A novel application of RNase H2-dependent quantitative PCR for detection and quantification of Grosmannia clavigera, a mountain pine beetle fungal symbiont, in environmental samples
Identifieur interne : 000B47 ( Pmc/Curation ); précédent : 000B46; suivant : 000B48A novel application of RNase H2-dependent quantitative PCR for detection and quantification of Grosmannia clavigera, a mountain pine beetle fungal symbiont, in environmental samples
Auteurs : Chandra H. Mcallister [Canada] ; Colleen E. Fortier [Canada] ; Kate R. St Onge [Canada] ; Bianca M. Sacchi [Canada] ; Meaghan J. Nawrot [Canada] ; Troy Locke [Canada] ; Janice E K. Cooke [Canada]Source :
- Tree Physiology [ 0829-318X ] ; 2018.
Abstract
Mountain pine beetle (
Url:
DOI: 10.1093/treephys/tpx147
PubMed: 29329457
PubMed Central: 5982843
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">A novel application of RNase H2-dependent quantitative PCR for detection and quantification of <italic>Grosmannia clavigera</italic>, a mountain pine beetle fungal symbiont, in environmental samples</title><author><name sortKey="Mcallister, Chandra H" sort="Mcallister, Chandra H" uniqKey="Mcallister C" first="Chandra H" last="Mcallister">Chandra H. Mcallister</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Fortier, Colleen E" sort="Fortier, Colleen E" uniqKey="Fortier C" first="Colleen E" last="Fortier">Colleen E. Fortier</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="St Onge, Kate R" sort="St Onge, Kate R" uniqKey="St Onge K" first="Kate R" last="St Onge">Kate R. St Onge</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Sacchi, Bianca M" sort="Sacchi, Bianca M" uniqKey="Sacchi B" first="Bianca M" last="Sacchi">Bianca M. Sacchi</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Nawrot, Meaghan J" sort="Nawrot, Meaghan J" uniqKey="Nawrot M" first="Meaghan J" last="Nawrot">Meaghan J. Nawrot</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Locke, Troy" sort="Locke, Troy" uniqKey="Locke T" first="Troy" last="Locke">Troy Locke</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Cooke, Janice E K" sort="Cooke, Janice E K" uniqKey="Cooke J" first="Janice E K" last="Cooke">Janice E K. Cooke</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">29329457</idno><idno type="pmc">5982843</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5982843</idno><idno type="RBID">PMC:5982843</idno><idno type="doi">10.1093/treephys/tpx147</idno><date when="2018">2018</date><idno type="wicri:Area/Pmc/Corpus">000B47</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000B47</idno><idno type="wicri:Area/Pmc/Curation">000B47</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000B47</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A novel application of RNase H2-dependent quantitative PCR for detection and quantification of <italic>Grosmannia clavigera</italic>, a mountain pine beetle fungal symbiont, in environmental samples</title><author><name sortKey="Mcallister, Chandra H" sort="Mcallister, Chandra H" uniqKey="Mcallister C" first="Chandra H" last="Mcallister">Chandra H. Mcallister</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Fortier, Colleen E" sort="Fortier, Colleen E" uniqKey="Fortier C" first="Colleen E" last="Fortier">Colleen E. Fortier</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="St Onge, Kate R" sort="St Onge, Kate R" uniqKey="St Onge K" first="Kate R" last="St Onge">Kate R. St Onge</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Sacchi, Bianca M" sort="Sacchi, Bianca M" uniqKey="Sacchi B" first="Bianca M" last="Sacchi">Bianca M. Sacchi</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Nawrot, Meaghan J" sort="Nawrot, Meaghan J" uniqKey="Nawrot M" first="Meaghan J" last="Nawrot">Meaghan J. Nawrot</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Locke, Troy" sort="Locke, Troy" uniqKey="Locke T" first="Troy" last="Locke">Troy Locke</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author><author><name sortKey="Cooke, Janice E K" sort="Cooke, Janice E K" uniqKey="Cooke J" first="Janice E K" last="Cooke">Janice E K. Cooke</name><affiliation wicri:level="1"><nlm:aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</nlm:aff><country>Canada</country><wicri:regionArea>Department of Biological Sciences, University of Alberta, Edmonton, AB</wicri:regionArea></affiliation></author></analytic><series><title level="j">Tree Physiology</title><idno type="ISSN">0829-318X</idno><idno type="eISSN">1758-4469</idno><imprint><date when="2018">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Abstract</title><p>Mountain pine beetle (<italic>Dendroctonus ponderosae</italic> Hopkins; MPB) is an economically and ecologically important pest of pine species in western North America. Mountain pine beetles form complex multipartite relationships with