8;yYkdR~8\ug+m'WUS+'m}jJ*c'4{>? 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