microbial partners, including the ophiostomoid fungi <italic>Grosmannia clavigera</italic> (Robinson-Jeffrey and Davidson) Zipfel, de Beer and Wingfield, <italic>Ophiostoma montium</italic> (Rumbold) von Arx, <italic>Grosmannia aurea</italic> (Robinson-Jeffrey and Davidson) Zipfel, de Beer and Wingfield, <italic>Leptographium longiclavatum</italic> (Lee, Kim, and Breuil) <italic>and Leptographium terebrantis</italic> (Barras and Perry). These fungi are vectored by MPB to new pine hosts, where the fungi overcome host defenses to grow into the sapwood. A tree’s relative susceptibility to these fungi is conventionally assessed by measuring lesions that develop in response to fungal inoculation. However, these lesions represent a symptom of infection, representing both fungal growth and tree defense capacity. In order to more objectively assess fungal virulence and host tree susceptibility in studies of host–pathogen interactions, a reliable, consistent, sensitive method is required to accurately identify and quantify MPB-associated fungal symbionts in planta. We have adapted RNase H2-dependent PCR, a technique originally designed for rare allele discrimination, to develop a novel RNase H2-dependent quantitative PCR (rh-qPCR) assay that shows greater specificity and sensitivity than previously published PCR-based methods to quantify MPB fungal symbionts in pine xylem and MPB whole beetles. Two sets of assay probes were designed: one that amplifies a broad range of ophiostomoid species, and a second that amplifies <italic>G. clavigera</italic> but not other MPB-associated ophiostomoid species. Using these primers to quantify <italic>G. clavigera</italic> in pine stems, we provide evidence that lesion length does not accurately reflect the extent of fungal colonization along the stem nor the quantity of fungal growth within this colonized portion of stem. The sensitivity, specificity, reproducibility, cost effectiveness and high-throughput potential of the rh-qPCR assay makes the technology suitable for identification and quantification of a wide array of pathogenic and beneficial microbes that form associations with plants and other organisms, even when the microbial partner is present in low abundance.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Adams, As" uniqKey="Adams A">AS Adams</name></author><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Adams, As" uniqKey="Adams A">AS Adams</name></author><author><name sortKey="Aylward, Fo" uniqKey="Aylward F">FO Aylward</name></author><author><name sortKey="Adams, Sm" uniqKey="Adams S">SM Adams</name></author><author><name sortKey="Erbilgin, N" uniqKey="Erbilgin N">N Erbilgin</name></author><author><name sortKey="Aukema, Bh" uniqKey="Aukema B">BH Aukema</name></author><author><name sortKey="Currie, Cr" uniqKey="Currie C">CR Currie</name></author><author><name sortKey="Suen, G" uniqKey="Suen G">G Suen</name></author><author><name sortKey="Raffa, Kf" uniqKey="Raffa K">KF Raffa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Addison, Al" uniqKey="Addison A">AL Addison</name></author><author><name sortKey="Powell, Ja" uniqKey="Powell J">JA Powell</name></author><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author><author><name sortKey="Moore, M" uniqKey="Moore M">M Moore</name></author><author><name sortKey="Bentz, Bj" uniqKey="Bentz B">BJ Bentz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alamouti, Sm" uniqKey="Alamouti S">SM Alamouti</name></author><author><name sortKey="Wang, V" uniqKey="Wang V">V Wang</name></author><author><name sortKey="Diguistini, S" uniqKey="Diguistini S">S Diguistini</name></author><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author><author><name sortKey="Bohlmann, J" uniqKey="Bohlmann J">J Bohlmann</name></author><author><name sortKey="Hamelin, Rc" uniqKey="Hamelin R">RC Hamelin</name></author><author><name sortKey="Feau, N" uniqKey="Feau N">N Feau</name></author><author><name sortKey="Breuil, C" uniqKey="Breuil C">C Breuil</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Almquist, C" uniqKey="Almquist C">C Almquist</name></author><author><name sortKey="Persson, L" uniqKey="Persson L">L Persson</name></author><author><name sortKey="Olsson, " uniqKey="Olsson ">Å Olsson</name></author><author><name sortKey="Sundstrom, J" uniqKey="Sundstrom J">J Sundström</name></author><author><name sortKey="Jonsson, A" uniqKey="Jonsson A">A Jonsson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arango Velez, A" uniqKey="Arango Velez A">A Arango-Velez</name></author><author><name sortKey="Gonzalez, Lmg" uniqKey="Gonzalez L">LMG González</name></author><author><name sortKey="Meents, Mj" uniqKey="Meents M">MJ Meents</name></author><author><name sortKey="El Kayal, W" uniqKey="El Kayal W">W El Kayal</name></author><author><name sortKey="Cooke, Bj" uniqKey="Cooke B">BJ Cooke</name></author><author><name sortKey="Linsky, J" uniqKey="Linsky J">J Linsky</name></author><author><name sortKey="Lusebrink, I" uniqKey="Lusebrink I">I Lusebrink</name></author><author><name sortKey="Cooke, Jek" uniqKey="Cooke J">JEK Cooke</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arango Velez, A" uniqKey="Arango Velez A">A Arango-Velez</name></author><author><name sortKey="Zwiazek, Jj" uniqKey="Zwiazek J">JJ Zwiazek</name></author><author><name sortKey="Thomas, Br" uniqKey="Thomas B">BR Thomas</name></author><author><name sortKey="Tyree, Mt" uniqKey="Tyree M">MT Tyree</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arango Velez, A" uniqKey="Arango Velez A">A Arango-Velez</name></author><author><name sortKey="El Kayal, W" uniqKey="El Kayal W">W El Kayal</name></author><author><name sortKey="Copeland, Ccj" uniqKey="Copeland C">CCJ Copeland</name></author><author><name sortKey="Zaharia, Li" uniqKey="Zaharia L">LI Zaharia</name></author><author><name sortKey="Lusebrink, I" uniqKey="Lusebrink I">I Lusebrink</name></author><author><name sortKey="Cooke, Jek" uniqKey="Cooke J">JEK Cooke</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ballard, Rg" uniqKey="Ballard R">RG Ballard</name></author><author><name sortKey="Walsh, Ma" uniqKey="Walsh M">MA Walsh</name></author><author><name sortKey="Cole, We" uniqKey="Cole W">WE Cole</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ballard, Rg" uniqKey="Ballard R">RG Ballard</name></author><author><name sortKey="Walsh, Ma" uniqKey="Walsh M">MA Walsh</name></author><author><name sortKey="Cole, We" uniqKey="Cole W">WE Cole</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bleiker, Kp" uniqKey="Bleiker K">KP Bleiker</name></author><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bleiker, Kp" uniqKey="Bleiker K">KP Bleiker</name></author><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bleiker, Kp" uniqKey="Bleiker K">KP Bleiker</name></author><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Boutigny, Al" uniqKey="Boutigny A">AL Boutigny</name></author><author><name sortKey="Beukes, I" uniqKey="Beukes I">I Beukes</name></author><author><name sortKey="Small, I" uniqKey="Small I">I Small</name></author><author><name sortKey="Zuhlke, S" uniqKey="Zuhlke S">S Zühlke</name></author><author><name sortKey="Spiteller, M" uniqKey="Spiteller M">M Spiteller</name></author><author><name sortKey="Van Rensburg, Bj" uniqKey="Van Rensburg B">BJ Van Rensburg</name></author><author><name sortKey="Flett, B" uniqKey="Flett B">B Flett</name></author><author><name sortKey="Viljoen, A" uniqKey="Viljoen A">A Viljoen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Boyle, B" uniqKey="Boyle B">B Boyle</name></author><author><name sortKey="Hamelin, Rc" uniqKey="Hamelin R">RC Hamelin</name></author><author><name sortKey="Seguin, A" uniqKey="Seguin A">A Seguin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cahoon, Ab" uniqKey="Cahoon A">AB Cahoon</name></author><author><name sortKey="Nauss, Ja" uniqKey="Nauss J">JA Nauss</name></author><author><name sortKey="Stanley, Cd" uniqKey="Stanley C">CD Stanley</name></author><author><name sortKey="Qureshi, A" uniqKey="Qureshi A">A Qureshi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chang, S" uniqKey="Chang S">S Chang</name></author><author><name sortKey="Puryear, J" uniqKey="Puryear J">J Puryear</name></author><author><name sortKey="Cairney, J" uniqKey="Cairney J">J Cairney</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cullingham, Ci" uniqKey="Cullingham C">CI Cullingham</name></author><author><name sortKey="Cooke, Jek" uniqKey="Cooke J">JEK Cooke</name></author><author><name sortKey="Dang, S" uniqKey="Dang S">S Dang</name></author><author><name sortKey="Davis, Cs" uniqKey="Davis C">CS Davis</name></author><author><name sortKey="Cooke, Bj" uniqKey="Cooke B">BJ Cooke</name></author><author><name sortKey="Coltman, Dw" uniqKey="Coltman D">DW Coltman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dobosy, Jr" uniqKey="Dobosy J">JR Dobosy</name></author><author><name sortKey="Rose, Sd" uniqKey="Rose S">SD Rose</name></author><author><name sortKey="Beltz, Kr" uniqKey="Beltz K">KR Beltz</name></author><author><name sortKey="Rupp, Sm" uniqKey="Rupp S">SM Rupp</name></author><author><name sortKey="Powers, Km" uniqKey="Powers K">KM Powers</name></author><author><name sortKey="Behlke, Ma" uniqKey="Behlke M">MA Behlke</name></author><author><name sortKey="Walder, Ja" uniqKey="Walder J">JA Walder</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Floren, A" uniqKey="Floren A">A Floren</name></author><author><name sortKey="Kruger, D" uniqKey="Kruger D">D Krüger</name></author><author><name sortKey="Muller, T" uniqKey="Muller T">T Müller</name></author><author><name sortKey="Dittrich, M" uniqKey="Dittrich M">M Dittrich</name></author><author><name sortKey="Rudloff, R" uniqKey="Rudloff R">R Rudloff</name></author><author><name sortKey="Hoppe, B" uniqKey="Hoppe B">B Hoppe</name></author><author><name sortKey="Linsenmair, Ke" uniqKey="Linsenmair K">KE Linsenmair</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Franceschi, Vr" uniqKey="Franceschi V">VR Franceschi</name></author><author><name sortKey="Krokene, P" uniqKey="Krokene P">P Krokene</name></author><author><name sortKey="Christiansen, E" uniqKey="Christiansen E">E Christiansen</name></author><author><name sortKey="Krekling, T" uniqKey="Krekling T">T Krekling</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Goodsman, Dw" uniqKey="Goodsman D">DW Goodsman</name></author><author><name sortKey="Lusebrink, I" uniqKey="Lusebrink I">I Lusebrink</name></author><author><name sortKey="Landhausser, Sm" uniqKey="Landhausser S">SM Landhausser</name></author><author><name sortKey="Erbilgin, N" uniqKey="Erbilgin N">N Erbilgin</name></author><author><name sortKey="Lieffers, Vj" uniqKey="Lieffers V">VJ Lieffers</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Grosdidier, M" uniqKey="Grosdidier M">M Grosdidier</name></author><author><name sortKey="Aguayo, J" uniqKey="Aguayo J">J Aguayo</name></author><author><name sortKey="Marcais, B" uniqKey="Marcais B">B Marçais</name></author><author><name sortKey="Ioos, R" uniqKey="Ioos R">R Ioos</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hacquard, S" uniqKey="Hacquard S">S Hacquard</name></author><author><name sortKey="Veneault Fourrey, C" uniqKey="Veneault Fourrey C">C Veneault-Fourrey</name></author><author><name sortKey="Delaruelle, C" uniqKey="Delaruelle C">C Delaruelle</name></author><author><name sortKey="Frey, P" uniqKey="Frey P">P Frey</name></author><author><name sortKey="Martin, F" uniqKey="Martin F">F Martin</name></author><author><name sortKey="Duplessis, S" uniqKey="Duplessis S">S Duplessis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hulcr, J" uniqKey="Hulcr J">J Hulcr</name></author><author><name sortKey="Dunn, Rr" uniqKey="Dunn R">RR Dunn</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Khadempour, L" uniqKey="Khadempour L">L Khadempour</name></author><author><name sortKey="Massoumi Alamouti, S" uniqKey="Massoumi Alamouti S">S Massoumi Alamouti</name></author><author><name sortKey="Hamelin, R" uniqKey="Hamelin R">R Hamelin</name></author><author><name sortKey="Bohlmann, J" uniqKey="Bohlmann J">J Bohlmann</name></author><author><name sortKey="Breuil, C" uniqKey="Breuil C">C Breuil</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Khadempour, L" uniqKey="Khadempour L">L Khadempour</name></author><author><name sortKey="Lemay, V" uniqKey="Lemay V">V Lemay</name></author><author><name sortKey="Jack, D" uniqKey="Jack D">D Jack</name></author><author><name sortKey="Bohlmann, J" uniqKey="Bohlmann J">J Bohlmann</name></author><author><name sortKey="Breuil, C" uniqKey="Breuil C">C Breuil</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Knufer, J" uniqKey="Knufer J">J Knüfer</name></author><author><name sortKey="Teshome Lopisso, D" uniqKey="Teshome Lopisso D">D Teshome Lopisso</name></author><author><name sortKey="Koopman, B" uniqKey="Koopman B">B Koopman</name></author><author><name sortKey="Karlovsky, B" uniqKey="Karlovsky B">B Karlovsky</name></author><author><name sortKey="Von Tiedeman, A" uniqKey="Von Tiedeman A">A von Tiedeman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lee, S" uniqKey="Lee S">S Lee</name></author><author><name sortKey="Kim, J J" uniqKey="Kim J">J-J Kim</name></author><author><name sortKey="Breuil, C" uniqKey="Breuil C">C Breuil</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lee, S" uniqKey="Lee S">S Lee</name></author><author><name sortKey="Kim, J J" uniqKey="Kim J">J-J Kim</name></author><author><name sortKey="Breuil, C" uniqKey="Breuil C">C Breuil</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lieutier, F" uniqKey="Lieutier F">F Lieutier</name></author><author><name sortKey="Yart, A" uniqKey="Yart A">A Yart</name></author><author><name sortKey="Salle, A" uniqKey="Salle A">A Salle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Llorente, B" uniqKey="Llorente B">B Llorente</name></author><author><name sortKey="Bravo Almonacid, F" uniqKey="Bravo Almonacid F">F Bravo-Almonacid</name></author><author><name sortKey="Cvitanich, C" uniqKey="Cvitanich C">C Cvitanich</name></author><author><name sortKey="Orlowska, E" uniqKey="Orlowska E">E Orlowska</name></author><author><name sortKey="Torres, Hn" uniqKey="Torres H">HN Torres</name></author><author><name sortKey="Flavia, Mm" uniqKey="Flavia M">MM Flaviá</name></author><author><name sortKey="Alonso, Gd" uniqKey="Alonso G">GD Alonso</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lu, M" uniqKey="Lu M">M Lu</name></author><author><name sortKey="Wingfield, Mj" uniqKey="Wingfield M">MJ Wingfield</name></author><author><name sortKey="Gillette, Ne" uniqKey="Gillette N">NE Gillette</name></author><author><name sortKey="Mori, Sr" uniqKey="Mori S">SR Mori</name></author><author><name sortKey="Sun, J H" uniqKey="Sun J">J-H Sun</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Malacrin, A" uniqKey="Malacrin A">A Malacrinò</name></author><author><name sortKey="Rassati, D" uniqKey="Rassati D">D Rassati</name></author><author><name sortKey="Schena, L" uniqKey="Schena L">L Schena</name></author><author><name sortKey="Mehzabin, R" uniqKey="Mehzabin R">R Mehzabin</name></author><author><name sortKey="Battisti, A" uniqKey="Battisti A">A Battisti</name></author><author><name sortKey="Palmeri, V" uniqKey="Palmeri V">V Palmeri</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Masuya, H" uniqKey="Masuya H">H Masuya</name></author><author><name sortKey="Kaneko, S" uniqKey="Kaneko S">S Kaneko</name></author><author><name sortKey="Yamaoka, Y" uniqKey="Yamaoka Y">Y Yamaoka</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Matsiakh, I" uniqKey="Matsiakh I">I Matsiakh</name></author><author><name sortKey="Solheim, H" uniqKey="Solheim H">H Solheim</name></author><author><name sortKey="Nagy, Ne" uniqKey="Nagy N">NE Nagy</name></author><author><name sortKey="Hietala, Am" uniqKey="Hietala A">AM Hietala</name></author><author><name sortKey="Kramarets, V" uniqKey="Kramarets V">V Kramarets</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mcdowell, Ng" uniqKey="Mcdowell N">NG McDowell</name></author><author><name sortKey="Beerling, Dj" uniqKey="Beerling D">DJ Beerling</name></author><author><name sortKey="Breshears, Dd" uniqKey="Breshears D">DD Breshears</name></author><author><name sortKey="Fisher, Ra" uniqKey="Fisher R">RA Fisher</name></author><author><name sortKey="Raffa, Kf" uniqKey="Raffa K">KF Raffa</name></author><author><name sortKey="Stitt, M" uniqKey="Stitt M">M Stitt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Millar, Ci" uniqKey="Millar C">CI Millar</name></author><author><name sortKey="Stephenson, Nl" uniqKey="Stephenson N">NL Stephenson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Plattner, A" uniqKey="Plattner A">A Plattner</name></author><author><name sortKey="Kim, J" uniqKey="Kim J">J Kim</name></author><author><name sortKey="Diguistini, S" uniqKey="Diguistini S">S Diguistini</name></author><author><name sortKey="Breuil, C" uniqKey="Breuil C">C Breuil</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Raffa, Kf" uniqKey="Raffa K">KF Raffa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Raffa, Kf" uniqKey="Raffa K">KF Raffa</name></author><author><name sortKey="Aukema, Bh" uniqKey="Aukema B">BH Aukema</name></author><author><name sortKey="Bentz, Bj" uniqKey="Bentz B">BJ Bentz</name></author><author><name sortKey="Carroll, Al" uniqKey="Carroll A">AL Carroll</name></author><author><name sortKey="Hicke, Ja" uniqKey="Hicke J">JA Hicke</name></author><author><name sortKey="Turner, Mg" uniqKey="Turner M">MG Turner</name></author><author><name sortKey="Romme, Wh" uniqKey="Romme W">WH Romme</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Reid, Rw" uniqKey="Reid R">RW Reid</name></author><author><name sortKey="Whitney, Hs" uniqKey="Whitney H">HS Whitney</name></author><author><name sortKey="Watson, Ja" uniqKey="Watson J">JA Watson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Reischer, Gh" uniqKey="Reischer G">GH Reischer</name></author><author><name sortKey="Lemmens, M" uniqKey="Lemmens M">M Lemmens</name></author><author><name sortKey="Farnleitner, A" uniqKey="Farnleitner A">A Farnleitner</name></author><author><name sortKey="Adler, A" uniqKey="Adler A">A Adler</name></author><author><name sortKey="Mach, Rl" uniqKey="Mach R">RL Mach</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rice, Av" uniqKey="Rice A">AV Rice</name></author><author><name sortKey="Langor, Dw" uniqKey="Langor D">DW Langor</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rice, Av" uniqKey="Rice A">AV Rice</name></author><author><name sortKey="Thormann, Mn" uniqKey="Thormann M">MN Thormann</name></author><author><name sortKey="Langor, Dw" uniqKey="Langor D">DW Langor</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Roe, Ad" uniqKey="Roe A">AD Roe</name></author><author><name sortKey="Rice, Av" uniqKey="Rice A">AV Rice</name></author><author><name sortKey="Bromilow, Se" uniqKey="Bromilow S">SE Bromilow</name></author><author><name sortKey="Cooke, Jek" uniqKey="Cooke J">JEK Cooke</name></author><author><name sortKey="Sperling, Fah" uniqKey="Sperling F">FAH Sperling</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Roe, Ad" uniqKey="Roe A">AD Roe</name></author><author><name sortKey="James, Pma" uniqKey="James P">PMA James</name></author><author><name sortKey="Rice, Av" uniqKey="Rice A">AV Rice</name></author><author><name sortKey="Cooke, Jek" uniqKey="Cooke J">JEK Cooke</name></author><author><name sortKey="Sperling, Fah" uniqKey="Sperling F">FAH Sperling</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Roe, Ad" uniqKey="Roe A">AD Roe</name></author><author><name sortKey="Rice, Av" uniqKey="Rice A">AV Rice</name></author><author><name sortKey="Coltman, Dw" uniqKey="Coltman D">DW Coltman</name></author><author><name sortKey="Cooke, Jek" uniqKey="Cooke J">JEK Cooke</name></author><author><name sortKey="Sperling, Fah" uniqKey="Sperling F">FAH Sperling</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Safranyik, L" uniqKey="Safranyik L">L Safranyik</name></author><author><name sortKey="Carroll, Al" uniqKey="Carroll A">AL Carroll</name></author><author><name sortKey="Regniere, J" uniqKey="Regniere J">J Regniere</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sanzani, Sm" uniqKey="Sanzani S">SM Sanzani</name></author><author><name sortKey="Li Destri Nicosia, Mg" uniqKey="Li Destri Nicosia M">MG Li Destri Nicosia</name></author><author><name sortKey="Faedda, R" uniqKey="Faedda R">R Faedda</name></author><author><name sortKey="Cacciola, So" uniqKey="Cacciola S">SO Cacciola</name></author><author><name sortKey="Schena, L" uniqKey="Schena L">L Schena</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Six, Dl" uniqKey="Six D">DL Six</name></author><author><name sortKey="Wingfield, Mj" uniqKey="Wingfield M">MJ Wingfield</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Solheim, H" uniqKey="Solheim H">H Solheim</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Solheim, H" uniqKey="Solheim H">H Solheim</name></author><author><name sortKey="Safranyik, L" uniqKey="Safranyik L">L Safranyik</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Solheim, H" uniqKey="Solheim H">H Solheim</name></author><author><name sortKey="Krokene, P" uniqKey="Krokene P">P Krokene</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Song, Z" uniqKey="Song Z">Z Song</name></author><author><name sortKey="Vail, A" uniqKey="Vail A">A Vail</name></author><author><name sortKey="Sadowsky, Mj" uniqKey="Sadowsky M">MJ Sadowsky</name></author><author><name sortKey="Schilling, Js" uniqKey="Schilling J">JS Schilling</name></author><author><name sortKey="Baldrian, P" uniqKey="Baldrian P">P Baldrian</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Stenlid, J" uniqKey="Stenlid J">J Stenlid</name></author><author><name sortKey="Oliva, J" uniqKey="Oliva J">J Oliva</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Strid, Y" uniqKey="Strid Y">Y Strid</name></author><author><name sortKey="Schroeder, M" uniqKey="Schroeder M">M Schroeder</name></author><author><name sortKey="Lindahl, B" uniqKey="Lindahl B">B Lindahl</name></author><author><name sortKey="Ihrmark, K" uniqKey="Ihrmark K">K Ihrmark</name></author><author><name sortKey="Stenlid, J" uniqKey="Stenlid J">J Stenlid</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tedersoo, L" uniqKey="Tedersoo L">L Tedersoo</name></author><author><name sortKey="Lindahl, B" uniqKey="Lindahl B">B Lindahl</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tsui, Ckm" uniqKey="Tsui C">CKM Tsui</name></author><author><name sortKey="Wang, B" uniqKey="Wang B">B Wang</name></author><author><name sortKey="Khadempour, L" uniqKey="Khadempour L">L Khadempour</name></author><author><name sortKey="Massoumi, S" uniqKey="Massoumi S">S Massoumi</name></author><author><name sortKey="Bohlmann, J" uniqKey="Bohlmann J">J Bohlmann</name></author><author><name sortKey="Murray, Bw" uniqKey="Murray B">BW Murray</name></author><author><name sortKey="Hamelin, Rc" uniqKey="Hamelin R">RC Hamelin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Villari, C" uniqKey="Villari C">C Villari</name></author><author><name sortKey="Tomlinson, Ja" uniqKey="Tomlinson J">JA Tomlinson</name></author><author><name sortKey="Battisti, A" uniqKey="Battisti A">A Battisti</name></author><author><name sortKey="Boonham, N" uniqKey="Boonham N">N Boonham</name></author><author><name sortKey="Capretti, P" uniqKey="Capretti P">P Capretti</name></author><author><name sortKey="Faccoli, M" uniqKey="Faccoli M">M Faccoli</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wallin, Kf" uniqKey="Wallin K">KF Wallin</name></author><author><name sortKey="Raffa, Kf" uniqKey="Raffa K">KF Raffa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Weed, As" uniqKey="Weed A">AS Weed</name></author><author><name sortKey="Ayres, Mp" uniqKey="Ayres M">MP Ayres</name></author><author><name sortKey="Hicke, Ja" uniqKey="Hicke J">JA Hicke</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Winder, Rs" uniqKey="Winder R">RS Winder</name></author><author><name sortKey="Macey, De" uniqKey="Macey D">DE Macey</name></author><author><name sortKey="Cortese, J" uniqKey="Cortese J">J Cortese</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wingfield, Mj" uniqKey="Wingfield M">MJ Wingfield</name></author><author><name sortKey="Brockerhoff, Eg" uniqKey="Brockerhoff E">EG Brockerhoff</name></author><author><name sortKey="Wingfield, Bd" uniqKey="Wingfield B">BD Wingfield</name></author><author><name sortKey="Slippers, B" uniqKey="Slippers B">B Slippers</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wong, Bl" uniqKey="Wong B">BL Wong</name></author><author><name sortKey="Berryman, Aa" uniqKey="Berryman A">AA Berryman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Woo, Kl" uniqKey="Woo K">KL Woo</name></author><author><name sortKey="Watson, P" uniqKey="Watson P">P Watson</name></author><author><name sortKey="Mansfield, Sd" uniqKey="Mansfield S">SD Mansfield</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yamaoka, Y" uniqKey="Yamaoka Y">Y Yamaoka</name></author><author><name sortKey="Hiratsuka, Y" uniqKey="Hiratsuka Y">Y Hiratsuka</name></author><author><name sortKey="Maruyama, P" uniqKey="Maruyama P">P Maruyama</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhu, Zx" uniqKey="Zhu Z">ZX Zhu</name></author><author><name sortKey="Zheng, L" uniqKey="Zheng L">L Zheng</name></author><author><name sortKey="Hsiang, T" uniqKey="Hsiang T">T Hsiang</name></author><author><name sortKey="Yang, Gl" uniqKey="Yang G">GL Yang</name></author><author><name sortKey="Zhao, Dl" uniqKey="Zhao D">DL Zhao</name></author><author><name sortKey="Lv, B" uniqKey="Lv B">B Lv</name></author><author><name sortKey="Chen, Yf" uniqKey="Chen Y">YF Chen</name></author><author><name sortKey="Huang, Jb" uniqKey="Huang J">JB Huang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zipfel, Rd" uniqKey="Zipfel R">RD Zipfel</name></author><author><name sortKey="Wilhelm De Beer, Z" uniqKey="Wilhelm De Beer Z">Z Wilhelm de Beer</name></author><author><name sortKey="Jacobs, K" uniqKey="Jacobs K">K Jacobs</name></author><author><name sortKey="Wingfield, Bc" uniqKey="Wingfield B">BC Wingfield</name></author><author><name sortKey="Wingfield, Mj" uniqKey="Wingfield M">MJ Wingfield</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Tree Physiol</journal-id><journal-id journal-id-type="iso-abbrev">Tree Physiol</journal-id><journal-id journal-id-type="publisher-id">treephys</journal-id><journal-title-group><journal-title>Tree Physiology</journal-title></journal-title-group><issn pub-type="ppub">0829-318X</issn><issn pub-type="epub">1758-4469</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">29329457</article-id><article-id pub-id-type="pmc">5982843</article-id><article-id pub-id-type="doi">10.1093/treephys/tpx147</article-id><article-id pub-id-type="publisher-id">tpx147</article-id><article-categories><subj-group subj-group-type="heading"><subject>Methods Paper</subject></subj-group></article-categories><title-group><article-title>A novel application of RNase H2-dependent quantitative PCR for detection and quantification of <italic>Grosmannia clavigera</italic>, a mountain pine beetle fungal symbiont, in environmental samples</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>McAllister</surname><given-names>Chandra H</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref></contrib><contrib contrib-type="author"><name><surname>Fortier</surname><given-names>Colleen E</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref></contrib><contrib contrib-type="author"><name><surname>St Onge</surname><given-names>Kate R</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref></contrib><contrib contrib-type="author"><name><surname>Sacchi</surname><given-names>Bianca M</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref></contrib><contrib contrib-type="author"><name><surname>Nawrot</surname><given-names>Meaghan J</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref></contrib><contrib contrib-type="author"><name><surname>Locke</surname><given-names>Troy</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid" authenticated="false">http://orcid.org/0000-0002-4990-628X</contrib-id><name><surname>Cooke</surname><given-names>Janice E K</given-names></name><xref ref-type="aff" rid="tpx147af1"></xref><xref ref-type="corresp" rid="tpx147cor1"></xref><pmc-comment>janice.cooke@ualberta.ca </pmc-comment>
</contrib></contrib-group><aff id="tpx147af1">Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada T6G 2E9</aff><author-notes><corresp id="tpx147cor1">Corresponding author (<email>janice.cooke@ualberta.ca</email>)</corresp><p>handling Editor Chung-Jui Tsai</p></author-notes><pub-date pub-type="collection"><month>3</month><year>2018</year></pub-date><pub-date iso-8601-date="2018-01-10" pub-type="epub"><day>10</day><month>1</month><year>2018</year></pub-date><pub-date pub-type="pmc-release"><day>10</day><month>1</month><year>2018</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>38</volume><issue>3</issue><fpage>485</fpage><lpage>501</lpage><history><date date-type="received"><day>16</day><month>5</month><year>2017</year></date><date date-type="rev-recd"><day>25</day><month>10</month><year>2017</year></date><date date-type="accepted"><day>30</day><month>10</month><year>2017</year></date></history><permissions><copyright-statement>© The Author 2018. Published by Oxford University Press.</copyright-statement><copyright-year>2018</copyright-year><license license-type="cc-by" xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><self-uri xlink:href="tpx147.pdf"></self-uri><abstract><title>Abstract</title><p>Mountain pine beetle (<italic>Dendroctonus ponderosae</italic> Hopkins; MPB) is an economically and ecologically important pest of pine species in western North America. Mountain pine beetles form complex multipartite relationships with microbial partners, including the ophiostomoid fungi <italic>Grosmannia clavigera</italic> (Robinson-Jeffrey and Davidson) Zipfel, de Beer and Wingfield, <italic>Ophiostoma montium</italic> (Rumbold) von Arx, <italic>Grosmannia aurea</italic> (Robinson-Jeffrey and Davidson) Zipfel, de Beer and Wingfield, <italic>Leptographium longiclavatum</italic> (Lee, Kim, and Breuil) <italic>and Leptographium terebrantis</italic> (Barras and Perry). These fungi are vectored by MPB to new pine hosts, where the fungi overcome host defenses to grow into the sapwood. A tree’s relative susceptibility to these fungi is conventionally assessed by measuring lesions that develop in response to fungal inoculation. However, these lesions represent a symptom of infection, representing both fungal growth and tree defense capacity. In order to more objectively assess fungal virulence and host tree susceptibility in studies of host–pathogen interactions, a reliable, consistent, sensitive method is required to accurately identify and quantify MPB-associated fungal symbionts in planta. We have adapted RNase H2-dependent PCR, a technique originally designed for rare allele discrimination, to develop a novel RNase H2-dependent quantitative PCR (rh-qPCR) assay that shows greater specificity and sensitivity than previously published PCR-based methods to quantify MPB fungal symbionts in pine xylem and MPB whole beetles. Two sets of assay probes were designed: one that amplifies a broad range of ophiostomoid species, and a second that amplifies <italic>G. clavigera</italic> but not other MPB-associated ophiostomoid species. Using these primers to quantify <italic>G. clavigera</italic> in pine stems, we provide evidence that lesion length does not accurately reflect the extent of fungal colonization along the stem nor the quantity of fungal growth within this colonized portion of stem. The sensitivity, specificity, reproducibility, cost effectiveness and high-throughput potential of the rh-qPCR assay makes the technology suitable for identification and quantification of a wide array of pathogenic and beneficial microbes that form associations with plants and other organisms, even when the microbial partner is present in low abundance.</p></abstract><kwd-group><kwd>bark beetles</kwd><kwd>blue-stain fungi</kwd><kwd>defense responses</kwd><kwd>forest biotechnology</kwd><kwd>forest pathology</kwd><kwd>pine</kwd></kwd-group><funding-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">Natural Science and Engineering Research Council of Canada (NSERC)</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">TRIA Network</named-content></funding-source><award-id>NET GP 434810-12</award-id></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">Alberta Agriculture and Forestry, fRI Research</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">Ministry of Natural Resources and Forestry</named-content><named-content content-type="funder-identifier">10.13039/100008138</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">Saskatchewan Ministry of Environment</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">West Fraser and Weyerhaeuser</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">NSERC Undergraduate Student Research Awards</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">University of Alberta Undergraduate Research Initiative Stipend</named-content></funding-source></award-group><award-group award-type="grant"><funding-source><named-content content-type="funder-name">NSERC Canada Graduate Scholarships-Master’s (CGS M)</named-content></funding-source></award-group></funding-group><counts><page-count count="17"></page-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd \
| NlmPubMed2Wicri -a MelampsoraV2
| This area was generated with Dilib version V0.6.38. |  |